CN103989667A - Drug for treating senile dementia - Google Patents

Abstract

The invention relates to a medicament for treating senile dementia. The active ingredient of the medicament is brilliant blue G, the molecular formula of the brilliant blue G is C ₄₇ H ₄₈ N ₃ NaO ₇ S ₂ , and the molecular weight is 854.02. Its structural formula is It can effectively relieve the symptoms of Alzheimer's disease, slow down the memory function decline of Alzheimer's disease, slow down the cognitive dysfunction of Alzheimer's disease, prevent the neuron damage caused by Alzheimer's disease, and has high efficiency and safety for the treatment of Alzheimer's disease .

Description

A medicine for treating Alzheimer's disease

technical field

The invention relates to the field of treating senile dementia, in particular to a medicament for treating senile dementia.

Background technique

The pathogenesis of senile dementia (also known as Alzheimer's disease) is still unclear. Drugs currently on the market are mainly drugs that increase cholinergic function, such as cholinesterase inhibitors galantamine, donepezil (Aricept), M receptor agonist Zanomelin, etc. These drugs can only delay the onset process , but it cannot cure Alzheimer's disease, and it is accompanied by cardiovascular and gastrointestinal side effects. Other commonly used drugs such as antidepressants, anti-anxiety, sedative-hypnotics, antipsychotics and other drugs have limited improvement in the symptoms of Alzheimer's disease, and have neurological side effects. In view of this, there is a need for a more efficient and safe agent for treating Alzheimer's disease.

In recent decades, the hypothesis that the P2X7 receptor signaling pathway is involved in Alzheimer's disease has been supported by some experimental evidence. Current research shows that Alzheimer's disease is closely related to the deposition of β-amyloid protein. Under the induction of β-amyloid protein, P2X7 receptors may cause neuroinflammation by excessively activating glial cells, thereby inducing neuron apoptosis, which promotes the process of neuron loss in Alzheimer's patients. Brilliant Blue G (BBG), as a commonly used stain, has been proven safe. At the same time, BBG can act as a highly effective inhibitor of P2X7 receptor (IC ₅₀ value is 10-200nM). At the same time, BBG has been confirmed to have the ability to penetrate the blood-brain barrier; and BBG can inhibit the neuroinflammatory response mediated by β-amyloid protein.

Contents of the invention

The technical problem to be solved by the present invention is to provide a drug that can effectively alleviate the symptoms of Alzheimer's disease, slow down the memory function decline of Alzheimer's disease, slow down the cognitive dysfunction of Alzheimer's disease, and prevent the neurological disorders caused by Alzheimer's disease. A drug for the treatment of Alzheimer's disease.

The technical solution adopted by the present invention to solve the problems of the technologies described above is:

A medicament for treating senile dementia, the active ingredient of the medicament is Brilliant Blue G, the molecular formula of Brilliant Blue G is C ₄₇ H ₄₈ N ₃ NaO ₇ S ₂ , the molecular weight is 854.02, and its structural formula is .

The aforementioned brilliant blue G is a pharmacologically acceptable salt or solvate.

The above-mentioned agents are used as drugs for alleviating memory function impairment and cognitive function impairment in senile dementia.

The above-mentioned agents are used as drugs having a neuroprotective effect on senile dementia.

The medicament for treating senile dementia of the present invention, its active ingredient is brilliant blue G, which can effectively alleviate the symptoms of senile dementia, slow down the memory function decline of senile dementia, slow down the cognitive dysfunction of senile dementia, prevent the Neuron damage caused by dementia is more efficient and safe for the treatment of Alzheimer's disease.

Description of drawings

Figure 1 is the effect of BBG treatment on spatial learning and memory in Alzheimer's disease model mice;

Figure 2 is the effect of BBG treatment on the cognitive function of Alzheimer's disease model mice;

Figure 3 is the effect of BBG treatment on neuron synapse damage in Alzheimer's disease model.

Detailed ways

The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

A medicament for treating senile dementia, the active ingredient of the medicament is Brilliant Blue G, the molecular formula of Brilliant Blue G is C ₄₇ H ₄₈ N ₃ NaO ₇ S ₂ , the molecular weight is 854.02, and its structural formula is .

The aforementioned brilliant blue G is a pharmacologically acceptable salt or solvate.

The above-mentioned agents are used as drugs for alleviating memory function impairment and cognitive function impairment in senile dementia.

The above-mentioned agents are used as drugs having a neuroprotective effect on senile dementia.

