Effect of different dosage regimens of a topical formulation of Hemantane on complete Freund`s adjuvant-induced nonspecific inflammation in rats

Цель исследования - оценка способности гимантана (N-(2-адамантил)-гексаметиленимина гидрохлорида) в наружной гелевой лекарственной форме ослаблять у крыс неспецифическую воспалительную реакцию, вызванную полным адъювантом Фрейнда (ПАФ). Методика. Воспалительный процесс моделировали субплантарным введением ПАФ в левую заднюю лапу аутбредных крыс. Оценивали степень отёчности лапы животных по изменению диаметра стопы и голеностопного сустава до и после введения ПАФ. Гимантан в виде 5% гелевой лекарственной формы и препарат сравнения диклофенак в виде 1% гелевой лекарственной формы наносили ежедневно на повреждённую конечность крыс в течение 14 сут. Эксперимент проводили в 2 этапа. На 1-м этапе исследования препараты начинали наносить за 1 сут до индукции воспаления; на 2-м этапе препарат наносили ежедневно спустя 2 нед после индукции воспаления. Во время 1-го этапа работы на 4-е сут после введения ПАФ у животных регистрировали «Opto-Varimex» (Columbus Instruments, США). Результаты. Установлено, что на 1-м этапе (на 3-и и 7-е сут после индукции воспаления) гель гимантана значимо снижает отёчность стопы и голеностопного сустава поврежденной лапы животных, по эффективности не уступая гелю диклофенака; на 14-е сут после введения ПАФ значимого влияния на выраженность отёка повреждённой конечности не зафиксировано. На 4-е сут после введения ПАФ у животных с моделью воспаления наблюдается снижение на 34,03% числа горизонтальных перемещений в актометре «Opto-Varimex»; 5% гель гимантана и 1% гель диклофенака улучшают координацию движений крыс соответственно на 52,35% и 45,72%. На 2-м этапе исследования установлено, что 5% гель гимантана уменьшает отёчность стопы животных с экспериментальным воспалением через 7 и 14 сут применения препарата, 1% гель диклофенака значимого влияния не оказывает. Заключение. Курсовое ежедневное применение гимантана в наружной гелевой лекарственной форме снижает выраженность отёчного синдрома у аутбредных крыс с неспецифической воспалительной реакцией, вызванной субплантарным введением в левую заднюю лапу животных флогогенного агента. Эффективность лечения гимантаном, начатого как за 1 сут до введения ПАФ, так и через 2 нед после индукции воспаления, не уступает диклофенаку, применяемому в наружной лекарственной форме. The aim of the study was to evaluate the ability of different once-daily treatment regimens with N-(2-adamantyl) hexamethyleneimine hydrochloride (hemantane) topical gel vs. diclofenac topical gel to reduce nonspecific inflammation in rats induced with complete Freund’s adjuvant. Methods. Inflammation was induced by a subplantar injection of complete Freund’s adjuvant into the left hind paw of outbred rats. Edema of the left hind paw was measured. 5% hemantane gel (provided by the Laboratory of Finished Dosage Forms, V.V. Zakusov Institute of Pharmacology) and 1% diclofenac gel (produced by Hemofarm) were applied daily to the affected rat limb for 14 days. The experiment was performed in two stages. At the first stage, the drugs in the form of topical gels were applied to the left hind paw daily starting one day before the induction of inflammation. At the second stage, the topical gel formulations were applied daily starting two weeks after the induction of inflammation. At the first stage of the investigation, on day 4 after the injection of complete Freund’s adjuvant, coordination disorder of rats induced by the experimental pathology was evaluated using an Opto-Varimex actometer (Columbus Instruments, USA). Results. At the first stage of the study, hemantane gel significantly reduced edema of the foot and ankle joint of the left hind paw on days 3 and 7 after the induction of inflammation and was non-inferior to diclofenac gel. On day 14 after the injection of complete Freund’s adjuvant into the rat hind paw, the drugs had no significant effect on edema of the paw. On day 4 after the injection, the number of horizontal movements decreased by 34.03% as indicated by the Opto-Varimex actometer; 5% hemantane gel and 1% diclofenac gel improved coordination in rats by 52.35% and 45.72%, respectively. At the second stage of the study, 5% hemantane gel reduced foot edema in the model of inflammation after 7 and 14 days of treatment, whereas 1% diclofenac gel exerted no significant effect. Conclusion. Hemantane as a topical formulation reduces edema in outbred rats with nonspecific inflammation induced by the subplantar injection of complete Freund’s adjuvant into the left hind paw when applied daily, starting either one day before the induction of inflammation or two weeks after the complete Freund’s adjuvant administration, and its efficacy was not inferior to diclofenac as a topical formulation.

