Polypeptide and its application in preparing drugs for treating neurodegenerative diseases

Abstract

The present invention relates to a polypeptide and its application. The polypeptide sequence is SEQ ID NO: 7. The polypeptide is used to prepare a drug for treating or preventing neurodegenerative diseases. The polypeptide of the present invention can treat or prevent neurodegenerative diseases and has good application prospects.

Description

[0001] This invention is a divisional application. The filing date of the original application is July 21, 2021, the application number is 202110827432.X, and the name is "Polypeptides and their applications in the preparation of anti-neurodegenerative disease drugs." Technical Field

[0002] The invention relates to a polypeptide and its application in preparing a drug for resisting neurodegenerative diseases. The polypeptide is one of a series of specific polypeptide fragments of DVL-2 protein and belongs to the technical field of biomedicine. Background Art

[0003] Neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease, etc.) are characterized by specific neuronal loss and are a type of progressive disease. Most neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease, have complex mechanisms of occurrence, and the specific causes are still unknown, and there is a lack of effective cures. There is an urgent need to research and develop methods and drugs that can effectively prevent and treat neurodegenerative diseases.

[0004] Parkinson's disease is one of the most common neurodegenerative diseases, which is more common in the elderly. The main pathological changes of Parkinson's disease include abnormal degeneration of dopaminergic neurons in the substantia nigra of the midbrain, which leads to a significant decrease in striatal dopamine and the formation of Lewy bodies. The clinical manifestations include movement disorders such as resting tremor, bradykinesia, muscle rigidity, and abnormal posture and gait, and even non-motor symptoms such as cognitive impairment. Chinese Parkinson's disease patients account for about half of the countries with a high incidence of Parkinson's disease in the world, and as the aging of the population continues to accelerate, the social burden caused by Parkinson's disease is also increasing.

[0005] Alzheimer's disease is the most common neurodegenerative disease, with the main clinical manifestations of cognitive function and learning disorders, memory loss, etc. With the accelerating aging of the population, the number of people suffering from Alzheimer's disease has been increasing sharply in recent years, and the social burden caused by it has also been increasing day by day, especially in China, where the number of Alzheimer's disease patients ranks first in the world, and the market demand for drugs to prevent and treat Alzheimer's disease is huge.

[0006] Dishevelled (DVL) is an important regulatory factor in the canonical Wnt signaling pathway. DVL in mammals mainly includes DVL-1, DVL-2 and DVL-3, all of which contain three highly conserved basic domains: DIX, PDZ and DEP. The inventors of the present invention have obtained research results related to DVL-2 after a lot of exploration, experiments and research work. Summary of the invention

[0007] The main purpose of the present invention is to propose a polypeptide that can be used to prepare drugs against neurodegenerative diseases based on the inventor's recent research results and to address the problems existing in the prior art. At the same time, the invention also provides the use of the polypeptide.

[0008] The technical solution of the present invention to solve the technical problem is as follows:

[0009] A polypeptide, characterized in that the amino acid sequence of the polypeptide is SEQ ID NO:7.

[0010] The polypeptide is one of a series of specific polypeptide fragments (SEQ ID NO: 1 to SEQ ID NO: 14) of the DVL-2 protein and can be used for preparing drugs for resisting neurodegenerative diseases.

[0011] A polypeptide, characterized in that it is composed of the polypeptide described above connected to a cell-penetrating peptide, wherein the cell-penetrating peptide is located at the N-terminus of the polypeptide; the amino acid sequence of the cell-penetrating peptide is one of SEQ ID NO: 15 to SEQ ID NO: 47.

[0012] A polypeptide, characterized in that it is formed by connecting the polypeptide described above to a cell-penetrating peptide via a connecting peptide, wherein the connecting peptide and the cell-penetrating peptide are both located at the N-terminus of the polypeptide; the amino acid sequence of the cell-penetrating peptide is one of SEQ ID NO:15 to SEQ ID NO:47, and the amino acid sequence of the connecting peptide is one of SEQ ID NO:48 to SEQ ID NO:72.

