Application of allose in preparation of medicine for treating Alzheimer's disease

Abstract

The invention discloses application of allose in preparation of a medicine for treating Alzheimer's disease, and relates to the technical field of biological medicine. An experiment is carried out on a fruit fly model with the Alzheimer's disease, and the allose shows that the movement function of the fruit fly with the Alzheimer's disease can be effectively improved, the service life of the fruit fly with the Alzheimer's disease is effectively prolonged, and death of brain neurons of the fruit fly with the Alzheimer's disease is reduced; therefore, the allose is proved to have a positive effect on disease control of patients with neurodegenerative diseases, and has important application potential.

Description

Technical Field

[0001] The present invention relates to the field of biological medicine technology, and particularly to the application of allose in the preparation of drugs for treating Alzheimer's disease. Background Art

[0002] Alzheimer's Disease (AD), as a globally prevalent neurodegenerative disease, its pathological features mainly include plaques formed by extracellular deposition of β-amyloid protein (Aβ) and neurofibrillary tangles (NFTs) caused by abnormal aggregation of intracellular tau protein. These lesions lead to the loss of normal functions of neurons, further causing brain tissue atrophy and significant degradation of cognition and memory. Although the etiology of AD has not been fully clarified at present, it is generally believed that it is affected by the combined action of multiple factors, including genetic factors, the aging process, metabolic disorders, and various environmental influences. At present, the clinical diagnosis of AD relies on detecting changes in the levels of Aβ and tau proteins in cerebrospinal fluid or through specific imaging methods. However, these methods are costly, complex to operate, and invasive, making it difficult to meet the extensive early screening needs. Although the detection of non-invasive markers in blood tests is constantly advancing, it is still vulnerable to other pathological interferences and its accuracy remains to be improved.

[0003] In the field of drug treatment for AD, interventions targeting the pathological features of Aβ and tau proteins have been continuously explored. Although the Aβ-targeted drug Aducanumab has been approved for marketing, significant side effects have been found in its actual clinical application, especially amyloid-related imaging abnormalities (ARIA) such as brain edema and intracerebral hemorrhage can be triggered. Drugs targeting tau protein aggregation such as HMTM have shown certain efficacy in clinical trials, but the safety of its long-term use remains to be further verified. In addition, the research directions of AD drugs also cover the regulation of neuroinflammation, the alleviation of oxidative stress, the maintenance of neurotransmitter homeostasis, etc. At present, only a few acetylcholinesterase inhibitors and NMDA receptor antagonists have been approved for clinical use. These drugs can only transiently relieve the symptoms of AD and are often accompanied by adverse reactions such as nausea, vomiting, and hepatotoxicity. Considering the particularity of AD patients and the limitations of existing treatment methods, it is particularly important to develop new treatment methods with neuroprotective effects. Summary of the Invention

[0004] The present invention deeply studies the mechanism of action of allose in neurodegenerative diseases and explores its possibility as a therapeutic drug for neurodegenerative diseases, opening up a new way for the treatment of neurodegenerative diseases.

[0005] Therefore, the present invention provides the application of allose in the preparation of drugs for treating neurodegenerative diseases.

[0006] Furthermore, the allose is used for preparing a drug for improving the motor ability of patients with neurodegenerative diseases.

[0007] Furthermore, the allose is used for preparing a drug for prolonging the lifespan of patients with neurodegenerative diseases.

[0008] Furthermore, the allose is used for preparing a drug for reducing the death of brain neurons in patients with neurodegenerative diseases.

[0009] Furthermore, the neurodegenerative disease includes Alzheimer's disease, and the allose is used for preparing a drug for inhibiting the vacuolization of brain neurons in patients with Alzheimer's disease.

[0010] Furthermore, the allose is the only active ingredient or one of the active ingredients in the drug.

[0011] Furthermore, the drug is a combination of the allose and its pharmaceutically acceptable carrier or diluent, or, the drug includes the allose and its pharmaceutically acceptable carrier or diluent; the dosage form of the drug is powder, tablet, capsule, oral liquid, injection or aerosol.

[0012] Furthermore, when the drug is an oral liquid, injection or aerosol, the concentration of the allose is 30 μg / mL - 60 μg / mL.

[0013] Furthermore, the drug includes the allose and its pharmaceutically acceptable diluent, and also includes other active ingredients; wherein, the concentration of the other active ingredients is 0.05 mg / mL - 0.15 mg / mL.

[0014] Furthermore, the drug is a mixture of the allose and water, and the concentration of the allose is 30 μg / mL - 60 μg / mL.

