Compound for treating neurodegenerative diseases and application thereof

Abstract

The present invention relates to a compound for the prevention and / or treatment of neurodegenerative diseases. On one hand, the compound provided by the invention can effectively promote the proliferation of neuronal cells, and on the other hand, the compound has a protection effect on neuronal cell toxicity induced by neurotransmitters such as glutamic acid caused by excessive accumulation of Abeta amyloid protein, so that the effect of treating neurodegenerative diseases is comprehensively exerted; meanwhile, the compound disclosed by the invention has no obvious cytotoxicity. Therefore, the compound has a good application prospect in medicines for preventing or treating neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease and the like. Besides, a parent structure with a key active function on the neurodegenerative disease and an important influencing group in the cynanchum otophyllum extract are defined, the basic structure-function relationship between the compound and the treatment of the neurodegenerative disease is defined, and a sufficient scientific basis is provided for research and development of subsequent related drugs.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to a compound for preventing and / or treating neurodegenerative diseases and its application. Background Art

[0002] Glutamate is one of the most important excitatory neurotransmitters in the central nervous system, crucially involved in synaptic plasticity, learning, and memory formation. However, excessive extracellular glutamate accumulation can lead to uncontrolled, continuous neuronal depolarization, ultimately leading to neuronal death. Glutamate is primarily cleared from the synaptic cleft in the CA1 region of the hippocampus via the excitatory amino acid transporter 2 (EAAT2) / glutamate transporter 1 (GLT-1), expressed in astrocytes. Recent studies have demonstrated that upregulating GLT-1 expression can reduce extracellular glutamate accumulation and the resulting excitotoxicity. Studies have shown that glutamate excitotoxicity, resulting in N-methyl-d-aspartate receptor (NMDA) overactivation, mitochondrial dysfunction, oxidative stress, neuroinflammation, synaptic loss, and neuronal death, is associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0003] Qingyang ginseng (Cynanchum otophyllum Schneid.) is a perennial herbaceous, twining vine of the Apocynaceae family, whose rhizomes are used as medicine. First recorded in the Illustrated Catalogue of Plant Names and Realities, Qingyang ginseng is also known as Baishi ginseng and dog poison. It is slightly warm in nature, sweet and slightly bitter in taste. A common folk medicine in Yunnan, it is known as "Bai Shou Wu" (white-shouwu) and is commonly used to treat uterine fibroids, lumbar muscle strain, traumatic injuries, low back pain, pulmonary and endometrial tuberculosis, and various forms of epilepsy. Modern pharmacological studies have found that Qingyang ginseng has anti-epileptic, immunomodulatory, anti-hepatitis, and anti-Meniere's syndrome effects. A Qingyang ginseng extract can improve learning and cognitive impairment in 3xTg-AD mice. Chemical composition studies have shown that the chemical components of Qingyang ginseng are structurally diverse, with C21 steroids considered to be the representative compounds and primary active ingredients in Qingyang ginseng. Recent studies have shown that C21 steroidal saponins extracted from Panax notoginseng exhibit significant anti-epileptic activity and strong anti-proliferative activity against various human tumor cell lines. Therefore, the study of compounds with anti-glutamate excitotoxicity and neuroprotective activity against hippocampal neuronal damage has important clinical implications for the prevention and treatment of neurodegenerative diseases such as Alzheimer's disease. Summary of the Invention

[0004] The purpose of the present invention is to solve the problems existing in the prior art for the treatment of neurodegenerative diseases, thereby providing a compound for preventing and / or treating neurodegenerative diseases. The compound is extracted and purified from the natural product Cynoglossum eryngii, has good safety, has a significant promoting effect on the proliferation of neuronal cells, and can inhibit glutamate-induced cytotoxicity, thereby having neuroprotective activity on neuronal cells.

[0005] To achieve the above object, the present invention is achieved by the following means:

[0006] The first aspect of the present invention provides a compound for preventing and / or treating neurodegenerative diseases, the structural formula of which is shown in Formula I:

[0007]

[0008] Wherein, R1 is selected from Any one of; R2 is selected from

[0009] Any one of; R3 is selected from O, Any one of, wherein S4 is selected from

[0010] Preferably, the neurodegenerative disease includes one or more of Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0011] Preferably, the senile dementia includes one or more of Alzheimer's disease, vascular dementia, Lewy body dementia and frontotemporal dementia.

