Rotarod Method Research Articles

Muscle relaxant activity of hydroalcoholic extract of Mimosa pudica whole plant in mice

Background: Mimosa pudica is a traditionally used folk medicine to treat various disorders such as bleeding disorders, dysentery, piles, convulsions, rheumatoid arthritis, muscular pain, and snake bite. Aims and Objectives: To evaluate the muscle relaxant activity of hydroalcoholic extract of M. pudica whole plant (HAEMPWP) in albino mice. Materials and Methods: HAEMPWP was prepared using Soxhlet apparatus. Acute toxicity tests were done using the extract which was given orally to albino mice in increasing doses up to 3200 mg/kg body weight. For pharmacological study, the extract in doses of 200, 400, and 800 mg/kg was orally administered to albino mice. The muscle relaxant action was evaluated by Chimney test and Rotarod method. Data analysis was done by unpaired t-test, one-way ANOVA, Chi-square test, and log-probit analysis using SPSS software version 16. Results: HAEMPWP showed a significant dose-dependent increase in muscle relaxant activity as compared to control. Conclusion: HAEMPWP possesses significant muscle relaxant activity and could be an effective treatment option for various spasmodic conditions.

Design, synthesis and evaluation of benzofuran-acetamide scaffold as potential anticonvulsant agent.

A series of N-(2-(benzoyl/4-chlorobenzoyl)-benzofuran- 3-yl)-2-(substituted)-acetamide derivatives (4a-l, 5a-l) was synthesized in good yield. All synthesized compounds were in agreement with elemental and spectral data. The anticonvulsant activity of all synthesized compounds was assessed against the maximal electroshock induced seizures (MES) model in mice. Neurotoxicity was evaluated using the rotarod method. The majority of compounds exhibited anticonvulsant activity at a dose of 30 mg kg-1 body mass during 0.5-4 h, indicating their ability to prevent seizure spread at low doses. Relative to phenytoin, [N-(2-(4-chlorobenzoyl)benzofuran-3-yl)-2-(cyclohexyl( methyl) amino)-acetamide] (5i) and [N-(2-(4-chlorobenzoyl)benzofuran-3-yl)-2-(4-methylpiperidin-1- yl)-acetamide] (5c) demonstrated comparable relative anticonvulsant potency of 0.74 and 0.72, respectively, whereas [(N-(2-(4-chlorobenzoyl)benzofuran-3-yl)-2-(4-(furan-2-carbonyl)-piperazin-1-yl)-acetamide] (5f) exhibited the lowest relative potency of 0.16. The ALD50 of tested compounds ranged from 1.604 to 1.675 mmol kg-1 body mass. The ED50 of synthesized compounds ranged from 0.055 to 0.259 mmol kg-1 (~23.4 to 127.6 mg kg-1) body mass. The pharmacophore mapping of the examined compounds on standard drugs (phenobarbital, phenytoin, ralitolin and carbamazepine) strongly suggests that these compounds may exert their anticonvulsant activity via the same established mechanism as that of known drugs.

Exploring new pharmacology and toxicological screening and safety evaluation of one widely used formulation of Nidrakar Bati from South Asia region.

