Author name:
رائد عبد شاكر محمود
Supervisor name:
عبد الكريم محمد علي جبر السامرائي
Abstract:
الهدف من هده الدراسة هو تقدير تاثير بعض مشتقات الثياديازول والثيازين الجديدة التحضير على فعالية كل من الانزيمين الكرياتين كاينيز و3 - هيدروكسي - 3 - مثيل كلوتاريل كو - انزيم اي ريدكتيز بالاضافة الى قياس صورة الدهون في مرضى ارتفاع الدهون والفئران المختبرية | The aim of this study is to evaluate the effect of some novel prepared derivatives of thiadiazole and thiazine on the activities of creatine kinase (CK) and 3 - hydroxy - 3 - methylglutaryl CoA reductase(HMGR) in addition to lipid profile in sera of hyperlipidemic patients and in mice induced hyperlipidemia by feeding cholesterol rich diet.The study includes two parts; in vitro study : Sixty individuals with age ranged between (40 - 60) years were enrolled in this study. They were divided into two groups; first group (G1) consists of 30 healthy individuals as a control group with body mass index (BMI) (25.67). The second group (G2)consists of 30 patients with hyperlipidemia and BMI (26.48) which diagnosed by physician. The patients attended the Ibn - Al Naphes hospital during November 2013 to February 2014. Patients with high blood viscosity, diabetes mellitus, renal failure as well as those who are under treatment with statins were excluded. The serum which obtained used in the determination of lipid profile[total cholesterol(Tch),triglyceride(TG), high density lipoprotein(HDL - c), very low density lipoprotein(VLDL - c)], fasting blood glucose(FBG), aspartate transaminase (AST), alanine transaminase(ALT) and C - reactive protein(CRP).Four organic compounds 3 - (4 - (dimethylamino) phenyl) - 2,3 - dihydro - 2 - (3 - nitrophenyl benzo[1,3 - e]thiazin - 4 - one[I], 5 - (4 - imethylamino)benzylideneamino) - 1,3,4 - thiadiazole - 2 - thiol[II], 2 - (4 - dimethylamino)phenyl) - 2,3 - dihydro - 3 - (5 - mercapto - 1,3,4 - thiadiazol - 2 - yl)benzo[1,3 - e]thiazin - 4 - one[III], and N - (4 - (dimethyl amino)benzylidene) - 5 - (isopropylthio) - 1,3,4 - thiadiazole - 2 - amine[IV] were used in this study to test their antihyperlipidaemic ability and their effect on CK and HMGR activities. The results revealed that compounds(III and IV)showed an activation effect in all concentrations on CK and HMGR activities, while compounds(I and II) showed an inhibitory effect in some concentrations for CK and in all concentrations for HMGR. Therefore, compounds (III and IV) were excluded from this study. The results showed that (10 - 4M) for compound I and (10 - 5M) for compounds II give the best inhibition percentage among the other concentrations on CK and HMGR activities which the kinetic study throughout with these concentrations for these compounds. Simvastatin, which considered as standard drug for lipid lowering, was used for comparsion with the potency of compounds I and II on HMGR activity in treatment of hyperlipidaemia. The results showed an inhibitory effect of simvastatin on HMGR activity with percentage inhibition 88%. The effect of compounds (I and II) on ALT and AST were examined in (10 - 4 M) for compound I and (10 - 5M) for compound II in vitro study. The results showed the inhibitory effect of compounds I in concentration (10 - 4 M) and compound II in concentration (10 - 5M) on ALT and AST activities.The Vmax, Km and type of inhibition for compounds I and II on CK and HMGR activities were studied by using Lineweaver - Burk plot. The results showed that also compound I at 10 - 4M was considered to be a noncompetitive inhibitor for CK activity with Vmax values (1000 and 344.82)U/L for uninhibited and inhibited enzyme respectively and Km value (10) mmol/L. The results also showed that compound II at concentration 10 - 5M was considered to be a competitive inhibitor for CK activity with Vmax value (588.23)U/L and Km values (5.51 and 4)mmol/L for the uninhibited and inhibited enzyme respectively.In vitro, the effect of compound (I) with concentration (10 - 4M) and compound (II) with concentration (10 - 5M) were examined in vivo study. The study was carried out with sixty male Wister mice aged seven to eight weeks and theirweight were (180 - 200 g) ,obtained from animal house , in College of Medicine, Baghdad University. The mice were grouped as follow : group one (12 mice) as control group, group(2) : consists of 48 mice in which the mice were daily administered cholesterol (25mg/k/day), in coconut oil 6% and creamy cheese for 28 days. Lipid profile were measured for twelve mice chosen randomly from G2 to diagnosis hyperlipidemia. Then group2 is subdivided into three groups as follows : group (2.A) : (12 mice) as positive control group in which the mice were daily administered simvastatin (40mg /day) as standard drug for hyperlipidemia, group 2B : (12 mice) in which the mice were daily treated with (10 - 4)M of compound (I)via drinking water for 20 days and Group(2.C) : (12 mice) in which the mice were daily treated with (10 - 5)M of compound II for 20 days. The results showed significant elevation in levels of Tch, TG, LDL - c and VLDL - c, while there is significant reduction in HDL - c levels in G2 comparing to control group(G1), after administration of fat rich diet. Simvastatin, compound I with concentration (10 - 4M) and compound II with concentration (10 - 5M) were administrated to G2A, G2B and G2C respectively. Also, the results showed that the activities of CK reduced for group G2B and G2C while it is increased for G2A. The results also showed that the activities of HMGR were reduced in the three treated groups. The results revealed that compounds I and II exhibit more potent antihyperlipidaemic effect than simvastatin. Also, compound I showed more potent antihyperlipidaemic effect than compound II.The results revealed that compounds I and II showed a noncompetitive inhibitor effect on CK with Vmax values(1000and 166.6) U/L for uninhibited and inhibited enzyme respectively and Km value (0.6) mmol/L for compound I, and with Vmax vales (1000 and 250)U/L for uninhibited and inhibited enzyme respectively and Km value (0.84) mmol/L for compound II.In conclusion, the novel synthetic compounds (I and II) seem to be of interest in the development of new antihyperlipidaemic agents that exhibit inhibition effect on CK while statins cause increase in this enzyme. Also these compounds exhibit inhibition effect on HMGR activity more than simvastatin, which is a key enzyme in cholesterol synthesis.
