Available online Research Article

Transcription

1 Avilble online Journl of Chemicl nd Phrmceuticl Reserch, 213, 5(5): Reserch Article ISSN : CODEN(USA) : JCPRC5 Evlution of the qulity nd in vivo therpeutic efficcy of generic Glimepiride mnufctured in Algeri, compred to the brnd nme drug Amrel on lloxn dibetic rts Meryem Ouzouz, Meriem Derrdj nd Cherif Henchiri * Lbortory of Applied Microbiology nd Biochemistry, Deprtment of Biochemistry, Bdji- Mokhtr University, Annb, Algeri ABSTRACT The present study ims to evlute physicochemicl chrcteristics nd the effectiveness of phrmcologicl effects of Glimepiride generic product, compred to its origintor Amrel sulfonylure used to tret type 2 dibetes mellitus. Two doses 2 nd 4 mg per dy were tested on mle lbino rts dibetics with lloxn. Body weights, glycemi, HbA1c, lipid profile, totl protein, the ctivity of certin enzymes nd some renl prmeters were determined. The results showed tht physicochemicl chrcteristics re complint with stndrds of the Americn Phrmcopei compred with the used reference substnce. However, the biologicl study reveled dosedependent effect of Amrel on body weights, blood glucose nd lipid profile of rts during nd fter the tretment, which ws not obtined with the generic. Totl protein nd enzymes ctivities of serum trnsminses, mylse nd lipse were lmost identicl in groups treted with 4 mg of origintor nd generic. The 2 mg dose hs not exerted the expected therpeutic effect. These results show tht the studied generic exhibited less efficcy thn the origintor despite complince of physicochemicl chrcteristics. A supply of the generic with the queous extrct of locl medicinl plnt (Zygophyllum cornutum) ws performed where the results of mesuring blood glucose before nd fter scrifice of rts were similr to those of helthy controls. No significnt difference ws recorded for renl prmeters in ll treted groups. Keywords: Generic; Brnded drug; Glimepiride; Alloxn dibetes. INTRODUCTION Type 2 dibetes, chronic widespred disese, is one of the mjor chllenges of the 21 st century in terms of helth nd development, WHO estimtes tht dibetes, hs reched criticl levels, ech yer, not less thn four million people die of dibetes nd bout ten million re suffering from other disbilities nd life-thretening complictions [1]. The gol of tretment is to trget glycemic control by mintining hemoglobin glycted (HbA1c) s close to norml s possible while voiding hypoglycemi. Glimepiride, n orl sulfonylure, indicted for the tretment of type 2 dibetes, it cts by stimulting the relese of insulin in β cells of the pncretic islets. This ction is not glucose-dependent, it cn led to hypoglycemi. Stimultion occurs by closing the potssium chnnels of the β cell membrne, llowing the opening of clcium chnnels nd the entry of clcium into the cell, thereby stimulting insulin secretion. Glimepiride lso increses the number of ctive glucose trnsport molecules, the sensitivity of extr-pncretic tissue is then rised (dipocytes, myocytes) nd the speed of ction of glucose slowed. Finlly, it decreses the uptke of insulin by the liver by inhibiting gluconeogenesis [2]. Although the generic of drug (origintor or originl molecule), re considered by the legisltion s mediction tht hs the sme qulittive nd quntittive composition in ctive ingredient, the sme phrmceuticl form, nd 31

