CN112675183B

CN112675183B - Application of PB in preparation of medicine for preventing or treating PILO-induced epilepsy

Info

Publication number: CN112675183B
Application number: CN202110098271.5A
Authority: CN
Inventors: 王信; 李莉; 刘天易; 师伟; 马传江; 杨培民
Original assignee: Affiliated Hospital of Shandong University of Traditional Chinese Medicine
Current assignee: Affiliated Hospital of Shandong University of Traditional Chinese Medicine
Priority date: 2021-01-25
Filing date: 2021-01-25
Publication date: 2021-11-16
Anticipated expiration: 2041-01-25
Also published as: CN112675183A

Abstract

The invention belongs to the field of pharmaceutical preparations, and particularly relates to application of balamin B (PB) in preparation of a medicament for preventing or treating Pilocarpine (PILO) induced epilepsy. The first discovery shows that the balaneboside B can obviously prolong the latency of pilocarpine-induced epilepsy, shorten the status time of epilepsy, reduce the epileptic seizure level and slow down the weight loss caused by epilepsy, thereby having good prevention and treatment effects on the PILO-induced epilepsy. Has important clinical application value in the aspect of pilocarpine-induced epilepsy and opens up new medicinal application for the active component of the balaneb B.

Description

Application of PB in preparation of medicine for preventing or treating PILO-induced epilepsy

Technical Field

The invention belongs to the field of pharmaceutical preparations, and particularly relates to application of balamin B (PB) in preparation of a medicament for preventing or treating Pilocarpine (PILO) induced epilepsy.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Epilepsy (epilepsy, EP), a chronic, repetitive, progressive, chronic disease caused by excessive abnormal discharge of brain nerve cells, is clinically manifested as sudden seizures accompanied by transient cerebral dysfunction and is classified as the second largest, central nervous system disease worldwide.

Currently, the clinical treatment of EP mainly includes taking Antiepileptic drugs (AEDs), but the AEDs cannot completely and effectively control the attack of about 30% of EP patients, and long-term taking of the AEDs can also cause adverse reactions such as decline of cognitive function and ataxia which seriously affect the quality of life. The traditional Chinese medicine has a long history of epilepsy treatment, has the advantages of small toxic and side effects and multiple action targets, has great excavation potential in the research and development of antiepileptic medicines taking the traditional Chinese medicine as a source, and becomes a hotspot of anti-EP medicine research in recent years. For example, the alpha-asarone in the rhizoma acori graminei can obviously improve the learning and memory ability of EP rats, prolong the incubation period of spontaneous and repeated attacks of EP, and reduce the attack frequency and grade; the rhynchophylline in the uncaria rhynchophylla can improve the activity of serum superoxide dismutase and reduce the expression of a Hippocampus tissue Toll-like receptor 4, and has a protection effect on rat brain injury caused by an EP (EP) continuous state; ligustrazine in rhizoma Ligustici Chuanxiong can reduce the expression of IL-2, IL-6 and TNF-alpha to resist EP attack of rats caused by pentaerythrine; piperine in pepper can remarkably prolong the eclamptic latency of EP mice, reduce the death rate and the like by influencing the functions of inhibitory amino acid and gamma-aminobutyric acid, and has important social and economic benefits by searching high-efficiency and low-toxicity EP control natural active ingredients in Chinese medical treasury.

However, the inventor researches and discovers that various bioactive components extracted from the existing animals and plants have certain prevention and prevention effects on EP. However, the causes of epilepsy include heredity, endocrine dyscrasia, sleep cause, chemical components and the like, and epilepsy caused by different causes needs to be prevented or treated by different methods in the actual prevention or treatment process.

Disclosure of Invention

In order to more accurately and efficiently prevent or treat epilepsy, the inventor finds that the paliperidone B (Parishin B, PB) can significantly prolong the latency of epilepsy induced by Pilocarpine (PILO), shorten the status time of epilepsy, reduce the epileptic seizure level, and slow down the weight loss caused by epilepsy, thereby having good prevention and treatment effects on the PILO-induced epilepsy.