[0013] The polypeptide described above is used for preparing a drug for treating or preventing neurodegenerative diseases.

[0014] Wherein, the neurodegenerative disease is Alzheimer's disease or Parkinson's disease.

[0015] The present invention also proposes:

[0016] A nucleic acid encoding the polypeptide described above.

[0017] A pharmaceutical composition containing the polypeptide or nucleic acid described above.

[0018] The pharmaceutical composition described above is used for preparing a pharmaceutical preparation for treating or preventing neurodegenerative diseases.

[0019] After extensive exploration, experiments and research, the inventors of the present invention conducted structure-activity relationship analysis based on DVL-2, designed several polypeptide sequences and conducted activity tests, and finally obtained a series of polypeptides with biological activity for preventing and treating neurodegenerative diseases; one of the series of polypeptides involved in the present invention can treat or prevent neurodegenerative diseases and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a schematic diagram of the results of Example 1 of the present invention. The ordinates of Figures A and B are both relative cell survival rates. In Figure A, the Control group is the control group, the Rotenone group is the model group, and the Rotenone+peptide (2, 5, 7, 8) group is the polypeptide group to be tested (the polypeptides to be tested are polypeptide SEQ ID NO: 2, polypeptide SEQ ID NO: 5, polypeptide SEQ ID NO: 7, and polypeptide SEQ ID NO: 8). In Figure B, the Control group is the control group, the Rotenone group is the model group, and the other experimental groups are polypeptide groups to be tested. The polypeptide to be tested in the Rotenone+Tat group is the cell penetrating peptide SEQ ID NO: 15, and the polypeptide to be tested in the Rotenone+Tat-peptide (2, 5, 7, 8) group is a second group of polypeptides containing polypeptide SEQ ID NO: 2, polypeptide SEQ ID NO: 5, polypeptide SEQ ID NO: 7, or polypeptide SEQ ID NO: 8.

[0021] Figure 2 The result of Example 2 of the present invention is shown in Figure 2. Hoechst labels the cell nucleus in blue, and PI labels apoptotic or necrotic cells in red. In the figure, the Control group is the control group, the Rotenone group is the model group, and the Rotenone+peptide 2 group is the polypeptide group to be tested (the polypeptide to be tested is polypeptide SEQ ID NO: 2).

[0022] Figure 3 The result of Example 3 of the present invention is shown in FIG. 3. In the figure, the Rotenone (-) Peptide 2 (-) group is the control group, the Rotenone (+) Peptide 2 (-) group is the model group, and the Rotenone (+) Peptide 2 (+) group is the test polypeptide group (the test polypeptide is polypeptide SEQ ID NO: 2).

[0023] Figure 4 The results of Example 4 of the present invention are shown in the figure. The vertical axis is the relative cell survival rate. The Control group is the control group, and Aβ 25-35 The model group, Aβ 25-35 The +peptide (2, 5, 7, 8) group is the polypeptide group to be tested (the polypeptides to be tested are polypeptide SEQ ID NO: 2, polypeptide SEQ ID NO: 5, polypeptide SEQ ID NO: 7, polypeptide SEQ ID NO: 8).

[0024] Figure 5This is a schematic diagram of the results of Example 5 of the present invention. Hoechst labels the cell nucleus, which appears blue; PI labels apoptotic or necrotic cells, which appears red. In the figure, the Control group is the control group, and Aβ 25-35 The model group, Aβ 25-35 +peptide 2 group is the polypeptide group to be tested (the polypeptide to be tested is polypeptide SEQ ID NO: 2).

