{"id":49458,"date":"2023-06-30T10:04:05","date_gmt":"2023-06-30T10:04:05","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=49458"},"modified":"2023-07-18T08:07:20","modified_gmt":"2023-07-18T08:07:20","slug":"evaluation-of-the-cerebroprotective-properties-of-ademol-gel-in-the-analysis-of-specific-indicators-in-the-open-field-test","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no2\/evaluation-of-the-cerebroprotective-properties-of-ademol-gel-in-the-analysis-of-specific-indicators-in-the-open-field-test\/","title":{"rendered":"Evaluation of the Cerebroprotective Properties of Ademol-gel in the Analysis of Specific Indicators in the Open Field Test"},"content":{"rendered":"\n

Introduction <\/strong> <\/p>\n\n\n\n

According to WHO data, more than 10 million people in\nworldwide have traumatic brain injury (TBI), of which 250-300 thousand people\ndie. In Europe, TBI is the leading cause of death among people under 35 years\nof age. Approximately in 10\u201315 years, the frequency of TBI increases by almost\n2 times.1<\/sup> TBI has a high and steadily growing prevalence and is\nmore common in young and middle-aged people (20-50 years) in the most\nable-bodied population group. Every 10\u201315 years, the frequency of TBI increases\nby almost 2 times.2<\/sup>  <\/sup>According\nto the literature, the number of deaths after cerebral hemorrhages ranges from\n52 to 82 %, and after subarachnoid hemorrhages (SAH) \u2013 from 32 to 64%.2<\/sup> Stroke is the main cause of disability in the adult\npopulation, and only 20% of survivors return to work. Given the above and the\nvariety of clinical manifestations and severity of TBI, it is necessary to\ndevelop strategies for evidence-based approaches to the pathogenetic treatment\nof TBI based on an in-depth study of various aspects of pathogenesis. The\nurgency of the problem of management of TBI and its consequences is associated\nwith significant economic losses, which are complemented by the persistent\ndisability of 10-12 % of victims.1  <\/sup>Contemporary medicine has a large range\nof drugs of pharmacological brain protection for the prevention and treatment\nof the consequences of TBI, but not all of them meet modern requirements.\nTherefore, the search and development of new highly effective and safe medicines\ncharacterized by polytropic pharmacodynamic effects and aimed at the simultaneous\ncorrection of various links in the pathogenesis of TBI, is one of the priority\nareas for modern fundamental and clinical neurology. <\/p>\n\n\n\n

Despite a\nmyriad of drugs affecting cerebral hemodynamics, treating these patients is not\neffective enough. This is due to the short duration of action of many drugs\n(Aminophylline, Vinpocetine, and Pentoxifylline), adverse changes in\nhemodynamics, the development of the intracerebral steal phenomenon. In recent\nyears, drugs from the class of calcium channel blockers, a prominent\nrepresentative of which is nimodipine, have been used to manage chronic\ndisorders of cerebral circulation.<\/p>\n\n\n\n

The main\nneuroprotective agents include the N- and L-subtype calcium channel blocker\nNimotop (3-(2-methoxyethyl) 5-propan-2-yl\n2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate) that\nprevents the launch of the glutamate-calcium cascade of neuronal damage,\nexhibits potent membrane stabilizing, fibrinolytic and antioxidant effects.\nClinical trials suggest that Nimotop reduces all-cause mortality and\nneurological deficit in subarachnoid hemorrhage, TBI, and ischemic stroke. 3<\/sup> Nimodipine has been shown\nto attenuate early brain damage after TBI with subarachnoid hemorrhage,\nneurological deficit, and cerebral edema in the experiment.4<\/sup><\/p>\n\n\n\n

Among many\nprotective mechanisms of some cerebroprotective drugs used to treat acute\ncerebral ischemia, particularly in conditions of hemorrhagic stroke (HS), the\nleading place is occupied by the drug’s ability to improve cerebral circulation\nby reducing reflex vasospasm. The leading place among the many protective\nmechanisms of action of cerebroprotective agents used to treat acute cerebral\nischemia, particularly under conditions of hemorrhagic stroke, is the ability\nof the drug’s ability to improve cerebral circulation by reducing reflex\nvasospasm. An experimental study of the anti-ischemic properties of a new molecule\ncalled Ademol (1-adamantyloxy-3-morpholino-2-propanol hydrochloride) showed\nthat after its parenteral introduction to rats with severe intracerebral\nhemorrhage (ICH) and SAH, mortality decreased to the level of that after the\nadministration of Mexidol, Citicoline and Nimodipine.5<\/sup><\/p>\n\n\n\n