Specifically, the invention is realized by the following technical scheme:

in a first aspect of the invention, the invention provides an application of the balaneb B in preparing a medicament for preventing and/or treating epilepsy.

In a second aspect of the invention, a medicament for preventing and/or treating epilepsy is provided, which comprises the balaneboside B.

One or more embodiments of the present invention have the following advantageous effects:

1) the invention discovers for the first time that the balaneb B can obviously prolong the latent period of the pilocarpine-induced epilepsy, shorten the duration time of the epilepsy, reduce the epileptic seizure grade and slow down the weight loss caused by the epilepsy, thereby having good prevention and treatment effects on the PILO-induced epilepsy.

2) By synthesizing PPI network, target enrichment analysis and other results, FGFR3, FGF-2, PTEN, MAPK12 and MTOR 5 proteins are selected as key targets, and PDB-ID of corresponding proteins is found through Protein Data Bank (http:// www.rcsb.org/PDB /). The SYBYL-X2.0 software Surflex-Dock module is used for carrying out molecular docking on PB and key target proteins, and the result shows that the PB and each target protein have certain interaction, wherein the PB and each target protein have better binding activity with FGFR3, MAPK12 and MTOR.

3) The research of the invention shows that the barbaloin B has the capacity of preventing or treating pilocarpine-induced epilepsy, has important clinical application value in the aspect of pilocarpine-induced epilepsy, and opens up new medicinal application for the active ingredient of the barbaloin B.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1 is a graph showing the effect of PB on body weight of EP mice in example 2 of the present invention;

FIG. 2 is a graph showing the effect of PB on the survival of EP mice in example 2 of the present invention;

FIG. 3 is a graph showing the effect of PB on the latency to epilepsy in EP mice in example 2 of the present invention;

FIG. 4 is a graph showing the effect of PB on the duration of epileptic seizure in EP mice in example 2 of the present invention;

FIG. 5 is a diagram of interaction network association of PB anti-EP core targets in embodiment 2 of the present invention;

FIG. 6 is a GO biofunctional annotation of the PB anti-EP core target in example 2 of the present invention;

FIG. 7 is a KEGG pathway enrichment analysis bubble map of PB anti-EP action target in example 2 of the present invention;

FIG. 8 is a diagram showing the docking pattern of PB and key target molecules in example 2 of the present invention.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

The molecular formula of the balaneboside B (Parishin B, PB) is as follows:

specifically, the invention is realized by the following technical scheme:

In one or more embodiments of the invention, the epilepsy is due to pilocarpine-induced priming.

In one or more embodiments of the invention, the balaneboside B can significantly prolong the epileptic latency of epileptic mice, shorten the status epilepticus, reduce the epileptic seizure level of epileptic mice, and inhibit the weight loss caused by epilepsy.

In one or more embodiments of the invention, the balancide B prolongs the epileptic latency of epileptic mice, shortens the status epilepticus, reduces the epileptic seizure grade of epileptic mice, and inhibits the weight reduction caused by epilepsia by combining with the target FGFR3, MAPK12 and MTOR.

The discovery of the combination of the new drug and the target is helpful to provide a basis for multi-target administration, avoid the competition of the drugs acting on the same target and reduce the drug effect.

The prevention and/or treatment of epilepsy is caused by pilocarpine-induced priming.

In one or more embodiments of the invention, the pharmaceutically effective concentration is 50-200 μmg/kg and above, preferably 200 mg/kg.

In one or more embodiments of the present invention, the medicament for preventing and/or treating epilepsy further comprises one or more pharmaceutically or dietetically acceptable excipients.

In one or more embodiments of the present invention, solid form preparations include powders, tablets, dispersible granules, capsules, pills, and suppositories; preferably, powders and tablets contain from about 0.1% to about 99.9% of the active ingredient; the solid adjuvant is selected from magnesium carbonate, magnesium stearate, pulvis Talci, sugar or lactose;

the liquid form of the preparation includes solutions, suspensions and emulsions, and preferably, the liquid form is an parenteral solution in water or water-propylene glycol, or an oral solution with added sweeteners and contrast agents.