[0012] Preferably, the compound is selected from one or more of compounds 1-15 represented by the following structural formulas:

[0013]

[0014]

[0015]

[0016] The second aspect of the present invention provides a pharmaceutical composition for preventing and / or treating neurodegenerative diseases, comprising one or more of the above-mentioned compounds, pharmaceutically acceptable salts thereof, and solvates thereof, and a pharmaceutically acceptable carrier.

[0017] Preferably, the neurodegenerative disease includes one or more of Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0018] Preferably, the senile dementia includes one or more of Alzheimer's disease, vascular dementia, Lewy body dementia and frontotemporal dementia.

[0019] Preferably, the pharmaceutically acceptable carrier includes one or more of a filler, a binder, a disintegrant, a solvent, a preservative, a lubricant, and a flavoring agent.

[0020] Preferably, the compound is selected from one or more of compounds 1-15 represented by the following structural formulas:

[0021]

[0022]

[0023]

[0024] The third aspect of the present invention provides the use of one or more of the above-mentioned compounds, pharmaceutically acceptable salts or solvates thereof in the preparation of products for preventing and / or treating neurodegenerative diseases.

[0025] Preferably, the neurodegenerative disease includes one or more of Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0026] Preferably, the senile dementia includes one or more of Alzheimer's disease, vascular dementia, Lewy body dementia and frontotemporal dementia.

[0027] Preferably, the compound is selected from one or more of compounds 1-15 represented by the following structural formulas:

[0028]

[0029]

[0030]

[0031] Preferably, the product includes one or more of medicines, health products, and foods.

[0032] The present invention has the following beneficial effects compared to the prior art:

[0033] On the one hand, the compounds provided by the present invention can effectively promote the proliferation of neuronal cells, and on the other hand, they can have a protective effect on neuronal cell toxicity induced by excessive accumulation of Aβ amyloid protein, such as neurotransmitters such as glutamate, thereby comprehensively playing the role of treating neurodegenerative diseases. At the same time, the compounds of the present invention have no obvious cytotoxicity. Therefore, they have good application prospects in drugs for preventing or treating neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, etc. In addition, the present invention found through in-depth analysis that the neuroprotective effect of the compounds of the present invention is closely related to the length of the sugar chain connected to its C-3, which is mainly manifested as 3-sugar chain>2-sugar chain>1-sugar chain>4-sugar chain>5-sugar chain. Accordingly, the parent structure and important influencing groups in the Qingyang ginseng extract that have key active functions for neurodegenerative diseases are clarified, and the basic structure-activity relationship between the compound and the treatment of neurodegenerative diseases is clarified, providing a sufficient scientific basis for the subsequent research and development of related drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 Schematic diagram of the cytotoxic activity of compound 1-15 on HT22 hippocampal neuronal cells.

[0035] Figure 2 Schematic diagram of the neuroprotective effect of compound 1-7 on glutamate-induced HT22 hippocampal neuronal cell damage.

[0036] Figure 3 Schematic diagram of the neuroprotective effect of compounds 8-15 on glutamate-induced HT22 hippocampal neuronal cell damage.

[0037] Figure 4 Schematic diagram of the percentage of live cells in the experiment of compound 4 inhibiting glutamate-induced apoptosis in HT22 cells.

[0038] Figure 5 Schematic diagram of the flow cytometry results of the experimental inhibition of glutamate-induced apoptosis of HT22 cells by compound 4.

[0039] Figure 6 Schematic diagram of the percentage of live cells in the experiment of compound 10 inhibiting glutamate-induced apoptosis in HT22 cells.

[0040] Figure 7 Schematic diagram of the flow cytometry results of the experimental inhibition of glutamate-induced apoptosis of HT22 cells by compound 10.

[0041] Figure 8 Schematic diagram of the effect of compound 4 on the expression of apoptosis-related genes in HT22 cells induced by glutamate.

