Application of combination of resolvin D1 and sodium valproate in preparation of medicine for treating epilepsy and medicine for treating epilepsy

Abstract

The invention provides application of combination of resolvin D1 and sodium valproate in preparation of a medicine for treating epilepsy and the medicine for treating epilepsy, and relates to the technical field of biomedicine. Research of the inventor finds that RvD1 and sodium valproate have a synergistic promotion effect on treatment of epilepsy, through synergistic cooperation of RvD1 and sodium valproate, epilepsy symptoms can be remarkably relieved, chronic epilepsy can be prevented, damage of epilepsy to nerves can be reduced, and RvD1 and sodium valproate can be used for preparing the medicine for treating epilepsy. According to the medicine for treating epilepsy provided by the invention, the active ingredients are RvD1 and sodium valproate, and the epilepsy can be effectively treated through the synergistic cooperation of RvD1 and sodium valproate.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to the use of resolvin D1 combined with sodium valproate in the preparation of a drug for treating epilepsy, and a drug for treating epilepsy. Background Art

[0002] Epilepsy is a neurological disorder characterized by the sudden onset of abnormal high-frequency activity in specific regions of the brain, which may spread to adjacent brain regions. This can lead to temporary disturbances in motor, sensory perception, consciousness, and autonomic nervous system functions. Among various types of epilepsy, temporal lobe epilepsy (TLE) is the most common, originating from the hippocampus or amygdala, accounting for 60% of all epilepsy cases. If left untreated, epilepsy can lead to severe neurological impairments, including cognitive comorbidities, manifested as hippocampus-dependent spatial learning and memory deficits. Current treatment methods, such as sodium valproate (VPA) and sodium channel blockers, aim to relieve symptoms by reducing these abnormal nerve discharges and manage the occurrence and progression of epileptic seizures, but fail to fundamentally address the underlying causes of epilepsy. However, approximately one-third of patients have drug-resistant epilepsy, and the types of epileptic seizures are diverse, highlighting the urgent need for innovative treatment strategies to address epilepsy itself and its related cognitive complications.

[0003] Dysregulation of glial cell function is increasingly recognized as a common hallmark feature in human epilepsy and experimental models. Among glial cells, astrocytes, as an important component of the central nervous system, have attracted attention due to their key role in shaping the dynamics of neural circuits. Recent studies have shown that astrocytes play an active role in the pathogenesis of epilepsy. For example, their activation can promote synaptic plasticity and enhance memory processes. In addition, astrocytes affect the epileptic pathogenesis by regulating the availability of neurotransmitters, gliotransmitters, synaptogenic factors, and intracellular signaling molecules. However, dysregulated astrocyte signaling may disrupt the balance of synapses and neural circuits, leading to chronic epileptic seizures and cognitive impairments. Therefore, treatment strategies are now beginning to focus on targeting pathways related to glial cell activation and the production of inflammatory factors, providing new prospects for anti-epileptic treatment.

[0004] Resolvin D1 (RvD1) is a metabolite of docosahexaenoic acid (DHA), known to promote the resolution of inflammation, restore tissue homeostasis, and alleviate diseases such as ischemic stroke, Parkinson's disease, and traumatic brain injury. However, the role of RvD1 in epilepsy has not been fully explored.

[0005] In view of this, the present invention is specifically proposed. Summary of the Invention

[0006] The first object of the present invention is to provide the application of resolvin D1 combined with sodium valproate in the preparation of a drug for treating epilepsy, so as to provide a new direction for the treatment of epilepsy.

[0007] The second object of the present invention is to provide a drug for treating epilepsy.

[0008] In order to achieve the above objects, the following technical solutions are specifically adopted:

[0009] In the first aspect, the present invention provides the application of resolvin D1 combined with sodium valproate in the preparation of a drug for treating epilepsy.

[0010] As a further technical solution, the treatment of epilepsy includes relieving epilepsy symptoms, preventing the occurrence of chronic epilepsy, and avoiding nerve damage.

