RU2600477C1 - Method of detecting anticonvulsant action of sodium citicoline and valproate when applying them together on model of acute generalized convulsions caused by pentylenetetrazole in wistar male rats - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to neurochemistry, pathophysiology, neurology and psychiatry. On the model of acute generalized convulsions, caused by pentylenetetrazole in Wistar male rats, the method of anticonvulsant action when applying sodium citicoline and sodium valproate is disclosed. Preparations are being intraperitoneally administered, at that, citicoline is being administered in a dose of 300 mg/kg one hour before the administration of pentylenetetrazole, sodium valproate is being administered in a dose of 70 mg/kg 10 minutes before the administration of pentylenetetrazole.

EFFECT: selected dose schedule of the preparations provides higher anticonvulsant action also related to the neuroprotective action of preparations, which reduces the neurodegeneration in the rat brain under conditions of acute generalized convulsions caused by pentylenetetrazole.

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Description

The invention relates to medicine, namely to neurochemistry, pathophysiology, neurology and psychiatry. It is associated with an increase in the effectiveness of the treatment of epilepsy. Epilepsy, a neuropsychic disease complex in etiology and pathogenesis, has a chronically progressive course and is characterized by the polymorphism of both paroxysmal manifestations and mental disorders. On the one hand, classic and new antiepileptic drugs can stop the increased convulsive activity of the brain, but, as a rule, do not have protective properties against neurodegeneration [1]. Thus, the problem of preventing brain damage due to seizures remains unresolved. On the other hand, the effect of most antiepileptic drugs is accompanied by a number of side effects, which include impaired basic cognitive functions - attention, memory, perception, etc. [5, 7, 10]. In connection with the foregoing, it seemed appropriate to study the effect of citicoline (an analogue of endogenous citicoline, which is currently widely used in the treatment of cognitive, sensitive, motor neurological disorders of degenerative and vascular etiology, and has neuroregenerative and neuroprotective properties) [6, 8, 9, 11, 12] and sodium valproate, an anticonvulsant drug [2, 3, 4].

A known method of preventing neurodegeneration of neurons in animals using piracetam in the conditions of PTZ-kindling [11]. It is known that piracetam is a nootropic drug that increases the brain's resistance to hypoxia, integrative work, positively affects memory and learning processes, improves blood circulation in the brain and does not have a sedative effect. Under the conditions of PTZ-kindling in mice, it has been shown that the use of piracetam leads to the prevention of neuronal injuries and a decrease in the deficit of previously acquired skills when training in animals.

The disadvantage of this method is the lack of evaluation of the anticonvulsant effect of piracetam in acute generalized seizures caused by pentylenetetrazole,

As the closest analogue (prototype), a method for enhancing the noopept anticonvulsant activity of valproate in mice was selected. Noopept is an original dipeptide drug with nootropic, neuroprotective and anxiolytic activity. In this method, in the model of seizures caused by corazole in mice, noopept (0.5 mg / kg ip) during either single or repeated exposure (10 or 35 days) did not enhance the convulsive effect of corazole, which suggests the absence of proconvulsive properties in the drug [3]. This is the advantage of noopept over piracetam, which increases convulsive readiness. With prolonged (5-week) preliminary exposure, noopept enhanced the anticonvulsant effect of valproate. The study can serve as an experimental justification for the advisability of course use of noopept together with valproate in order to enhance the anticonvulsant properties of the latter, as well as to have a beneficial effect on cognitive functions and slow the progression of neurodegenerative processes.

The disadvantage of this method is the need for a long five-week exposure to noopept to enhance the anticonvulsant effect of valproate, as well as the fact that the use of noopept and valproate does not reduce their doses when combined to obtain an anticonvulsant effect.

The objective of the invention is to develop a method that allows you to identify the anticonvulsant activity of citicoline and sodium sodium valproate in acute generalized seizures caused by PZT in male rats of the Wistar strain.

The problem is solved due to the fact that the method for detecting the anticonvulsant effect of citicoline and sodium valproate when used together provides the following differences: 3 series of experiments were conducted to reliably evaluate the spectrum and degree of anticonvulsant effect of citicoline and sodium valproate. The preparations were administered intraperitoneally to experimental animals so that the peak of their action coincided: citicoline was administered at a dose of 300 mg / kg 1 hour before the PTZ test, and sodium valproate at a dose of 70 mg / kg 10 minutes before the PTZ test. The essence of the proposed method was that with the combined use of citicoline and valproate in minimally effective doses on the model of acute generalized seizures caused by PZT, the effect of both drugs was enhanced. The latent periods of the onset of the first convulsive manifestations and the latent period of the phase of clonic convulsions significantly increased by 66.70% and 63.36%, respectively, compared with the group of control animals, and in 12.5% there was no clonic and tonic phase of seizures of experimental animals. In these rats, under conditions of acute generalized seizures caused by PTZ, the anticonvulsant effect of citicoline and sodium valproate was more pronounced when used together.

There is a causal relationship between the set of essential features of the claimed object and the achieved technical results: the combined administration of citicoline and sodium valproate expands their spectrum and increases the degree of their anticonvulsant effect in Wistar rats against acute pentylenetetrazole seizures.

New features of the claimed method for detecting the anticonvulsant effect of citicoline and sodium valproate when used together on the model of acute generalized seizures caused by PZT in rats of males of the Wistar line are:

1. The presence of an anticonvulsant effect after a single administration of citicoline at a dose of 500 mg / kg one hour before the administration of PTZ, an increase in convulsive thresholds was noted: the severity of seizures was 33.18% lower, the life expectancy of animals increased 3.26 times.