1. Experimental animals:

Male ICR mice weighing 25 to 30 g were used in the experiment. Divided into 4 groups, normal control group, β-amyloid (Aβ) group, BBG administration group, Aβ and BBG administration group, 8 rats in each group. The daily test time is fixed at 9:00AM-4:30PM. The experimental operation followed the NIH guidelines for the use of experimental animals (NIH publications No.80-23, 1996). BBG is bright blue G.

2. Establishment of Alzheimer's disease animal model:

Inject the mice with chloral hydrate at a dose of 10 mg/kg, anesthetize them, fix them on a stereotaxic instrument, cut off the head hair, expose the skin, start from the midpoint of the line connecting the two eyes, and connect the rear edges of the two ears Cut an incision at the midpoint of the end point to expose the anterior bregma. The anterior bregma is the coordinate origin, retreat 1.5 mm, and open 1.2 mm in the left and right sides. These two points are the coordinates of the CA1 area of the hippocampus. Drill the hole and insert the needle 1.5 mm. Aβ1 ug was injected, and normal saline was injected into the control group and BBG administration group, the skin was sutured, disinfected, and put back into the cage for feeding.

3. Administration:

After the model was established, the control group was injected with normal saline, and BBG was administered by intraperitoneal injection of BBG 50 mg/kg, administered once every two days for 14 days, and then behavioral tests were performed.

4. Behavioral test of learning and memory ability (water maze test):

The water maze device is a round black pool with a diameter of 110cm and a height of 50cm. It is filled with water, the water depth is 24cm, the water temperature is 22-24°C, and there is a platform with a diameter of 8cm at 1cm below the water surface. There are black curtains around the pool. Visible markers were hung to allow the mice to remember the position and direction. The water maze was artificially divided into 4 quadrants. The mice entered the water from different quadrants each time they were trained. The end of the 4 quadrants was a training cycle. The mice swam freely in the water until Find the platform, if the mouse cannot find the platform within 90 seconds, then artificially guide the mouse to the platform and allow it to stay on the platform for 15 seconds. Each mouse was trained 8 times a day (2 cycles) for 4 consecutive days. On the 5th day, the underwater platform was removed, and mice were put into the opposite quadrant of the original platform, and allowed to swim freely in the water maze device for 60 seconds. The video surveillance system is used to track the recordings. After the experiment, the software analyzes the mouse movement trajectory.

5. Test of cognitive ability (new object recognition):

The new object recognition experiment was carried out in a self-made white plastic behavior box. The length, width and height of the behavior box were 50*50*30cm respectively. The whole experiment was completed in 3 stages and 3 days. In the first stage, the adaptation period on the first day, the mice were put into the behavior box and allowed to explore freely for 5 minutes. At this stage, no data was collected. In the second stage, the training period on the second day, two identical objects were put into the behavior box, the distance between the object and the behavior box was about 5cm, and the distance between the two objects was 20cm, and the mouse was put into the mouse and allowed to explore freely for 5 minutes. We defined exploratory behavior: the mouse's nose was less than 2 cm from the object or it touched the object with its nose. Walking around or crouching near an object is not an act of exploration. During this phase, the time spent exploring each object by the mice was recorded, and after the end, the mice were immediately returned to their original cages. In the third stage, the test period on the third day, the mice were put back into the original behavior box to explore freely for 5 minutes, and one of the objects was replaced with a new object. The cognitive function of mice was measured by resolution index, resolution index = time to explore new objects/(time to explore new objects + time to explore old objects).

6. Establishment of Alzheimer's disease cell model:

  The hippocampal tissue of newborn mice was used for primary neuron culture. The hippocampus was isolated from neonatal mice, put into trypsin solution, digested in a 37°C incubator for 12 minutes, and then added DMEM medium containing 10% serum to terminate the digestion. Inoculate into disposable petri dishes after gently pipetting and culture in a cell culture incubator at 37°C. The medium was completely changed 3 hours after inoculation, and the Neurobasal medium was changed 24 hours later. On the 5th day, cytarabine with a final concentration of 5 μM was added to inhibit the growth of glial cells. Hippocampal neurons were cultured in vitro until day 5, and transfected with GFP-actin and F-GFP plasmids to mark the cytoskeleton. At DIV14, the model group was incubated with 500nM Aβ for 3h, and the BBG treatment group was incubated with 1μM BBG for 24h.

7. Statistical analysis and processing

The data in this part are expressed in the format of mean±SEM (mean±standard error). The data were analyzed by one-way ANOVA with one-way multilevel design. P<0.05 was regarded as statistically significant difference.

