Traditional Chinese medicine composition for preventing and treating Alzheimer disease as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention provides a traditional Chinese medicine composition for preventing and treating Alzheimer disease as well as a preparation method and application thereof. The traditional Chinese medicine composition is prepared from chrysanthemum, mulberry, gastrodia elata, cistanche, codonopsis pilosula, dogwood, poria cocos and prepared rehmannia root. The invention aims to solve the problem that the effectiveness and the safety of the conventional Alzheimer disease treatment medicine are greatly limited due to the influence of action targets and mechanisms.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a traditional Chinese medicine composition for preventing or treating Alzheimer's disease, a preparation method and application thereof. Background Art

[0002] The pathogenesis of Alzheimer's disease is complex and remains incompletely elucidated. Current research focuses on amyloid deposition, abnormal tau protein phosphorylation, oxidative stress, cholinergic system degeneration, genetic susceptibility, and mitochondrial dysfunction. Currently, the main medications used to treat AD in my country are cholinesterase inhibitors, glutamate receptor antagonists, the China National Medical Products Administration-approved drug mannitol sodium, and immunotherapy targeting amyloid deposition. However, these drugs have significant limitations in both efficacy and safety, depending on their targets and mechanisms of action. Summary of the Invention

[0003] The purpose of the present invention is to provide a traditional Chinese medicine composition for preventing and treating Alzheimer's disease, as well as its preparation method and application, so as to solve the problem that the current therapeutic drugs for Alzheimer's disease are greatly limited in effectiveness and safety due to the influence of their targets and mechanisms of action.

[0004] To solve the above problems, the present invention first provides a traditional Chinese medicine composition for preventing and treating Alzheimer's disease, comprising the following raw materials: chrysanthemum, mulberry, gastrodia elata, cistanche deserticola, codonopsis pilosula, cornus officinalis, poria, and rehmannia root.

[0005] Furthermore, in parts by weight, the weight portions of each raw material are: 5-10 parts of chrysanthemum, 5-15 parts of mulberry, 8-15 parts of gastrodia, 5-12 parts of cistanche, 10-20 parts of codonopsis, 8-15 parts of cornus, 10-20 parts of poria and 10-25 parts of rehmannia.

[0006] Furthermore, in parts by weight, the weight parts of each raw material are: 5 parts of chrysanthemum, 10 parts of mulberry, 10 parts of gastrodia elata, 10 parts of cistanche, 10 parts of codonopsis pilosula, 10 parts of cornus officinalis, 15 parts of poria and 15 parts of rehmannia root.

[0007] The present invention also provides a traditional Chinese medicine preparation for preventing and treating Alzheimer's disease, which is composed of the traditional Chinese medicine composition described in the above technical solution and pharmaceutical excipients.

[0008] Furthermore, the dosage forms of the Chinese medicine preparation include oral liquid, ointment, tablet, capsule, granule, granule, pill, powder, tea bag, beverage or injection.

[0009] The present invention also provides a preparation method of the traditional Chinese medicine preparation described in the above technical solution, comprising the following steps: an ethanol heating reflux extraction method, grinding the raw materials of the traditional Chinese medicine composition into fine powder, adding the powder into a round-bottom flask containing 70% ethanol, placing the powder into a water bath, connecting the flask to cold water, and reflux heating at 70°C for 1.5 hours; or an ultrasonic-assisted extraction method, grinding the raw materials of the traditional Chinese medicine composition into fine powder, placing the powder into a conical flask, adding 70% ethanol into the container, vortex stirring, and extracting the powder in an ultrasonic cleaner at 60°C for 40 minutes; or a surfactant-assisted ultrasonic enzyme method, grinding the raw materials of the traditional Chinese medicine composition into fine powder, uniformly mixing the powder with a non-ionic surfactant and 0.2% complex enzyme in a PTFE container, wherein the content of cellulase to pectinase in the complex enzyme is 4:6; sealing the container, placing the powder in a constant temperature water bath at 60°C, incubating the powder for enzymatic hydrolysis for 90 minutes, and then placing the reaction compound in an ultrasonic cleaner for ultrasonic treatment for 40 minutes.