[0015] Compared with the prior art, the present invention has at least the following beneficial effects:

[0016] The application of allose provided by the present invention in the preparation of drugs for treating neurodegenerative diseases. Through experiments on Drosophila models of Alzheimer's disease, allose has been shown to be able to effectively improve the motor function of Drosophila with Alzheimer's disease, effectively prolong the lifespan of Drosophila with Alzheimer's disease, and reduce the death of brain neurons in Drosophila with Alzheimer's disease, thus confirming that allose has a positive effect on the control of the condition of patients with neurodegenerative diseases, especially on the control of the condition of patients with Alzheimer's disease, and has important application potential. And allose, as a natural monosaccharide, has low toxicity and good metabolic properties, and shows excellent clinical potential in terms of safety and tolerance. Brief Description of the Drawings

[0017] Figure 1 The effects of the drug containing 50 μg / mL allose under the culture condition of 29°C provided by the embodiments of the present invention on the motor ability of female F1 generation Aβ-type and wild-type (WT) Drosophila

[0018] Figure 2 The effects of the drugs containing 25 μg / mL and 50 μg / mL allose under the culture condition of 25°C provided by the embodiments of the present invention on the motor ability of female F1 generation Aβ-type and wild-type (WT) Drosophila

[0019] Figure 3 The effects of the drug containing 50 μg / mL allose under the culture condition of 29°C provided by the embodiments of the present invention on the lifespan of female F1 generation Aβ-type and wild-type (WT) Drosophila

[0020] Figure 4 The effects of the drugs containing 25 μg / mL and 50 μg / mL allose under the culture condition of 25°C provided by the embodiments of the present invention on the lifespan of female F1 generation Aβ-type and wild-type (WT) Drosophila

[0021] Figure 5 The effects of the drug containing allose provided by the embodiments of the present invention on the H&E staining results of female F1 generation Aβ-type and wild-type (WT) Drosophila

[0022] Figure 6 The effects of the drug containing allose provided by the embodiments of the present invention on the number of vacuoles in the brains of female F1 generation Aβ-type and wild-type (WT) Drosophila Detailed implementation manners

[0023] To better understand the above technical solutions, the technical solutions of the embodiments of the present application will be described in detail below through the accompanying drawings and specific embodiments. It should be understood that the specific features in the embodiments of the present application and the embodiments are detailed descriptions of the technical solutions of the embodiments of the present application, rather than limitations on the technical solutions of the present application. Without conflict, the technical features in the embodiments of the present application and the embodiments can be combined with each other.

[0024] The embodiments of the present invention provide the application of allose in the preparation of drugs for treating neurodegenerative diseases.

[0025] In a feasible implementation manner, allose is used to prepare a drug for improving the motor ability of patients with neurodegenerative diseases. Allose is used to prepare a drug for improving the motor dysfunction caused by the excessive accumulation of Aβ protein in patients with neurodegenerative diseases. The neurodegenerative diseases include but are not limited to Alzheimer's disease.

[0026] In a feasible embodiment, allose is used to prepare a drug for prolonging the lifespan of patients with neurodegenerative diseases. Allose is used to prepare a drug for improving the shortened lifespan caused by the toxicity of Aβ protein in patients with neurodegenerative diseases. The neurodegenerative diseases include, but are not limited to, Alzheimer's disease.

[0027] In a feasible embodiment, allose is used to prepare a drug for reducing neuronal death in the brains of patients with neurodegenerative diseases. Allose is used to prepare a drug for reducing neuronal injury and death caused by the excessive accumulation of Aβ protein in patients with neurodegenerative diseases. The neurodegenerative diseases include, but are not limited to, Alzheimer's disease.

[0028] Furthermore, allose is used to prepare a drug for treating neuronal vacuolization in the brains of patients with neurodegenerative diseases. Allose is used to prepare a drug for inhibiting neuronal vacuolization in the brains of Alzheimer's disease patients.

[0029] In a feasible embodiment, allose is the sole active ingredient or one of the active ingredients in a drug for treating neurodegenerative diseases. Optionally, allose is D-allose or L-allose.

[0030] In a feasible embodiment, the drug for treating neurodegenerative diseases is a combination of allose and its pharmaceutically acceptable carrier or diluent. Allose combined with its pharmaceutically acceptable carrier or diluent is used to improve the stability and absorbability of allose. Optionally, allose is anhydrous allose. Optionally, the preparation method of the drug for treating neurodegenerative diseases is to mix allose with its pharmaceutically acceptable excipients to prepare a dosage form suitable for patients to take.

