Pharmaceutical composition comprising donepezil and red ginseng kyung-ok-ko for preventing or treating dementia

Abstract

The present invention relates to a pharmaceutical composition comprising donepezil and red ginseng Kyung-ok-ko. When the composition of the present invention is used, donepezil and red ginseng Kyung-ok-ko are co-administered to improve memory and cognitive function, and thus can be used for preventing or treating dementia.

Description

Pharmaceutical composition for preventing or treating dementia containing donepezil and Hongsamgyeongokgo

[0001] The present invention relates to a pharmaceutical composition for preventing or treating dementia, comprising donepezil and Hongsamgyeongokgo.

[0002] The prevalence of dementia among those aged 65 and older in Korea is 11%. Furthermore, according to the National Dementia Center at the Korea Medical Center, the number of dementia patients among those aged 65 and older in Korea is projected to exceed 1 million by 2023. Furthermore, the number is projected to continue increasing, reaching 1.42 million in 2023, 2.26 million in 2040, and 3.15 million in 2050.

[0003] Dementia is a disease in which a person who was living a normal life experiences a general and continuous decline in cognitive function compared to before due to damage to brain function for various reasons. Pathologically, it is a brain disease in which severe impairments in memory, concentration, language, and cognition gradually progress over a long period of time due to damage and loss of nerve cells, ultimately leading to death.

[0004] Although Alus Alzheimer reported the first case of Alzheimer's disease in 1906, it wasn't until 1993 that the first Alzheimer's treatment, tacrine, appeared. Following this, donepezil, rivastigmine, galantamine, and memantine were developed. However, these drugs, centered around donepezil, only partially improved the clinical symptoms of Alzheimer's dementia. Therefore, the need for treatments that can deliver more significant effects than donepezil alone continues to grow.

[0005] One aspect of the present invention is to provide a pharmaceutical composition for preventing or treating dementia, comprising donepezil and Hongsamgyeongokgo.

[0006] One aspect of the present invention is to provide a pharmaceutical composition for preventing or treating dementia, comprising donepezil and Hongsamgyeongokgo.

[0007] According to one specific example, the Hongsamgyeongokgo may be composed of red ginseng, Poria cocos, raw Rehmannia glutinosa, and honey.

[0008] According to one specific example, the red ginseng, Poria cocos, raw Rehmannia glutinosa, and honey may be in a weight ratio of 1:1:5:5 to 1:3:10:10.

[0009] According to one specific example, the weight ratio of donepezil and Hongsamgyeongokgo may be 1:100 to 1:2,000.

[0010] According to one specific example, the dementia may be Alzheimer's dementia or vascular dementia.

[0011] The pharmaceutical composition comprising donepezil and Hongsamgyeongokgo according to the present invention improves memory and cognitive function by co-administering donepezil and Hongsamgyeongokgo, and thus can be used for the prevention or treatment of dementia such as Alzheimer's dementia and vascular dementia.

[0012] Figure 1 shows the survival rate of neurons treated with amyloid beta (Aβ) and then treated with various concentrations of herbal medicines (Golsaebo, Gwonbaek, Palgakhoehyang, and Sagan) and 5uM donepezil. Control cells (Control) were untreated and Aβ25 cells treated only with amyloid beta were used.

[0013] Figure 2 is a drawing showing the survival rate of nerve cells treated with amyloid beta (Aβ) and then treated with various concentrations of herbal medicines (pilbal, samsam, danggui, and baekdanhyang) and 5 μM of donepezil.

[0014] Figure 3 is a drawing confirming the survival rate of nerve cells when white sandalwood (white), red ginseng (salt ginseng), and donepezil were used together.

[0015] Figure 4 is a drawing confirming the survival rate of nerve cells when sandalwood (white), star anise (pal), and donepezil were used together.

[0016] Figure 5 is a drawing confirming the survival rate of nerve cells when Hongsamgyeongokgo (KOK) and donepezil were used together.

[0017] Figure 6 is a diagram showing the crossover rate in the Y maze experiment of mice orally administered Hongsamgyeongokgo, donepezil, and Hongsamgyeongokgo and donepezil.