[0025] Figure 6 It is a schematic diagram of the results of Example 6 of the present invention. In the figure, the Control group is the control group, the MPTP group is the model group, and the other experimental groups are the tested polypeptide groups. The tested polypeptide in the MPTP+Tat group is the cell penetrating peptide SEQ ID NO: 15, and the tested polypeptides in the MPTP+Tat-peptide (2, 5, 7, 8) group are 4 polypeptides containing polypeptide SEQ ID NO: 2, polypeptide SEQ ID NO: 5, polypeptide SEQ ID NO: 7 or polypeptide SEQ ID NO: 8, respectively.

[0026] Figure 7 It is a schematic diagram of the results of Example 7 of the present invention. Among them, Figure A is the detection of TH expression in the substantia nigra of each group of experimental mice, and Figure B is the detection of TH expression in the striatum of each group of experimental mice. In the figure, the Con group is the control group, the MPTP-Non group is the model group, and the other experimental groups are the tested polypeptide groups. The tested polypeptide in the MPTP+Tat group is the cell penetrating peptide SEQ ID NO: 15, and the tested polypeptide in the MPTP+Tat-peptide (2, 5, 7, 8) group is 4 polypeptides containing polypeptide SEQ ID NO: 2, polypeptide SEQ ID NO: 5, polypeptide SEQ ID NO: 7 or polypeptide SEQ ID NO: 8. DETAILED DESCRIPTION

[0027] The present invention will be described in further detail below with reference to the accompanying drawings and in combination with embodiments, but the present invention is not limited to the examples given.

[0028] Example 1

[0029] This example is to detect the effect of each polypeptide on the death of nerve cells in a Parkinson's disease-like model.

[0030] The polypeptides to be tested in this example are as follows:

[0031] (1) The first group of polypeptides: polypeptide of SEQ ID NO: 2, polypeptide of SEQ ID NO: 5, polypeptide of SEQ ID NO: 7, polypeptide of SEQ ID NO: 8.

[0032] (2) The second group of polypeptides: ① polypeptide SEQ ID NO: 2 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15; ② polypeptide SEQ ID NO: 5 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15; ③ polypeptide SEQ ID NO: 7 + connecting peptide SEQ ID NO: 57 + cell penetrating peptide SEQ ID NO: 15; ④ polypeptide SEQ ID NO: 8 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15. Note: The cell penetrating peptide SEQ ID NO: 15 is the cell penetrating peptide Tat.

[0033] The specific experimental process of this embodiment is as follows:

[0034] Peptide group: SH-SY5Y cells were induced to differentiate with 10 μM retinoic acid for 6 days, and then differentiated SH-SY5Y cells in the logarithmic growth phase were selected and cultured at 3×10 4 The cells were inoculated at a density of 100 μL / mL in a 96-well plate (100 μL per well) and cultured in a cell culture incubator at 37°C and 5% CO2. After 24 hours of culture, Rotenone (0.1 μM) (Note: Rotenone is rotenone) was added, and the peptide to be tested (40 μM) was added, and 6 replicate wells were set. After 24 hours of culture, 10% of the volume of the culture medium was added to each well. The cells were incubated in a 37°C constant temperature cell culture incubator for 4 hours, the supernatant was discarded, 150 μL DMSO was added to each well, and the plate was shaken at 500 r / min for 10 minutes on an ELISA plate oscillator to completely dissolve the blue-purple crystalline formazan. After the oscillation, the OD value of each well was measured using a microplate reader with a detection wavelength of 570 nm and a reference wavelength of 630 nm, and the cell survival was calculated.

[0035] Compared with the above-mentioned test polypeptide group, the control group did not add Rotenone and the test polypeptide, and the model group did not add the test polypeptide.

[0036] The results are as follows Figure 1 As shown, compared with the model group, both the first group of peptides and the second group of peptides can alleviate the death of neuronal cells in the Parkinson's disease-like model, while the cell penetrating peptide Tat alone did not show significant cell protective effect.

[0037] Example 2

[0038] This example is to detect the effect of the polypeptide on apoptosis of nerve cells in a Parkinson's disease-like model.

[0039] The polypeptide to be tested in this example is polypeptide SEQ ID NO:2.

