Ginseng small molecule peptide and application of ginseng small molecule peptide in senile dementia related protein kinase inhibitor

Abstract

The invention provides a ginseng small molecule peptide and application thereof in an inhibitor for treating senile dementia related protein kinase, and relates to the field of bioactive peptides. The ginseng small-molecule peptide is a small-molecule peptide with DAPK1 potential inhibitory activity, the peptide sequence of the ginseng small-molecule peptide is TNHAK, and the application of the small-molecule peptide with the peptide sequence being TNHAK in preparation of a death-related protein kinase 1 inhibitor is provided. Through a virtual screening method, the small molecule peptide possibly having death-related protein kinase 1 inhibitory activity in the ginseng is obtained. Experiments prove that the TNHAK small molecule peptide has the potential of serving as a death-related protein kinase 1 inhibitor, and has important significance on treatment of Alzheimer's disease.

Description

Technical Field

[0001] The present invention relates to the field of bioactive peptides, in particular to ginseng small molecule peptides and their application in treating Alzheimer's disease-related protein kinase inhibitors. Background Art

[0002] Alzheimer's disease (AD) is the most common type of dementia in the elderly and a major clinical challenge in an aging society. The neuropathological characteristics of AD include neurofibrillary tangles composed of hyperphosphorylated tau protein, senile plaques produced by β-amyloid protein deposition, brain atrophy caused by neuronal loss, and synaptic dysfunction. Death-associated protein kinase 1 (DAPK1) is a serine / threonine protein kinase that participates in multiple cell death pathways, such as apoptosis, autophagy, and necrosis. It is ubiquitously expressed in the central nervous system. Dysregulation of DAPK1 has been shown to cause a variety of neurological diseases, including Alzheimer's disease.

[0003] DAPK1 plays multiple roles in the pathogenesis of Alzheimer's disease (AD). DAPK1 is involved in the neurotoxic process induced by Aβ and promotes the formation of neurofibrillary tangles by phosphorylating tau protein. DAPK1 has become an emerging target for the development of AD treatment methods. An increasing number of DAPK1 inhibitors are being developed for the treatment of various neurological diseases, including synthetic compounds, natural product-derived chemicals, and peptide-based modulators. The development of DAPK1 inhibitors with balanced efficacy, safety, and good pharmacological characteristics for further preclinical and clinical studies is of great significance for the intervention and treatment of AD. Summary of the invention

[0004] 1. Technical issues to be resolved

[0005] In view of the deficiencies of the prior art, the present invention provides ginseng small molecule peptides and their application in the treatment of Alzheimer's disease-related protein kinase inhibitors. The method uses a simpler and more feasible research method to screen DAPK1 inhibitory peptides. Virtual screening is to use a computer to perform preliminary screening of compound molecules according to preset conditions before biological activity screening to identify small molecules that are most likely to bind to the target. It can significantly reduce the number of compounds that need to be screened and improve the efficiency of discovering lead compounds. It uses computer technology to simulate the hydrolysis process of trypsin and predict the affinity of the generated small peptides. It can be used as a more feasible and efficient method than traditional enzymatic screening of DAPK1 inhibitory peptides.

[0006] (II) Technical solution

[0007] To achieve the above objectives, the present invention is implemented through the following technical scheme: a ginseng small molecule peptide and its application in treating Alzheimer's disease-related protein kinase inhibitors, the ginseng small molecule peptide is a small molecule peptide with potential DAPK1 inhibitory activity, and its peptide sequence is TNHAK. The ginseng small molecule peptide and its application in treating Alzheimer's disease-related protein kinase inhibitors involve using computer virtual screening technology to obtain a small molecule peptide TNHAK with DAPK1 inhibitory activity from ginseng.

[0008] Preferably, the application of the ginseng small molecule peptide in the treatment of Alzheimer's disease-related protein kinase inhibitors involves using a cell tau phosphorylation detection experiment to verify the small molecule peptide whose peptide sequence is TNHAK, thereby verifying that the small molecule peptide has the potential to serve as a death-related protein kinase 1 inhibitor.

[0009] Preferably, the small molecule peptide plays a role in the process of preparing death-associated protein kinase 1 inhibitor.

[0010] (III) Beneficial effects The present invention provides a ginseng small molecule peptide and its application in treating Alzheimer's disease-related protein kinase inhibitors. It has the following beneficial effects: 1. The present invention provides ginseng small molecule peptides and their application in treating Alzheimer's disease-related protein kinase inhibitors. Compared with ordinary small molecule inhibitors, peptide-based inhibitors are highly selective and specific to protein targets and their substrates. At the same time, peptide-based inhibitors can be easily modified to increase inhibitory efficacy and biodiversity, and the metabolites of peptides are amino acids with low cytotoxicity. This method uses a simpler and more feasible research method to screen DAPK1 inhibitory peptides. Virtual screening is to use computers to conduct preliminary screening of compound molecules according to preset conditions before biological activity screening to identify small molecules that are most likely to bind to the target. It can significantly reduce the number of compounds that need to be screened and improve the efficiency of discovering lead compounds. Using computer technology to simulate the hydrolysis process of trypsin and predict the affinity of the generated small peptides can be used as a more feasible and efficient method than traditional enzymatic screening of DAPK1 inhibitory peptides.