[0010] The present invention also provides an application of the Chinese medicine composition described in the above technical solution, which is used as a medicine, food or health product for treating or preventing Alzheimer's disease and related cognitive impairment diseases.

[0011] According to the present invention, a traditional Chinese medicine composition for preventing and treating Alzheimer's disease, its preparation method and application are provided. The traditional Chinese medicine composition has multiple components, multiple targets and multiple pathways to jointly exert an overall effect. The prescription that can simultaneously strengthen the spleen, nourish the kidney and calm the liver is selected, and has a significant therapeutic effect on Alzheimer's disease. In addition, the traditional Chinese medicine has few toxic and side effects, and long-term use can treat both the symptoms and the root cause. It is suitable for daily health care use of middle-aged and elderly people. DETAILED DESCRIPTION

[0012] In order to make the above-mentioned objects, features and advantages of the present invention more clearly understood, the following is a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0013] With the widespread development of traditional Chinese medicine (TCM) and advancements in extraction technology for its active ingredients, the unique advantages of TCM, through its multi-component, multi-target, and multi-pathway approach, have been further explored, achieving even more remarkable results in preventing and treating multi-etiological and complex diseases. my country boasts a rich natural resource of Chinese herbal medicines, and studies have shown that many of these herbs have significant effects in both the prevention and treatment of Alzheimer's disease. These herbs also have minimal toxicity and side effects, and long-term use can address both the underlying cause and the underlying disease, making them suitable for daily health care use among middle-aged and elderly people.

[0014] Alzheimer's disease, or AD for short, falls under the category of "dementia" and "forgetfulness" in Traditional Chinese Medicine (TCM). The core TCM location of Alzheimer's disease lies in the brain, and its pathological evolution often involves imbalances in the function of multiple internal organs. The core pathogenesis can be summarized as gradual depletion of brain marrow and dysfunction of the spirit. From the perspective of syndrome differentiation between deficiency and excess, deficiency is primarily characterized by kidney-essence deficiency and deficiency of both the heart and spleen. Kidney-essence deficiency leads to insufficient brain orifices; spleen-yang deficiency results in insufficient Qi and blood production, impairing brain nourishment. Liver depression and Qi stagnation are common in the elderly, leading to insufficient liver function and dispersal. This can easily transform Yang into fire and generate wind, disturbing the clear orifices and exacerbating brain malnutrition. Clinical syndromes are not singular; they can manifest as either pure deficiency or excess, or more often present as complex combinations of both deficiency and excess, such as spleen-kidney deficiency with hyperactivity of liver Yang. Therefore, the present invention selects a prescription that can simultaneously strengthen the spleen, nourish the kidney, and calm the liver, with the aim of providing a traditional Chinese medicine composition with better prevention and treatment effects on Alzheimer's disease.

[0015] This embodiment provides a Chinese medicine composition, a Chinese medicine preparation and a preparation method thereof for preventing and treating Alzheimer's disease, and demonstrates their therapeutic effects through experiments.

[0016] 1. Ingredients and preparation methods of Chinese medicine compositions

[0017] The Chinese medicine composition of this embodiment comprises chrysanthemum, mulberry, gastrodia, cistanche, codonopsis, cornus officinalis, poria, and rehmannia root, all of which are known to those skilled in the art. A preferred composition comprises 5g of chrysanthemum, 10g of mulberry, 10g of gastrodia, 10g of cistanche, 10g of codonopsis, 10g of cornus officinalis, 15g of poria, and 15g of rehmannia root.

[0018] The Chinese medicine preparation can be supported by the Chinese medicine composition and pharmaceutical excipients, and its dosage forms include but are not limited to oral liquid, ointment, tablet, capsule, granule, granule, pill, powder, tea bag, beverage or injection.

[0019] The extraction methods of combined Chinese herbal medicine extracts or Chinese herbal medicine preparations include the following three methods: the raw materials are weighed, mixed, and then crushed into fine powder. The final extract is named X8:

[0020] Ethanol heating reflux extraction (HRE): The fine powder of the composition was added to a round-bottom flask containing 450 mL of 70% ethanol, which was then placed in a water bath connected to cold water and refluxed at 70° C. for 1.5 h.

