Study of In-vivo Analgesic Activity in Animals

Study of In-vivo Analgesic occupation in AnimalsA) ANIMALSSwiss albino mice (20-25 g) and wistar rats (150-200 g) of either sex were use for study of in-vivo moderating activity. Animals were unploughed under stock laboratory conditions i.e. temprature is 24 2C and coition humidity is 60-70%. The study protocol was approved by the institutional animal ethics committee (IAEC) before experiment (Approval No. 1452/PO/a/11/CPCSEA). Albino-Swiss mice were taken from Laboratory Animal House, Devsthali Vidyapeeth College of Pharmacy, Lalpur, Rudrapur (U. S. Nagar) and utilise for the study. The animals were procured from IVRI, B areilly (U.P.) The animals were kept in polypropylene cages and maintained on balanced ration with relax access to clean drinking water. all experimental procedures were conducted in uniformity with the guide for Care and use of laboratory animals and in accordance with the topical anaesthetic animal care and use committee. Paddy husk was provided as su pply material, which was cleaned every day. The cages were maintained clean. any of the animals were left for 2 days in the laboratory for getting utilise to before the day of experiment and on the last day they were inclined water only. Minimum of 6 animals were utilize in each free radical.B) ACUTE TOXICITY STUDIESThe acute oral toxicity studies were carried out to study the acute toxic effects and to deter minutee token(prenominal) toxic superman of the synthesized compounds. For the study swiss albino mice of either sex calculation 20-25 g were used. The aqueous solution of compounds were administered orally to different separates of over dark fasted mice at the doses of 30, 100, 300, 1000 and 3000 mg/kg body weight. After system of the compounds, animals were observed continuously for any toxic manifestation for the first triplet hours. There after, observations were made at regular intervals for 24 hrs. Further the animals were under investigation up to a period o f single week.I) ANALGESIC natural processFor the study of analgesisc activity two rules were used.(A) Hot Plate method(B) acetic caid induced writhing methodA) method 1 Hot habitation method186,187,188,189 By applying heat pain is inced to animals. All the animals one by one are kept in the hot plate maintain at constant temperature (55C) and there reactions was noted i.e. paw licking or jump response.Work planAlbino rats of either sex (150-200 g) were selected and divided into four groups of cardinal animals each. All the animals were fasted for 24 hrs. before the start of the experiment and water was devoted adlibitum. The animals were inured as follows concourse 1 Control group veritable 0.5% sodium CMC (1mg/kg) orally.Group 2 Diclofenac sodium 50mg/kg were administered orally.Group 3 new(a) benzimidazole substituted pyrazolidine 3,5 dione derivative in dose take aim of 50mg/kg was administered orally.Group 4 refreshing 2-quinolone substituted pyrazolidine 3,5 d ione derivative in dose level of 50mg/kg was administered orally.hither Group 1 is the control, group 2 is active quantity and group 3 and group 4 are test.Experimental expoundThe hot plate method is based on the fact that anodyne compounds increases the response metre. This method was first described by Eddy Leimbach, where a cut off period of 15 sec is observed to neutralise damage to the paw. All the synthesized compounds were dissolved in the CMC (0.5% suspension). After administration of control, meter and test compounds the animals were kept at the hot plate and their reaction time were note at 15, 30, 60 120 min interval. All the doses were disposed(p) orally to animals. Diclofenac Sodium at dose of 50 mg/kg was used standard drug for comparison. The results so obtained were tabulated in Table 10, 12, 14 and 16 and figure 07, 09, 11 and 13. Results were expressed as means S.E.M. statistical conditional relation was analyzed using the two-part anova analysis of di vergency followed by Tukeys Multiple similarity Test where p B) Method 2 Acetic Acid Induced Writhing Method186,187,188,189 In this method pain is induced by intraperitoneal (I.P) administration of 0.6% (0.1 ml/10g) acetic vinegarish in mice. Analgesic activity was determined by calculating radical number of writhings.Work planAlbino mice of either sex (25-30 g) were used for the study. All the animals were fasted for 24 hrs. before the start of the experiment and water was minded(p) adlibitum. The animals were treated as follows Group 1 Control group reliable 0.5% sodium CMC (1mg/kg) orally.Group 2 Diclofenac sodium 20mg/kg were administered orally.Group 3 Novel benzimidazole substituted pyrazolidine 3,5 dione derivative in dose level of 20mg/kg was administered orally.Group 4 Novel 2-quinolone substituted pyrazolidine 3,5 dione derivative in dose level of 20mg/kg was administered orally. here(predicate) Group 1 is the control, group 2 is active standard and group 3 and gr oup 4 are test.Experimental DetailsAll the synthesized compounds were administered intraperitonealy (0.5 ml) as a suspension in sterile 0.9% DMSO solution as vehicle. Diclofenac Sodium at dose of 20 mg/kg was used standard drug for comparison. Acetic acid solution was intraperitonealy administered 30 min after administration of the compounds. 