- Email: [email protected]
322 Substational of hepatoprotective action of biomelan preparation in experiment
s88
Poster Session P12. Drug toxicology
extracts was studied by brine shrimp method. The results showed that ethylacetate extract was most cytotoxic (LD50 = 150 µg/cc), the alcoholic extract had LD50 = 205 µg/ cc and the ether extract didn’t show cytotoxocity. 321
STUDY ON ADSORPTION OF ET-743 TO MATERIALS USED IN INTRAVENOUS RODENT INFUSION STUDIES
J. Verbeeck 1 , A. Looszova 1 , T. Verhaeghe 1 , L. Diels 1 , L. Lammens 1 , R. De Coster 1 , I. Manzanares 2 , P. Aviles 2 , W. Coussement 1 . 1 Johnson and Johnson Pharmaceutical Research and Development, a division of Janssen Pharmaceutica N.V., GPCD/PCDE, Beerse, Belgium 2 PharmaMar, Madrid, Spain Yondelis™ (trabectedin, ET-743) is a marine derived anti-cancer agent with a unique mode of action that is administered intravenously in low concentrations and, in rodent infusion studies, at low flow rates (∼ 5 ml/kg b.w./h). Adsorption of the compound to the infusion materials used in these studies becomes problematic. The purpose of the presented work was to determine and assess the extent of adsorption of Yondelis™ to different catheter and syringe materials and to select appropriate materials for future rodent infusion studies. LC-MS/MS was the method of choice to analyze the formulation for Yondelis™. Firstly, adsorption to different materials was tested after overnight instillation of an Yondelis™ formulation. Adsorption was slightly higher for polyethylene than for glass syringes, but remained acceptable. Polyethylene and silicone tubing showed clearly more adsorption than polyurethane tubing. Secondly, polyurethane tubing, in combination with a polyurethane catheter, was tested in a mock-up infusion setting. A polyethylene syringe was used, as adsorption to this type of syringe was acceptable. Infusion was performed using a low concentration (0.67 µg/ml, 0.75 ml/h) over 24 hours and a high concentration (3.33 µg/ml, 1.5 ml/h) over 3 hours. In the first setting, adsorption was clearly present in the beginning of the infusion but decreased (saturation). Total adsorption was between 10 to 13% which is considered acceptable. Infusing a formulation at the high concentration over 3 hours caused a total adsorption of about 9%. In vitro adsorption experiments indicate that adhesion of Yondelis™ to polyethylene and silicone infusion lines is clearly present. Yondelis™ is compatible with polyurethane infusion lines, which will be used as the material of choice in future rodent infusion studies. The impact on clinical studies is negligible since higher flow rates and/or concentrations are used in these studies. 322
in acetaminophen intoxication. Melanins are stable polyradical, contain some semiquinone radicals and accumulate the active oxygen species and electrophyl toxic compounds. Some thise properties determine the antitoxic activity of biomelan preparation. The revealed biomelan hepatoprotective activity makes it possible to be applied as an effective antidote therapy under acetaminophen toxicity.