[0021] Ultrasound-assisted extraction (UAE): The fine powder of the composition was placed in a conical flask, 450 mL of 70% ethanol was added, and after vortex stirring, the mixture was extracted in an ultrasonic cleaner at a temperature of 60° C. and an ultrasonic power of 465 W for 40 minutes.

[0022] Surfactant assisted ultrasound enzyme method (EUASE): The fine powder of the composition was mixed with 450 mL of Triton X-100 solution (0.24 mmol / L) and 0.2% complex enzyme [m (cellulase) : m (pectinase) = 4:6] in a PTFE container. The container was covered and incubated for enzymatic hydrolysis in a constant temperature water bath at 60°C for 90 min. The reaction mixture was then placed in an ultrasonic cleaner and ultrasonically treated at 465 W for 40 min.

[0023] 2. Experimental methods and conclusions

[0024] 2.1 Experimental animals

[0025] One hundred and fifty 8-week-old SPF Sprague-Dawley rats weighing 200-220 g were selected, half male and half female. The animal room was controlled at 22-25°C, with natural light and alternating light and dark cycles of 12 h each, and free access to food and water.

[0026] 2.2 Drugs and reagents

[0027] Dissolve 1 mg of Aβ1-42 in PBS (phosphate-buffered saline). Aβ1-42, a 42-amino acid polypeptide fragment produced by enzymatic cleavage of amyloid precursor protein (APP), is dissolved in PBS. Incubate at 37°C for 72 hours to produce an oligomeric Aβ1-42 solution.

[0028] Prepare materials and equipment, including: chrysanthemum, mulberry, gastrodia, cistanche, codonopsis, cornus officinalis, poria, rehmannia root; isoflurane; methanol, acetonitrile, formic acid; ultrapure water, and all other reagents are analytical grade; KQ-600E ultrasonic cleaner; 1 / 100,000 electronic analytical balance; electric constant temperature water bath; micro plant sample grinder; and microplate reader.

[0029] Extract X8 was prepared separately by the above three extraction methods for later use.

[0030] 2.3 Experimental Procedure

[0031] 2.3.1 Construction of AD rat model

[0032] 150 rats were divided equally into a blank group, a control group, a HRE group, a UAE group, and an EUASE group. All rats were anesthetized with isoflurane inhalation. The skin was cut open along the midline of the rat brain to expose the skull. A 1.0 mm drill hole was drilled 3.6 mm posterior to the bregma and 2 mm lateral to the midline. The specific location was determined by referring to the "Rat Brain Stereotaxic Atlas." A 5 μL Aβ1-42 solution (2 g / L) was injected into the bilateral hippocampus of the rat brain at a rate of 0.1 μL / min. The blank group received no treatment. After injection, the skin was sutured and the rats were allowed to recover for 7 days.

[0033] 2.3.2 Identification of AD rats

[0034] Three weeks after modeling, Morris water maze navigation test was performed on each group to detect the escape latency of the rats. The average escape latency of the blank group rats trained four times was used as the control (A), and the average escape latency of the remaining rats on the same day was tested (B). If (BA) / B>20%, the AD rat model was successfully established.

[0035] 2.3.3AD rat intervention

[0036] AD rats in the HRE, UAE, and EUASE groups were gavaged daily with 75 mg / kg of X8 extracted using the corresponding extraction method. AD rats in the blank and control groups were given an equal volume of 0.9% sodium chloride solution. All five groups received gavages starting on day 7 after successful model establishment, with daily gavages for 8 consecutive weeks.

[0037] 2.3.4 Learning and memory behavior test of rats

[0038] Prepare experimental equipment in advance, including a water tank, a camera, a monitor, and a computer image acquisition and analysis system. After oral administration, analyze the rats' behavior using the Morris water maze (MWM) in sequence: a visual platform test, a navigation test, and a spatial exploration test. Record the results.