[0042] Figure 9Schematic diagram of the effect of compound 10 on the expression of apoptosis-related genes in HT22 cells induced by glutamate.

[0043] Figure 10 Schematic diagram of the effect of compound 4 on the expression of apoptosis-related proteins in HT22 cells induced by glutamate.

[0044] Figure 11 Schematic diagram of the effect of compound 10 on the expression of apoptosis-related proteins in HT22 cells induced by glutamate.

[0045] Figure 12 Schematic diagram of the effect of compound 4 on autophagic flux in cells stably expressing tf-LC3.

[0046] Figure 13 Schematic diagram of the quantitative analysis results of the effect of compound 4 on autophagic flux in cells stably expressing tf-LC3.

[0047] Figure 12 Schematic diagram of the effect of compound 4 on autophagic flux in cells stably expressing tf-LC3.

[0048] Figure 14 Schematic diagram of the effect of compound 10 on autophagic flux in cells stably expressing tf-LC3.

[0049] Figure 15 Schematic diagram of the quantitative analysis results of the effect of compound 10 on autophagic flux in cells stably expressing tf-LC3. DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solution and effect of the present invention clearer and more specific, the present invention is further described in detail with reference to the following examples. 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.

[0051] Unless otherwise specified, compounds 1-15 listed in the context of the present invention were extracted from Panax notoginseng using conventional methods in the prior art, and 1 H NMR and / or 13C NMR was used for structural identification. The cell lines including HT22 used in the context of the present invention were cultured according to the ATCC guidelines. All cell lines were identified by short tandem repeat analysis of the China Center for Type Culture Collection (Wuhan), and PCR detection kits (Shanghai Biothrive Sci) were used to verify the presence of mycoplasma contamination, and were simultaneously frozen in liquid nitrogen and used for subsequent experiments. The reagents, consumables, etc. used in the present invention are all commercially available or configured according to a conventional method. The experimental methods used in the present invention, such as cell culture, cell proliferation experiments, apoptosis experiments, flow cytometry, etc., are all conventional methods and techniques in the art. The instruments and equipment used in the present invention are all commercially available, and wherein the microplate reader is a U.S. BioTEK Synergy H1 Hybrid Multi-ModeReader; the flow cytometer is a U.S. BECKMAN COULTER, CytoFLEX S.

[0052] Representative results from replicates are presented in the accompanying figures. Data are presented as mean ± SD as indicated in the figures. All experiments were repeated at least three times. Data were analyzed using GraphPad Prism 8.0. Means between two or more groups were compared using t-tests or analysis of variance. A p value of less than 0.05 was considered significant.

[0053] Example 1 Extraction and separation of compounds

[0054] (1) 15 kg of dried Panax notoginseng root was extracted with 95% ethanol-water (30 L) reflux four times for 2 h each time, and then concentrated under reduced pressure to obtain a crude extract (2.25 kg).

[0055] (2) 400 g of the crude extract was separated by silica gel open column chromatography using a dichloromethane-methanol gradient elution (100:0→3:1, V / V) to obtain 13 fractions (YS.1-YS.13).

[0056] (3) Fraction YS.3 (36.2 g) was subjected to silica gel open column chromatography and dichloromethane / methanol gradient elution (100:0→80:1, V / V) to obtain 5 sub-fractions (YS.3-1 to YS.3-5). Fraction YS.3-4 (23.1 g) was subjected to ODS open column chromatography and methanol / ultrapure water gradient elution (40%→100%, V / V) to obtain 6 sub-fractions (YS.3-4-1 to YS.3-4-6). Fraction YS.3-4-4 (8.93 g) was subjected to preparative HPLC (75% methanol in water, flow rate 3.0 mL / min) to obtain compound 2 (t R =22.6min, 29.0mg), compound 3 (t R=29 min, 62.4 mg), compound 4 (t R =35min, 29.0mg), compound 5 (t R =59.2 min, 29.0 mg).