A stimulating\neffect of Ademol on cerebral circulation in the basin of the internal carotid\nartery and its modulating action on microcirculation in the cerebral cortex in\nSAH has been demonstrated as well as its modulating action on central\nhemodynamics and intracranial pressure, the factors that form stable cerebral\nblood flow, have been established in various types of acute cerebral ischemia.6<\/sup><\/sup><\/sup><\/sup> Recently, intranasal dosage forms of\nneuroprotectors have been intensively developed as delivery of drugs to the\nbrain is important for the effective prevention and management of brain\ndiseases. The intranasal delivery route has several advantages and may be used\nto introduce active ingredients manufactured as solutions for injections. The\nintranasal dosage form provides the highest neuroavailability of the active\npharmaceutical ingredient (both due to rapid absorption in the epithelium of\nthe nasal cavity and due to axonal transport) and fast onset of action. After the\nintranasal introduction, the drug reaches its main site of action in the brain within\n5 min, whereas typical oral and intravenous routes have limitations in drug\ndelivery to the brain. Besides, this non-invasive way does not require the\nparticipation of medical personnel or special training. It can be used not only\nin the acute period of the disease but also later on an outpatient basis, which\nin turn has certain economic advantages and high compliance with treatment.7<\/sup> Therefore, considering all\nthe positive pharmacological effects of adamantane derivatives in TBI and the\nprospect of nasal drug administration, a nasal form of Ademol was developed and\na study of its cerebroprotective properties was conducted on the open field\ntest. <\/p>\n\n\n\n

Material and methods<\/strong><\/p>\n\n\n\n

The method\nof synthesis of the Ademol substance was developed by the Head of Technology\nTransfer, Innovation and Intellectual Property Department of Institute of\nOrganic Chemistry NAS of Ukraine Yu.V. Korotkyi. The composition and properties\nof 1 % Ademol-gel was developed as a fragment of the Research work of the\nNational Pirogov Memorial Medical University, Vinnytsya in collaboration with\nthe Department of Medicines Technology of the ZSMU. Various approaches are\ncommonly used for formulation development: empirical, based on the\nsubstantiation of the dosage form components due to the experience of the\nresearcher; the methods of mathematical planning of the experiment, when a\nsingle or multifactorial experiment allows to select the components of the\nformulation by determining the influence of the factor (factors) on the\nselected optimization parameter; use of software in the form of an expert\nsystem that combines in its functionality the possibilities of an empirical\napproach, methods of mathematical planning of an experiment and built-in\nmachine learning models.8<\/sup>–<\/sup>10<\/sup> <\/sup><\/p>\n\n\n\n

In silico<\/em> research on the choice of the rational composition of the formulation of the nasal form with Ademol was carried out using the ExpSys Nasalia expert system developed at the Departments of Medical and Pharmaceutical Informatics and Advanced Technologies, and Pharmacology and Medical Formulation with Course of Normal Physiology of ZSMU. The system contains machine learning models (blender [random forest, extra tree]) that is a complex model of machine learning, which includes random forest extra tree models; and blender [catboost, lightgbm, xgboost] that is a complex machine learning model containing catboost, lightgbm, xgboost models) and allows for predicting the compatibility of formulation ingredients. <\/p>\n\n\n\n

Ingredients\nof the nasal form formulation were introduced in pairs into the system\ninterface using the Simplified Molecular Input Line Entry System (SMILES), and\ntheir compatibility was checked. A platinum-platinum-rhodium thermocouple to\nheat the samples in aluminum crucibles (from\n15 to 250 \u00ba<\/sup>\u0421<\/em>)and \u03b1-Al2<\/sub>O3<\/sub>\nas a reference substance(the heating rate was 10\u00ba<\/sup> \u0421\/min<\/em>)\nwere used for thermogravimetric studies (Shimadzu DTG-60, Japan) to confirm the\ncompatibility of the formulation ingredients. <\/p>\n\n\n\n