[0039] (1) Visual platform experiment: The platform is exposed to the water surface. After the rat is placed in the swimming pool, if it can swim to the platform without difficulty, the subsequent experiment can be started to eliminate the influence of the rat's swimming ability and vision.

[0040] (2) Positioning and navigation experiment: Rats were placed in a swimming pool for daily training. They were required to enter the water from four different entry points with their heads facing the walls of the pool. The training was conducted at a fixed time every day, with each rat trained four times with an interval of 240 seconds between each training session. The training lasted for 7 days, and the behavior and swimming direction of the rats were observed. In this experiment, the platform was hidden below the water surface, and it was defined as a successful settlement if the rat found the platform during the swimming process and stayed on it for 5 seconds. Within 120 seconds, the rat successfully found the hidden platform and landed on it to stay. The time spent in this process was defined as the escape latency and recorded. If the rat failed to find the platform or could not successfully land on it and stay for 5 seconds within 120 seconds, it was considered a settlement and landing failure. At this time, the rat was guided to learn and remember, and was guided to stay on the platform for 10 seconds, and the escape latency was recorded as 120 seconds.

[0041] (3) Spatial exploration experiment: The trained rats were placed back into the pool on the last day of the experiment. The hidden platform under the pool was removed, and the rats were allowed to enter the water at any point along the quadrant of the original platform. The swimming trajectory of the rats in the pool was observed, and the number of times the rats passed through the original quadrant within 120 s and the time they stayed in each quadrant were recorded.

[0042] 2.4 Statistical analysis

[0043] SPSS 26.0 software was used for statistical analysis. Data were expressed as mean ± standard deviation. Homogeneity of variance was tested. When the variances were equal, the LSD-t test was used for comparison between two groups. When the variances were unequal, the Bunnett's T3 test was used for comparison between two groups. One-way analysis of variance was used for comparison between multiple groups. P < 0.05 was considered statistically significant.

[0044] 2.5X8 improves cognitive ability in AD model rats

[0045] The results of the navigation experiment showed that the escape latency of rats in the blank, control, HRE, UAE, and EUASE groups decreased in a time-dependent manner, with significant differences beginning on the first day of the experiment. The escape latency of rats in the HRE, UAE, and EUASE groups was significantly shorter than that in the control group (P < 0.05), as shown in Table 1. The escape latency of rats in the EUASE group was significantly shorter than that in the HRE and UAE groups (P < 0.01).

[0046] In the spatial exploration experiment, compared with the control group, the latency to reach the platform and the number of crossings in the target area in the HRE, UAE and EUASE groups were statistically significant (P < 0.05). In the comparison between the HRE, UAE and EUASE groups, the latency of the EUASE group rats to explore the target platform was significantly reduced (P < 0.05), as shown in Table 2; the number of crossings in the target area was significantly increased, and the difference was statistically significant (P < 0.05), suggesting that the combined Chinese herbal extract X8 has the effect of improving the cognition and memory of experimental rats, and the X8 obtained by the EUASE purification method has the best effect.

[0047]

[0048]

[0049] Table 1 Results of the Morris water maze navigation test (mean ± SD)

[0050]

[0051] Table 2 Morris water maze spatial exploration test results (mean ± SD)

[0052] AD is a common neurodegenerative disease in the elderly. Its pathological hallmarks are the formation of large amounts of Aβ and its deposition in the hippocampus and cortex, forming senile plaques, accompanied by oxidation of lipids, proteins, and nucleic acids in neurons. Neurons have a higher metabolic rate than other cells and are therefore more susceptible to oxidative damage. Oxidative stress in the context of AD is primarily associated with the destruction of biomolecules and membrane integrity in neuronal membranes. Oxidative stress can trigger a vicious cycle in the body, as the production of reactive oxygen species (ROS) can damage biomolecules, leading to increased ROS accumulation. Given the complex pathogenesis of AD and the interplay between related indicators, multi-component combination therapy to modulate multiple indicators is an effective approach for treating AD. Traditional Chinese medicine (TCM) utilizes multiple components to simultaneously regulate multiple targets and multiple links in the disease network, achieving high efficacy while minimizing the potential toxic side effects of single-target chemical drugs. It is an important source of new anti-AD drugs.