[0011] As a further technical solution, the dosage of resolvin D1 is 250 ng / kg / day;

[0012] The dosage of sodium valproate is 300 mg / kg / day.

[0013] In the second aspect, the present invention provides a drug for treating epilepsy, and the active ingredients of the drug include resolvin D1 and sodium valproate.

[0014] As a further technical solution, the treatment of epilepsy includes relieving epilepsy symptoms and protecting nerves.

[0015] As a further technical solution, the dosage of resolvin D1 is 250 ng / kg / day;

[0016] The dosage of sodium valproate is 300 mg / kg / day.

[0017] As a further technical solution, the drug further includes pharmaceutical excipients.

[0018] As a further technical solution, the pharmaceutical excipients include at least one of fillers, binders, disintegrants, lubricants, flavoring agents or preservatives.

[0019] As a further technical solution, the dosage form of the drug includes oral preparations.

[0020] As a further technical solution, the oral preparations are selected from tablets, capsules, granules and pills.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The inventors have found through research that resolvin D1 (RvD1) and sodium valproate have a synergistic promoting effect in the treatment of epilepsy. Through their synergistic cooperation, they can significantly relieve epileptic symptoms, prevent the occurrence of chronic epilepsy, and reduce the nerve damage caused by epilepsy, and can be used for the preparation of drugs for treating epilepsy.

[0023] The drug for treating epilepsy provided by the present invention has RvD1 and sodium valproate as its active ingredients. Through the synergistic cooperation of RvD1 and sodium valproate, the treatment of epilepsy can be effectively achieved. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0025] Figure 1 Showing the effects of different treatment groups on the latency, duration and severity of PTZ-induced epileptic seizures;

[0026] Figure 2 Showing the anticonvulsant effects of different treatment groups on the formation of pilocarpine-induced epileptic seizures;

[0027] Figure 3 Showing the effects of different treatment groups on the survival rate of pilocarpine-induced epileptic seizures;

[0028] Figure 4 Showing the immunofluorescence staining results of different treatment groups;

[0029] Figure 5 For Figure 4 the proportion of neuron+ fluorescent cells in the unit area in DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] The following will describe the implementation schemes of the present invention in detail in combination with the embodiments and examples. However, those skilled in the art will understand that the following embodiments and examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention. Those not specifying specific conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. Those reagents or instruments not indicating the manufacturer can be obtained as conventional products through commercial purchase.

[0031] In a first aspect, the present invention provides the application of resolvin D1 combined with sodium valproate in the preparation of drugs for treating epilepsy.

[0032] The inventors have found through research that resolvin D1 and sodium valproate have a synergistic promoting effect in the treatment of epilepsy. Through their synergistic cooperation, they can significantly relieve epileptic symptoms, reduce the damage of epilepsy to nerves, and can be used in the preparation of drugs for treating epilepsy.

[0033] In some alternative embodiments, the treatment of epilepsy includes relieving epileptic symptoms, preventing the occurrence of chronic epilepsy, and avoiding nerve damage.

[0034] In some alternative embodiments, the dosage of resolvin D1 is 250 ng / kg / day;

[0035] The dosage of sodium valproate is 300 mg / kg / day.

[0036] In a second aspect, the present invention provides a drug for treating epilepsy, and the active ingredients of the drug include resolvin D1 and sodium valproate.

[0037] The drug for treating epilepsy provided by the present invention has active ingredients of RvD1 and sodium valproate, and through the synergistic cooperation of RvD1 and sodium valproate, the treatment of epilepsy can be effectively achieved.

[0038] In some alternative embodiments, the treatment of epilepsy includes relieving epileptic symptoms and protecting nerves.

[0039] In some alternative embodiments, the dosage of resolvin D1 is 250 ng / kg / day;

[0040] The dosage of sodium valproate is 300 mg / kg / day.

[0041] In some alternative embodiments, the drug further includes pharmaceutical excipients.

[0042] In the present invention, no specific limitation is imposed on the pharmaceutical excipients, and the well-known pharmaceutical excipients in the art can be used.