In addition\nto assessing the compatibility of ingredients, this method made it possible to\nadditionally characterize the temperature regime for manufacturing the\nformulation. The individual API were studied including Ademol and other\ningredients, such as glycerin, hydroxyethylcellulose (HEC), benzalkonium\nchloride, and the manufactured final nasal dosage form without active\ningredients, and dosage form with Ademol. The mass of the studied samples\nranged from 16.08 mg to 73.29 mg.<\/p>\n\n\n\n

Rheological\nstudies of the nasal form with Ademol were conducted using an MCR 302 rheometer\n(Anton Paar GmbH). As a measuring device, coaxial cylinders CC27\/T200\/SS were\nused at a temperature of 29-\u00b1 0.5\u00b0\u0421 33 \u00b1 0.5\u00b0 \u0421, 37 \u00b1 0.5\u00b0 \u0421, which makes it\npossible to determine the characteristics of the structural and mechanical\nproperties of the nasal form as well as offer rational dosage form packaging.\nAs a result, the prescription for the extemporaneous preparation of an\nintranasal dosage form for the cerebroprotective agent Ademol was created. In\naddition to the API, it also included the plasticizer glycerin, the\nmucoadhesive component HEC and the antimicrobial component benzalkonium\nchloride.7<\/sup> The research was carried out at the Department of\nExperimental Pathophysiology and Functional Morphology in the Training Medical\nand Laboratory Center of ZSMU.<\/p>\n\n\n\n

The study\nwas carried out on Wistar rats of both sexes, aged 10-12 weeks, weighing\n170-230 g (n=29), obtained from the nursery of the Institute of Pharmacology\nand Toxicology NAMS of Ukraine. The cages with the animals were kept in\nseparate rooms with a standard lighting regime: 12 hrs of light and 12 hrs of\ndarkness. Animals were excluded from the study during quarantine, if they did\nnot meet the standard criteria. The duration of the quarantine (acclimatization\nperiod) was 14 days for all animals. During quarantine, each animal was\nexamined daily (behavior and general condition), and twice a day rats were\nobserved in cages (morbidity and mortality). Before the start of the study,\nanimals that met criteria for inclusion in the experiment were divided into\ngroups using the randomization method. Animals not meeting the criteria were\nexcluded from the study during quarantine. The experimental animals were kept\non the same diets under normal vivarium conditions. All manipulations were\ncarried out in accordance with the regulation on the use of animals in\nbiomedical experiments (Strasbourg, 1986, as amended in 1998-2001).<\/p>\n\n\n\n

To\nfacilitate subsequent experimental studies (open\nfield test<\/em>), animals were kept in hands for 2-3 min for 5 days before the\nexperiment. Anesthesia with Thiopental sodium (40 mg\/kg) intravenously into the\ntail vein was carried out by a 26G needle. <\/p>\n\n\n\n

Experimental\nanimals were subjected to suboccipital puncture under general anesthesia. The modified\nSAH model with a single injection of autologous blood based on the method of\nDudhani R.V. et al. 11<\/sup> was developed and used. Blood\n(0.2 ml) was taken from the tail vein with a heparin syringe. Then a large\noccipital cistern was punctured and 0.2 ml of autologous blood was injected\ninto the subarachnoid space. The hole was closed with dental wax. Sham-operated\nanimals underwent anesthesia and other surgical procedures without the\nintroduction of autologous blood. The investigated agents were administered\nonce a day at the same time: Ademol \u2013intranasally at a conditionally effective\ndose of 2 mg\/kg for 7 days; Nimodipine (Nimotop\u00ae, Bayer, Germany) \u2013 intraperitoneally\n30 mcg\/kg for 7 days. Animals in the control groups received equivolume quantities\nof solvents. <\/p>\n\n\n\n

The open\nfield test, based on subjecting an animal to an unknown environment whose\nescape is prevented by surrounding walls, was used to evaluate locomotor and\nsearch activity in animals on the 7th day after the SAH using the arena of our\nown production with dimensions of 80x80x35 cm, as indicated earlier.12<\/sup> <\/p>\n\n\n\n

The\nexperimental rat was seated with its muzzle against the wall and was given free\naccess to movement (8 min). The total distance that the rat walked (cm), the\nsum of the activity of all movements (cm2<\/sup>\/sec), the structure of\nactivity (inactivity, low activity, high activity, %), the number of ascents\nand visits to the center, approaches to the center, the distance traveled near\nthe wall and to the central area of the arena were recorded (cm, %); movement\nspeed, delay time before entering the center, number of grooming acts and\nnumber of defecation acts were estimated. <\/p>\n\n\n\n