[0018] Figure 7A is a diagram showing the time to reach the shock chamber during training and testing in a passive avoidance experiment of mice orally administered Hongsamgyeongokgo, donepezil, and Hongsamgyeongokgo and donepezil, and Figure 7B shows the difference in time to reach the shock chamber during training and testing.

[0019] Figure 8 is a drawing showing the location of the platform used in the underwater maze experiment and the area inside the tank divided into four quadrants.

[0020] Figure 9A shows the time it took for mice to reach a platform in a water maze experiment that was orally administered Hongsamgyeongokgo, donepezil, and Hongsamgyeongokgo and donepezil for 1 to 4 days, Figure 9B shows a bar graph showing the time it took for mice to reach a platform in a water maze experiment on the 4th day, and Figure 9C is a diagram showing the time each group stayed in each quadrant and platform (ZONE P).

[0021] One aspect of the present invention is to provide a pharmaceutical composition for preventing or treating dementia, comprising donepezil and Hongsamgyeongokgo.

[0022] The above donepezil is a dementia treatment that is effective in treating Alzheimer's disease, and helps improve cognitive function by ensuring that acetylcholine, a neurotransmitter that plays an important role in memory and cognitive function in the brain, is maintained normally.

[0023] In one specific example, the Hongsamgyeongokgo may be composed of red ginseng, Poria cocos, raw Rehmannia glutinosa, and honey.

[0024] In one specific example, the red ginseng, Poria cocos, raw Rehmannia glutinosa, and honey may be in a weight ratio of 1:1:5:5 to 1:3:10:10.

[0025] In one specific example, the weight ratio of donepezil and Hongsamgyeongokgo may be 1:5 to 1:40, 1:5 to 1:50, 1:10 to 1:40, or 1:15 to 1:40. Preferably, the weight ratio of donepezil and Hongsamgyeongokgo may be 1:20 to 1:40.

[0026] In one specific example, the dementia may be Alzheimer's dementia or vascular dementia.

[0027] These Hongsamgyeongokgo and donepezil can be formulated in advance as a mixture and administered simultaneously, or formulated separately and administered separately. If Hongsamgyeongokgo and donepezil are administered separately, Hongsamgyeongokgo can be administered 1 to 60 minutes after donepezil administration.

[0028] The pharmaceutical composition of the present invention may additionally contain other ingredients, in addition to the Hongsamgyeongokgo and donepezil, as long as they do not impair the effects aimed at by the present invention. For example, it may include conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and flavorings, or carriers.

[0029] The route of administration of the above pharmaceutical composition may be oral, intravenous, intramuscular, transdermal, intraarterial, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal.

[0030] The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

[0031] In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined according to factors including the type and severity of the individual, age, sex, type of disease, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by taking all of the above factors into consideration, and it can be easily determined by a person skilled in the art.

[0032] The above pharmaceutical composition may contain one or more active ingredients having the same or similar function as the above donepezil and Hongsamgyeongokgo. The above composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, external preparations, suppositories, and sterile injectable solutions, each according to a conventional method.

[0033] Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are provided for illustrative purposes only and the scope of the present invention is not limited to these examples.

[0034]

[0035] Example 1: Manufacturing method of Hongsamgyeongokgo

[0036] Hongsam Gyeongokgo was obtained from Dongguk University International Hospital (Ilsan, Korea), and its preparation method is as follows. 240 g of ground raw Rehmannia glutinosa and its juice, 60 g of Poria cocos, 30 g of Korean Red Panax ginseng powder, and 200 g of boiled and filtered honey were mixed. The mixture was placed in a jar and sealed with five sheets of oiled paper and one cotton cloth. The sealed jar was placed in a copper vessel and hung in water with the mouth of the jar above the waterline. It was boiled in an electric water heater for three days, with more water added to compensate for evaporation. After removing the jar, the wax paper was resealed, and it was left in the water in a dark place for one day. After that, the jar was removed again, placed in a pot, and boiled for one day. The moisture was drained and used. 10 mg / mL of Hongsam Kyungokgo was extracted with ethanol, water, or dimethyl sulfoxide (DMSO) at room temperature with low-speed stirring for 30 min. The extract was filtered through a 0.2 μM filter using a Buchner apparatus and stored at -20°C until analysis.