[0011] 2. The present invention provides a ginseng small molecule peptide and its application in treating Alzheimer's disease-related protein kinase inhibitors. Through a virtual screening method, a small molecule peptide in ginseng that may have death-related protein kinase 1 inhibitory activity is obtained. Experimental verification shows that the TNHAK small molecule peptide has the potential to be an inhibitor of death-related protein kinase 1. The present invention is of great significance for the treatment of Alzheimer's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1This is a schematic diagram of the tau protein phosphorylation inhibitory activity of the small molecule peptide TNHAK of the present invention; Figure 2 It is a schematic diagram of the molecular docking results of the polypeptide TNHAK of the present invention. DETAILED DESCRIPTION

[0013] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. In the description of this application, it should be noted that the terms used here are only for describing specific implementations, and are not intended to limit the exemplary implementations according to the present application. For ease of description, the sizes of the various parts shown in the drawings are not drawn according to the actual proportional relationship. The techniques, methods and devices known to ordinary technicians in the relevant fields may not be discussed in detail, but where appropriate, the techniques, methods and devices should be regarded as part of the authorization specification. In all examples shown and discussed here, any specific value should be interpreted as merely exemplary, not as a limitation. Therefore, other examples of the exemplary embodiments may have different values. It should be noted that similar reference numerals and letters represent similar items in the following drawings, so once an item is defined in one drawing, it does not need to be further discussed in subsequent drawings.

[0014] Embodiment 1: like Figure 1-2 As shown, the embodiment of the present invention provides ginseng small molecule peptides and their application in treating Alzheimer's disease-related protein kinase inhibitors. First, all ginseng protein sequences were obtained from the National Center for Biotechnology Information of the United States. According to the principle of trypsin hydrolysis, all ginseng peptides were subjected to computer-simulated trypsin hydrolysis. Trypsin mainly acts on lysine and arginine residues in the peptide chain, hydrolyzing the C-terminal peptide bonds of these residues, thereby decomposing the protein into smaller peptide segments. Secondly, the ConPLex tool was used to predict the affinity of all tripeptides, tetrapeptides, pentapeptides and hexapeptides obtained by hydrolysis. The affinity prediction results are shown in the following table:

[0015] Then, the selected peptides were subjected to molecular docking analysis using the AlphaFold Server provided by Alpha Fold 3 to predict their binding sites on the target protein. The binding sites of the peptides and DAPK1 are as follows: Figure 2 As shown; The final results combined with the prediction results of AlphaFold3 can provide a detailed understanding of the interaction between peptides and proteins. TNHAK is a non-competitive site inhibitor.

[0016] Embodiment 2: like Figure 1-2 As shown, the embodiment of the present invention provides ginseng small molecule peptides and their application in treating Alzheimer's disease-related protein kinase inhibitors, and the ginseng small molecule peptide TNHAK cell tau phosphorylation detection experiment is carried out, and the experimental verification steps are as follows: S1: The experimental system for tau protein phosphorylation detection consists of SK-N-BE cells overexpressing GFP-Tau, small molecule peptide solution and culture medium. The cells were cultured in growth medium supplemented with 10% fetal bovine serum until they reached a confluence of approximately 90%. Subsequently, the cells were treated with small molecule peptides at different concentrations and incubated for 30 minutes; S2: Prepare small molecule peptide solutions of different concentrations according to the following standards: 10 μM, 50 μM, 100 μM and 200 μM; S3: The cells should be treated at a constant temperature of 37°C. After incubation for 30 minutes, the cells were exposed to RIPA cell lysis buffer. After full lysis for 30 minutes, the supernatant was separated by centrifugation at 12,000 rpm for 10 minutes for protein extraction. After protein extraction, the proteins were separated by 10% SDS-PAGE gel electrophoresis, then transferred to a polyvinylidene fluoride membrane (PVDF membrane) and detected using pSer396 antibody. Finally, the level of phosphorylated tau protein was detected by enhanced chemiluminescence. S4: Cellular tau phosphorylation detection experiment of ginseng small molecule peptide TNHAK, the results are as follows Figure 1 This small molecule peptide has a certain inhibitory effect on tau protein phosphorylation and is a potential inhibitor of DAPK1.

[0017] The present invention simulates trypsin hydrolysis by computer technology, and uses the Complex tool to predict the affinity of the peptides obtained by hydrolysis. The AlphaFold Server provided by Alpha Fold 3 is used to perform molecular docking analysis on the screened peptides to determine the interaction mode and binding site between the peptides and proteins. The cell tau protein phosphorylation detection experiment verifies that the small molecule peptide TNHAK has the potential to be an inhibitor of death-associated protein kinase 1.

[0018] The above describes the preferred embodiments of the present invention, which are however not intended to limit the present invention. Those skilled in the art may make improvements and changes to the embodiments disclosed herein without departing from the scope and spirit of the present invention.

[0019] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Ginseng small molecule peptide, characterized by: The ginseng small molecule peptide is a small molecule peptide with potential DAPK1 inhibitory activity, and its peptide sequence is TNHAK.

2. The ginseng small molecule peptide according to claim 1, characterized in that: The ginseng small molecule peptide and its application in treating Alzheimer's disease-related protein kinase inhibitors involve obtaining a small molecule peptide TNHAK with DAPK1 inhibitory activity from ginseng using computer virtual screening technology.

3. The use of the ginseng small molecule peptide in treating Alzheimer's disease-related protein kinase inhibitor according to claim 1, characterized in that: The application of the ginseng small molecule peptide in the treatment of Alzheimer's disease-related protein kinase inhibitors involves the use of cell tau phosphorylation detection experiments to verify the small molecule peptide whose peptide sequence is TNHAK, thereby verifying that the small molecule peptide has the potential to serve as a death-related protein kinase 1 inhibitor.

4. The use of the ginseng small molecule peptide in the treatment of Alzheimer's disease-related protein kinase inhibitors according to claim 3, characterized in that: The small molecule peptide plays a role in the process of preparing death-associated protein kinase 1 inhibitor.

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