[0031] Furthermore, the drug for treating Alzheimer's disease is a combination of allose and its pharmaceutically acceptable carrier or diluent. Allose combined with its pharmaceutically acceptable carrier or diluent is used to improve the stability and absorbability of allose. Optionally, allose is anhydrous allose. Optionally, the preparation method of the drug for treating Alzheimer's disease is to mix allose with its pharmaceutically acceptable excipients to prepare a dosage form suitable for patients to take.

[0032] In a feasible embodiment, the drug for treating neurodegenerative diseases has a dosage form of powder, tablet, capsule, oral liquid, injection or aerosol, and the form of the pharmaceutical preparation is adapted to the administration method.

[0033] Furthermore, the drug for treating Alzheimer's disease has a dosage form of powder, tablet, capsule, oral liquid, injection or aerosol, and the form of the pharmaceutical preparation is adapted to the administration method.

[0034] In a feasible embodiment, when the drug for treating neurodegenerative diseases is an oral liquid, an injection or an aerosol: the drug for treating neurodegenerative diseases is a combination of allose and its pharmaceutically acceptable diluent, or, the drug for treating neurodegenerative diseases comprises the allose and its pharmaceutically acceptable carrier or diluent; wherein, the concentration of allose is 30 μg / mL - 60 μg / mL, and preferably the concentration of allose is 50 μg / mL.

[0035] Further, when the drug for treating Alzheimer's disease is an oral liquid, an injection or an aerosol: the drug for treating Alzheimer's disease is a combination of allose and its pharmaceutically acceptable diluent, or, the drug for treating Alzheimer's disease comprises the allose and its pharmaceutically acceptable carrier or diluent; wherein, by evaluating the effect of allose on rescuing the motor ability of Aβ flies, the concentration of allose is screened, and it is obtained that the concentration of allose is 30 μg / mL - 60 μg / mL, and preferably the concentration of allose is 50 μg / mL.

[0036] In a feasible embodiment, the drug for treating neurodegenerative diseases comprises: allose, a pharmaceutically acceptable diluent of allose, and other active ingredients; wherein, the concentration of the other active ingredients is 0.05 mg / mL - 0.15 mg / mL, and preferably the concentration of the other active ingredients is 0.1 mg / mL.

[0037] Further, the drug for treating Alzheimer's disease comprises: allose, a pharmaceutically acceptable diluent of allose, and other active ingredients; wherein, the concentration of the other active ingredients is 0.05 mg / mL - 0.15 mg / mL, and preferably the concentration of the other active ingredients is 0.1 mg / mL.

[0038] In a feasible embodiment, the drug for treating neurodegenerative diseases is a mixture of allose and water, the concentration of allose is 30 μg / mL - 60 μg / mL, and preferably the concentration of allose is 50 μg / mL. The preparation method is to dissolve allose in an appropriate amount of water to obtain it.

[0039] Further, the drug for treating Alzheimer's disease is a mixture of allose and water, the concentration of allose is 30 μg / mL - 60 μg / mL, and preferably the concentration of allose is 50 μg / mL. The preparation method is to dissolve allose in an appropriate amount of water to obtain it.

[0040] Embodiments of the present invention provide a new use of allose in the preparation of drugs for treating neurodegenerative diseases. Specifically, allose is used to prepare drugs for improving the motor ability of patients with neurodegenerative diseases, drugs for prolonging the lifespan of patients with neurodegenerative diseases, and / or drugs for reducing neuronal death in the brains of patients with neurodegenerative diseases. Neurodegenerative diseases include Alzheimer's disease, and allose can effectively inhibit the "vacuolization" phenomenon of brain neurons.

[0041] Allose (D-Alulose) is a natural hexose monosaccharide with the chemical formula C 6 H 12 O 6 , with a molecular weight of 180.16. It is a white, odorless crystal with high water solubility and low calorie characteristics, and thus has broad application potential in food and drug preparation. The structural formula is as follows:

[0042]

[0043] In the treatment research of neurodegenerative diseases, including Alzheimer's disease (AD), allose exhibits significant neuroprotective effects. Relevant experiments in this application show that allose can significantly reduce the mortality rate of neurons in the brains of AD model flies. Through experiments in the fly model, it is found that allose can effectively improve motor ability and prolong its lifespan. Specifically, allose effectively inhibits the "vacuolization" phenomenon of brain neurons in AD model flies. Brain "vacuolization" refers to the round cavity structure observed in the brain tissue sections of flies, representing the degeneration or death of neurons. This phenomenon is one of the typical pathological characteristics in both AD patients and AD models.