[0057] (4) Fraction YS.5 (30.0 g) was subjected to silica gel open column chromatography and petroleum ether / ethyl acetate gradient elution (5:1→1:6, V / V) to obtain 8 sub-fractions (YS.5-1 to YS.5-8). Fraction YS.5-3 (25.0 g) was subjected to silica gel open column chromatography and petroleum ether / ethyl acetate gradient elution (3:1→1:2, V / V) to obtain 9 sub-fractions (YS.5-3-1 to YS.5-3-9). Fraction YS.5-3-3 (2.45 g) was subjected to preparative HPLC (75% methanol in water, flow rate 3.0 mL / min) to obtain compound 1 (t R =13.8min, 458.3mg); fraction YS.5-3-6 (3.26g) was purified by preparative high performance liquid chromatography (80% methanol in water, flow rate 3.0mL / min) to give compound 11 (t R =12.1min, 293.8mg); fraction YS.5-3-7 (2.80g) was purified by preparative high performance liquid chromatography (75% methanol in water, flow rate 3.0mL / min) to give compound 9 (t R =15.2min, 221.0mg), compound 10 (t R =18.2min, 276.8mg). Fraction YS.5-6 (10.0g) was subjected to silica gel open column chromatography and petroleum ether / ethyl acetate gradient elution (2:1→2:5, V / V) to obtain 8 sub-fractions (YS.5-6-1 to YS.5-6-8). Fraction YS.5-6-4 (2.61g) was subjected to preparative high performance liquid chromatography (72% methanol in water, flow rate 3.0mL / min) to obtain compound 15 (t R =23.6min, 44.2mg), compound 14 (t R =27 min, 78.8 mg).

[0058] (5) Fraction YS.9 (24.7 g) was subjected to silica gel open column chromatography and ethyl acetate / methanol gradient elution (40:1→8:1, V / V) to obtain 8 sub-fractions (YS.9-1 to YS.9-8). Fraction YS.9-2 (7.45 g) was subjected to preparative HPLC (80% methanol in water, flow rate 3.0 mL / min) to obtain 8 sub-fractions (YS.9-2-1 to YS.9-2-8), of which fraction YS.9-2-4 was compound 7 (t R=39 min, 103 mg). Fraction YS.9-2-5 (169.5 mg) was purified by preparative HPLC (80% methanol in water, flow rate 3.0 mL / min) to give compound 6 (t R =39min, 25.8mg). Fraction YS.9-4 (4.76g) was subjected to preparative high performance liquid chromatography (80% methanol in water, flow rate 3.0mL / min) to obtain 3 sub-fractions (YS.9-4-1 to YS.9-4-3). Fraction YS.9-4-1 (575.7mg) was subjected to preparative high performance liquid chromatography (68% methanol in water, flow rate 3.0mL / min) to obtain 7 sub-fractions (YS.9-4-1-1 to YS.9-4-1-7). Fraction YS.9-4-1-6 (68mg) was subjected to preparative high performance liquid chromatography (68% methanol in water, flow rate 3.0mL / min) to obtain compound 13 (t R =80min, 34.3mg); fraction YS.9-4-2 is compound 8 (t R =33 min, 1.04 g).

[0059] (6) Fraction YS.12 (20.36 g) was purified by preparative HPLC (75% methanol in water, flow rate 3.0 mL / min) to obtain compound 12 (t R =11.8 min, 37.5 mg).

[0060] use 1 H NMR and / or 13 Compounds 1-15 were identified by C NMR, and their structural formulas are shown below:

[0061]

[0062]

[0063]

[0064] Example 2 Neuronal cytotoxicity experiment

[0065] (1) HT22 cells in the logarithmic growth phase were seeded in a 96-well plate at a density of 3000 cells / well and cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotics. Three replicate wells were set up for each group.

[0066] (2) After 24 h of culture, different concentrations of compounds 1-15 (10 -6 M~10 -5 M) Culture was continued for 48 h; Group C served as the control group, to which an equal volume of culture medium was added.

[0067] (3) After 48 h, the medium containing the compound was removed, and 10 μL of DMEM medium containing CCK-8 was added to each well. The cells were incubated in a 5% CO2, 37°C incubator for 1 h.

[0068] (4) Oscillate at room temperature for 10 seconds and measure the absorbance (OD) of each well at 450 nm using a microplate reader.