In the\ncourse of the study, we excluded (chamber of conditioned reflexes) the\ninfluence on the rat all possible stimuli as well as the presence of the\nexperimenter during the period of registration. Registration of rat movements\nin the open field was performed using a SSC-DC378P color video camera (Sony,\nJapan). <\/p>\n\n\n\n

The data\nwere analyzed using the Smartv 3.0 program (Harvard Apparatus, USA).\nStatistical analysis of the experimental data was performed using the Microsoft\nExcel 2016 program with the AtteStat 12 statistical processing package. To\nassess the significance of differences in the study groups, the Kruskal-Wallis\ntest with the Dunn’s post hoc correction was used. Differences were considered\nsignificant at p<0.05.<\/p>\n\n\n\n

Results and Discussion<\/strong><\/p>\n\n\n\n

Simulation of SAH led to a decrease in the\ntotal activity of animals (by 3 times) and a decrease in the total path of\nmovement (by 2 times). The experimental SAH also increased the duration of the\npath along the periphery (near walls and corners) by 2.3 times, and reduced the\ndistance traveled before crossing the periphery-center (shadow-light) border by\n2.43 times whereas the distance traveled in the illuminated center decreased by\n3.8 times. Animals were characterized by inactivity, anxious-aggressive\nbehavior, and poor orientation in space.<\/p>\n\n\n\n

The simulation of the SC led to a significant increase in\ninertia when moving from the dark field of the arena to the illuminated part by\n2 times indicating a decrease in locomotor and\nsearch activity as well as an increase in anxiety and fear. On the 7th\nday after SAH the duration of inactivity near the wall increased by 1.67 times\nindicating inhibition of exploratory activity and depression development. In\nanimals after SAH, the number of rearings, a common measure of activity and\nexploratory behavior, increased by 3 times, immobility increased by 2.46 times when\npassing from the periphery to the center of the arena (which can be regarded as\nan increase in anxiety, fear and disorientation in animals with SAH). The speed\nof passing the path in the illuminated center of the arena decreased by 5.86\ntimes.<\/p>\n\n\n\n

The number of free rearings did not change when modeling\nthe ICG, but the number of rearings at the wall increased. A decrease in the number\nof short grooming acts against the background of a constant number of long-term\ngrooming testified to increased anxiety, excitability and irritability of the\nanimals, as well as their discomfort and depression. A decrease in short\ngrooming by 7 times and defecation acts by 3.5 times also testified to a\ndecrease in high activity. The decrease in high activity noted above in the\nanimals of the control group testified to the low emotionality and excitability\nof the animals. The introduction of Nimotop immediately after the animals came\nout of anesthesia as well as intranasal application of a new dosage form of the\nneuroprotector Ademol-gel had different effects on behavioral reactions,\ncognitive and exploratory functions, as well as emotional status of animals\n(table).<\/p>\n\n\n\n

Ademol-gel reduced the distance traveled in the\nborder-periphery zone, and significantly exceeded the effect of nimodipine in\nthis indicator (by 2.3 times). Ademol-gel increased the number of short\ngrooming and defecation acts by 3.3 and 2.67 times, respectively, and reduced\nthe number of rearing by 1.75 times. All this indicated a decrease in anxiety,\ndepressive behavior, and aggressiveness, and an increase in empathy in\nexperimental animals. <\/p>\n\n\n\n

Table 1: The effects of the investigated drugs on the behavior and exploratory activity of rats<\/strong> in the open field test on the 7th day after SAH<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Characteristics<\/strong><\/p>\n<\/td>\n

