CN101194899A - Use of bocchiol in the preparation of medicines for preventing and treating senile dementia - Google Patents

Abstract

The invention discloses the usage of patchouli alcohol in the process of preparing medicament for preventing and curing senile dementia, belonging to the medical field. The invention has the advantages that the invention is extensively and deeply researched, patchouli alcohol is unexpectedly found to have the functions of neural protection and nutrition in the process of screening medicament which is anti senile dementia. Further study finds that patchouli alcohol has a promoting effect to the study and memory of a senile dementia mould mouse. Thereby patchouli alcohol can be used to prepare a medicament which can prevent and cure senile dementia. The medicament which utilizes patchouli alcohol to be active component may have significant effects in clinic, thereby bringing out large social and economic benefits.

Description

Use of bocchiol in the preparation of medicines for preventing and treating senile dementia

technical field

The invention relates to the use of bocicol in the preparation of medicaments for preventing and treating senile dementia, and belongs to the field of medicine.

Background technique

Patchouli, Pogostemon cablin (Blanco) Benth, is a plant of the genus Patchouli of the Labiatae family, a perennial herb, and the whole herb is used for medicine. Its taste is pungent, slightly warm, and it returns to the three meridians of spleen, stomach, and lung. The medicinal ingredients of patchouli are mainly its volatile oil, which is called patchouli oil, which has the functions of regulating digestive tract function, relieving spasm of gastrointestinal smooth muscle and antibacterial effect. Its main component is patchouli alcohol (patchoulialcohol, buccal alcohol), which has calcium ion antagonistic effect (Chem Pharm Bull, 1989, 37 (2): 345-348). Patchouli alcohol is a tricyclic sesquiterpene compound, crystallizable, melting point 55-56%, boiling point 140°C (1.06kPa), optical rotation -97.4° (c=24, chloroform), insoluble in water, soluble in alcohol, ethers and common organic solvents. The molecular formula is C15H26O, the CAS registration number is 5986-55-0, the molecular weight is 222.37, and the chemical name is (1 R-(1 alpha, 4beta, 4-alpha, 6beta, 8-alpha))-Octahydro-4, 8a, 9, 9-tetramethyl-1

, 6-methano-1(2H)-naphthol, the structural formula is as follows:

Alzheimer's disease is a chronic central nervous system degenerative disease with progressive cognitive decline as the main clinical symptom. Its incidence rate rises sharply with the increase of age. In my country, the prevalence rate of the elderly over 65 years old is 1% to 5%, and that over 85 years old reaches 20% to 40%. The pathological changes of senile dementia include extensive neuronal loss, numerous neurofibrillary tangles (neurofibrillary tangles) and senile plaques in the brain. Currently marketed drugs for the treatment of senile dementia, such as Aricept and tacrine, can improve or alleviate the symptoms of dementia in mild patients to a certain extent, but cannot prevent or reverse the further development of lesions.

Contents of the invention

The technical problem to be solved by the present invention is to provide the use of bokuli alcohol in the preparation of medicaments for preventing and treating senile dementia.

Another technical problem to be solved by the present invention is to provide a pharmaceutical composition for preventing and treating senile dementia.

To achieve the above object, the present invention adopts the following technical solutions:

Application of bocicol in the preparation of medicaments for preventing and treating senile dementia.

A pharmaceutical composition for preventing and treating senile dementia, which consists of active ingredient bocicol and pharmaceutically acceptable auxiliary materials.

The pharmaceutically acceptable excipients include but not limited to pharmaceutically acceptable diluents, preservatives, solvents, emulsifiers, adjuvants and/or carriers.

The medicament or pharmaceutical composition for preventing and treating senile dementia is in the form of oral preparation, injection, transpulmonary, nasal, transdermal or other administration forms.

The oral dosage forms include but are not limited to capsules, tablets, pills, lozenges, granules, liposomes and the like.

The bucchini alcohol in the present invention is purchased from China National Institute for the Control of Pharmaceutical and Biological Products, with a purity of over 98%. At the cellular level, flow cytometry was used to detect the anti-apoptosis effect of β-amyloid (Aβ) on PC12 and the promotion of axon growth in primary hippocampal neurons. At the animal level, the mouse platform test and Morris water maze test were used to observe its promoting effect on the learning and memory of scopolamine-induced Alzheimer's model mice. The results showed that bocicol has protective and nutritional effects on nerve cells, and can improve the learning and memory ability of senile dementia model mice.