[0044] Embodiments of the present invention show that allose, as a natural low-toxicity component, has great application potential, providing new possibilities for the development of safe and effective drugs for neurodegenerative diseases, especially drugs for the treatment of Alzheimer's disease.

[0045] Example 1

[0046] 1.1 Construction of the fly model: In this embodiment, the Drosophila strain Elav-Gal4 capable of specifically expressing a specific gene in brain neurons is combined with the Drosophila strain UAS-Aβ42 overexpressing amyloid-β protein (Aβ42) for hybridization to construct an Alzheimer's disease (AD) fly model. After hybridization, the F1 generation Elav>Aβ42 flies are obtained to overexpress Aβ42 in brain neurons and simulate the pathological characteristics of AD. At the same time, the Drosophila strain Elav-Gal4 is hybridized with the wild-type Drosophila W1118 to produce the F1 generation Elav>W1118 flies as the control group to facilitate the comparison of the effects of different treatments on AD flies.

[0047] 1.2 Preparation of allose drug: First, dissolve allose in an appropriate amount of water to prepare a stock solution. Subsequently, mix the stock solution with Drosophila food at a volume ratio of 1:1000 and dilute it to different working concentrations to prepare Drosophila food containing allose. The specific preparation scheme is shown in Table 1.

[0048] Table 1 Working concentrations of allose drug

[0049] Drug Mother liquor concentration Working concentration Allose 50mg / mL 50μg / mL

[0050] 1.3 Drug administration treatment: Raise the F1 generation of wild-type Drosophila (Elav>W1118) and Drosophila overexpressing Aβ42 (Elav>Aβ42) separately in food containing different concentrations of allose, and control the breeding temperatures at 25°C and 29°C respectively. To ensure the effectiveness of the experiment, replace the fresh allose-treated food every three days to avoid abnormal death of Drosophila caused by food spoilage.

[0051] 1.4 Experimental methods

[0052] Measurement of locomotor ability: Raise the F1 generation of wild-type Drosophila and Elav>Aβ42 Drosophila separately in the corresponding drug-administered food and breed them at 25°C and 29°C for 2 - 3 weeks. During measurement, pour each group of Drosophila into a graduated Drosophila tube, tap the tabletop to make the Drosophila gather at the bottom of the tube, and start timing. Within 5 seconds, count the Drosophila that have climbed over and reached the specified scale line, and calculate the proportion of Drosophila that have crossed the line to the total number of Drosophila. Each group should conduct at least 3 independent replicate experiments, with a total of 60 Drosophila.

[0053] Measurement of lifespan: Raise the F1 generation of wild-type Drosophila and Elav>Aβ42 Drosophila separately in the corresponding drug-administered food and breed them at 25°C and 29°C until all Drosophila die. Replace the fresh food every 3 days and record the number of Drosophila deaths each time. Finally, calculate their survival rates and plot graphs. Each group should conduct at least 5 independent replicate experiments, with a total of 100 Drosophila.

[0054] H&E staining of the brain: Raise the F1 generation of wild-type Drosophila and Elav>Aβ42 Drosophila separately in the corresponding drug-administered food. After breeding at 29°C for 2 - 3 weeks, perform H&E staining of the brain. Clamp the Drosophila side by side in a Drosophila clamp, with about 20 Drosophila in each group. After fixation and gradient dehydration, perform paraffin embedding. After completing the paraffin sections, perform dewaxing, gradient hydration, hematoxylin and eosin staining, alcohol dehydration, xylene transparency, and neutral gum mounting. Finally, observe under an ordinary microscope and record the number of brain vacuolizations in each group. At least 10 complete brain sections should be counted for each group of experiments.

[0055] 1.5 Statistical analysis: T-test was used for variance comparison analysis, and all values were expressed as mean ± standard error. When *P < 0.05, **P < 0.01, ***P < 0.001, the data differences were considered significant.