BackgroundNidrakar Bati (NKB) is an herbal remedy consisted with seven medicinal herbs widely used to cure Somnifacient (sleeping aid) in South Asia as Ayurvedic medicinal system. In the present study, pharmacological and toxicological effects of this medicine was investigated in mice to validate the safety and efficacy of the herb.MethodsOrganic solvent extracts NKB were prepared using maceration method. Effect of extracts on the central nervous system was evaluated using hypnotic activity assay. Effect of the extracts on metabolic activity, assessing involvement of thyroid was conducted using hypoxia test. analgesic and anti-inflammatory activities were assessed in mice using acetic acid induced writhing, formalin induced paw edema, xylene induced ear edema assays. Anxiolytic activity was performed using plus maze, climbing out and forced swimming tests. Effect of the extracts on psychopharmacological effect was carried out using locomotor activity tests (open field, Hole-board and Hole-cross tests). Neuropharmacological effect of the extracts was performed using motor coordination (rotarod test). Toxicological potential of the extract was evaluated using gastro-intestinal activity (gastric emptying and gastrointestinal motility tests).ResultsThe studied formulation reduced the CNS stimulant effects dose independently. In the hypoxia test, only a dose of 100 mg/kg of NKB decreased the survival time. Orally administration of the NKB (200 and 400 mg/kg) produced significant inhibition (P < 0.01) of the acetic acid-induced writhing in mice and suppressed xylene induced ear edema and formalin-induced licking response of animals in both phases of the test. NKB showed locomotor activity (p < 0.05) both in higher and lower doses (100 and 400 mg/kg). NKB increased the total ambulation dose dependently (p < 0.05). NKB, at all tested doses (100, 200 and 400 mg/kg) increased some locomotion activity parameters (ambulation, head dipping and emotional defecation) in hole board test. At higher doses (200 and 400 mg/kg), NKB showed a significant increase in hole cross test. NKB showed an increase in the time on the open arms of the maze at low to medium doses (100 and 200 mg/kg). When using the Rotarod method, NKB showed a considerable increase on motor coordination of the mice. NKB produced marked gastric emptying effect and decreased gastrointestinal motility in mice at low dose.ConclusionsNKB demonstrated various pharmacological effects and toxicological effects due to presence of several herbs in the formulation those are not closely fit for the effect of CNS depressants.

Design, synthesis and pharmacological evaluation of N-[4-(4-(alkyl/aryl/heteroaryl)-piperazin-1-yl)-phenyl]-carbamic acid ethyl ester derivatives as novel anticonvulsant agents

A series of alkyl/aryl/heteroaryl piperazine derivatives (37–54) were designed and synthesized as potential anticonvulsant agents. The target compounds are endowed with satisfactory physicochemical as well as pharmacokinetic properties. The synthesized compounds were screened for their in vivo anticonvulsant activity in maximal electroshock (MES) and subcutaneous pentylenetetrazole (sc-PTZ) seizure tests. Further, neurotoxicity evaluation was carried out using rotarod method. Structure activity relationship studies showed that compounds possessing aromatic group at the piperazine ring displayed potent anticonvulsant activity. Majority of the compounds showed anti-MES activity whereas compounds 39, 41, 42, 43, 44, 50, 52, and 53 exhibited anticonvulsant activity in both seizure tests. All the compounds except 42, 46, 47, and 50 did not show neurotoxicity. The most active derivative, 45 demonstrated potent anticonvulsant activity in MES test at the dose of 30mg/kg (0.5h) and 100mg/kg (4h) and also delivered excellent protection in sc-PTZ test (100mg/kg) at both time intervals. Therefore, compound 45 was further assessed in PTZ-kindling model of epilepsy which is widely used model for studying epileptogenesis. This compound was effective in delaying onset of PTZ-evoked seizures at the dose of 5mg/kg in kindled animals and significantly reduced oxidative stress better than standard drug phenobarbital (PB). In result, compound 45 emerged as a most potent and safer anticonvulsant lead molecule.

Synthesis and antiepileptic activity of schiff's bases of dialkylamino alkoxy isatin derivatives.

In the present work, some new 5-[2(3)-dialkylamino alkoxy] Indole 3-thiosemicarbazone 2-ones and 5-[2(3)-dialkylamino alkoxy] Indole 3-hydrazone 2-one were prepared from 5-hydroxy isatin. The structures of the products were characterized by IR, NMR, and MASS Spectral studies. All the compounds were examined for antiepileptic activity by maximal electroshock seizure (MES) and pentylenetetrazole (PTZ) induced convulsion method. These compounds were also evaluated for their neurotoxicity study by rotarod method. Some of these compounds showed good antiepileptic activity when compared with standard drug Phenytoin and all the compounds showed less neurotoxicity when compared with standard drug Diazepam.