In one or more embodiments of the invention, the medicine is prepared into small water injection for injection, freeze-dried powder injection for injection, large infusion solution or small infusion solution.

The present invention is described in further detail below with reference to specific examples, which are intended to be illustrative of the invention and not limiting.

Instrument and reagent

KQ-250 ultrasonic cleaner (Kunshan ultrasonic instruments Co., Ltd.); an AV412 model tenths electronic balance (aohaus corporation, usa); model EP214C ten thousandth electronic balance (mettler-toledo instruments ltd, switzerland); balisin B (batch number: PRF7070743, mass fraction not less than 98%) purchased from Dougeny scientific development Co., Ltd; pilocarpine hydrochloride (batch number: 066k1730, mass fraction not less than 98%) purchased from Sigma company of America; 0.9% physiological saline (batch No. 20190720) obtained from Shandong Tupu bioengineering, Inc.; atropine sulfate injection (batch No. 1902251) and diazepam injection (batch No. 1903101) were purchased from Tianjin Jinyao pharmaceutical Co., Ltd.

Laboratory animal

SPF grade C57BL/6J male mouse, 6 weeks old, 20.0-23.0 g weight, purchased from experimental animal breeding Limited of Jinaponi, production license number: SCXK (lu) 20140007. A breeding environment: animal experiment center of affiliated hospital of Shandong Chinese medicine university, barrier environment is raised, uses license serial number: SYXK (lu) 20180015. The experiment was approved by the ethical committee of experimental animals of the subsidiary hospital of Shandong Chinese medicine university.

Example 1 experimental part

Establishment of EP model

After C57BL/6J mice are adaptively fed for 1 week, the mice are taken and injected with atropine sulfate (1mg/kg) in the abdominal cavity to antagonize peripheral cholinergic reaction, and PILO (280mg/kg) is injected in the abdominal cavity after 15min, and the reaction is stopped by injecting diazepam (10mg/kg) in the abdominal cavity 60min after EP symptoms appear. And (4) evaluating according to Racine grades, and judging that the model preparation is successful when the attack grade reaches 4-5 grades. The specific indexes of the Racine grading are as follows: level 1: facial twitching; and 2, stage: pointing the head; and 3, level: onset of unilateral forelimb clonus; 4, level: rising, onset of bilateral forelimb clonus; and 5, stage: bilateral anterior limb clonic attacks accompany it.

2. Animal grouping and administration

15 mice of C57BL/6J are randomly selected as blank groups, and the mice successfully prepared by the model are randomly divided into a model group and PB low, medium and high dose administration groups, wherein each group comprises 15 mice. Blank group: the normal saline is injected into the stomach and the normal saline is injected into the abdominal cavity; model group: the normal saline is given by intragastric administration and the PILO is injected into the abdominal cavity; PB administration group: the PB was gavaged + intraperitoneal PILO. The dose of the PILO intraperitoneal injection is 280mg/kg, 1 time per 2 days, and the total time is 28 days. The low, medium and high dosage of PB is 50, 100, 200mg/kg, 1 time/1 day, 28 days. The model group and PB administration group were administered by intragastric administration for 30min, and were administered by intraperitoneal injection of atropine sulfate (1mg/kg) to antagonize peripheral cholinergic reaction, and after 15min, by intraperitoneal injection of PILO, mice were administered by intraperitoneal injection of diazepam (10mg/kg) 60min after epileptic symptoms.

Effect of PB on weight, survival and epileptic behavior in epileptogenic mice

Weighing the weight of the mice before gavage on the 1 st and 28 th days, and recording and calculating the average weight of each group of mice; the death of each group of mice was recorded daily and the survival rate was calculated; epileptic behaviors such as the incubation period (min), the attack duration(s), the attack grade and the like of EP attacks of the mice are closely observed and recorded after the PILO is injected into the abdominal cavity on the 2 nd and the 28 th days, the attack grade is evaluated according to the Racine grades, and the successful preparation of the model is regarded as if the attack grade reaches the grade of 4-5. The specific indexes of the Racine grading are as follows: level 1: facial twitching; and 2, stage: pointing the head; and 3, level: onset of unilateral forelimb clonus; 4, level: rising, onset of bilateral forelimb clonus; and 5, stage: bilateral anterior limb clonic attacks accompany it.