[0069] Test results such as Figure 1 The results showed that the cell viability after treatment with compound 1-15 was not significantly different from that of the control group C (p>0.05), indicating that the compounds of the present invention had no obvious cytotoxicity to neuronal cells and had good safety.

[0070] Example 3 Study on the neuroprotective effect of hippocampal neurons

[0071] (1) HT22 cells in the logarithmic growth phase were seeded in a 96-well plate at a density of 3000 cells / well and cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotics. Three replicate wells were set up for each group.

[0072] (2) After culturing for 24 h, different concentrations of compounds 1-15 (10 -6 M~10 -5 M) Culture was continued for 24 h; the NC group served as the blank group, to which an equal volume of culture medium was added; the C group served as the control group, to which an equal volume of culture medium was added.

[0073] (3) After 24 h, 15 mM glutamate was added to each group. The NC group was not treated and cultured for another 24 h.

[0074] (4) After 24 h, the medium containing the compound was removed, and 10 μL of DMEM medium containing CCK-8 was added to each well. The cells were incubated in a 5% CO 2 , 37° C. incubator for 1 h.

[0075] (5) Oscillate at room temperature for 10 seconds and measure the absorbance (OD) of each well using a microplate reader (450 nm).

[0076] Test results such as Figure 2-3 The results showed that the hippocampal neuron injury model was established by adding 15mM glutamate. After treatment with compounds 1-15, the cell damage rate was significantly reduced, indicating that the compounds of the present invention have significant neuroprotective activity against glutamate-induced hippocampal neuron injury, and the difference compared with group C was statistically significant (*p<0.05, ***p<0.001).

[0077] Example 4 Hippocampal neuron apoptosis inhibition experiment

[0078] (1) Take HT22 cells in the logarithmic growth phase and plate them at 8×10 4 The cells were seeded at a density of 100 cells / well in 6-well plates and cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotics.

[0079] (2) After 24 h of culture, different concentrations (0.1, 1, 5, 10, μM) of compound 4 or compound 10 were added and cultured for another 12 h; the NC group was a blank group, and an equal volume of culture medium was added; the C group was a control group, and an equal volume of culture medium was added.

[0080] (3) After 12 h, 15 mM glutamate was added to each group. The NC group was not treated and cultured for another 24 h.

[0081] (4) After 24 h, flow cytometry apoptosis detection was performed using the Annexin V-FITC / PI cell apoptosis detection kit (Meilunbio, China) and a Beckman flow cytometer (CytoFLEX S model) according to conventional methods.

[0082] Test results such as Figure 4-7 The results showed that by adding 15mM glutamate to establish a hippocampal neuron injury model, after treatment with compound 4, the cell survival rate was significantly increased and the cell apoptosis rate was significantly decreased by flow cytometry, indicating that compound 4 has significant protective activity against glutamate-induced hippocampal neuron apoptosis and can inhibit the apoptosis of hippocampal neurons. The difference with group C was statistically significant (**p<0.01, ***p<0.001) (see Figure 4-5 A hippocampal neuron injury model was established by adding 15 mM glutamate. After treatment with compound 10, flow cytometry analysis revealed a significant increase in cell survival rate and a significant decrease in cell apoptosis rate, indicating that compound 4 has significant protective activity against glutamate-induced hippocampal neuron apoptosis and can inhibit the apoptosis of hippocampal neurons. The difference from group C was statistically significant (*p<0.05, ***p<0.001) (see Figure 6-7 ).

[0083] Example 5 Detection of genes related to inhibition of hippocampal neuron apoptosis

[0084] (1) Take HT22 cells in the logarithmic growth phase and plate them at 8×10 4 The cells were seeded at a density of 100 cells / well in 6-well plates and cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotics.

[0085] (2) After 24 h of culture, different concentrations (1, 5, 10 μM) of compound 4 or compound 10 were added and cultured for another 12 h; the NC group was a blank group, and an equal volume of culture medium was added; the C group was a control group, and an equal volume of culture medium was added.

[0086] (3) After 12 h, 15 mM glutamate was added to each group. The NC group was not treated and cultured for another 24 h.