Analgesic, anti-inflammatory and hypouricemic effects of GT1 film-coated tablets on experimental animals

Aim: To evaluate pain relief, anti-inflammatory and hypouricemic effects of GT1 tablets on experimental animals. Method: GT1 at the doses of 22.32 g/kg/day and 66.96 g/kg/day were evaluated for its analgesic effect in three models (hot plate, pain threshold, and acetic acid-induced writhing), its chronic anti-inflammatory effect in the granulomatous reaction model, and its hypouricemic effect in potassium oxonate-induced hyperuricemic mice. Acute anti-inflammatory effects of GT1 at the doses of 11.16 g/kg/day and 33.48 g/kg/day were evaluated in rats with two models: carrageenin-induced paw edema and peritonitis. Results: GT1 prolonged the temperature reaction time on the hot plate (22.73 s and 20.37 s at both doses of 22.32 g/kg and 66.96 g/kg, respectively, compared to 16.96 s in control group), reduced the number of acid acetic-induced writhing effects, decreased the weight of granulomas, and decreased the level of acid uric in blood and urine (p < 0.05). GT1 caused a significant reduction in paw edema after subplantar injection of carrageenan in rats (p < 0.05). Moreover, there was a substantial decline of GT1 at the dose of 11.16 g/kg/day in terms of the volume and the quantity of protein in the inflammation fluid of the peritonitis model (p < 0.05). Conclusion: GT1 at both doses of 11.16 g/kg/day and 33.48 g/kg/day posed acute anti-inflammatory effects on rats. GT1 at both doses of 22.32 g/kg/day and 66.96 g/kg/day exerted analgesic, chronic anti-inflammatory and hypouricemic effects on mice.

Study to Investigate the anti-inflammatory effect of Codelac® Broncho with Thymus Serpyllum (elixir) in comparison with reference drug Fenspiride (syrup) using accute Carrageenan-induced Paw Inflammation Model

Introduction: Evaluation of anti-inflammatory action of Codelac® Broncho with Thymus Serpyllum (elixir) in comparison with Fenspiride was carried out on the model of acute carrageenan inflammation of the paws in rats. Materials and methods: Edema was caused by subplantar injection of 0.1 ml of 1% λ- carrageenan gel into the hind paw. The severity of edema was assessed by using 37140 plethysmometer (UGO BASILE, Italy). The measurements were performed before edema induction and 1, 2, 4, 12, 24, 48, 72, 96 and 120 hours afterwards. Anti-inflammatory activity of the drugs was also evaluated based on the analysis of rats’ blood, C-reactive protein concentration and histological examination results. Results and discussion: A decrease in the paw volume increment was revealed in the group with the studied drug in comparison with the group with the carrageenan edema model (control) 4, 12, 24 hours after injection of carrageenan (p&lt;0.05). As a result of plethysmometry, a more pronounced anti-inflammatory effect of the studied drug than that of Fenspiride was revealed. There was a significant decrease in the levels of leukocytes (p&lt;0.05), lymphocytes (p&lt;0.05), monocytes (p&lt;0.05) and neutrophils (p&lt;0.05) in the group with the studied drug compared to those the the control 48 hours after the initiation of edema, while in the group with Fenspiride, there was only a decrease in the levels of leukocytes (p&lt;0.05) and lymphocytes (p&lt;0.05). There were no differences in the concentration of C-reactive protein between the groups. Conclusion: The obtained data indicate a more pronounced anti-inflammatory activity of Codelac® Broncho with Thymus Serpyllum in comparison with Fenspiride, on the model of acute carrageenan inflammation of the paw in rats.