[0043] In some alternative embodiments, the pharmaceutical excipients include but are not limited to fillers, binders, disintegrants, lubricants, flavoring agents, or preservatives.

[0044] For the types of fillers, binders, disintegrants, lubricants, flavoring agents, and preservatives, those skilled in the art can make selections according to needs.

[0045] The dosage form of the drug can be various. In some alternative embodiments, the dosage form of the drug includes but is not limited to oral dosage forms.

[0046] In some alternative embodiments, the oral dosage forms are selected from tablets, capsules, granules, and pills.

[0047] The present invention will be further described below through specific embodiments. However, it should be understood that these embodiments are only for more detailed description and should not be construed as limiting the present invention in any way.

[0048] Embodiment

[0049] 1. Animals and epilepsy models

[0050] In this experiment, male mice (C57BL / 6 strain, weighing 20 - 25 g, 6 - 8 weeks old) were provided by GemPharmatech. The mice were housed in a sterile isolator with the environmental temperature controlled at 22°C, humidity at 50%, and a 12 - hour light - dark cycle. A pilocarpine - induced epilepsy mouse model was used. The specific operation was as follows: Before injecting 300 mg / kg of pilocarpine into each mouse, 1 mg / kg of atropine was first given by intraperitoneal injection, and pilocarpine injection was carried out 30 minutes after pretreatment. Epileptic seizures were classified according to the following grading criteria: I - standing still or showing wet dog shakes; II - rhythmic head nodding and chewing movements; III - unilateral forelimb clonic convulsions; IV - bilateral forelimb clonic convulsions accompanied by generalized spasms when standing; V - retreating, rolling, accompanied by generalized tonic - clonic seizures. The success of the status epilepticus (SE) model was marked by the mice reaching grade IV - V within 30 minutes and having continuous seizures. For mice that did not reach this severity, 50 mg / kg of pilocarpine was supplemented and injected every 30 minutes until the mice showed grade IV - V epileptic seizures, with a maximum of 350 mg / kg of pilocarpine given to each mouse. If the epilepsy lasted for 60 minutes, diazepam (10 mg / kg, intraperitoneal injection) was injected to terminate the seizure. The success of modeling was marked by the mice having limb spasms within 30 minutes and showing epilepsy within 2 hours.

[0051] 2. Animal grouping and systemic administration of RvD1

[0052] A total of 40 mice were used in this experiment. The 37 mice with successfully induced SE (success rate 87.5%) were randomly divided into a model group (ddH2O + NS group, 8 mice), a Resolvin D1 group (RvD1 group, 9 mice), a sodium valproate group (VPA group, 8 mice), and an RvD1 + VPA group (RvD1 + VPA group, 10 mice).

[0053] Among them, the ddH2O + NS group was: continuously intragastrically administered distilled water and intraperitoneally injected 0.9% NaCl to the mice with successfully induced SE for 7 days;

[0054] The RvD1 group was as follows: RvD1 was dissolved in 0.9% NaCl to obtain an injection solution, and mice induced with SE were intraperitoneally injected with RvD1 (0.25 μg / kg) and gavaged with distilled water continuously for 7 days;

[0055] The VPA group was as follows: Mice induced with SE were gavaged with sodium valproate (300 mg / kg) and intraperitoneally injected with 0.9% NaCl continuously for 7 days;

[0056] The RvD1 + VPA group was as follows: RvD1 was dissolved in 0.9% NaCl to obtain an injection solution, and mice induced with SE were intraperitoneally injected with RvD1 (0.25 μg / kg) and gavaged with sodium valproate (300 mg / kg) continuously for 7 days.

[0057] 3. Detection of epilepsy susceptibility

[0058] The survival rates of the mice in the above 4 groups after treatment were statistically analyzed, and the results were as Figure 3 shown. After treatment, there were 5 remaining in the ddH2O + NS group, 6 remaining in the RvD1 group, 6 remaining in the VPA group, and 8 remaining in the RvD1 + VPA group.