Screening of action targets of PB component

Selecting a human gene 'Homo sapiens' as a research object by using a Swiss Target Prediction (http:// www.swisstargetprediction.ch /) database, and predicting a potential action Target point of a PB component; and searching PB related action target points through an SEA database (http:// SEA. bkslab. org /), merging the compound target points searched by 2 databases, and deleting repeated genes to obtain the PB component action target point gene.

EP disease target screening

And (3) utilizing OMIM (https:// www.omim.org /), GeneCards (http:// www.genecards.org /) and ETCM (http:// www.nrc.ac.cn:9090/ETCM/Index. php/Home/Index. html) databases to respectively search EP disease targets by taking the epilesys as a keyword, and merging the search results of the 3 databases to obtain the target gene related to the EP disease.

Construction of PB component target-EP disease target interaction network (PPI) and screening of core target

The PB component target point-EP disease target point PPI is constructed on an STRING (https:// STRING-db.o-rg) platform, the species is set as ' Homo sapiens ', the lowest interaction threshold value is set as Medium confidence ' 0.4, and the rest parameters are kept at default settings. And (3) carrying out topological attribute analysis on the PPI network by using Cytoscape 3.7.1, and screening a core target point in the PPI network. Screening conditions are as follows: selecting a core node of the PPI by taking a median of 2 times of the node connectivity (degree) as a stuck value; on the basis, taking the median of 'degree', 'node compactness' and 'node betweenness' as the card value, and selecting the target point which meets 3 card values at the same time as the core target point.

7. GO biological function annotation and KEGG pathway enrichment analysis of core target

And performing GO biological function annotation and KEGG pathway enrichment analysis on the screened core targets by using a DAVID database (https:// DAVID. nciff. gov/summary. jsp), wherein the GO biological function annotation comprises a Biological Process (BP), a Molecular Function (MF) and a Cellular Component (CC). The name of the species is set as 'Homo sapiens', and P is set to be less than or equal to 0.05.

8. Molecular docking

And (3) integrating the results of PPI network, target enrichment analysis and the like, performing molecular docking verification on PB and the key target by using a SYBYL-X2.0 software Surflex-Dock module, predicting the possible interaction between PB molecules and target protein, and evaluating the binding activity between PB components and the key target according to the docking scoring Total score result.

9. Statistical method

Statistical analysis was performed using SPSS 20.0 statistical software. The measurement data is expressed by x +/-s, the single-factor analysis of variance is adopted for the comparison among multiple groups, the SNK test is adopted for uniform variance, the Dunnet test is adopted for irregular variance, and P is less than or equal to 0.05, so that the difference has statistical significance.

EXAMPLE 2 discussion of results

Effect of PB on EP mice body quality and survival Rate

No significant difference (P is more than or equal to 0.05) exists in the body mass of each group of mice in the experiment of day 1. On the 28 th day, compared with the blank group, the body mass of the mice in the model group is remarkably reduced (P is less than or equal to 0.05), and the long-term EP attack is prompted to have a certain inhibiting effect on the body mass growth of the mice; compared with the model group, the body mass of the mice in the low-dose group has no significant change (P is more than or equal to 0.05), and the body mass of the EP mice can be significantly increased (P is less than or equal to 0.05) in the medium-dose group and the high-dose group, which indicates that the PB administration can inhibit the body mass reduction trend of the EP mice; compared with the low-dose group, the mice in the medium-dose group have no significant difference in body mass change (P is more than or equal to 0.05), and the mice in the high-dose group have significant difference in body mass change (P is less than or equal to 0.05), so that the PB administration has certain dose dependence on the trend of improving the reduction of the body mass of the EP mice, and specific results are shown in figure 1.