SUBSTATIONAL OF HEPATOPROTECTIVE ACTION OF BIOMELAN PREPARATION IN EXPERIMENT
A. Shayakhmetova, G. Saifetdinova, V. Kovalenko, O. Voloshina. Institute of Pharmacology and Toxicology, Kyiv, Ukraine, Pharmacological Centre, Ministry of Health, Kyiv, Ukraine Hepatoprotective efficacy of a new original sorbent biomelan preparation was studied under acetaminophen toxicity using Wistar rats. Acetaminophen was administered intragastrically for two days in the dose 1.25 mg/kg. The biomelan preparation (it built on a basis of melanin from the biomass of fungal producent Cladosporidium cladosporioides) was administered intragastrically, treatment-andprophylactic in the dose 10 mg/kg. After treatment of acetaminophen the cytochrome P-450 and b5 level in liver microsomes decreased by 2 fold, the liver protein SH-groups contents and catalase activity decreased by 31% and 15%. We observed a decrease of cholesterol level by 19% and an increase of AlAT by 2.5 fold in serum of blood as compared to intact animals. It was shown biomelan preparation administration as treatment or prophylaxis significantly attenuates analgesic toxicity enhancing liver detoxication via concerving both cytochrome P-450 and b5 content. Biomelan administration prevents electrophylic acetaminophen metabolites binding with the free protein SH-groups, normalized the cytochrome P-450 and b5 level, increases enzyme antioxidant catalase activity by 2 fold as compared to acetaminophen treated rats, corrects lipid turnover as well as limits the phenomenon of hepatocytolysis. Administration of biomelan preparation to rats exerted significant normalizing influence on liver function exceeding efficacy of metionine, which is use as an antidote
323
CYTOTOXICITY AND ANTIMICROBIALBIOL ACTIVITY OF EUPHORBIA HEBECARPA BOISS
R. Miri 1 , K. Javidnia 1 , A. Jafari 2 . 1 Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran, 2 Research center of Natural Resource and Animal Husbandry, Yasuje, Iran Euphorbia is a family of plants spread in all parts of the world exept in cold regions. Some reports have been shown that the genus Euphorbia has cytotoxicity and anti tumor activity. Euphorbia hebecarpa Boiss ia an endemic plant of Iran, which in literature review no reports can be found on its pharmacological activities. Four extracts (petroleum ether, ether, ethylacetate and methanolic) of the plant were prepared by maceration method. Antibacterial and antifungal activities of the extracts were studied by Disc Diffusion method. The ethylacetate and methanolic extract showed a moderate antibacterial and antifungal activity. Biological activities of four extracts were studied by brine shrimp method. The results showed that methanolic extract was most cytotoxic (LD50 = 11 µg/cc). Cytotoxic activity of methanolic extract was studied on four different cell lines by MTT assay. The methanolic extract was mostly active against K562 and vero cell lines. 324
ANALGESIC ACTIVITY OF IRANIAN HYPERICUM PERFORATUM
K. Morteza-Semnani 1 , M. Mahmoudi 2 , M. Saeedi 1 , A. Javanmardi 1 . 1 Department of Medicinal Chemistry, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran, 2 Department of Pharmacology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Hypericum is a genus of about 400 species, wide-spread in warm temperature areas throughout the world and well represented in the mediterranean area. Some species of this genus are used in folk medicine as anthelmintics, diuretics, on wounds, scalds and herpes. One of the most important species of this genus is Hypericum perforatum L. (Hypericaceae). The analgesic effect of Iranian Hypericum perforatum extract, a native plant of Iran, was studied using formalin, hot plate and writhing tests. The aerial parts of plant were dried in the shade and powdered. Powder was Soxhlett-extracted with methanol for 16 h., then this extract was evaporated to dryness and weighed (24 g, 24%). Just prior to use, the dried methanolic extract was dissolved in a mixture of propylene glycol and water (1:4). We have also tested the analgesic doses of extract for its effect on motor coordination by rotarod test. In the formalin test, the extract (25–250 mg/kg, i.p.) caused graded inhibition of both phases of formalin-induced pain (P<0.001). In the hot plate test, the i.p. administration of the extract at the doses of 25- 250 mg/kg significantly raised the pain threshold at an observation time of 30 min in comparison with the control group (P<0.001). In the writhing test, the extract at doses of 25 mg/kg (P<0.05), 50, 75, 100 and 150mg/kg (P<0.001) produced a significant decrease in the number of writhing in comparison with the control group. The extract, at antinociceptive doses, did not affect motor coordination of animals when assessed in the rotarod model. It seems that the extract relieved pain through both central and peripheral mechanisms. Based on the results of this study, we suggest that the anti-nociceptive effect of this extract may be attributed to inhibition of prostaglandin synthesis or release and other mediators.