10 min after intraperitoneal shooting of acetic acid solution, the number of writhings per animal was recorded for 20 min. Control animals receive an equal masses of vehicle. Results were expressed as means S.E.M. statistical significance was analyzed using the two-way anova analysis of pas seul followed by Tukeys Multiple Comparison Test where p II) ANTI-PYRETIC ACTIVITY STUDIESxclFor antipyretic activity yeast induced pyrexia model was used for the study.Work planAlbino rats of either sex (150-200 g) were selected and divided into four groups of sise animals each. All the animals were fasted for 24 hrs. before the start of the experimen t and water was given adlibitum. The animals were treated as follows Group 1 Control group receive 0.5% sodium CMC (1mg/kg) orally.Group 2 Peracetamol 100mg/kg were administered orally.Group 3 Novel benzimidazole substituted pyrazolidine 3,5 dione derivative in dose level of 100mg/kg was administered orally.Group 4 Novel 2-quinolone substituted pyrazolidine 3,5 dione derivative in dose level of 100mg/kg was administered orally.Here Group 1 is the control, group 2 is active standard and group 3 and group 4 are test.Experimental DetailsFor instauration of fever in rats, 20% w/v of brewers yeast in distilled water was administered by subcutaneous injection. All animals which were used for study, were induced pyrexia by injection of 10 ml/kg of brewers yeast solution under the skin in between the shoulder blades. The place of the injection was massaged in order to overspread the suspension beneath the skin. Basal rectal temperature was measured before the injection of yeast, by inserting digital clinical thermometer to a depth of 2 cm into the rectum. The rise in rectal temperature was recorded after 19 hours of yeast injection. The rectal temperature was taken after 30, 60, 120, 180 and 300 minutes direct treatment. If a drug is having antipyretic effect then there is a fall in the rectal temprature. Results were expressed as means S.E.M. Statistical significance was analyzed using the two-way anova analysis of variance followed by Tukeys Multiple Comparison Test where p III) anti-inflammatory drug ACTIVITY 186,187,188,189 For anti-inflammatory activity carrageenin-induced rat paw oedema method was used.Work planAlbino rats of either sex (150-200 g) were selected and divided into four groups of half a dozen animals each. All the animals were fasted for 24 hrs. before the start of the experiment and water was given adlibitum. The animals were treated as follows Group 1 Control group received sterile normal saline (0.85% NaCl) orally.Group 2 Ibuprofen 20mg/kg were administered orally.Group 3 Novel benzimidazole substituted pyrazolidine 3,5 dione derivative in dose level of 50mg/kg was administered orally.Group 4 Novel 2-quinolone substituted pyrazolidine 3,5 dione derivative in dose level of 50mg/kg was administered orally.Here Group 1 is the control, group 2 is active standard and group 3 and group 4 are test.Experimental DetailsThis method was described by Winter et al. in 1962. The experimental animals were divided into ten groups, each containing five animals. After 30 min of administration of test compounds, 0.1 ml of 1% (w/v) carrageenin was injected subcutaneously in the subplantar region of the left hind paw. The right paw served as a reference to non inflammed paw for comparison. The initial paw volume was measured within 30 sec of the carrageenin injection by plethysmometer. The relative increase in paw volume was measured in control, standard and test compounds at 1, 2, 3, 4, 5, 6, 7 and 8 h after the carrageenin inj ection. The discrepancy between initial and final readings was taken as the volume of oedema and the percentage inhibition by the compounds was calculated using the formula-where dt is the difference in paw volume in the test compound-treated group and dc the difference in paw volume in the control group. Results were expressed as means S.E.M. Statistical significance was analyzed using the two-way anova analysis of variance followed by Tukeys Multiple Comparison Test where p Uttarakhand Technical University, Dehradun 1