[0040] The specific experimental process of this embodiment is as follows:

[0041] Peptide group: SH-SY5Y cells were induced to differentiate with 10 μM retinoic acid for 6 days, and then differentiated SH-SY5Y cells in the logarithmic growth phase were selected and cultured at 5×10 4 The cells were inoculated at a density of 1000 μL per well in a 6-well plate at 1000 μL / mL and cultured in a cell culture incubator at 37°C and 5% CO2. After 24 hours of culture, Rotenone (0.1 μM) was added, and the peptide to be tested (40 μM) was added, and 3 replicate wells were set up. After 24 hours of culture, the culture medium in the 6-well plate was discarded, and the cells were washed 3 times with PBS buffer, each time for 3 minutes; 1000 μL of culture medium containing Hoechst 33342 (7 mM) was added to each well and incubated in a 37°C incubator for 25 minutes. The cells were washed twice with PBS buffer, and 1000 μL of culture medium containing PI (2 mM) was added to each well, and incubated at room temperature in the dark for 15 minutes. After staining, the cells were washed 3 times with PBS buffer, and observed and photographed under an inverted fluorescence microscope after washing.

[0042] Compared with the above-mentioned test polypeptide group, the control group did not add Rotenone and the test polypeptide, and the model group did not add the test polypeptide.

[0043] The results are as follows Figure 2 As shown, Hoechst labels the cell nucleus, showing blue; PI labels apoptotic or necrotic cells, showing red. Compared with the model group, the polypeptide SEQ ID NO: 2 can effectively reduce the apoptosis of nerve cells in the Parkinson's disease-like model.

[0044] Example 3

[0045] This example is to detect the effect of the polypeptide on the abnormal pathological changes of nerve cells in a Parkinson's disease-like model.

[0046] The polypeptide to be tested in this example is polypeptide SEQ ID NO:2.

[0047] The specific experimental process of this embodiment is as follows:

[0048] Peptide group: SH-SY5Y cells were induced to differentiate with 10 μM retinoic acid for 6 days, and then differentiated SH-SY5Y cells in the logarithmic growth phase were selected and cultured at 5×10 4The cells were inoculated in a 6-well plate (1000 μL per well) at a density of 1000 μL / mL and cultured in a cell culture incubator at 37°C and 5% CO2. After 24 hours of culture, Rotenone (0.1 μM) was added, and the peptide to be tested (40 μM) was added, and 3 replicate wells were set. After 24 hours of culture, the cell protein was extracted and the sample was prepared. The specific process was as follows: the culture medium in the 6-well plate was discarded, the cells were washed 3 times with PBS buffer, the cells in the wells were collected in an EP tube, 300 μL RIPA lysis buffer (a mixture of protease inhibitors and phosphatase inhibitors was added at a ratio of 1:100), and lysis was performed on ice for 30 minutes. Centrifuge at 4°C 12000r / min for 20 minutes, collect the supernatant, use the BCA protein concentration determination kit to determine the protein concentration, and use PBS buffer to adjust the protein concentration to the same level. 1 / 4 of the total volume of 5×SDS loading buffer was added to the protein solution, and after sufficient mixing, it was placed in boiling water and heated for 10 minutes and stored in a -20°C refrigerator. The protein samples were analyzed by western blotting, and the specific process was as follows: SDS-PAGE gel was prepared, placed in an electrophoresis tank, electrophoresis buffer was added, and the protein sample was added to the loading well for constant voltage electrophoresis. When the bromophenol blue band migrated to the bottom of the gel plate, the electrophoresis was stopped, and the target protein band was transferred to a PVDF membrane using a semi-dry transfer apparatus, blocked with 3% skim milk powder for 2 hours, incubated with anti-TH antibody at 4°C overnight, and incubated with the corresponding horseradish peroxidase-coupled secondary antibody, and finally exposed using a chemiluminescent reagent.

[0049] Compared with the above-mentioned test polypeptide group, the control group did not add Rotenone and the test polypeptide, and the model group did not add the test polypeptide.

