PHARMACODYNAMIC EFFECTS OF CIMETIDINE AND METRONIDAZOLE ON THE BLOOD GLUCOSE LOWERING EFFECT OF GLYBURIDE IN RATS.

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1 Journal of Pharmaceutical and Allied Sciences 3 (1) (2006) Journal of Pharmaceutical and Allied Sciences PHARMACODYNAMIC EFFECTS OF CIMETIDINE AND METRONIDAZOLE ON THE BLOOD GLUCOSE LOWERING EFFECT OF GLYBURIDE IN RATS. OKONTA, J.M., *UDEOGARANYA, P.O. AND EZEAGU, CA. DEPARTMENT OF CLINICAL PHARMACY AND PHARMACY MANAGEMENT, UNIVERSITY OF NIGERIA, NSUKKA. ABSTRACT The effects of the two imidazole derivatives (cimetidine and metronidazole) on the control of blood glucose level by Glyburide were determined in normal and hyperglycemic rats. Both imidazole derivatives insignificantly reduced the blood glucose lowering ability of glyburide in both normal and hyperglycemic rats after a single dose administration. However, on the chronic administrations for 3 weeks, there were no observed significant variations in blood glucose levels of all the treated rats. Although the drugs did not significantly affect the control of blood glucose levels by glyburide, precautions must be taken when the need to co administer these drugs arises especially in a prolonged therapy situation. Keywords: Glucose lowering, normoglycemic, hyperglycemic, imidazole derivatives and `Glyburide INTRODUCTION Diabetes mellitus is a metabolic disease caused by either impairment of pancreatic insulin secretion or impaired tissue responsiveness to insulin or through both ways(1).the dysfunction of pancreatic ß- cells may be due to invasion by viruses, chemicals, toxins or autoimmune antibodies (2). This metabolic disease affects diverse organs of the body systems producing its long-term complications such as micro- and macroangiopathies, atherosclerosis, congestive *Correspondence author okojema@yahoo.com Journal of Pharmaceutical and Allied Sciences Vol 3 No 1 (2006) ISSN: Website: heart failure, hypertension, diarrhoea and spontaneous abortion (3) and neuropathy (4, 5). Most diabetic patients with prolonged exposure to this disease are prone to gastrointestinal disturbances such as bacteria overgrowth - induced diarrhoea and gastroparesis (6). These complications, most times engender multidrug therapy that when not properly administered and monitored may lead to adverse drug interactions. Gastrointestinal disorders in diabetic patients most times, warrant the co administration of the imidazole drugs (metronidazole and cimetidine) and sulphonylureas, the primary oral hypoglycemic drugs used in the management of diabetes. These imidazole drugs are known to inhibit certain enzymes responsible for the metabolism of some drugs thereby prolonging the 283

2 pharmacological effects of these drugs in the patient (7) and may affect the pharmacokinetics of such drugs (8). Glyburide is metabolized in the liver by cytochrome P 450 enzymes, which are often times inhibited by the imidazole drugs, cimetidine and metronidazole (9). Glyburide, a potent oral hypoglycemic agent, can precipitate severe hypoglycemia when its action is prolonged by enzyme inhibition. These agents that can inhibit the metabolism of glyburide to inactive metabolites could lead to its adverse effect hypoglycemia. Although most diabetic patients suffering from gastrointestinal tract disorders and infectious diarrhoea are often treated with metronidazole or cimetidine concurrently with oral hypoglycemic agents, no documented study on their pharmacodynamic interactions has been reported in recent literature. This study therefore has been designed to understudy the pharmacodynamic effects of these imidazole drugs on the control of blood glucose level by glyburide, using animal model. MATERIALS Drugs Glyburide, 5 mg (Daonil, NGC, Nigeria) tablets were weighed, crushed and dissolved in Tween-20 to give 5 mg/ml. Cimetidine 200 mg tablets (Cemtab Fidson, Nigeria) and Metronidazole (Amebanil Dana, Nigeria) 200 mg were similarly treated. All drugs were administered immediately after dissolution to avoid degradation. Alloxan monohydrate (Sigma, USA) was administered 120 mg/kg body weight intraperitoneally (ip). Animals White albino rats of both sexes ( g body weight) bred and housed in the animal house of the Department of Pharmacology and Toxicology, University of Nigeria, Nsukka were used. The animals were kept to acclimatize with laboratory condition for 7 days with free access to water and food before the experiments. Effects of cimetidine and metronidazole on the control of blood glucose level by glyburide in normal rats Twelve albino rats of g body weights were fasted for 12 h. Blood sample was withdrawn from the tail vein of the rats and blood glucose level determined using the One Touch Glucose Meter (Lifescan USA). The rats were divided into 3 groups of four rats per group. Group 1 received 5 mg/kg of Glyburide only; group 2 received 25 mg/kg Cimetidine and 5 mg/kg glyburide, while group 3 received 25 mg/kg metronidazole and 5 mg/kg glyburide. The cimetidine and metronidazole were orally administered, 30 minutes before the administration of glyburide. Blood samples were collected at 0, 1, 2, 4 and 8 hr from the rats and blood glucose levels determined using One-Touch Glucose meter kit. Effects of Cimetidine and Metronidazole on the control of blood glucose level by glyburide in alloxanized rats Twelve albino rats of g body weight of both sexes were used. The rats were made hyperglycemic by injecting alloxan monohydrate (120 mg/kg) intraperitoneally. The rats were fed for 8 days but on day 9, they were fasted for