Synthesis and Pharmacological Evaluation of Novel 1′‐[2‐(Difluoromethoxy)benzyl]‐2′H,5′H‐spiro[8‐azabicyclo[3.2.1]octane‐3,4′‐imidazolidine]‐2′,5′‐diones and Their Derivatives

A series of novel 1'-[2-(difluoromethoxy)benzyl]-2'H,5'H-spiro[8-azabicyclo[3.2.1]octane-3,4'-imidazolidine]-2',5'-dione substituted hydantoins (5-32) were synthesized using an appropriate synthetic route and characterized by elemental analysis and spectral data. The novel molecules were screened for anticonvulsant activity in mice by maximal electroshock (MES) and subcutaneous pentylenetetrazol (ScPTZ)-induced seizure tests. The neurotoxicity was assessed using the rotarod method. Compounds 9, 10, 18, 30, and 31 exhibited anticonvulsant potency against MES seizure and in the ScPTZ model, with lesser neurotoxicity. Some title compounds showed lesser central nervous system depression compared to phenytoin.

New 6-bromoimidazo[1,2-a]pyridine-2-carbohydrazide derivatives: synthesis and anticonvulsant studies

In the present work, we report the facile synthesis and anticonvulsant study of new imidazo[1,2-a]pyridines carrying biologically active hydrazone functionality (3a–3e) and suitably substituted 1,2,4-triazole moieties (4, 5a–5d, 6, and 7a–7d). The newly synthesized intermediates and final compounds were characterized by various spectral techniques such as FTIR, 1H NMR, 13C NMR, and mass spectral and elemental analysis studies. The in vivo anticonvulsant study of the target compounds were carried out following maximal electroshock seizure and subcutaneous pentylene tetrazole methods, while their toxicity study was performed following rotarod method by taking 20, 40, and 100 mg/kg dose levels. Most of the new compounds displayed remarkable anticonvulsant properties at these doses. Particularly, compounds 3b and 4 carrying hydrogen bond donor groups, viz. hydroxyl and amine moieties respectively, exhibited complete protection against seizure and their results are comparable to that of standard drug diazepam. Further, the motor impairment study revealed that all the compounds are nontoxic upto 100 mg/kg.

Synthesis of Pyrazine Substituted 1,3,4-Thiadiazole Derivatives and Their Anticonvulsant Activity

The synthesis of new pyrazine substituted 1,3,4-thiadiazole derivatives was carried out in good yield by the reaction of pyrazine substituted 1,3,4-thiadiazoles with various sulfonyl chlorides. A chemical structure of all the new compounds was confirmed by ¹H NMR and mass spectral data. The new compounds were screened for their anticonvulsant activity against maximal electroshock (MES) seizure method. Rotarod method was employed to determine the neurotoxicity. Few compounds showed significant changes in anticonvulsant activity. The same compounds showed no neurotoxicity at the maximum dose administered (100&#x2009;mg/kg).

ChemInform Abstract: Facile Synthesis of New Imidazo[1,2‐a]pyridines Carrying 1,2,3‐Triazoles via Click Chemistry and Their Antiepileptic Studies.

Abstract The present article reports the synthesis and anticonvulsant studies of new 2-arylimidazo[1,2- a ]pyridines carrying suitably substituted 1,2,3-triazoles as well as their intermediates. The structures of newly synthesized compounds were confirmed by various spectroscopic techniques. The anticonvulsant study was carried out by MES and scPTZ screening methods, while their toxicity study was performed following Rotarod method. The active compounds showed enhanced seizure control in scPTZ method when compared with that of MES method. Compounds 3f , 4c , 4f , 5k , 5p and 5w carrying active pharmacophores exhibited complete protection against seizure and their results were comparable with standard drug diazepam. Majority of new compounds were found to be non-toxic, while few of them showed toxicity at 100 mg/kg. The c log P values of target compounds are in the range of 3.5–5.3, which confirm their lipophilic nature.

Facile synthesis of new imidazo[1,2-a]pyridines carrying 1,2,3-triazoles via click chemistry and their antiepileptic studies

The present article reports the synthesis and anticonvulsant studies of new 2-arylimidazo[1,2-a]pyridines carrying suitably substituted 1,2,3-triazoles as well as their intermediates. The structures of newly synthesized compounds were confirmed by various spectroscopic techniques. The anticonvulsant study was carried out by MES and scPTZ screening methods, while their toxicity study was performed following Rotarod method. The active compounds showed enhanced seizure control in scPTZ method when compared with that of MES method. Compounds 3f, 4c, 4f, 5k, 5p and 5w carrying active pharmacophores exhibited complete protection against seizure and their results were comparable with standard drug diazepam. Majority of new compounds were found to be non-toxic, while few of them showed toxicity at 100mg/kg. The clogP values of target compounds are in the range of 3.5–5.3, which confirm their lipophilic nature.