\n

Subarachnoid hemorrhage (Control) (n=5)<\/strong><\/p>\n<\/td>\n

\n

Nimodipine<\/strong><\/p>\n

(n=7)<\/strong><\/p>\n<\/td>\n

\n

Ademol-<\/strong><\/p>\n

gel<\/strong><\/p>\n

(n=7)<\/strong><\/p>\n<\/td>\n

\n

False-<\/strong><\/p>\n

operated<\/strong><\/p>\n

animals (n=10)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

General activity, cm2<\/sup>\/s<\/p>\n<\/td>\n

\n

12836.20 \u00b1 <\/strong>1354.021<\/sup><\/p>\n<\/td>\n

\n

13736.57 \u00b1 <\/strong>1843,071<\/sup><\/p>\n<\/td>\n

\n

23497.03\u00b1 <\/strong>9883.61<\/p>\n<\/td>\n

\n

39592,10 \u00b1 <\/strong>3692,40<\/p>\n<\/td>\n<\/tr>\n

\n

Short grooming acts<\/p>\n<\/td>\n

\n

1.00 \u00b1 0.001<\/sup><\/p>\n<\/td>\n

\n

2.29 \u00b1 <\/strong>0.761*<\/sup><\/p>\n<\/td>\n

\n

3.29 \u00b1 <\/strong>1.701<\/sup>*<\/p>\n<\/td>\n

\n

7.10 \u00b1 <\/strong>0.57<\/p>\n<\/td>\n<\/tr>\n

\n

Defecation<\/p>\n<\/td>\n

\n

0.80 \u00b1 0.841<\/sup><\/p>\n<\/td>\n

\n

1.71 \u00b1 <\/strong>0.76*<\/p>\n<\/td>\n

\n

2.14 \u00b1 <\/strong>1.07*<\/p>\n<\/td>\n

\n

2.80 \u00b1 <\/strong>0.42<\/p>\n<\/td>\n<\/tr>\n

\n

Distance the border-periphery zone, cm<\/p>\n<\/td>\n

\n

480.42 \u00b1 125.471<\/sup><\/p>\n<\/td>\n

\n

553.32 \u00b1 <\/strong>190.971<\/sup><\/p>\n<\/td>\n

\n

280.66 \u00b1 <\/strong>53.12*2<\/sup><\/p>\n<\/td>\n

\n

207.36 \u00b1 <\/strong>50.63<\/p>\n<\/td>\n<\/tr>\n

\n

Distance from the periphery to the center, cm<\/p>\n<\/td>\n

\n

1684.07 \u00b1 739.231<\/sup><\/p>\n<\/td>\n

\n

1792.67 \u00b1 <\/strong>836.271<\/sup><\/p>\n<\/td>\n

\n

2618.32 \u00b1 1662.85<\/p>\n<\/td>\n

\n

4103.54 \u00b1 <\/strong>555.48<\/p>\n<\/td>\n<\/tr>\n

\n

Distance in the center, cm<\/p>\n<\/td>\n

\n

66.84 \u00b1 19.451<\/sup><\/p>\n<\/td>\n

\n

120.32 \u00b1 <\/strong>13.571<\/sup>*<\/p>\n<\/td>\n

\n

161.21 \u00b1 <\/strong>54.58*<\/p>\n<\/td>\n

\n

249.67 \u00b1 <\/strong>41.27<\/p>\n<\/td>\n<\/tr>\n

\n

Total distance, cm<\/p>\n<\/td>\n

\n

2026.04 \u00b1 696.531<\/sup><\/p>\n<\/td>\n

\n

2495.38 \u00b1 <\/strong>557.791<\/sup><\/p>\n<\/td>\n

\n

2751.73 \u00b1 <\/strong>356.561<\/sup><\/p>\n<\/td>\n

\n

4148.40 \u00b1 <\/strong>507.26<\/p>\n<\/td>\n<\/tr>\n

\n

Duration of high activity, %<\/p>\n<\/td>\n

\n

9.24 \u00b1 <\/strong>1.401<\/sup><\/p>\n<\/td>\n

\n

12.94 \u00b1 <\/strong>4.33<\/p>\n<\/td>\n

\n

17.44 \u00b1 <\/strong>2.71*<\/p>\n<\/td>\n

\n

18.79 \u00b1 <\/strong>2.16<\/p>\n<\/td>\n<\/tr>\n

\n

Duration of low activity %<\/p>\n<\/td>\n

\n

40.70 \u00b1 <\/strong>9.881<\/sup><\/p>\n<\/td>\n

\n

37.13 \u00b1 <\/strong>13.621<\/sup><\/p>\n<\/td>\n

\n

50.56 \u00b1 <\/strong>9.10<\/p>\n<\/td>\n

\n

61.99 \u00b1 <\/strong>7.95<\/p>\n<\/td>\n<\/tr>\n

\n

Total duration of immobility (sec)<\/p>\n<\/td>\n

\n

230.25 \u00b1 <\/strong>73.991<\/sup><\/p>\n<\/td>\n

\n

108.33 \u00b1 <\/strong>30.211<\/sup>*<\/p>\n<\/td>\n

\n

76.73 \u00b1 <\/strong>20.93*<\/p>\n<\/td>\n

\n

50.30 \u00b1 <\/strong>9.24<\/p>\n<\/td>\n<\/tr>\n

\n

Maximum moving speed in the center (cm2<\/sup>\/s)<\/p>\n<\/td>\n

\n