Poster Session P12. Drug toxicology
extracts was studied by brine shrimp method. The results showed that ethylacetate extract was most cytotoxic (LD50 = 150 µg/cc), the alcoholic extract had LD50 = 205 µg/ cc and the ether extract didn’t show cytotoxocity. 321
STUDY ON ADSORPTION OF ET-743 TO MATERIALS USED IN INTRAVENOUS RODENT INFUSION STUDIES
J. Verbeeck 1 , A. Looszova 1 , T. Verhaeghe 1 , L. Diels 1 , L. Lammens 1 , R. De Coster 1 , I. Manzanares 2 , P. Aviles 2 , W. Coussement 1 . 1 Johnson and Johnson Pharmaceutical Research and Development, a division of Janssen Pharmaceutica N.V., GPCD/PCDE, Beerse, Belgium 2 PharmaMar, Madrid, Spain Yondelis™ (trabectedin, ET-743) is a marine derived anti-cancer agent with a unique mode of action that is administered intravenously in low concentrations and, in rodent infusion studies, at low flow rates (∼ 5 ml/kg b.w./h). Adsorption of the compound to the infusion materials used in these studies becomes problematic. The purpose of the presented work was to determine and assess the extent of adsorption of Yondelis™ to different catheter and syringe materials and to select appropriate materials for future rodent infusion studies. LC-MS/MS was the method of choice to analyze the formulation for Yondelis™. Firstly, adsorption to different materials was tested after overnight instillation of an Yondelis™ formulation. Adsorption was slightly higher for polyethylene than for glass syringes, but remained acceptable. Polyethylene and silicone tubing showed clearly more adsorption than polyurethane tubing. Secondly, polyurethane tubing, in combination with a polyurethane catheter, was tested in a mock-up infusion setting. A polyethylene syringe was used, as adsorption to this type of syringe was acceptable. Infusion was performed using a low concentration (0.67 µg/ml, 0.75 ml/h) over 24 hours and a high concentration (3.33 µg/ml, 1.5 ml/h) over 3 hours. In the first setting, adsorption was clearly present in the beginning of the infusion but decreased (saturation). Total adsorption was between 10 to 13% which is considered acceptable. Infusing a formulation at the high concentration over 3 hours caused a total adsorption of about 9%. In vitro adsorption experiments indicate that adhesion of Yondelis™ to polyethylene and silicone infusion lines is clearly present. Yondelis™ is compatible with polyurethane infusion lines, which will be used as the material of choice in future rodent infusion studies. The impact on clinical studies is negligible since higher flow rates and/or concentrations are used in these studies. 322
in acetaminophen intoxication. Melanins are stable polyradical, contain some semiquinone radicals and accumulate the active oxygen species and electrophyl toxic compounds. Some thise properties determine the antitoxic activity of biomelan preparation. The revealed biomelan hepatoprotective activity makes it possible to be applied as an effective antidote therapy under acetaminophen toxicity.