The 28-day survival rate of the model group is 60.0 percent, and is obviously reduced compared with the survival rate of the blank group (P is less than or equal to 0.05), which indicates that the survival rate of the mice can be reduced by long-term EP attack; the survival rates of the low-dose group and the medium-dose group in 28 days are 66.7 percent and 73.3 percent respectively, the survival rates are not significantly different (P is more than or equal to 0.05) compared with the survival rate of the model group, the survival rate of the high-dose group in 28 days is 93.3 percent, the survival rate is significantly improved (P is less than or equal to 0.05) compared with the survival rate of the model group, the survival rate of the EP mice can be improved by the appropriate dose of PB administration, and specific results are shown in figure 2.

Note: p is less than or equal to 0.05 compared with the blank group; compared with the model group, # P is less than or equal to 0.05, compared with the low dose group, # P is less than or equal to 0.05

Effect of PB on Ep mouse epileptic behavior Change

In the experiment of 2 days, the EP attack latency and the sustained attack time of each group of mice have no significant difference (P is more than or equal to 0.05), and the attack grade is 4-5. On the 28 th day, compared with the model group, the low-dose group EP mice have no significant difference in onset latency and sustained onset time (P is more than or equal to 0.05), and the onset grade is 4-5 grade; compared with a model group and a low dose group, the medium dose group and the high dose group can obviously prolong the epileptic seizure latency (P is less than or equal to 0.05) of the EP mice and shorten the sustained seizure time (P is less than or equal to 0.05), and the seizure grade is 3-4; compared with the medium-dose group, the high-dose group EP mice have significant differences in seizure latency and seizure duration (P is less than or equal to 0.05), and the results indicate that the appropriate dose of PB administration can significantly improve the epileptic behavior of the EP mice, and the specific results are shown in fig. 3 and fig. 4.

PB component action target

After searching through an SEA database and a Swiss Target Prediction database, 63 PB potential action targets are obtained after repeated values are removed, and the results are shown in Table 1.

TABLE 1 PB component potential action target

Table 1 Potential targets of PB

EP disease targets

After the repeated values are removed, 719 potential action targets of the EP disease are obtained by searching OMIM, GeneCards and ETCM databases.

Construction of PB component target-EP disease target PPI and core target

The PB component target and the EP disease target are primarily screened in a STRING database to obtain 304 targets, then the PB component target and the EP disease target are introduced into Cytoscape-v 3.7.1 software to carry out topology analysis screening, and finally 52 core targets are obtained, specifically shown in Table 2, and PPI is shown in FIG. 5.

TABLE 2 PB anti-EP core target and its topology parameters

Table 2 Key targets of PB in antiepileptic and their topological parameters

6. GO biological function annotation and KEGG pathway enrichment analysis of core target

And importing 52 target information acquired by the item of 'PB component target-EP disease target PPI construction and core target' into a DAVID database for GO enrichment analysis. The results show that 27 entries are related to BP, including angiogenesis, cell proliferation, neuronal synaptic plasticity, cell division, cellular hypoxia response, central nervous system myelination, protein phosphorylation, intracellular sodium homeostasis, etc.; there are 3 items associated with MF, including growth factor activity, protein binding, ATP binding; there are 13 entries associated with CC, including golgi, neuronal synapses, plasma membranes, exosomes, dendrites, axons, microtubules, etc., see fig. 6.

59 biological pathways are enriched by KEGG pathway annotation analysis, 15 pathways related to EP are obtained by screening according to a threshold value that P is less than or equal to 0.05, the results of the pathway related target protein genes are shown in Table 3, and a pathway enrichment bubble map is shown in FIG. 7. The result analysis shows that the PB anti-EP mechanism can be related to a VEGF signaling pathway, a MAPK signaling pathway, a PI3K-Akt signaling pathway, a FoxO signaling pathway, a Rap1 signaling pathway, a Ras signaling pathway and the like. In addition, the research also finds that PB has certain effects on glioma, renal cell carcinoma, endometrial carcinoma and the like.