29.59 \u00b1 <\/strong>13.281<\/sup><\/p>\n<\/td>\n

\n

178.31 \u00b1 <\/strong>40.09*<\/p>\n<\/td>\n

\n

199.52 \u00b1 <\/strong>45.16*<\/p>\n<\/td>\n

\n

169.08 \u00b1 <\/strong>33.82<\/p>\n<\/td>\n<\/tr>\n

\n

The first delay when entering the center, sec<\/p>\n<\/td>\n

\n

92.04 \u00b1 <\/strong>8.171<\/sup><\/p>\n<\/td>\n

\n

79.39 \u00b1 <\/strong>13.871<\/sup><\/p>\n<\/td>\n

\n

68.12 \u00b1 <\/strong>12.62*<\/p>\n<\/td>\n

\n

47.42 \u00b1 <\/strong>6.01<\/p>\n<\/td>\n<\/tr>\n

\n

The duration of inactivity in the internal periphery, %<\/p>\n<\/td>\n

\n

52.00 \u00b1 <\/strong>4.531<\/sup><\/p>\n<\/td>\n

\n

40.14 \u00b1 <\/strong>5.49*<\/p>\n<\/td>\n

\n

33.43 \u00b1 <\/strong>6.21*<\/p>\n<\/td>\n

\n

31.40 \u00b1 <\/strong>3.50<\/p>\n<\/td>\n<\/tr>\n

\n

Duration of inactivity periphery border \u2013 center, %<\/p>\n<\/td>\n

\n

32.00 \u00b1 <\/strong>5.051<\/sup><\/p>\n<\/td>\n

\n

38.14 \u00b1 <\/strong>24.75<\/p>\n<\/td>\n

\n

19.86 \u00b1 <\/strong>5.30<\/p>\n<\/td>\n

\n

13.80 \u00b1 <\/strong>9.91<\/p>\n<\/td>\n<\/tr>\n

\n

Rearing<\/p>\n<\/td>\n

\n

832.35 \u00b1 69.911<\/sup><\/p>\n<\/td>\n

\n

734.17 \u00b1 <\/strong>78.341<\/sup><\/p>\n<\/td>\n

\n

476.00 \u00b1 <\/strong>158.95*<\/p>\n<\/td>\n

\n

278.81 \u00b1 <\/strong>50.41<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Note<\/em>: * \u2013 statistically significant difference (p<0.05) compared with the control\ngroup; <\/p>\n\n\n\n

1<\/sup> \u2013\nstatistically significant difference (p<0.05) compared with the\ngroup of false-operated animals<\/p>\n\n\n\n

Intranasal administration of Ademol-gel to animals with\nSAH led to the restoration of the active component of the research activity.\nThis was evidenced by an increase (significantly by 1.88times) in the duration\nof high activity of animals such as examination of objects above the floor of\nthe arena and the number of jerks near the wall. This fact indicated the\npositive effect of Ademol-gel on the cognitive functions of the central nervous\nsystem.<\/p>\n\n\n\n

Animals treated with Ademol-gel for 7 days after SAH also\nshowed better spatial orientation and freer movement in the illuminated part of\nthe arena. Thus, the speed of their movement in the illuminated center\nincreased by 6.8 times. The time of general immobility in the Ademol group also\nsignificantly (by 67 %) decreased, while the indicators of general activity and\nthe total distance traveled did not change significantly in the nimodipine\ngroup.<\/p>\n\n\n\n

The results of the research showed that experimental SAH\nled to a significant violation of orienting-exploratory behavior, as evidenced\nby the indicators of the open field test. Thus, in animals after SAH modeling,\na decrease in high and low activity was noted, which indicated the suppression\nof the exploratory function of the central nervous system, as well as the\nformation of anxiety and excitability. The lack of movements aimed at mastering\na new environment, a decrease in high and low activity in animals may indicated\na decrease in cognitive abilities.<\/p>\n\n\n\n