Anti-inflammatory and analgesic activity of ointment based on dense ginger extract (Zingiber officinale)

Introduction: Zingiber officinale (Zingiberaceae family) is traditionally used in alternative medicine to reduce pain from rheumatoid arthritis and osteoarthritis. Ginger is also often applied for stomach and chest pain, toothaches and as anti-inflammatory agent. The aim of this study is to investigate analgesic and anti-inflammatory activities of Z. officinale dense extract after its transdermal delivery using allyl isothiocyanate (AITC) induced model with further discussion of possible action mechanism of ginger phytoconstituents. Methods: Inflammation was induced by subplantar injection to the plantar fasciitis (aponeurosis) of the hind limb of rats using 30 µL AITC solution (100 µg/limb) in 1,2-propyleneglycol. The dynamics of changes of inflammatory process was evaluated before addition of the inflammation inducer and after 1, 2, 3, 4, 6 and 24 hours of its injection for measuring the volume and the thickness of affected limb. Analgesic activity of ointments with ginger extract was examined using the model of AITC-induced pain. Results: The most effective inhibition of the development of inflammation process was 0.025% ointment with ginger extract, and the highest anti-nociceptive effect was observed at the application of 0.05% ointment 10 minutes before pain inducer agent. Conclusion: Zingiber officinale dense extract was revealed to possess significant antinociceptive and anti-inflammatory actions after its transdermal delivery. Since the pharmacological effects of ginger extract have been investigated on AITC-induced model, we may suggest the vital role of phytoconstituents binding to TRPA1 and TRPV1 ion channels as possible mechanism of action.

Quantitative Characterization of the Hemorrhagic, Necrotic, Coagulation-Altering Properties and Edema-Forming Effects of Zebra Snake (Naja nigricincta nigricincta) Venom

This study was designed to investigate the cytotoxicity and haemotoxicity of the Western barred (zebra) spitting cobra (Naja nigricincta nigricincta) venom to help explain atypical and inconsistent reports on syndromes by Namibian physicians treating victims of human ophidian accidents. Freeze-dried venom milked from adult zebra snakes was dissolved in phosphate buffered saline (PBS) for use in this study. Haemorrhagic and necrotic activity of venom were studied in New Zealand albino rabbits. Oedema-forming activity was investigated in 10-day-old Cobb500 broiler chicks. Procoagulant and thrombolytic activity was investigated in adult Kalahari red goat blood in vitro. The rabbit skin minimum hemorrhagic dose (MHD) for N. n. nigricincta was 9.8μg. The minimum necrotizing dose (MND) for N. n. nigricincta venom was 12.2μg. The N. n. nigricincta venom showed linear dose-dependent procoagulant activity on goat blood (p<0.05). Likewise, N. n. nigricincta venom showed linear dose-dependent thrombolytic activity on goat blood (p<0.05, n = 6). Subplantar injection of N. n. nigricincta venom (25μg, 50μg, 75μg, and 100μg) into chick paw resulted in peak oedema of 35.5%, 38.5%, 42.9%, and 47.5%, respectively, two hours after injection. Paw oedema subsided within five hours to a mean volume ranging from 5% (25μg venom) to 17.6% (100μg venom). In conclusion, though N. n. nigricincta belongs to the genus Elapidae, the current study has shown its venom to possess potent hemorrhagic, necrotic (cytotoxic), and paradoxically, both procoagulant and thrombolytic activity. The authors propose further work to fractionate, isolate, and elucidate the structure of the various N. n. nigricincta venom toxins as a prelude to the development of an antivenom.