[0037]

[0038] Example 2: Confirmation of the neuroprotective effects of herbal medicine and donepezil.

[0039] 2-1. Neuronal cell culture and experimental methods

[0040] SH-SY5Y cells, a human neuroblastoma cell line, were used as neural cells. Cells were cultured at 37°C in an incubator with 5% CO2 using DMEM (Dulbecco's Modified Eagle Medium) medium supplemented with 10% FBS (fetal bovine serum), 100 units / mL of penicillin, and 100 μg / mL of streptomycin. When the cell density reached 80% or more, they were treated with trypsin-EDTA, cultured in suspension, and used for experiments.

[0041] To confirm the protective effect on nerve cells, 25 μM amyloid beta (Aβ) was injected into the SH-SY5Y cells to damage nerve cells, and the neuroprotective effect was confirmed by treating them with herbal medicine and donepezil.

[0042]

[0043] 2-2. Confirmation of the neuroprotective effects of herbal medicine and donepezil

[0044] SY-SY5Y cells treated with amyloid beta were injected with 10 ug / ml, 50 ug / ml, and 200 ug / ml (hereinafter, units are omitted as '10', '50', and '200') of Golsaebo, Gwonbaek, Palgakhoehyang, Sagan, Pilbal, Dansam, Angelica, and Baekdanhyang, and 5uM of donepezil was injected to check cell viability.

[0045] As a result, as shown in Figure 1, it was confirmed that cell viability increased in Golsaebom 200, Gwonbaek 200, Palgakhoehyang 50, and Palgakhoehyang 200, and in the case of Palgakhoehyang, it was confirmed that the effect was more significant than donepezil and more significant than the control group that was not treated with amyloid beta.

[0046] In addition, as shown in Fig. 2, it was confirmed that cell viability increased with 200 μM of Dansam, 200 μM of Angelica, 50 μM of Baekdanhyang, 200 μM of Baekdanhyang, and 5 μM of Donepezil, and it was confirmed that 200 μM of Baekdanhyang was more effective than the control group that was not treated with amyloid beta.

[0047]

[0048] 2-3. Confirmation of the synergistic effects of sandalwood, red ginseng, and donepezil.

[0049] The cell viability of SH-SY5Y cells treated with amyloid beta was confirmed by combining 50 ug / ml and 200 mg / ml of sandalwood and red ginseng, which had neuroprotective effects as single substances, with 5uM of donepezil.

[0050] As a result, as shown in Fig. 3, only when Baekdanhyang 200 and Salvia miltiorrhiza uralensis 50 were combined, a cytoprotective effect was observed, and other combinations failed to inhibit neuronal death caused by amyloid beta. In addition, when Baekdanhyang 200 and Salvia miltiorrhiza uralensis 50 were treated with 5uM donepezil together, the cell viability was confirmed to be lower than that of cells treated with donepezil alone, confirming that Baekdanhyang, Salvia miltiorrhiza uralensis, and donepezil do not exhibit a synergistic effect on neuronal protection.

[0051]

[0052] 2-4. Confirmation of the synergistic effects of sandalwood, star anise, and donepezil.

[0053] Among the herbal medicines that had a neuroprotective effect as a single substance, a synergistic effect was confirmed by combining 50 ug / ml of white sandalwood, 200 ug / ml of star anise, and 5 uM of donepezil.

[0054] As a result, as shown in Figure 4, when sandalwood and star anise were used together, a significant neuroprotective effect was observed, but when used together with donepezil, the effect was confirmed to be reduced to the level of the neuroprotective effect of donepezil.

[0055] Therefore, it was confirmed that the combined use of sandalwood, star anise, and donepezil did not show a synergistic effect on neuronal protection.

[0056]

[0057] 2-5. Confirmation of the synergistic effect of Hongsamgyeongokgo and donepezil.