[0087] (4) After 24 h, RNA was extracted using the TRIzol method, and q-PCR experiments were performed according to conventional methods to detect the mRNA expression levels of the corresponding indicators.

[0088] Test results such as Figure 8-9 The results showed that by adding 15mM glutamate to establish a hippocampal neuron injury model, the expression of Bax, Bax / Bcl-2 and caspase3 genes was significantly increased compared with the NC group, and the difference was statistically significant (***p<0.001); after treatment with compound 4, the expression of Bax, Bax / Bcl-2 and caspase3 genes was significantly decreased compared with group C, indicating that compound 4 inhibited glutamate-induced hippocampal neuron apoptosis by mediating the Bcl-2 / Bax / Caspase-3 signaling pathway and exerted neuroprotective activity, and the difference was statistically significant compared with group C (**p<0.01, ***p<0.001) (see Figure 8 ). A hippocampal neuron injury model was established by adding 15 mM glutamate. The expression of Bax, Bax / Bcl-2, and caspase3 genes was significantly increased compared with the NC group, and the difference was statistically significant (***p<0.001). After treatment with compound 10, the expression of Bax, Bax / Bcl-2, and caspase3 genes was significantly decreased compared with group C, indicating that compound 10 inhibited glutamate-induced hippocampal neuron apoptosis by mediating the Bcl-2 / Bax / Caspase-3 signaling pathway and exerted neuroprotective activity. The difference was statistically significant compared with group C (*p<0.05, **p<0.01, ***p<0.001) (see Figure 9 ).

[0089] Example 6 Detection of Expression of Proteins Related to Apoptosis Inhibition in Hippocampal Neurons

[0090] (1) Take HT22 cells in the logarithmic growth phase and plate them at 8×10 4 The cells were seeded at a density of 100 cells / well in 6-well plates and cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotics.

[0091] (2) After 24 h of culture, different concentrations (1, 5, 10 μM) of compound 4 or compound 10 were added and cultured for another 12 h; the NC group was a blank group, and an equal volume of culture medium was added; the C group was a control group, and an equal volume of culture medium was added.

[0092] (3) After 12 h, 15 mM glutamate was added to each group. The NC group was not treated and cultured for another 24 h.

[0093] (4) After 24 hours, cells were collected, proteins were extracted, and Western blot analysis was performed. The antibodies used were Bax (#T40051, abmart), Bcl-2 (#T40056, abmart), caspase3 (#T40044, abmart), and β-tubulin (#ab7291, abcam).

[0094] Test results such as Figure 10-11 The results showed that by adding 15mM glutamate to establish a hippocampal neuron injury model, the expression of Bax and caspase3 proteins increased compared with the NC group, and the expression of Bcl-2 protein decreased compared with the NC group; after treatment with compound 4, the expression of Bax and caspase3 proteins decreased compared with the C group, and the expression of Bcl-2 protein increased compared with the C group, indicating that compound 4 inhibits glutamate-induced hippocampal neuron apoptosis by mediating the Bcl-2 / Bax / Caspase-3 signaling pathway and exerts neuroprotective activity (see Figure 10 A hippocampal neuron injury model was established by adding 15 mM glutamate. The expression of Bax and caspase3 proteins increased compared with the NC group, while the expression of Bcl-2 protein decreased compared with the NC group. After treatment with compound 10, the expression of Bax and caspase3 proteins decreased compared with the C group, while the expression of Bcl-2 protein increased compared with the C group, indicating that compound 10 inhibits glutamate-induced hippocampal neuron apoptosis by mediating the Bcl-2 / Bax / Caspase-3 signaling pathway and exerts neuroprotective activity (see Figure 11 ).

[0095] Example 7 Autophagy flux effect experiment

[0096] (1) 293T cells stably expressing tf-LC3 were constructed according to conventional methods in the art. Cells in the logarithmic growth phase were taken and 5×10 3 The cells were seeded at a density of 1000 cells / well in a 4-well glass bottom dish and cultured in DMEM medium containing 10% FBS (fetal bovine serum) and 1% antibiotics.

[0097] (2) After culturing for 24 h, compound 4 or compound 10 was added at a concentration of 10 μM. The NC group was used as a control and an equal volume of DMSO was added. The cells were cultured for another 12 h.