[0050] The results are as follows Figure 3 As shown, compared with the model group, the polypeptide SEQ ID NO: 2 can increase the TH expression level of the nerve cells in the Parkinson's disease-like model and alleviate the pathological abnormalities of the nerve cells in the Parkinson's disease-like model.

[0051] Example 4

[0052] This example is to detect the effect of each polypeptide on the death of neural cells in the Alzheimer's disease-like model.

[0053] The polypeptides to be tested in this embodiment are 4 polypeptides: polypeptide SEQ ID NO: 2, polypeptide SEQ ID NO: 5, polypeptide SEQ ID NO: 7, and polypeptide SEQ ID NO: 8.

[0054] The specific experimental process of this embodiment is as follows:

[0055] Peptide group to be tested: SH-SY5Y cells in the logarithmic growth phase were selected and 3×10 4The cells were seeded at a density of 100 μL / mL in a 96-well plate (100 μL per well) and cultured in a 37°C, 5% CO2 cell culture incubator. Aβ was added after 24 h of culture. 25-35 (0.05μM) (Note: Aβ 25-35 i.e., β-amyloid protein fragment), and add the peptide to be tested (40 μM), set up 6 duplicate wells. After 24 hours of culture, add 10% of the volume of culture medium to each well. 5mg / mL MTT solution, incubate in a 37°C constant temperature cell culture incubator for 4 hours, discard the supernatant, add 150 μL DMSO to each well, and oscillate at 500r / min on an ELISA plate oscillator for 10 minutes to completely dissolve the blue-purple crystalline formazan. After the oscillation is completed, place the plate in an ELISA instrument with 570nm as the detection wavelength and 630nm as the reference wavelength, measure the OD value of each well, and calculate the cell survival.

[0056] Compared with the above-mentioned peptide groups, the control group did not add Aβ 25-35 and the peptide to be tested, while the model group did not add the peptide to be tested.

[0057] The results are as follows Figure 4 As shown, compared with the model group, the four peptides can alleviate the death of neurons in the Alzheimer's disease-like model.

[0058] Example 5

[0059] This example is to detect the effect of the polypeptide on apoptosis of neural cells in an Alzheimer's disease-like model.

[0060] The polypeptide to be tested in this example is polypeptide SEQ ID NO:2.

[0061] The specific experimental process of this embodiment is as follows:

[0062] Peptide group to be tested: SH-SY5Y cells in the logarithmic growth phase were selected and cultured at 5×10 4 The cells were inoculated at a density of 1000 μL / mL in a 6-well plate (1000 μL per well) and cultured in a 37°C, 5% CO2 cell culture incubator. Aβ was added after 24 h of culture. 25-35 (0.05μM), and add the peptide to be tested (40μM), set up 3 replicates. After 24h of culture, discard the culture medium in the 6-well plate, wash the cells 3 times with PBS buffer, 3min each time; add 1000μL of culture medium containing Hoechst 33342 (7mM) to each well and incubate in a 37℃ incubator for 25min. Wash the cells twice with PBS buffer, add 1000μL of culture medium containing PI (2mM) to each well, and incubate at room temperature in the dark for 15min. After staining, wash the cells 3 times with PBS buffer, and observe and photograph them under an inverted fluorescence microscope after washing.

[0063] Compared with the above-mentioned peptide groups, the control group did not add Aβ25-35 and the peptide to be tested, while the model group did not add the peptide to be tested.

[0064] The results are as follows Figure 5 As shown, Hoechst labels the cell nucleus, showing blue; PI labels apoptotic or necrotic cells, showing red. Compared with the model group, the polypeptide SEQ ID NO: 2 can effectively reduce the apoptosis of neural cells in the Alzheimer's disease-like model.

[0065] Example 6

[0066] This example is to detect the effects of various polypeptides on Parkinson's disease-like behavioral abnormalities in model mice.