The advantages of the present invention are: after extensive and in-depth research, the present invention unexpectedly finds that bocicaster has neuroprotective and nutritional effects in the screening of anti-senile dementia drugs. Further studies have found that it can promote learning and memory in Alzheimer's model mice. Therefore, the buccal alcohol can be used to prepare medicines for preventing and treating senile dementia. Drugs with potocio as active ingredient may have clinically significant curative effect, thereby generating huge social and economic benefits.

The present invention will be further described below in conjunction with the specific embodiments, which is not a limitation of the present invention. According to the prior art known in the art, the embodiments of the present invention are not limited thereto. Therefore, all equivalents in the field made according to the present disclosure Replacement all belong to the protection scope of the present invention.

Detailed ways

Example 1: Anti-apoptosis effect of bucchini alcohol

1. Materials

1. Baiqiu Li alcohol, purchased from China Institute for the Control of Pharmaceutical and Biological Products, with a purity of over 98%.

2. Aβ _25-35 , purchased from Sigma.

3. Annexin V-FITC, FITC-labeled annexin V, was purchased from Jingmei Bioengineering Co., Ltd.

4. PI solution, propidium iodide solution.

5. Rat pheochromocytoma cell (PC12) cell line, passaged and preserved in our laboratory.

2. Method

Flow cytometry was used to detect the anti-apoptosis of rat pheochromocytoma cells (PC12) induced by Aβ _25-35 . The experimental groups were as follows: control group; Aβ _25-35 treatment group; different doses of drug groups.

Take 100 μl of cell suspension in a flow tube, add 5 μl Annexin V-FITC and 10 μl PI solution (20 μg/ml) and mix well, incubate at room temperature in the dark for 15 minutes, add 400 μl PBS, and analyze by flow cytometry.

3. Results

Compared with the normal group, the apoptosis rate of the model group treated with Aβ _25-35 was significantly increased (P<0.01); the apoptosis rate of each drug-dosed group was significantly lower than that of the model group (P<0.01). Li alcohol has the effect of anti-apoptosis of PC12 cells induced by Aβ _25-35 .

Table 1: Anti-apoptotic effect of buccal alcohol on PC12 cells induced by Aβ _25-35

group Dose (μg·mL ^-1 ) Apoptosis rate (%) Control group Aβ _25-35 group Buocicol low-dose group Buocicol medium-dose group Bucichol high-dose group -100.252.525 2.3±1.90 30.79±2.66 ^** 22.49±1.02 ^* 20.28±1.63 ^* 13.94±1.79 ^*

Note: ^** P<0.01 compared with the control group; ^* P<0.01 compared with the model group

Example 2: Buocchiol promotes axonal growth of primary cultured hippocampal neurons

1. Materials

1. Baiqiu Li alcohol, purchased from China Institute for the Control of Pharmaceutical and Biological Products, with a purity of over 98%.

2. Trypsin, the concentration is 0.25%, purchased from Sigma company

3. DMEM/F-12 medium, purchased from Gibco.

4. Nerve growth factor (NGF), purchased from Beijing Shengri Pharmaceutical Technology Development Co., Ltd.

5. Fetal bovine serum, purchased from Gibco.

6. N2, purchased from Gibco.

7. B27, purchased from Gibco.

8. Cytarabine, purchased from Harbin Bolai Pharmaceutical Co., Ltd.

9. SD rat suckling mice, within 12 hours after birth, were given both ♀♂ and divided into control group and drug groups with different doses, purchased from the Experimental Animal Center of Academy of Military Medical Sciences.