Evaluation of Antidepressant, Motor Coordination and Locomotor Activities of Ethanolic Root Extract of Clitoria ternatea

Clitoria ternatea (CT) , a perennial herbaceous plant commonly known as 'butterfly pea', has been used for centuries as a traditional Ayurvedic medicine. The aim of present study was to investigate the anti depressant (by Tail suspension test and Forced swimming test), motor coordination (by Rota-rod method) and locomotor (with an actophotometer) activities of ethanolic root extract of Clitoria ternatea Linn. in experimental animals. The study revealed that the extracts (150 and 300 mg/kg, p.o.) were able to reduce the immobility time of rats in a dose-dependent manner when subjected to both Tail suspension and Forced swim tests, and the results were similar to that of standard drug imipramine (15 mg/kg, p.o.). Clitoria ternatea ethanolic root extract (300 mg/kg) showed mild reduction in locomotor and motor coordination activity. The results indicated that ethanolic root extract of Clitoria ternatea has significant anti depressant activity (150 mg/kg (p<0.01) and 300 mg/kg (p<0.001)) when compared with standard imipramine (p<0.001) with a mild sedative effect (300 mg/kg only). The results of the sedative effect are not statistically significant, and the sedative effect may be due to higher dose. Therefore, Clitoria ternatea may be served as a potential resource for natural psychotherapeutic agent against depression and mood disorders.

The role of the angiotensin AT2 receptor on the diurnal variations of nociception and motor coordination in rats

Phasic pain demonstrates significant diurnal variation in rats. Angiotensin II modulates pain transmission and the diurnal variation in nociception in several rodent pain models. The participation of AT2 receptors in the diurnal regulation of nociception is not yet elucidated. In the present study we investigated the effects of selective peptide AT2 agonist CGP 42112A and the nonpeptide AT2 receptor antagonist PD 123319 on the nociception, motor coordination and arterial blood pressure. Male Wistar 12 weeks old rats were used. CGP 42112A was injected at single doses of 1 and 5μg/rat intracerebroventricularly (ICV) and infused chronically ICV at a dose of 12μg/rat/day during 14 days by osmotic minipumps. PD123319 was injected at single doses of 1 and 5μg/rat, ICV and chronically subcutaneously at a dose of 10mg/kg/day/14 days. Nociception was assessed by an analgesimeter, arterial blood pressure (ABP) was measured by tail cuff method, and motor coordination by Rota-rod method. Single doses of CGP 42112A (1 and 5μg/rat) provoked a short lasting antinociception. Unlike acute injection, chronic CGP 42112A infusion increased nociception at the beginning and the end of light phase thus attenuating the diurnal variations observed in the controls. Moreover, it produced an increase of ABP and improved motor coordination. Both acute (1μg/rat) and chronic PD 123319 treatment resulted in a decrease of pain threshold and chronic treatment attenuated its diurnal fluctuation. Our data support a role for Ang II type 2 receptors in the control of diurnal variations of nociception in rats.

Anticonvulsant activity of the essential oil and methanolic extract of Bunium persicum (Boiss). B. Fedtsch.

Ethnopharmacological relevanceBunium persicum is an endemic plant to Iran which its seeds have a long history of medicinal uses. Aim of the studyThis work aimed to study the anticonvulsant effect of the essential oil and methanolic extract of the plant. Materials and methodsThe essential oil and methanolic extract of the plant were studied against pentylenetetrazole (PTZ) and maximal electroshock (MES) induced convulsions in mice in different doses. The neurotoxicity of the essential oil and methanolic extract was investigated using rotarod method. ResultsThe essential oil and methanolic extract prolonged the onset of clonic and tonic seizures in PTZ. The tonic seizures were prevented by essential oil in both methods at dose of 1mL/kg and higher doses. The methanolic extract inhibited PTZ-convulsions at dose 3g/kg and was ineffective against MES induced convulsion. ConclusionsThe essential oil of the plant might be useful to control absence and grand mal seizures at dose 1mL/kg. This activity might be due to its content of monoterpenes.