[0067] The polypeptides to be tested in this embodiment are 4 polypeptides: ① polypeptide SEQ ID NO: 2 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15; ② polypeptide SEQ ID NO: 5 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15; ③ polypeptide SEQ ID NO: 7 + connecting peptide SEQ ID NO: 57 + cell penetrating peptide SEQ ID NO: 15; ④ polypeptide SEQ ID NO: 8 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15.

[0068] The specific experimental process of this embodiment is as follows:

[0069] Eight-week-old male C57BL / 6 mice were raised in a feeding room with a temperature of 22℃~24℃, a humidity of 50%, and a daily light exposure time of 12h. The mice had free access to food and water. The experimental mice were randomly divided into 7 groups: Control group, MPTP group, MPTP+Tat group, MPTP+Tat-peptide 2 group, MPTP+Tat-peptide 5 group, MPTP+Tat-peptide 7 group, and MPTP+Tat-peptide 8 group. The peptide solution to be tested (1μg / g, once a day) was injected intraperitoneally on days 1 to 6, and MPTP (20mg / kg; 2h interval, a total of 4 times) was injected intraperitoneally on day 2. The control group was injected with an equal amount of saline. The open field test was performed on day 8. After the behavioral experiment, the mice were killed and brain tissues were collected.

[0070] The open field experiment device consists of an open field reaction box and an automatic data collection and processing system. The bottom side of the mouse open field reaction box is 50 cm long, the height of the reaction box is 35 cm, the inner wall is painted in light colors, and it is divided into four equal small squares by partitions. A camera is set up 1 to 2 meters above, and its field of view can cover the entire open field. The experiment was carried out in a quiet environment. First, open the open field experiment supporting software, set the parameters according to the experimental requirements, and then gently place the mouse in the center of the bottom of the reaction box, while taking pictures and collecting data. The mouse freely explores the environment in the reaction box for 5 minutes, and observes and records the activities of the mouse in the open field within 5 minutes.

[0071] The results are as follows Figure 6 As shown, compared with the model group mice, the four peptides can improve the autonomous activity and spatial exploration ability of Parkinson's disease model mice and alleviate the Parkinson's disease-like behavioral abnormalities in the model mice, while the cell penetrating peptide Tat alone did not show significant improvement.

[0072] Example 7

[0073] This example is to detect the effects of various polypeptides on pathological abnormalities of substantia nigra and striatum in Parkinson's disease model mice.

[0074] The polypeptides to be tested in this embodiment are 4 polypeptides: ① polypeptide SEQ ID NO: 2 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15; ② polypeptide SEQ ID NO: 5 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15; ③ polypeptide SEQ ID NO: 7 + connecting peptide SEQ ID NO: 57 + cell penetrating peptide SEQ ID NO: 15; ④ polypeptide SEQ ID NO: 8 + connecting peptide SEQ ID NO: 48 + cell penetrating peptide SEQ ID NO: 15.

[0075] The specific experimental process of this embodiment is as follows:

[0076] The mouse feeding and treatment methods are the same as those in Example 6. After killing the mice, the substantia nigra and striatum of the mouse brain were separated on ice and placed in a weighed EP tube, and RIPA tissue lysis solution was added at 10 μL / mg (a mixture of protease inhibitors and phosphatase inhibitors was added at a ratio of 1:100). Homogenize on ice using a tissue homogenizer and place on a vortex instrument for full oscillation and lysis, once every 5 minutes, for a total of 5 times, and then use a high-speed refrigerated centrifuge at 4°C, 12000r / min centrifugation for 10 minutes to take the supernatant. The protein concentration was determined using a BCA protein concentration assay kit and the protein concentration was adjusted to the same level using PBS buffer. 1 / 4 of the total volume of 5×SDS loading buffer was added to the protein solution, fully mixed and placed in boiling water for heating for 10 minutes, and stored in a -20°C refrigerator. The protein samples were analyzed by protein immunoblotting, using the same method as in Example 3.