2. Method

After the rat suckling mice were sterilized with 75% ethanol by volume fraction, they were decapitated and placed in a plate containing dissecting solution, the whole brain was isolated, and the hippocampal tissue was isolated under a dissecting microscope. Place it in 0.25% trypsin and mince it with scissors. Aspirate the trypsin into the serum-containing centrifuge tube every 4 minutes, and add fresh trypsin until there are no visible tissue pieces. After filtering with a 200-mesh metal screen, centrifuge at 1000rpm·min ^-1 for 10min. It was resuspended in DMEM/F-12 seeding solution with 10% fetal bovine serum, inoculated on a 24-well plate pre-coated with poly-lysine, cultured overnight and replaced with 3% fetal bovine serum, 1% N2, 1 %B27 and 3ng·mL ^-1 cytarabine in DMEM/F-12 culture solution. The final concentration mass fractions were 0.05, 0.5, 5 μg·mL ^-1 buccalol, and 2 ng·mL ^-1 nerve growth factor (NGF); the control group was only added 2 ng·mL ^-1 NGF. After 2 days, photographs were taken under a fluorescent microscope, and the axon length was counted. The method was 10 pictures for each group, and the axon length of each neuron was measured on each picture, and the sum was divided by the total number of neurons.

3. Results

Buocchiol itself did not promote the axon growth of primary hippocampal neurons, but in the presence of 2ng·mL ^-1 NGF it could significantly promote the axon growth, and the axon length was significantly longer than that of the control group (P<0.01). The results showed that in the presence of very low concentration of NGF, the concentration range of buccal alcohol in the range of 0.05-5 μg·mL ^-1 could promote the axon growth of primary hippocampal neurons (Table 2).

Table 2: Promoting effect of buccal alcohol on axonal growth of primary hippocampal neurons

group Concentration (μ g·mL ^-1 ) Axon length (μm) Control group Buocchiol low-dose group Buocchiol middle-dose group Buocchiol high-dose group -0.050.55 19.3±343.4±10 ^* 59.0±9 ^* 63.2±11 ^*

Note: n=10 ^* P<0.01 compared with the control group

Example 3: The promoting effect of bocicol on the learning and memory of senile dementia model mice

1. Materials

1. Baiqiu Li alcohol, purchased from China Institute for the Control of Pharmaceutical and Biological Products, with a purity of over 98%.

2. Scopolamine, produced by Xuzhou Lane Pharmaceutical Co., Ltd.

3. Kunming mice, weighing 14-16g, female, were divided into normal control group, scopolamine group and drug groups with different doses. The mice were randomly divided into groups according to the principle of weight balance, with 10 mice in each group. They were purchased from the Experimental Animal Center of Academy of Military Medical Sciences .

2. Method

The mouse platform test and the Morris water maze test were used to observe the promoting effect of bocicol on the learning and memory of scopolamine-induced senile dementia model mice. The mice were randomly grouped according to the principle of weight balance. After 7 days of administration, the model was established and tested.

1. Mouse platform test:

Place the animals on the platform, adapt to the environment for 3 minutes, and then pass through the voltage of 36V for 5 minutes of training. Take the test after 24 hours, record the latency period of jumping off the platform for the first time, the number of errors within 5 minutes and the total electric shock time, and use this as the memory score.

2. Morris water maze test:

The day before the experiment, the mice were put into the water maze to swim freely for 2 minutes to adapt to the environment. Scopolamine 5 mg·kg ^-1 was intraperitoneally injected 15 minutes before training in the morning every day, once a day. On the first day, the mice were trained 3 times, each time from 3 different water entry points, the mice were put into the water to swim facing the container wall, if the mice could not find the underwater platform within 90s, they were placed directly on the platform and Stay for 30s. From the next day, train once a day in the morning and afternoon, and record the time it took for the mice to climb up the platform after entering the water, and the trajectory they passed as indicators for detecting the learning of space and cues for the mice. After 24 hours, retest step by step, and a total of training 3 days.

The brain tissues of 4 mice in each group were taken for immunohistochemistry to measure the level of cholinergic M ₁ receptors. Two slices were made for each mouse. SABC method was used for immunohistochemistry, DAB was used for color development, and nuclear counterstaining was performed with hematoxylin at last. One slice was taken from each group as a negative control (the primary antibody was replaced by PBS). During data analysis, 5 fields of view were selected for each slice according to the same parts of the hippocampus and cortex, and the results were processed with MIAS medical image analysis management system 4.0 to analyze the average optical density and integral optical density.

3. Results

1. Mouse platform test:

The platform jumping results showed that compared with the normal group, the number of errors within 5 minutes in the model group increased significantly, and the latency period was significantly shortened. Compared with the model group, the number of errors in the potcine group was significantly reduced, but the latency period had a tendency to increase, but there was no statistical difference (see Table 3).