3 hr and their blood glucose levels determined. The animals with blood glucose levels above 160 mg/dl were grouped into three of four animals per group and dosed as in the normoglycemic animal experiment on day 9. The blood samples were collected at intervals of 0, 1, 2, 4 and 8 h and blood glucose levels determined as before. The collections were continued at weekly interval for 3 weeks according to the method described by Okonta (1). Statistical analysis DISCUS Mean blood glucose levels were expressed as mean ± SEM, and the significance of difference between the blood glucose levels at zero time and other time intervals in each treatment group determined using student s t-test (p=0.05) and ANOVA to compare the various groups. RESULTS From the effect of the different imidazole derivatives (cimetidine and metronidazole) on the control of mean fasting blood glucose levels of both normoglycemic and hyperglycemic rats by glyburide, the glyburide cimetidine combination caused 15.6% and 61.6% respectively, and glyburide metronidazole combination caused 16.1% and 65.7% respectively on normoglycemic and hyperglycemic rats. From the results, the effects of the combinations on the normoglycemic rats were insignificant (Table 1). In hyperglycemic rats, glyburidecimetidine and glyburide metronidazole combinations lowered the mean fasting blood glucose levels from 196.6±18.3 mg/dl (0h) to 75.5±20.1 mg/dl (4 h) and 220±36.0 mg/dl (0 h) to 41.02±1.54 mg/dl (8 h) respectively. These combinations caused lower percentage maximal reduction of mean fasting blood glucose level than Glyburide alone though the percentage maximal reductions were not statistically significant (Table 2). From Fig.1, the chronic administration of the glyburide -cimetidine and glyburidemetronidazole for 3 weeks showed no significant difference in the mean fasting blood glucose level at the 21 st day from that of glyburide alone. DISCUSSION Glyburide is a potent oral hypoglycemic agent that lowers blood glucose level through pancreatic and extra pancreatic ways especially in hyperglycemic patients with partially viable beta cells (10, 11). In the normoglycemic rats, the beta cells were viable but the percentage maximal reduction of blood glucose level caused by glyburide was not significantly altered by co administration of glyburide and the imidazole derivatives cimetidine and metronidazole. In the alloxanized rats however, glyburide alone caused significant percentage maximal reduction in blood glucose level while the co administration of glyburide with cimetidine and metronidazole did not enhance the percentage maximal reduction of blood glucose level by glyburide. These findings corroborate the findings of Kubacka and associates( 12) who observed that the co administration of H 2 -receptor antagonists, cimetidine and ranitidine, and glibenclamide led to unexpected higher plasma glucose levels than the sulphonylurea alone; and plasma insulin concentrations were significantly elevated when H 2 - receptor antagonists and glibenclamide were administered concurrently. The effects of the H 2-285

4 receptor antagonists and metronidazole on the blood glucose lowering activity of glyburide may be particularly resident on derivatives are known to be 2C9 cytochrome isoform inhibitors hence could inhibit the glyburide metabolism TABLE 1: Effects of Cimetidine and Metronidazole on glyburide treated normoglycemic rats Mean fasting blood sugar level (mg/dl) (MFBSL) Group 0 h 1 h 2 h 4 h 8 h % Max. Red ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± Grp. 1. Glyburide (5 mg/kg); Grp. 2. Glyburide (5 mg/kg) + cimetidine (25 mg/kg); Grp. 3. Glyburide (5 mg/kg) + metronidazole (25 mg/kg). Values are expressed as mean ± SEM; n=4 the imidazole moiety since all the imidazole derivatives co-administered with glyburide caused increase in plasma glucose levels although not significantly and cause its prolonged hypoglycemic action. In our present study however, the reverse seems to be the case as the coadministration of these imidazole TABLE 2: Effect of Cimetidine and Metronidazole on glyburide treated hyperglycemic rats Mean fasting blood sugar level (mg/dl) (MFBSL) Group 0 h 1 h 2 h 4 h 8 h % Max. Red ± ± ± ± ± ± ± ±58.7* 75.5±20.1* 84.5±24.7* ± ±20.7* 90.3±33.3* 85.5±32.2* 75.4±5.8* 65.7 Grp. 1 Glyburide (5 mg/kg); Grp. 2 Glyburide (5 mg/kg) + cimetidine (25 mg/kg); Grp. 3 Glyburide (5 mg/kg) + metronidazole (25 mg/kg). Values are expressed as mean ± SEM; n=4 *P<0.05 vs. 0 h different from that of glyburide alone. Glyburide metabolism is dominantly influenced by cytochrome CYP2C9 (13). Cimetidine and other imidazole derivatives caused less reduction of the maximal percentage reduction of blood glucose level compared to that caused by glyburide. 286