Molecular Modelling, Design, Synthesis and Biochemical Evaluation of a Range of Non-Steroidal Thiosemicarbazone-Based Inhibitors of Estrone Sulfatase (ES) - A Potential Insight into the Action of a Range of Allosteric Inhibitors of ES

Results of the modelling of thiosemicarbazone-based inhibitors of ES suggest that these compounds interact with the calcium ion whilst undergoing hydrogen bonding interaction with the active site - the extent of H-bonding determining the overall inhibitory activity. Novel inhibitors were designed, synthesised and evaluated and found to possess potent inhibitory activity. Keywords: Allosteric inhibitors, Breast cancer, Estrone sulfatase, Estrone sulfatase, Non-steroidal inhibitors, Thiosemicarbazones, estrogens, coumarin-based compounds, inhibitory activity, phenacyl bromides, sulfamate-based inhibitors, cancer, Elkay, diaryl ketoximes, American Type Culture Collection (ATCC), Rota rod method

Thiadiazole Derivatives as Potential Anticonvulsant Agents

A series of thiadiazole derivatives were synthesized with differently substituted benzoic acids which were cyclized to give differently substituted thiazolidin-4-one. Elemental analysis, IR, <TEX>$^1H$</TEX> NMR, <TEX>$^{13}C$</TEX> NMR and mass spectral data confirmed the structure of the synthesized compounds. The derivatives of these moieties were evaluated for anticonvulsant activity by MES model and neurotoxicity by rotarod method. The synthesized compounds showed good potential for anticonvulsant activity besides this, the compounds also showed neurotoxic effect. It was observed that compounds with <TEX>$OCH_3$</TEX> at 3, 4 position of phenyl ring [5(a-l)] showed less protection against convulsions as compared to compounds having unsubstituted phenyl ring [4(a-l)].

CNS Activity of Aqueous Extract of Root of Cissus Quadrangularis Linn. (Vitaceae)

ABSTRACT.In Ayurveda, Cissus quadrangularis Linn. is used to treat anorexia, asthma, sickle cell, colds, pain, and malaria. Aqueous C. quadrangularis extract was evaluated in vivo for its antiepileptic activity by using the maximal electroshock and isonicotinic hydrazide acid models, for its analgesic activity by using the hot plate method, and for its smooth muscle relaxant activity by using the rotarod method. Adult male Swiss mice were used for this study and animals were divided into 6 animals per group. Doses of 250 mg/kg body weight and 500 mg/kg body weight protected the mice against maximal electroshock seizure, and delayed the onset time of seizures induced by isonicotinic hydrazide acid. Prominent analgesic activity was observed using the hot plate method. The paw licking time was delayed significantly. The extract also displayed prominent smooth muscle relaxant activity. The results suggest that the aqueous extracts of C. quadrangularis roots possess anticonvulsant, analgesic, and smooth muscle relaxant properties.

Anxiolytic and hypnotic effect of Crocus sativus aqueous extract and its constituents, crocin and safranal, in mice

Saffron stigma (Crocus sativus L.) is used for insomnia and anxiety in traditional medicine. In this study, the anxiolytic and hypnotic effects of saffron aqueous extract and its constituents, crocin and safranal, were studied in mice. Agents were administered intraperitoneally in mice before the experiments for the evaluation of hypnotic activity (induced by sodium pentobarbital, 30 mg/kg, i.p.), anxiolytic activity (elevated plus maze test), locomotor activity (open field test) and motor coordination (Rotarod test). The aqueous extract reduced the locomotor activity dose dependently. At low doses, saffron showed a significant increase in the time on the open arms of the maze. When using the Rotarod method, the aqueous extract showed considerable effect on motor coordination of the mice. In the hypnotic test, only a dose of 0.56 g/kg of saffron increased the total sleep. Crocin showed no anxiolytic, hypnotic or myorelaxation effects. Safranal, in higher doses, 0.15 and 0.35 mL/kg, showed anxiolytic effects. Safranal increased the total sleep time dose dependently. This constituent at lower doses (0.05 and 0.15 mL/kg) decreased some locomotion activity parameters. Safranal demonstrated no effects on motor coordination. The results showed that saffron aqueous extract and safranal have anxiolytic and hypnotic effects.