[0058] The synergistic effect was confirmed by combining 50ug / ml, 100ug / ml, and 200ug / ml of Hongsamgyeongokgo prepared in Example 1 with 5uM of donepezil in SY-SY5Y cells treated with amyloid beta.

[0059] As a result, as shown in Figure 5, it was confirmed that when donepezil and Hongsamgyeongokgo were used together, a synergistic effect was shown compared to the existing Hongsamgyeongokgo and donepezil.

[0060]

[0061] Example 3: Preparation of experimental animals

[0062] 3-1. Mouse Acclimatization

[0063] A total of 20 healthy, 6-week-old male C57BL / 6 mice, weighing 18–21 g, were acclimated for 7 days and used. The mice were housed in groups of four and housed in an environment with a temperature of 20–25°C and a humidity of 50–55%. The light–dark cycle was 12 h, and food and water were always available. Animal experiments were conducted in compliance with national regulations on the use and welfare of laboratory animals and were approved by the Animal Experiment Ethics Committee of Dongguk University.

[0064]

[0065] 3-2. Mouse classification and experimental material administration

[0066] A total of 20 mice were divided into groups of 4 each: normal group (Normal), scopolamine-administered control group (Control), Hongsamgyeongokgo-treated group (KOK), Hongsamgyeongokgo and donepezil-treated group (KOK+D), and donepezil-treated group (D). Administration was performed as shown in the following table.

[0067] Scopolamine was administered intraperitoneally once, and Hongsamgyeongokgo and donepezil were administered orally once a day for one month.

[0068] Group name Administration details Normal group (Normal) Untreated control group (Control) 1mg / kg scopolamine administered intraperitoneally Hongsam Gyeongokgo treatment group (KOK) 1mg / kg scopolamine administered intraperitoneally, then 1000mg / kg Hongsam Gyeongokgo administered orally Hongsam Gyeongokgo and donepezil treatment group (KOK+D) 1mg / kg scopolamine administered intraperitoneally, then 1000mg / kg Hongsam Gyeongokgo and 1mg / kg donepezil administered orally Donepezil treatment group (D) 1mg / kg scopolamine administered intraperitoneally, then 1mg / kg donepezil administered orally

[0069]

[0070] Example 4: Y-maze experiment

[0071] After 30 days of drug administration, a Y-maze experiment was performed on mice. The Y-shaped maze has three branches, each 50 cm long, 10 cm wide, and 25 cm high, and the angle of the branches is 120 degrees. The three branches are designated A, B, and C, and all experimental groups are carefully placed so that they start from the end of one branch equally. The location where the experimental animals move during the 7-minute movement is recorded and continuous movements are filmed. A score was given only when the animals entered A, B, C once in a row, such as A, B, C or B, C, A or C, A, B (excluding cases such as A, A, B or B, B, A or C, C, C).

[0072]

[0073] As a result, as shown in Figure 6, when Hongsamgyeongokgo and donepezil were administered together, the crossover rate increased compared to the Hongsamgyeongokgo treatment group and the donepezil treatment group, confirming that cognitive ability was improved.

[0074]

[0075] Example 5: Passive avoidance experiment

[0076] 5-1. Manual avoidance experimental device

[0077] The experimental apparatus consisted of a shuttle box (53 cm W × 44 cm H × 33 cm D) with two chambers inside, each with a central guillotine door that served as a door between the two chambers. One chamber contained a very bright light bulb to create an environment that the experimental animals found unpleasant, while the other contained a device capable of delivering an electric shock (scrambled foot-shock) to the entire grid floor.

[0078]

[0079] 5-2. Training trial

[0080] Mice were placed one at a time in one room and given 15 seconds of exploration time. After being stimulated with light and noise from the top of the box, the mice were encouraged to pass through a guillotine door into another quiet, dark room (electric shock room). When the mouse entered the electric shock room, the guillotine door automatically closed and an electric shock was delivered. The shock was delivered with a current of 0.3 mA for 3 seconds. After the mouse received the foot shock, it was removed from the box and returned to its cage. The electric shock was delivered to the experimental and control groups in the same way. Mice that did not enter the shock room within 120 seconds were excluded from the experiment.