The Study of Activity of the Sum of Triterpene Acids From Fruits of Sea Buckthorn and Cranberry on the Model of Chronic Inflammation

Mandatory stage of the study of new compounds with strong pharmacological activity is the study of possible mechanisms of action. The purpose of this study was to investigate anti-inflammatory activity the amount of triterpene acids when they are used in the medical scheme in terms of adjuvant-induced arthritis. The studies were performed on rats male Wistar weighing 210-230 gr. Model of arthritis was reproduced by subplantar injection of 0.1 ml Freind&#180;scomplete adjuvant (Sigma, USA) into the right hind paw of rats. The researched objects (the triterpenoids buckthorn and cranberry) were administered orally once a day in the form of an aqueous suspension at a dose of 100 mg/kg of the medical scheme (for 12 days, starting 14 days after adjuvant injection). The diclofenac at a dose of 8 mg/kg was used as the comparison drug. The intensity of the inflammatory response was recorded by ecometrics data. The authors found that the introduction of diclofenac was accompanied by a decrease in the intensity of primary and secondary inflammatory response. Triterpenoids cranberry reduced the primary reaction and had a slight influence on the formation of secondary inflammation. The use of triterpenoids buckthorn in these experimental conditions had no reliably significant effect on the intensity of the inflammatory process.

Antiinflammatory Efficacy of Extracts of Latex ofCalotropis proceraAgainst Different Mediators of Inflammation

The latex of the plantCalotropis procerahas been reported to exhibit potent antiinflammatory activity against carrageenin and formalin that are known to release various mediators. In the present study, we have evaluated the efficacy of extracts prepared from the latex ofC proceraagainst inflammation induced by histamine, serotonin, compound 48/80, bradykinin (BK), and prostaglandin E(PGE) in the rat paw oedema model. The paw oedema was induced by the subplantar injection of various inflammagens and oedema volume was recorded using a plethysmometer. The aqueous and methanol extracts of the dried latex (DL) and standard antiinflammatory drugs were administered orally 1 hour before inducing inflammation. The inhibitory effect of the extracts was also evaluated against cellular influx induced by carrageenin. The antiinflammatory effect of aqueous and methanolic extracts of DL was more pronounced than phenylbutazone (PBZ) against carrageenin while it was comparable to chlorpheniramine and PBZ against histamine and PGE, respectively. Both extracts produced about 80%, 40%, and 30% inhibition of inflammation induced by BK, compound 48/80, and serotonin. The histological analysis revealed that the extracts were more potent than PBZ in inhibiting cellular infiltration and subcutaneous oedema induced by carrageenin. The extracts of DL exert their antiinflammatory effects mainly by inhibiting histamine and BK and partly by inhibiting PGE.

Platypus Venom and Envenomation.

Curiosity and controversy have surrounded the function of the crural system of the platypus since its discovery in the late 18th century. Early work on the venom confused rather than clarified the biological significance of the crural system. Many experiments gave conflicting results, especially concerning the coagulation effects of intravenous injections of venom extracts, although consistent observations were made of general vasodilation following intravenous injection into rabbits. More recent studies have shown that crural (venom) gland activity is seasonal and in synchrony with the breeding season. Secretion from the crural glands shows proteolytic activity and contains at least three major proteins, one of which has hyaluronidase activity. Subcutaneous injection of venom produced mild toxic effects whereas intravenous doses (75-90mg protein/kg) in mice were lethal. Whole venom induced local oedema after subplantar injection in rats and a 4.2kD peptide isolated from the venom caused relaxation of rat uterus in vitro. At least 16 incidents of envenomation by the platypus have been recorded in humans but no fatalities have been reported. In most human cases, envenomation resulted in immediate and severe local pain and oedema, sometimes associated with nausea, cold sweats, dull gastric pain and vomiting, hyperaesthesia and swelling of the axillary lymph nodes. Significant functional impairment of the upper limb for some weeks or months has been observed. Various treatments have been used to alleviate the symptoms of envenomation with differing successes. Envenomation has also been recorded in platypuses and dogs. The effects of these envenomations will be discussed.