An automated rotarod method for quantitative drug-free evaluation of overall motor deficits in rat models of parkinsonism

A variety of tests have been proposed for evaluation of motor deficits in rat models of hemiparkinsonism. The most widely used test, the “rotational behaviour” test, is based upon motor asymmetry induced by drug administration, leading to problems of sensitization, conditioning and priming. In addition, lesion-induced rotational behaviour is sometimes overcompensated by treatments (grafts) while other deficits are unaffected. However, rotational behaviour is reproducible and easy to quantify. On the contrary, most drug-free tests have proved of limited applicability because of subjective scaling, excessive interaction between the experimenter and animals, motivational effects, excessively complex tasks, etc., making it difficult to compare results from different laboratories. In this paper, we present a rotarod based method for automated drug-free evaluation of overall motor deficits in the rat model of hemiparkinsonism, and for identifying treatment-induced changes. Animals are pre-trained on the rotarod and then tested at a series of increasing rod speeds; from this set of values a unique score for each animal is computed (the overall rotarod performance, ORP) summarizing its performance at different speeds. This value is easy to compute and greatly simplifies statistical comparisons. Major advantages of the test are: (i) it does not require drugs, but is nevertheless highly objective, reproducible and easy to quantify; and (ii) falling of animals from rotating rod seems to depend on a combination of lesion-induced deficits which become more evident when rats are forced to move at faster speeds. Since the test is not based exclusively on motor asymmetry, it may also be useful for characterization of bilaterally lesioned animals, for which drug-induced rotational behaviour tests cannot be used.

Pharmacodynamics of Chlorzoxazone in Rats

Pharmacological effect–time profiles of a centrally acting muscle relaxant, chlorzoxazone, were evaluated by both rotarod and crossed extensor reflex (CER) methods. Drug response was measured by percent change in the remaining time on the rod for the former method and in intensity of isometric contraction of the muscle for the latter method. The drug response was quantitated using the logarithmic–logistic function and then biophase levels were calculated. The results indicated that the maximum response was observed 10 min after drug administration for both methods at every dose studied. However, the values of ED50 indicated that the CER method was more sensitive than the other. Pharmacodynamic analysis offered the results that the dose-normalized biophase levels for each dose obtained from the CER method were coincident with each other and that the logarithmic–logistic function was proven to be of use for the pharmacodynamic modeling which could simply assess the bioavailability. On the other hand, all dose-normalized values obtained from the rotarod method did not scatter around the same line. This evidence indicated a violation of assumption for the pharmacodynamic analysis. From the above data, the CER method proved to be of use for assessing the drug response based on the dose-independent kinetics with high sensitivity.

Drug interaction of imipramine hydrochloride to the pharmacodynamics and pharmacokinetics of oxazepam

In this report, we studied drug interaction between oxazepam and imipramine in rats. Oxazepam (20 mg/kg) and imipramine (20 or 50 mg/kg) were administrated orally. The oxazepam concentration in plasma, brain and liver were measured by the method of HPLC. The concomitant use of imipramine induced extension of the elimination half life (T 1/2 beta) and an increase of the area under the concentration time-curve (AUC) on the plasma concentration of oxazepam. With the concomitant use of imipramine, the AUC of oxazepam brain concentration increased approximately 1.42 to 1.56 in contradistinction to oxazepam alone. The anti-pentylenetetrazol effect of oxazepam at 1 hr after administration was increased by the concomitant use of imipramine, but there were no combination effects at 4 hr. The motor incoordination effect of oxazepam and diazepam was measured by the rotarod method. Oxazepam has little effect on the motor incoordination as compared with diazepam. The plasma protein binding of oxazepam was not changed by the combined use of imipramine both in vitro and in vivo. The pharmacodynamic effects of oxazepam were increased by the concomitant use of imipramine, and these effects were in reasonably good agreement with the change in brain concentration of oxazepam.