Our experimental results do not contradict the data of\nother researchers on the molecular and biochemical mechanisms of\ncognitive-mnestic disorders of the CNS after TBI and subarachnoid hemorrhage,\ncerebral stroke. Experimental SAH leads to persistent disturbances in memory,\norientation, research and cognitive activity of animals, the appearance of\nirritability, lethargy, fear, anxiety, disorientation, aggressiveness.13<\/sup>–<\/sup>14<\/sup> <\/p>\n\n\n\n

Cognitive-mnestic disorders after SAH are formed in\nresponse to transmitter autocoidosis, hyperactivity of neuronal nitric oxides synthase (nNOS) and inducible  nitric\noxides synthase (iNOS), increased production of reactive oxygen species,\nenergy deficiency, lactic acidosis, mitochondrial dysfunction, increased\nFe-dependent reactions of free-radical oxidation, leading to the initiation of\napoptosis of neurons in the zone hippocampus and sensorimotor cortex.15<\/sup> Experimental animals treated with ademol-gel after SAH\nwere significantly more mobile, and showed increased interest in the\nenvironment. Animals after SAH treated with Ademol-gel showed fewer signs of\nfear and anxiety compared to the control group, as evidenced by a decrease in\nthe time of the first delay when entering the center by 26 %.<\/p>\n\n\n\n

The pharmacological effects of Ademol are associated with\nits ability to reduce excitotoxicity and maintain the functional activity of\nhippocampal and sensorimotor cortex neurons by reducing the increased\nexcitability of NMDA receptors and modulating the NMDA polyamine site.16<\/sup> Professor A.A. Khodakovsky and co-authors also\ndemonstrated the presence of anxiolytic properties in Ademol due to its ability\nto increase affinity for GABA receptors, leading to a decrease in fear and\nanxiety.17<\/sup>–<\/sup>18<\/sup> Ademol can provide endotheliotropic action by reducing\nplatelet aggregation, increasing the expression of nitric oxide synthases. It\nwas also revealed that adamantane derivatives had mitoprotective and\nenergotropic action.19<\/sup> <\/sup>A less\npronounced positive effect of pharmacotherapy on the psycho-emotional behavior\nand motor-search activity of rats with SAH treated with nimodipine was noted.\nAnimals treated with nimodipine (Nimotop\u00ae, Bayer, Germany) were characterized\nby inactivity, greater anxiety and aggressiveness. In the nimodipine group, no\nsignificant changes in motor and search activity were recorded compared to the\ncontrol group. Some indicators of anxiety were worse, and indicators of\nexploratory activity were significantly lower in the Nimotop group than in the\ngroup of animals after SAH treated with Ademol-gel.<\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

The behavior of animals after SAH was characterized by a significant decrease in total activity by 3 times, a decrease in the distance traveled by 2 times and an increase in inertia when moving from the dark field to the illuminated part of the arena by 2 times, which indicated a significant decrease in motor and search activity. The number of unsupported rearings (in which the animal freezes without touching the walls of the arena) did not change, but the number of unsupported rearing (in which the animal leans on the walls of the arena) increased; the number  of short groomings decreased against the background of a constant number of long groomings. Such behavior indicated an increase in excitability, anxiety, irritability of animals, the development of severe discomfort and depression in them.<\/p>\n\n\n\n

In animals with severe neurological deficit after SAH Ademol-gel 2 mg\/kg intranasally  had a positive effect on the emotional status and behavior of animals in the open field test, as evidenced by a decrease in anxiety, indicators of aggressive and depressive behavior against the background of an increase in comfort and the formation of empathy. The motor and exploratory activity of animals with SAH also quickly returned to normal after the administration of Ademol-gel.<\/p>\n\n\n\n

In rats in the acute period of SAH, Nimodipine (Nimotop\u00ae, Bayer, Germany) intraperitoneally at a dose of 30 \u00b5g\/kg, as a primary neuroprotector, realized its action aimed solely at maintaining the viability of neurons, without having a significant effect on animal behavior and cognitive deficits.<\/p>\n\n\n\n

The experimental results obtained confirm the prospectivity and effectiveness of administration of a new dosage form – Ademol intranasal gel in the management of TBI to accelerate the elimination of cognitive deficits and restoration of motor activity.<\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

The authors would like to\nacknowledge the Zaporizhzhia State Medical University for providing some\nfacilities in carrying out the research.<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

The authors declare no conflict of interest, financial or otherwise with this work.<\/p>\n\n\n\n

References <\/strong> <\/p>\n\n\n\n

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