2. The manifestation of the anticonvulsant effect of sodium valproate at a dose of 100 mg / kg 10 minutes before the administration of PTZ led to a 50.7% reduction in severity of seizures and prevented the death of animals.

3. With the combined use of citicoline at a dose of 300 mg / kg and sodium valproate at a dose of 70 mg / kg when used together (citicoline an hour before the administration of PTZ and sodium valproate 10 minutes before the administration of PTZ), significantly latent periods of the appearance of the first convulsive manifestations and the latent period of the phase of clonic seizures increased by 66.70% and 63.36%, respectively, compared with the group of control animals, and 12.5% observed the absence of the clonic and tonic phase of seizures of experimental animals.

The method is developed based on the following experiments.

1. The experimental technique.

The experiments were performed on rats of males of the Wistar strain (n = 116) weighing 160-190 g. The animals were kept under standard vivarium conditions with free access to water and food. All procedures and experiments on animals were carried out in accordance with the "Rules of laboratory practice in the Russian Federation", approved by order of the Ministry of Health of the Russian Federation No. 267 of 06/19/2003.

The effectiveness of the combined use of sodium valproate and citicoline was studied in a model of acute generalized seizures caused by intraperitoneal administration of PTZ at a dose of 70 mg / kg. The latent periods of the first convulsive manifestations, the clonic and tonic phases (falling of the animal on its side) of the generalized convulsive reaction, the duration of these phases, the severity of the convulsive reaction, life expectancy and mortality were determined. The severity of the convulsive reaction was evaluated in points: 1 - myoclonic tremors; 2 - clonic cramps of the forelimbs (pose "kangaroo"); 3 - repeated clonic convulsions; 4 - tonic-clonic convulsions with the fall of the animal on its side; 5 - repeated tonic-clonic convulsions and / or death of the animal.

Conducted 3 series of experiments. In the 1st and 2nd series, the minimal anticonvulsant effect of citicoline (Ceraxon, Nicomed Ferrer Internacional, S.A.) and sodium valproate (Convulex) (Convulex, Gerot Pharmazeutika GmbH) on acute generalized convulsions was determined. For this purpose, citicoline was administered intraperitoneally at doses of 300, 500 mg / kg 1 hour before the administration of PTZ, and sodium valproate was administered at doses of 70 and 100 mg / kg 10 minutes before the administration of PTZ. In the 3rd series, the efficacy of the combined use of citicoline and sodium valproate in minimally effective anticonvulsant doses for acute generalized seizures in animals was investigated. In all series, the control was animals that were injected with saline under similar experimental conditions.

2. In control animals, in all series of experiments, the introduction of PTZ led to the development of generalized epileptic activity: after 43-50 s the first convulsive manifestations appeared in the form of trembling of the head or individual muscles of the body, which turned into clonic tremors of the whole body. Then, after 52-57 seconds, clonic cramps of the forelimbs followed with the raising of the animal to its hind legs (“kangaroo” pose). Convulsions ended in a tonic-clonic seizure with the animal falling on its side and a phase of post-attack depression. The severity of seizures in animals averaged 4.33-4.53 points, and mortality - 53-87%.

In the 1st series of experiments, the administration of citicoline at a dose of 500 mg / kg exerted an anticonvulsant effect, which was expressed in an increase of 42.09% in the latent period of the first convulsive manifestations and 47.43% of clonic convulsions compared with the group of control animals. The severity of seizures was 33.18% lower. The life expectancy of animals increased by 3.26 times. The number of dead animals was 80.61% lower than in the control group. The administration of citicoline at a dose of 300 mg / kg in animals caused only an increase in the latent period of the onset of tonic seizures and did not affect other indicators of convulsive activity compared with the group of control animals.

In the 2nd series of experiments, the anticonvulsant effect of sodium valproate at a dose of 100 mg / kg was expressed in a 50.77% reduction in the severity of seizures and the prevention of animal death. The dose of sodium valproate 70 mg / kg was less effective: the severity of seizures was 24.25% lower, and mortality - 46.42% compared with the group of control animals.

Thus, citicoline and sodium valproate at doses of 300 and 70 mg / kg individually exerted a less pronounced anticonvulsant effect and were selected for further research.

In the 3rd series of experiments, the combined use of sodium valproate at a dose of 70 mg / kg and citicoline at a dose of 300 mg / kg led to an increase in the anticonvulsant properties of both drugs. Thus, the latent periods of the appearance of the first convulsive manifestations and the latent period of the phase of clonic seizures increased by 66.70 and 63.36%, respectively, compared with the group of control animals. In 12.5% of the experimental animals, the absence of the clonic and tonic phase of seizures was observed.

Thus, the invention allows to identify both the spectrum of the anticonvulsant effect of the combined use of citicoline and sodium valproate and the degree of their pronounced anticonvulsant effect in animals on the model of acute generalized seizures caused by PZT, which is associated with an increase in the anticonvulsant effect of citicoline and sodium valproate when used together.

Literature

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Claims (1)

A method of anticonvulsant exposure when citicoline and sodium valproate are used together in a model of acute generalized seizures caused by pentylenetetrazole in rats of Wistar male rats, namely, the drugs are administered intraperitoneally, while citicoline is administered at a dose of 300 mg / kg an hour before pentylenetetrazole is administered, and sodium valproate is administered at a dose of 70 mg / kg 10 minutes before the administration of pentylenetetrazole