[0081]

[0082] 5-3. Memory test (Test trial)

[0083] Mice that received the above training remembered the previous day's shock when receiving an electric shock and thus avoided entering the shock chamber. The longer it took to reach the shock chamber, the better the learning and memory effects of passive avoidance. 24 hours after the above learning test, the mice were given an electric shock, and the time it took to reach the shock chamber (step-through latency time) was measured up to 300 seconds (cut-off time).

[0084]

[0085] As a result, as shown in Figure 7A, the Hongsamgyeongokgo and donepezil treatment group, which was administered a combination of Hongsamgyeongokgo and donepezil, reached the shock chamber after 90 seconds. As shown in Figure 7B, the difference in the time the mice reached the shock chamber on the first day and the time they reached the shock chamber on the next day was almost the same as the time the control group, which was administered only scopolamine, first reached the shock chamber, and it was confirmed that the mice administered a combination of Hongsamgyeongokgo and donepezil had a significantly increased memory compared to the mice administered only Hongsamgyeongokgo and the mice administered only donepezil.

[0086]

[0087] Example 6: Water maze experiment

[0088] A circular water tank (diameter 180 cm, height 65 cm) was filled with water at 22±2℃ to a depth of 45 cm, and ink was added to make the water black. A platform (diameter 4.5 cm, height 43.5 cm) was placed in the center of one of the four sectors of the entire water tank, with the top of the platform located 1.5 cm below the water surface. Learning tests were conducted three times a day for four consecutive days. When the mouse found the platform, it was allowed to remain on the platform for approximately 10 seconds and then returned to its original cage. The next learning test was performed 60 minutes later. If the mouse did not find the platform within 60 seconds, the mouse was left on the platform for 10 seconds and then returned to its original cage. The next learning test was performed 60 minutes later. A probe trial was conducted 24 hours after the last learning test on mice that had completed learning. The test was conducted by removing the platform from the water tank and measuring the time (latency) it took for the platform to remain in the four-minute circle it was placed in for 60 seconds. The experimental results were recorded and analyzed using equipment (video recorder / tracking device) installed on the ceiling above the water tank. The platform's position is shown in Figure 8.

[0089]

[0090] As a result, as shown in FIGS. 9A and 9B, the Hongsamgyeongokgo and donepezil treatment group, which was administered in combination with Hongsamgyeongokgo and donepezil, showed a significant decrease in the speed of finding the quadrant with the platform on the 4th day compared to the control group, and reached the quadrant with the platform as quickly as the untreated group. In addition, as shown in FIG. 9C, it was confirmed that the time spent in Zone 1, the quadrant with the platform, significantly increased in the Hongsamgyeongokgo and donepezil treatment group compared to the donepezil treatment group and the Hongsamgyeongokgo treatment group, and the time spent in Zone P was confirmed to be longer than that of the normal group.

[0091]

[0092]

[0093] The present invention has been described above, focusing on preferred embodiments thereof. Those skilled in the art will appreciate that the present invention can be implemented in modified forms without departing from its essential characteristics. Therefore, the disclosed embodiments should be considered illustrative rather than limiting. The scope of the present invention is set forth in the claims, not the foregoing description, and all differences within the scope equivalent thereto should be construed as being encompassed by the present invention.

Claims

1. A pharmaceutical composition for preventing or treating dementia containing donepezil and Hongsamgyeongokgo.

2. In paragraph 1, The above Hongsamgyeongokgo is composed of red ginseng, white poria, raw rehmannia glutinosa, and honey. A pharmaceutical composition for preventing or treating dementia.

3. In paragraph 2, The above red ginseng, white poria cocos, raw rehmannia glutinosa, and honey are in a weight ratio of 1:1:5:5 to 1:3:10:

10. A pharmaceutical composition for preventing or treating dementia.

4. In paragraph 1, The above donepezil and Hongsamgyeongokgo are in a weight ratio of 1:100 to 1:2,000. A pharmaceutical composition for preventing or treating dementia.

5. In paragraph 1, The above dementia is Alzheimer's type dementia or vascular dementia. Pharmaceutical composition.

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