5 Although the hypoglycemic effect Blood glucose conc.(mg/dl) Glyburide,5mg/kg glyburide,5mg/kg+cimetidine,25mg/kg Glyburide5mg/kg+metronidazole 25mg/kg Time(days) Fig.1: Effects of chronic administration of cimetidine and metronidazole on blood glucose lowering effect of glyburide in hyperglycemic rats of glyburide may not be significantly affected by cimetidine and metronidazole co administration as indicated in this study, precaution must be taken on individual basis when the need to co administer these combinations arises. REFERENCES 1. Okonta, J.M. (2005). Pharmacodynamic drug interaction study: Effect of Sertraline on the hypoglycemic action of Glibenclamide in rats. Bio-Research 3 (1): Mycek, J.M., Richard, A.H. and Pamella, C. C. (2000) Insulin and Oral hypoglycemic drugs, In: Lippincott s Illustrated Reviews Pharmacology, 2 nd ed. pp Yusuf, O., Meli Altan, V and Nurray, Y. (1996). Effects of Experimental Diabetes and Insulin on Smooth Muscle Functions. Pharm. Rev. 48(1), Frier, B.M., Truswell, A.S., and Shepherd, J.; Adeloyi A (1999). Diabetes Mellitus and Nutritional and Metabolic disorders In: Davidson s Principles of Medicine. Harcourt Pub. Edinburgh pp

6 5. Judd, Okonta, R.L. and J. M. et Raman, al.,/journal P. of (1999). Pharmaceutical and Allied Sciences 3 (1) (2006) Pharmacological Management of Type 2 Diabetes Mellitus: Current and Future 10. Wilkinson, M.G. (2000). Detecting therapy. Pharm. Times Oct Depression and Anxiety. African 6. Matthew, J O. (1998). Diabetes Mellitus and related disorders In: The Health 8: Zongo, M.G., Tolfo, L. and Draghi, E. Washington Manual of Medical (1994). Hypoglycemia caused by Therapeutics.Carey, CF et al Ed; 29th ed. Lippincott Williams & Wilkinson p.414. Maprotiline in a patient taking Oral Antidiabetics. Ann. Pharmacotherap. 28: Ojewole, J.O.A. (1999). Drug 12. Kubacka, R.T., Antal, F.I. and Juhl, R.P. interactions In: Therapeutic Basis of (1987). The paradoxical effects of Clinical Pharmacy in the Tropics, cimetidine and ranitidine on C.N.Aguwa Ed; 2nd ed. Optimal glibenclamide pharmacokinetics and Pub.Enugu, pp pharmacodynamics.br.j.clin.pharmacol 8. Garba, M., Odunola, M.T. and Bukhari,.23 (6): K.A. (1998). Single dose salivary 13. Yin, O. Q, Tomlinson, B. and Chow MS pharmacokinetic study on the effect of metronidazole on Paracetamol in healthy subjects. J.Pharm.Res.and Development 3(2): (2005). CYP2C9, but not CYP2C19, polymorphisms affect the pharmacokinetics and pharmacodynamics of glyburide in 9. Goodman, L and Gilman, A. (2001). The Pharmacological Basis of Therapeutics; 10 th edn. Macmillan, New York, p Chinese subjects.clin.pharmacol.therap.78 (4):

7 90 80 Blood glucose conc.(mg/dl) Time(days) Glyburide,5mg/kg glyburide,5mg/kg+cimetidine,25mg/kg Glyburide5mg/kg+metronidazole 25mg/kg Fig.1: Effects of chronic administration of cimetidine and metronidazole on blood glucose lowering effect of glyburide in hyperglycemic rats 1. INTRODUCTIONair-dried and 289

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