TABLE 3 PB antiepileptic KEGG enrichment related pathway target protein genes

Table 3 Genes enriched by PB inantiepileptic-associated KEGG pathway

7. Molecular docking validation

By synthesizing PPI network, target enrichment analysis and other results, FGFR3, FGF-2, PTEN, MAPK12 and MTOR 5 proteins are selected as key targets, and PDB-ID of corresponding proteins is found through Protein Data Bank (http:// www.rcsb.org/PDB /). The results of molecular docking of PB and key target proteins using the Surflex-Dock module of SYBYL-X2.0 software are shown in Table 4 and FIG. 8. The literature indicates that Total score is more than or equal to 4.25; the molecules and the target points have certain interaction; total score is more than or equal to 7.0, which indicates that the molecule has better binding activity with the target, and the docking result shows that PB has certain interaction with each target protein, wherein the PB has better binding activity with FGFR3, MAPK12 and MTOR.

TABLE 4 molecular docking score values of PB component with Key target proteins

Table 4 Docking scores of PB with its key targets

Example 3

Based on the early-stage wine-roasted gastrodia elata related research basis, the invention firstly discusses the preliminary intervention effect of the PB component on PILO-induced EP mouse epileptic behavior. The Shen nong Ben Cao Jing records that Tian Ma is pungent and warm in flavor. … …, long-term administration of the medicine can tonify qi, nourish yin, strengthen body constitution, and prolong life. The research shows that the PB administration can inhibit the reduction trend of the body mass of the EP mouse, has certain dose dependence, can improve the survival rate of the EP mouse, prolong the latent period of EP attack, shorten the duration time and reduce the attack grade, has certain improvement effect on the epileptic behavior of the EP mouse, and lays a research foundation for the development of PB antiepileptic innovative medicaments.

The cognitive function of the brain is damaged by EP attack or EP continuous state, and the improvement of the cognitive dysfunction caused by the EP attack, the learning and memory impairment and other brain function injuries is one of the hotspots of the current antiepileptic drug research, for example, the alpha-mangostin component in the mangosteen shell has a therapeutic effect on EP mice ignited by pentaerythrine and can improve the learning and memory abilities of the EP mice; the Ganoderma triterpene component in Ganoderma has effects of improving learning and memory injury of EP rats ignited by pentaerythrite, and improving nerve cell injury caused by EP. The study shows that the gastrodia elata has an improvement effect on learning and memory disorders of models such as rats with vascular dementia, chronic unpredictable mild stress rats, mice with memory acquisition, consolidation and recurrence disorder, rats with A beta 1-42 dementia and the like, and whether the PB component has the improvement effect on learning and memory abilities of EP mice induced by PILO (platelet activating hormone oxidase) is further investigated in the later stage of the study.

The research adopts a network pharmacology technical method to discuss the potential action mechanism of the PB component against EP, and PPI network analysis and screening obtains 52 core targets including VEGFA, FGF2, HRAS, PTEN, MTOR, SLC2A1, IL2, ESR1, CAV1, GRM5, SCN2A and the like. Wherein VEGFA is the most main vascular endothelial growth factor, VEGF can regulate EP intracerebral synaptic nerve transmission and cerebrovascular abnormality, improve angiogenetic edema, promote hippocampal nerve regeneration after EP continuous state and reverse cognitive deficiency, and the system participates in EP pathogenesis; FGF-2 has the function of neurotransmitter or neuromodulation, is a main mechanism for forming learning and memory, and can enhance the endogenous neuroprotective function of EP rats; the expression of PTEN can prolong the incubation period of EP rats, reduce the EP attack frequency, inhibit the apoptosis of cerebral neurons, reduce neuroinflammatory reaction, and target PTEN can play a key role in EP generation; MTOR regulates neuron development and synaptic plasticity, participates in regulation of oxidative stress and autophagy, plays an important role in the formation and generation of epilepsy, the analysis shows that PB treatment EP has the action characteristics of multiple targets, multiple links and multiple paths, related reports on PB anti-EP action mechanism research are not seen at present, and other molecular docking research results show that FGF-2, PTEN, MTOR and other target proteins and PB have certain binding activity and can become potential action targets for PB anti-EP research.