SUBSTATIONAL OF HEPATOPROTECTIVE ACTION OF BIOMELAN PREPARATION IN EXPERIMENT
A. Shayakhmetova, G. Saifetdinova, V. Kovalenko, O. Voloshina. Institute of Pharmacology and Toxicology, Kyiv, Ukraine, Pharmacological Centre, Ministry of Health, Kyiv, Ukraine Hepatoprotective efficacy of a new original sorbent biomelan preparation was studied under acetaminophen toxicity using Wistar rats. Acetaminophen was administered intragastrically for two days in the dose 1.25 mg/kg. The biomelan preparation (it built on a basis of melanin from the biomass of fungal producent Cladosporidium cladosporioides) was administered intragastrically, treatment-andprophylactic in the dose 10 mg/kg. After treatment of acetaminophen the cytochrome P-450 and b5 level in liver microsomes decreased by 2 fold, the liver protein SH-groups contents and catalase activity decreased by 31% and 15%. We observed a decrease of cholesterol level by 19% and an increase of AlAT by 2.5 fold in serum of blood as compared to intact animals. It was shown biomelan preparation administration as treatment or prophylaxis significantly attenuates analgesic toxicity enhancing liver detoxication via concerving both cytochrome P-450 and b5 content. Biomelan administration prevents electrophylic acetaminophen metabolites binding with the free protein SH-groups, normalized the cytochrome P-450 and b5 level, increases enzyme antioxidant catalase activity by 2 fold as compared to acetaminophen treated rats, corrects lipid turnover as well as limits the phenomenon of hepatocytolysis. Administration of biomelan preparation to rats exerted significant normalizing influence on liver function exceeding efficacy of metionine, which is use as an antidote
323
CYTOTOXICITY AND ANTIMICROBIALBIOL ACTIVITY OF EUPHORBIA HEBECARPA BOISS
R. Miri 1 , K. Javidnia 1 , A. Jafari 2 . 1 Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran, 2 Research center of Natural Resource and Animal Husbandry, Yasuje, Iran Euphorbia is a family of plants spread in all parts of the world exept in cold regions. Some reports have been shown that the genus Euphorbia has cytotoxicity and anti tumor activity. Euphorbia hebecarpa Boiss ia an endemic plant of Iran, which in literature review no reports can be found on its pharmacological activities. Four extracts (petroleum ether, ether, ethylacetate and methanolic) of the plant were prepared by maceration method. Antibacterial and antifungal activities of the extracts were studied by Disc Diffusion method. The ethylacetate and methanolic extract showed a moderate antibacterial and antifungal activity. Biological activities of four extracts were studied by brine shrimp method. The results showed that methanolic extract was most cytotoxic (LD50 = 11 µg/cc). Cytotoxic activity of methanolic extract was studied on four different cell lines by MTT assay. The methanolic extract was mostly active against K562 and vero cell lines. 324
ANALGESIC ACTIVITY OF IRANIAN HYPERICUM PERFORATUM
K. Morteza-Semnani 1 , M. Mahmoudi 2 , M. Saeedi 1 , A. Javanmardi 1 . 1 Department of Medicinal Chemistry, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran, 2 Department of Pharmacology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Hypericum is a genus of about 400 species, wide-spread in warm temperature areas throughout the world and well represented in the mediterranean area. Some species of this genus are used in folk medicine as anthelmintics, diuretics, on wounds, scalds and herpes. One of the most important species of this genus is Hypericum perforatum L. (Hypericaceae). The analgesic effect of Iranian Hypericum perforatum extract, a native plant of Iran, was studied using formalin, hot plate and writhing tests. The aerial parts of plant were dried in the shade and powdered. Powder was Soxhlett-extracted with methanol for 16 h., then this extract was evaporated to dryness and weighed (24 g, 24%). Just prior to use, the dried methanolic extract was dissolved in a mixture of propylene glycol and water (1:4). We have also tested the analgesic doses of extract for its effect on motor coordination by rotarod test. In the formalin test, the extract (25–250 mg/kg, i.p.) caused graded inhibition of both phases of formalin-induced pain (P<0.001). In the hot plate test, the i.p. administration of the extract at the doses of 25- 250 mg/kg significantly raised the pain threshold at an observation time of 30 min in comparison with the control group (P<0.001). In the writhing test, the extract at doses of 25 mg/kg (P<0.05), 50, 75, 100 and 150mg/kg (P<0.001) produced a significant decrease in the number of writhing in comparison with the control group. The extract, at antinociceptive doses, did not affect motor coordination of animals when assessed in the rotarod model. It seems that the extract relieved pain through both central and peripheral mechanisms. Based on the results of this study, we suggest that the anti-nociceptive effect of this extract may be attributed to inhibition of prostaglandin synthesis or release and other mediators.