[0053] The Morris water maze experiment showed that after AD rats were treated with the combined Chinese medicine extract X8, the escape latency was significantly shortened, the time they stayed in the target quadrant and the number of times they crossed the platform were significantly increased. It can be seen that the learning, memory and spatial exploration abilities of AD rats were significantly improved.

[0054] These results demonstrate that the disclosed Chinese herbal composition significantly improves the spatial learning and exploration abilities of Alzheimer's rats, demonstrating a significant therapeutic effect. Chinese herbal compound extracts are emerging as a new model for drug development, and this study provides further experimental support for the future development of Chinese herbal compound extracts for the treatment of AD.

[0055] The Chinese medicine composition provided in this embodiment can be used to treat or prevent Alzheimer's disease and related cognitive impairment diseases, including the preparation of medicines, foods or health products.

[0056] Although the present invention is disclosed as above, the present invention is not limited thereto. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the scope defined by the claims.

[0057] Finally, it should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or device comprising a series of elements includes not only those elements, but also other elements not explicitly listed, or elements inherent to such process, method, article, or device. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of additional identical elements in the process, method, article, or device comprising the element.

[0058] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0059] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one 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 present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A Chinese medicine composition for preventing and treating Alzheimer's disease, characterized in that: The invention comprises the following raw materials: chrysanthemum, mulberry, gastrodia elata, cistanche deserticola, codonopsis pilosula, cornus officinalis, poria cocos and rehmannia root.

2. The Chinese medicine composition according to claim 1, characterized in that In parts by weight, the weight portions of the raw materials are: 5-10 parts of chrysanthemum, 5-15 parts of mulberry, 8-15 parts of gastrodia, 5-12 parts of cistanche, 10-20 parts of codonopsis, 8-15 parts of cornus, 10-20 parts of poria and 10-25 parts of rehmannia.

3. The Chinese medicine composition according to claim 2, characterized in that In parts by weight, the weight parts of each raw material are: 5 parts of chrysanthemum, 10 parts of mulberry, 10 parts of gastrodia elata, 10 parts of cistanche, 10 parts of codonopsis pilosula, 10 parts of cornus officinalis, 15 parts of poria and 15 parts of rehmannia root.

4. A Chinese medicine preparation for preventing and treating Alzheimer's disease, characterized in that: The invention is composed of the traditional Chinese medicine composition according to claim 1 or 2 and pharmaceutical excipients.

5. The Chinese medicine preparation according to claim 4, characterized in that The dosage forms of the Chinese medicine preparation include oral liquid, ointment, tablet, capsule, granule, pill, powder, tea bag, beverage or injection.

6. A method for preparing the Chinese medicine preparation according to claim 4, characterized in that: The process includes the following steps: Ethanol heating reflux extraction method: grind the raw materials of the traditional Chinese medicine composition into fine powder, add it into a round-bottom flask containing 70% ethanol, place it in a water bath, connect it to cold water, and reflux it at 70°C for 1.5h; Alternatively, ultrasonic-assisted extraction method, the raw materials of the Chinese medicine composition are crushed into fine powder, placed in a conical flask, 70% ethanol is added to the container, vortex-stirred, and then extracted in an ultrasonic cleaner at 60°C for 40 minutes; Alternatively, a surfactant-assisted ultrasonic enzymatic method is used, wherein the raw materials of the traditional Chinese medicine composition are pulverized into fine powder, and the powder is uniformly mixed with a non-ionic surfactant and 0.2% complex enzyme in a PTFE container, wherein the content of cellulase to pectinase in the complex enzyme is 4:6; the container is sealed, and the enzymatic hydrolysis is incubated in a constant temperature water bath at 60°C for 90 minutes, and then the reaction compound is placed in an ultrasonic cleaner for ultrasonic treatment for 40 minutes.

7. Use of the Chinese medicine composition according to any one of claims 1 to 3 as a medicine, food or health product for treating or preventing Alzheimer's disease and related cognitive disorders.

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