[0056] 1.6 Experimental results

[0057] 1.6.1 Effect of allose on the locomotor ability of AD Drosophila: Compared with normal food (NF), after treatment with 50 μg / mL allose, the locomotor ability of Aβ Drosophila was significantly improved, indicating that allose has a positive effect on delaying the lifespan of Drosophila models of Alzheimer's disease. As Figure 1 shown, Aβ Drosophila cultured at 29 °C showed significant locomotor retardation, and its locomotor ability was significantly decreased compared with wild-type Drosophila. After feeding with food containing 50 μg / mL allose, the locomotor ability of Aβ Drosophila was significantly improved. As Figure 2 shown, the experiment was repeated at 25 °C using 25 μg / mL and 50 μg / mL allose. 25 μg / mL allose had no rescue effect on locomotion, indicating that 50 μg / mL allose had a better effect than 25 μg / mL allose treatment. Specifically, Aβ Drosophila in the allose treatment group showed faster crawling speed and higher crawling ratio in the locomotion test. This result indicates that allose can effectively improve the locomotor ability of Drosophila with Alzheimer's disease, thereby reversing the locomotor dysfunction caused by overexpression of Aβ protein to a certain extent.

[0058] 1.6.2 Effect of allose on the lifespan of AD Drosophila: Compared with normal food (NF), after treatment with allose, the lifespan of Aβ Drosophila was significantly extended, indicating that allose has a positive effect on delaying the lifespan of Drosophila models of Alzheimer's disease. As Figure 3 shown, under the condition of culturing at 29 °C, when the lifespan curve reached 50% survival rate, the average survival days of wild-type Drosophila were about 26 days, while the average survival days of Aβ Drosophila were about 21 days, showing that the lifespan of Aβ Drosophila was significantly lower than that of wild-type Drosophila. After treatment with allose, the survival days of Aβ Drosophila were significantly extended. At 50% survival rate, its average survival days increased by about 4 days. This indicates that the administration of allose can effectively extend the lifespan of Drosophila with Alzheimer's disease and offset the lifespan shortening effect caused by the toxicity of Aβ protein. Similarly, as Figure 4 , under the culture condition of 25 °C, the experiment was repeated with 25 μg / mL and 50 μg / mL allose, and 50 μg / mL allose had a better effect than 25 μg / mL allose treatment.

[0059] 1.6.3 Effect of allose on neuronal death in the brains of AD Drosophila: As Figure 3 and Figure 4As shown, through the H&E staining results, it can be seen that the brain tissue of Aβ fruit flies shows obvious vacuolization, and compared with wild-type fruit flies, a large number of their brain neurons die. After treatment with allose, the vacuolization in the brains of Aβ fruit flies is significantly reduced, indicating that the neuronal death has been effectively improved. This result suggests that allose has a protective effect in preventing neuronal damage and death caused by overexpression of Aβ protein.

[0060] It is easy for those skilled in the art to understand that, on the premise of no conflict, the above advantageous ways can be freely combined and superimposed. The above are only the preferred embodiments of the present application and are not intended to limit the present application. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present application shall be included in the protection scope of the present application. The above is only the preferred implementation manner of the present application. It should be noted that for those of ordinary skill in the art, without departing from the technical principle of the present application, several improvements and variations can be made, and these improvements and variations should also be regarded as the protection scope of the present application.

Claims

1. Application of allose in the preparation of drugs for treating neurodegenerative diseases.

2. The use according to claim 1, characterized in that: The allose is used for preparing a medicine for improving the motor ability of patients with neurodegenerative diseases.

3. The use according to claim 1, characterized in that: The allose is used for preparing a medicine for prolonging the life span of patients with neurodegenerative diseases.

4. The use according to claim 1, characterized in that: The allose is used for preparing a medicine for reducing the death of brain neurons in patients with neurodegenerative diseases.

5. The use according to any one of claims 1 to 4, characterized in that: The neurodegenerative disease includes Alzheimer's disease, and the allose is used to prepare a drug for inhibiting the cavitation phenomenon of neurons in the brain of patients with Alzheimer's disease.

6. The use according to any one of claims 1 to 4, characterized in that: The allose is the only active ingredient or one of the active ingredients in the medicine.

7. The use according to any one of claims 1 to 4, characterized in that: The drug is a combination of the allose and a pharmaceutically acceptable carrier or diluent, or the drug includes the allose and a pharmaceutically acceptable carrier or diluent; The dosage form of the medicine is powder, tablet, capsule, oral solution, injection or aerosol.

8. The use according to claim 7, characterized in that: When the drug is an oral solution, an injection or an aerosol, the concentration of allose is 30 μg / mL-60 μg / mL.

9. The use according to claim 8, characterized in that: The medicine also includes other active ingredients; wherein the concentration of the other active ingredients is 0.05 mg / mL-0.15 mg / mL.

10. The use according to claim 8, characterized in that: The drug is a mixture of allose and water.

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