GO enrichment analysis shows that PB anti-EP mainly relates to biological processes such as angiogenesis, cell proliferation, neuronal synaptic plasticity, cell hypoxia reaction, CNS myelination, protein phosphorylation and the like, molecular functions such as growth factor activity and protein combination and cell components such as exosomes and microtubules. Studies report that activation of neurosynaptic plasticity can inhibit EP morpho-activity; central nervous system dysmyelination can cause EP attack, and central nervous system white matter demyelination change caused by repeated EP attack can promote further EP attack; the growth of nerve axons can be inhibited by inhibiting the expression of the CRMP-2 protein and promoting the phosphorylation of the protein, so that the anti-EP effect can be exerted; the Tau protein is over-phosphorylated to lose the biological function of promoting microtubule assembly, and the reduction of the phosphorylation degree of the Tau protein can improve the cognitive function reduction caused by EP onset. By combining with the analysis of the effects of FGF-2, MTOR and other targets, the later PB anti-EP mechanism research can be deeply carried out from the aspects of regulating and controlling cell oxidative stress, autophagy, influence of microtubule assembly on nerve growth factor activity, neuronal synaptic plasticity and the like.

The enrichment analysis of the KEGG pathway shows that PB anti-EP mainly relates to signal pathways such as VEGF, MAPK, PI3K-Akt, FoxO, Rap1, Ras and the like. Research reports that the latent period after the immature cerebral convulsion persists is a key period influencing the formation of EP, and inhibition of VEGF pathway expression in the latent period can improve pathological damage of hippocampus and prevent the formation of EP; the MAPK signal pathway plays a key role in EP pathogenesis, and the p38MAPK inhibitor can effectively shorten the epileptic seizure time of an EP rat and improve the cognitive function of the rat; the adenosine A2A receptor blocker SCH58261 can play a certain role in protecting hippocampal neuron injury caused by EP sustained state by inhibiting MAPK pathway; miR129 inhibits c-Fos expression by inhibiting MAPK signal path, and influences proliferation and apoptosis of hippocampal neurons, thereby inhibiting generation and development of EP; h₂S can change the expression ratio of apoptosis protein Bcl-2 and Bax by activating PI3K/Akt channel, thereby playing the role of inhibiting EP and protecting hippocampal neurons; grape seed procyanidin can inhibit Caspase-3 expression by activating PI3K/Akt signal pathway so as to inhibit apoptosis of EP mouse hippocampal nerve cells and protect mitochondrial function; the EP persistence state can lead to a decrease in autophagy activity of hippocampal neurons, necrosis and apoptosis of neurons, and curcumin can protect neuronal damage in EP rats by up-regulating PI3K/AKt/mTOR expression. The above shows that at present, many researches are carried out on EP related channels such as MAPK, PI3K/Akt and the like, the research on the anti-epileptic action mechanism based on aspects of reducing the neuronal inflammatory response and regulating the neuronal autophagy of the signal channels PB such as MAPK, PI3K/Akt/mTOR and the like can be explored and developed in the later period by integrating the enrichment analysis of target spots and the docking results of related target gene molecules of the MAPK channel PI 3K-Akt.

The research discusses the anti-EP activity and potential action targets and action mechanisms of PB components through a combined research mode of primary pharmacodynamic evaluation, network pharmacological analysis, molecular docking and literature information mining, the research result indicates that the PB can play the anti-EP pharmacological action through multiple targets and multiple paths, and the potential key targets and paths obtained through prediction analysis are tested and verified in the next step, so that a research basis is provided for the deep development of the PB anti-epileptic innovative medicine.

Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The application of the balaneboside B as the only active component in preparing the medicine for preventing and/or treating epilepsy;

the epilepsy is caused by pilocarpine-induced initiation.

2. The use according to claim 1, wherein the Balisenside B is capable of significantly prolonging the latency to epilepsy in an epileptic mouse, shortening the duration of status epilepticus, reducing the level of epileptic seizures in an epileptic mouse, and/or inhibiting the weight loss resulting from epilepsy.

3. The use according to claim 1, wherein the balancide B, by binding to the target FGFR3, MAPK12, MTOR, prolongs the latency of epilepsy in epileptic mice, shortens the duration of status epilepticus, reduces the grade of epileptic seizures in epileptic mice, and/or inhibits weight loss due to epileptic production.