Experimental results:

(1) BBG treatment improves spatial learning and memory in Alzheimer's model mice (as shown in Figure 1):

On the 4th day of the training period, the time for the mice to escape from the water to the platform was measured, and it took longer for the mice to find the platform after the injection of Aβ. After intraperitoneal injection of BBG (50 mg/kg), the time for mice to find the platform was significantly shortened (P<0.005, part A in Figure 1). During the exploration period on day 5, we removed the platform of the second quadrant to test the memory ability of the mice. The results showed that the distance of Aβ mice in the second quadrant was significantly less than that of the normal control group (p<0.01), and the BBG intraperitoneal injection group spent more time in the second quadrant than the Aβ injection group after Aβ localization (p<0.01, Fig. Part B of 1). The statistics of the number of times the mice crossed the original platform position found that after stereotaxic injection of Aβ, the number of times the mice crossed the original platform position was significantly less than that of the normal group (p<0.01), after intraperitoneal injection of BBG, the mice crossed the original platform The number of positions was significantly increased (p<0.01, part C in Fig. 1).

(2) BBG treatment improves the cognitive function of Alzheimer's model mice (as shown in Figure 2):

The ratio of the time for mice to explore new and old objects was tested by the new object recognition experiment. Resolution index = time to recognize object 1/(time to recognize object 1 + time to recognize object 2), and parts A and B in Figure 2 represent the mouse’s ability to recognize the same object (object 1 and 2 are two identical objects) during the familiarization period Resolution index and total exploration time. Parts C and D in Figure 2 represent the resolution index and total exploration time of mice in the test period to new objects (object 1 was replaced with a new object that had not been touched before, and object 2 remained unchanged). Statistics were made on the time spent exploring two identical objects in each group of mice during the familiarization period, and it was found that the resolution index and total exploration time of the mice exploring two identical objects were basically equal, and there was no significant difference between the groups (Figure 2A, Part B). During the test period, the resolution index of Aβ mice was lower than that of the control group, and the difference was statistically significant; the resolution index of BBG-treated mice was significantly higher than that of Aβ mice (part C in Figure 2). There was a statistically significant difference between each group and the Aβ localization group; the resolution index of the Aβ localization group after intraperitoneal injection of BBG was higher than that of the Aβ control group, and there was no significant difference in the total exploration time of the mice in each group (Part D in Figure 2).

(3) Aβ treatment significantly reduced the dendritic spine density of hippocampal neurons, and BBG treatment reversed the loss of dendritic spine density caused by Aβ (as shown in Figure 3):

After the hippocampal neurons of DIV14 rats were incubated with 500 nM Aβ for 3 h, the density of dendritic spines of hippocampal neurons was significantly lower than that of the control group (parts A and B in Figure 3). Dendritic spine density of neurons in control group and Aβ treatment group were 35.87 ± 2.55/ 100 μm, n=10 and 20.66 ± 1.22/ 100 μm, n=10, P < 0.005. At the same time, 500 nM Aβ was added to hippocampal neurons of DIV14 rats while 1 μM BBG was added for 24 h. The results showed that the dendritic spine density of neurons in the BBG pre-incubation group was 37.96±2.45/ 100 μm, n=10, and BBG reversed Aβ-mediated loss of dendritic spines in hippocampal neurons, P < 0.005 (Part C in Figure 3).

Conclusion: BBG can effectively improve memory loss and cognitive impairment in Alzheimer's disease, and has a protective effect on neuron damage in Alzheimer's disease. Therefore, the medicament for treating senile dementia of the present invention has significant clinical significance and application value.

The preferred embodiment of the present invention has been illustrated, and various changes or modifications may be made by those skilled in the art without departing from the scope of the present invention.

Claims (4)

1. a medicament that is used for the treatment of senile dementia, is characterized in that: the effective ingredient of described medicament is light blue G, and the molecular formula of described light blue G is C ₄₇h ₄₈n ₃naO ₇s ₂, molecular weight is 854.02, its structural formula is .

2. a kind of medicament that is used for the treatment of senile dementia according to claim 1, is characterized in that: described light blue G is receptible salt or solvate in pharmacology.

3. a kind of medicament that is used for the treatment of senile dementia according to claim 2, is characterized in that: described medicament goes down as the memory function of alleviating senile dementia and the drug use of cerebral damage.

4. a kind of medicament that is used for the treatment of senile dementia according to claim 3, is characterized in that: described medicament is as senile dementia being had to the drug use of neuroprotective.