[0059] Pentylenetetrazol (PTZ)-induced epilepsy is caused by inhibiting the GABA neurotransmitter, thereby causing convulsive symptoms. Accordingly, on the 8th day (the treatment of the four treatment groups was recorded as the 1st day to the 7th day), the remaining mice in the above 4 groups were intraperitoneally injected with PTZ (40 mg / kg), and were observed within 60 min after administration. The epileptic seizure scores were performed according to the Racine grading standard (grades I - V were recorded as 1 - 5 points), and the duration of epileptic seizures, the latency of epileptic seizures (the time from injection of PTZ to the onset of epileptic seizures), the proportion of tonic - clonic epileptic seizures (T - C seizure), and the proportion of mice without epileptic seizures (seizure - free) were statistically analyzed.

[0060] The results of the epileptic seizure severity scores of the mice in each group were as Figure 1 shown in B, the duration of epileptic seizures was as Figure 1 shown in A, the results of the latency of epileptic seizures were as Figure 1 shown in C, the proportion of tonic - clonic epilepsy (T - C seizure) seizures was as Figure 2 shown in A, and the proportion of mice without epileptic seizures was as Figure 2 shown in B. It can be seen from the figure that RvD1 and VPA have a synergistic effect on the treatment of epilepsy and can significantly relieve the onset of epilepsy.

[0061] 3. Immunofluorescence staining

[0062] The above four groups of mouse brains were taken and post-fixed with paraformaldehyde at 4°C, and then immersed in 30% sucrose solution until complete sedimentation. Subsequently, the brains were cut into 60-μm thick coronal sections using a cryostat. For immunofluorescence analysis, the sections were incubated overnight in phosphate-buffered saline (PBS) containing 0.3% Triton X-100, and then incubated with the secondary antibody for 2 hours at room temperature. The primary antibodies used included: Neuron (1:200, Abcam, ab104224); the secondary antibody was goat anti-mouse Alexa 488 (1:500, Jackson ImmunoResearch). In addition, the sections were counterstained with DAPI and analyzed using a confocal laser scanning microscope (LSM700, Zeiss). The specificity of immunofluorescence labeling was verified by omitting the primary antibody and replacing it with normal serum (negative control).

[0063] The results are as Figure 4 and Figure 5 shown. It can be seen from the figure that the combination of 250 ng / kg of RvD1 and 300 mg / kg of sodium valproate has a significant protective effect on neuron loss in the pilocarpine-induced epilepsy model. Among them, Figure 5 shows the proportion of neuron+ fluorescent cells in the unit area counted by imageJ Figure 4 in.

[0064] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of resolvin D1 combined with sodium valproate in the preparation of drugs for treating epilepsy.

2. The use according to claim 1, characterized in that: The treatment of epilepsy includes relieving epileptic symptoms, preventing the occurrence of chronic epilepsy and avoiding nerve damage.

3. The use according to claim 1, characterized in that: The dosage of the resolvin D1 is 250 ng / kg / day; The dosage of the sodium valproate is 300 mg / kg / day.

4. A drug for treating epilepsy, characterized in that: The active ingredients of the drug include resolvin D1 and sodium valproate.

5. The drug according to claim 4, characterized in that The treatment of epilepsy includes relieving epileptic symptoms and protecting nerves.

6. The drug according to claim 4, characterized in that The dosage of the resolvin D1 is 250 ng / kg / day; The dosage of the sodium valproate is 300 mg / kg / day.

7. The drug according to claim 4, characterized in that The medicine also includes pharmaceutical excipients.

8. The drug according to claim 7, characterized in that The pharmaceutical excipients include at least one of a filler, a binder, a disintegrant, a lubricant, a flavoring agent or a preservative.

9. The drug according to claim 4, characterized in that The dosage form of the drug includes oral dosage form.

10. The drug according to claim 9, characterized in that The oral preparation is selected from the group consisting of tablets, capsules, granules and pills.

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