[0077] The results are as follows Figure 7 As shown, compared with the model group mice, the four peptides can increase the expression of TH in the substantia nigra and striatum of Parkinson's disease model mice and alleviate the Parkinson's disease-like pathological abnormalities in the model mice, while the cell penetrating peptide Tat alone did not show significant improvement.

[0078] In the present invention, the polypeptide may also be any sequence other than SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO: 7, and SEQ ID NO: 8 in the following table.

[0079]

[0080]

[0081] The cell penetrating peptide may also be a sequence other than SEQ ID NO: 15 in the following table.

[0082] SEQ ID NO sequence 15 YGRKKRRQRRR 16 GRKKRRQRRR 17 GRKKRRQRRRPPQ 18 CYGRKKRRQRRR 19 RKKRRQRRR 20 RRRQRRKKRGY 21 RRRR 22 RRRRRR 23 RRRRRRR 24 RRRRRRRR 25 RRRRRRRRR 26 RRRRRRRRRR 27 RRRRRRRRRRRR 28 RRRRRRRRRRRRRRR 29 RQIKIWFQNRRMKWKK 30 YARAAARQARA 31 LSTAADMQGVVTDGMASGLDKDYLKPDD 32 KETWWETWWTEWSQPKKRKV 33 CVKRGLKLRHVRPRVTRMDV 34 DAATATRGRSAASRPTERPRAPARSASRPRRPVD 35 RGGRLSYSRRRFSTSTGR 36 RRIRPRPPRLPRPRPRPLPFPRPG 37 PIEVCMYREP 38 VPTLK 39 CHHHHHRRRRRRRRRHHHHHC 40 MVTVLFRRLRIRRACGPPRVRV 41 Ac-GLWWRLWWRLRSWFRLWFRA-Cya 42 LLIILRRRIRKQAHAHSK 43 CRWRWKCCKK 44 KWRRKLKKLRPKKKRKV 45 MASIWVGHRG 46 RRRKIKR 47 YGRKKRRQRRRLLRAALRKAAL

[0083] The connecting peptide may also be a sequence other than SEQ ID NO:48 and SEQ ID NO:57.

[0084]

[0085]

[0086] In addition to the above embodiments, the present invention may also have other implementation modes. Any technical solution formed by equivalent replacement or equivalent transformation falls within the protection scope required by the present invention.

Claims

1. A polypeptide, characterized in that: The amino acid sequence of the polypeptide is SEQ ID NO:

7.

2. A polypeptide characterized in that: It is formed by connecting the polypeptide of claim 1 with a cell-penetrating peptide, wherein the cell-penetrating peptide is located at the N-terminus of the polypeptide; the amino acid sequence of the cell-penetrating peptide is one of SEQ ID NO: 15 to SEQ ID NO:

47.

3. A polypeptide characterized in that: The polypeptide according to claim 1 is connected to a cell-penetrating peptide via a connecting peptide, wherein the connecting peptide and the cell-penetrating peptide are both located at the N-terminus of the polypeptide; the amino acid sequence of the cell-penetrating peptide is one of SEQ ID NO: 15 to SEQ ID NO: 47, and the amino acid sequence of the connecting peptide is one of SEQ ID NO: 48 to SEQ ID NO:

72.

4. Use of the polypeptide according to any one of claims 1 to 3 for preparing a medicament for treating or preventing a neurodegenerative disease.

5. The use according to claim 4, characterized in that: The neurodegenerative disease is Alzheimer's disease.

6. The use according to claim 4, characterized in that: The neurodegenerative disease is Parkinson's disease.

7. A nucleic acid encoding the polypeptide according to any one of claims 1 to 3.

8. A pharmaceutical composition comprising the polypeptide according to any one of claims 1 to 3 or the nucleic acid according to claim 7.

9. Use of the pharmaceutical composition according to claim 8 for preparing a pharmaceutical preparation for treating or preventing neurodegenerative diseases.

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