[0098] (3) After 12 h, the culture medium was removed, the cells were rinsed twice with PBS, fixed with 4% paraformaldehyde for 30 min, stained with DAPI, and images were acquired using a confocal microscope.

[0099] Test results such as Figure 12-15 The results showed that compared with the control group (NC group), the number of red fluorescent spots in 293T cells stably expressing tf-LC3 increased significantly after treatment with 10 μM compound 4, indicating that 10 μM compound 4 can effectively promote the formation of autophagy lysosomes, and the difference is statistically significant (**p<0.01) (see Figure 12-13 Compared with the control group (NC group), the number of red fluorescent spots in 293T cells stably expressing tf-LC3 increased significantly after treatment with 10 μM compound 10, indicating that 10 μM compound 10 can effectively promote the formation of autophagy lysosomes, and the difference is statistically significant (*p<0.05) (see Figure 14-15 ).

[0100] In summary, the compounds provided by the present invention can, on the one hand, effectively promote the proliferation of neuronal cells, and on the other hand, have a protective effect on neuronal cell toxicity induced by excessive accumulation of Aβ amyloid protein, such as neurotransmitters such as glutamate, thereby comprehensively playing the role of treating neurodegenerative diseases. At the same time, the compounds of the present invention have no obvious cytotoxicity. Therefore, they have good application prospects in drugs for preventing or treating neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease. In addition, the present invention found through in-depth analysis that the neuroprotective effect of the compounds of the present invention is closely related to the length of the sugar chain connected to its C-3, which is mainly manifested as 3-sugar chain>2-sugar chain>1-sugar chain>4-sugar chain>5-sugar chain. Accordingly, the parent structure and important influencing groups in the Qingyang ginseng extract that have key active functions for neurodegenerative diseases are clarified, and the basic structure-activity relationship between the compound and the treatment of neurodegenerative diseases is clarified, providing sufficient scientific basis for the subsequent research and development of related drugs.

[0101] The above detailed description of the analytical methods involved in the present invention provides a detailed introduction. It should be noted that the above description is intended solely to help those skilled in the art better understand the methods and concepts of the present invention, and is not intended to limit the relevant content. Without departing from the principles of the present invention, those skilled in the art may make appropriate adjustments or modifications to the present invention, and such adjustments and modifications shall also fall within the scope of protection of the present invention.

Claims

1. A compound for preventing and / or treating neurodegenerative diseases, characterized in that: Its structural formula is shown in Formula I: Wherein, R1 is selected from Any one of; R2 is selected from Any one of; R3 is selected from O, Any one of, wherein S4 is selected from 2. The pharmaceutical composition according to claim 1, characterized in that The neurodegenerative diseases include one or more of Alzheimer's disease, Parkinson's disease, and Huntington's disease.

3. The compound for preventing and / or treating neurodegenerative diseases according to claim 2, characterized in that The senile dementia includes one or more of Alzheimer's disease, vascular dementia, Lewy body dementia and frontotemporal dementia.

4. The compound for preventing and / or treating neurodegenerative diseases according to claim 1, characterized in that The compound is selected from one or more of compounds 1-15 represented by the following structural formulas:

5. A pharmaceutical composition for preventing and / or treating neurodegenerative diseases, characterized in that: Comprising the compound for preventing and / or treating a neurodegenerative disease according to any one of claims 1 to 4, one or more of its pharmaceutically acceptable salts and solvates, and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, characterized in that The compound is selected from one or more of compounds 1-15 represented by the following structural formulas:

7. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutically acceptable carrier includes one or more of a filler, a binder, a disintegrant, a solvent, a preservative, a lubricant, and a flavoring agent.

8. Use of one or more of the compound for preventing and / or treating neurodegenerative diseases according to any one of claims 1 to 4, or a pharmaceutically acceptable salt or solvate thereof, in the preparation of a product for preventing and / or treating neurodegenerative diseases.

9. The use according to claim 8, characterized in that The compound is selected from one or more of compounds 1-15 represented by the following structural formulas:

10. The use according to claim 8, characterized in that The product includes one or more of medicines, health products, and foods.

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