Table 3: Effects of buccal alcohol on the platform jumping performance of scopolamine mice with learning and memory impairment model

Group Dose (mg·kg ^-1 ) Number of errors (n) Latency period (s) Normal control group Scopolamine group Bukitschyl alcohol low dose group Bukitschyl alcohol high dose group -550100 0.2±0.45.0±1.7 ^*** 1.5±1.2#1.0±1.1## 259.9±95.28.3±2.3 ^*** 110.0±94.4141.2±137.8

Note: Compared with the normal group, ^*** P<0.001; compared with the model group: #<0.05, ##P<0.01.

2. Morris water maze test:

The results of the Morris water maze showed that with the increase of training times, the latency period for the mice in each group to find the platform was gradually shortened. Compared with the control group, the model group (scopolamine 5mg·kg ^-1 ) showed significant differences in each training performance. From the 5th training session, the Buocchiol group showed a significant difference with the model group. The results are shown in Table 4:

Table 4: Effect of buccal alcohol on the stage latency of learning and memory impairment in scopolamine mice in the water maze

Incubation period (s) control group Scopolamine Pocket alcohol (100mg·kg ^-1 ) Pocket alcohol (50mg·kg ^-1 )   1st time 2nd time 3rd time 4th time 5th time 6th time 7th time 60.10±29.5340.32±31.0340.43±27.3024.87±10.4119.80±12.0415.22±7.4917.19±12.86 86.94±9.49 ^* 89.08±1.85 ^*** 77.73±26.37 ^** 76.52±28.31 ^*** 78.43±1.74 ^*** 83.30±20.15 ^*** 60.02±30.74 ^** 81.53±20.5870.50±31.2555.42±31.2252.28±33.5332.07±16.53###31.77±23.97###28.02±10.58## 87.38±6.6482.46±11.2369.87±22.1659.81±24.9044.49±33.24##34.03±10.58###32.70±19.24#

Note: Compared with the normal group: ^* P<0.05, ^** P<0.01, ^*** P<0.001; compared with the model group: #P<0.05, ##P<0.01, ###P<0.001.

The 1st-3rd time is the training result of the first day, the 4th and 5th time are the training results of the second day, and the 6th and 7th time are the training results of the third day.

The average optical density results showed that compared with the normal control group, the average optical density value of the model control group decreased significantly, indicating that the degree of cell staining decreased and the level of cholinergic M1 receptors decreased. However, the average optical density value of the Bukitchenol group was higher than that of the model group, indicating that the drug may increase the level of M1 receptors. The results of integrated optical density also showed that the model group was significantly lower than the normal control group, while the pottyrae alcohol group was significantly higher than the model group. It shows that the overall level of staining level of positive cells in the drug group is increased, that is, the total level of antigen is increased. The results are shown in Table 5.

Table 5: Effects of buccal alcohol on _M1 receptor levels in the brain tissue of mice with acquired learning and memory impairment caused by scopolamine

group   Average Optical Density Integral optical density Normal control scopolamine model Bukitchenol 100mg·kg ^-1 Bukitschyl alcohol 50mg·kg ^-1 0.21±0.040.15±0.04***0.18±0.05##0.17±0.03#   1077.70±612.46261.24±103.28***858.88±165.84###673.44±171.66###

Note: Compared with the normal group, ^*** P<0.001; compared with the model group: #P<0.05, ##P<0.01, ###P<0.001.

Claims (5)

1. patchouli alcohol is prevented and treated purposes in the medicine of senile dementia in preparation.

2. a pharmaceutical composition of preventing and treating senile dementia is characterized in that: be made up of the adjuvant that active component patchouli alcohol and pharmacy are accepted.

3. a kind of pharmaceutical composition of preventing and treating senile dementia according to claim 2 is characterized in that: the adjuvant that described pharmacy is accepted is pharmaceutically acceptable diluent, antiseptic, solvent, emulsifying agent, adjuvant and/or carrier.

4. a kind of pharmaceutical composition of preventing and treating senile dementia according to claim 2 is characterized in that: described pharmaceutical composition is oral formulations, injection, through lung, nose agent, percutaneous or other mode form of administration.

5. a kind of pharmaceutical composition of preventing and treating senile dementia according to claim 4 is characterized in that: described peroral dosage form is capsule, tablet, pill, lozenge, granule or liposome.