2 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): which the bioequivlence with the brnded nme product is demonstrted by biovilbility studies [3]. The effectiveness of these drugs is still questioned by physicins, s well s by ptients; however, their prescription is incresing worldwide [4]. In ddition, due to the high cost of drugs, Algeri, like ll developing countries, hs opted for generic drugs, by importing or producing these drugs, insted of originl medicines protected by ptents in developed countries [5]. This study ims to verify the ntidibetic effect of glimepiride generic product, mnufctured by ntionl phrmceuticl compny nd mrketed in Algeri, nd to evlute its effectiveness in two different doses, recommended in glimepiride dosge, 2 nd 4 mg per dy, compred to the brnd Amrel. This, leds to confirm the supposed effect of generic nd to eliminte doubt on their supposed lck of efficcy compred to the origintor [6], it will permit doctors to prescribe nd ptients to use generic drugs insted originl drugs with confidence nd void unnecessry costs, which will llow ccess to tretment for the poorest ptients. EXPERIMENTAL SECTION Phrmceuticls In this study, we used Amrel the brnded product of the molecule glimepiride nd one of its generic mnufctured in ntionl Algerin compny, the two products were commercilly purchsed. Biologicl mteril Wistr lbino mle rts (18-2 g) were purchsed from Psteur institute of Algiers. They were seprted nd divided into groups of 6 rts per cge with ccess d libitum to wter nd stndrd diet for rts (UAB: Ntionl unit of niml feed, Beji). They were lso kept nd mintined under lbortory conditions of temperture nd light (24 ± 1 C nd 12 h light/drk cycle) respectively nd strict hygiene for few dys, for cclimtiztion. The experiments were performed ccording to the guidelines of the Guide for Cre nd Use of Lbortory Animls [7]. Lbortory qulity control testing The determintion of the physicochemicl properties of the two drugs, Amrel nd the generic, were performed ccording to the Americn Phrmcopoei (Active ingredient ssy, dissolution test nd disintegrtion time) [8, 9]. Assy: The nlysis ws performed on Wters e2695 HPLC system equipped by binry pump, n UV/Vis Wters 2489 Detector nd column (4mm x 12.5 cm, pcking L1). The retention time of the mjor pek in the chromtogrm of the Assy preprtion corresponds to tht in the chromtogrm of the Stndrd preprtion, s obtined in the Assy. Mobile phse consisted of phosphte buffer t ph of 2.1 to 2.7 nd cetonitrile t equl volumes. The diluent ws mixture of cetonitrile nd wter (9:1). A stndrd solution of glimepiride reference stndrd (RS) t.1 mg/ml concentrtion ws used s reference. Assy preprtion: five Tblets were trnsferred into suitble volumetric flsk to prepre solution of pproximtely.1 mg/ml of glimepiride. 1 % of the volume of the flsk ws filled with wter. About 7 % of the volume of the flsk of cetonitrile ws dded. The smples were sonicted in wter bth for 5 to 1 minutes with occsionl shking. Acetonitrile ws dded to volume nd the smple ws mixed nd filtered. 1 µl of stndrd solution nd smples were injected seprtely into the chromtogrph t 1 ml/min flow rte nd the eluent ws mesured t 228 nm. The percentge of the lbeled mount of glimepiride (C24H34N4O5S) in the portion of Tblets tken ws clculted by the formul: 1(C S / C U )(R U / R S ). In which C S : the concentrtion, in mg/ml, of glimepiride in the Stndrd solution; C U : the concentrtion of glimepiride in the smple; nd R U nd R S re the pek responses for glimepiride of the smple nd the Stndrd solution, respectively. In vitro dissolution test: The dissolution test ws crried out ccording to USP Dissolution Test 1. The tblets were disposed seprtely in ph 7.8 phosphte buffer; 9 ml for 15 min t 37±.5 C t 75 rpm, using SOTAX AT 7 Smrt Dissolutest. The stndrd solution ws prepred s recommended in USP monogrphs to get solution of.75 µg/ml of glimepiride RS. After 15 min 1 ml of smple ws withdrwn from the dissolution medium nd ws centrifuged for 5 min t 25 rpm. To 3ml of the superntnt, 7ml of mixture of methnol nd wter (1:1) ws dded to get the smple solution. The mobile phse ws prepred s directed in the ssy. 5µl of stndrd solution nd the smple solution were injected seprtely in the sme conditions of ssy. Disintegrtion time test: Disintegrtion were crried out using SOTAX DT 2, disintegrtion tester using distilled wter s disintegrtion medium t 37 ± 2 C, six tblets were exmined. 311

3 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): Biologicl Study We evluted the ntidibetic effects of the two medictions, brnd nd generic glimepiride product, on Wistr lbino rts dibetics with lloxn. For the two drugs, the sme btches were used in the qulity control nlysis, s well s in the biologicl study. Dibetes induction The rts were treted by single intrperitonel injection of 15 mg / kg body weight of lloxn monohydrte (Sigm-Aldrich Co., USA) [1]. To ensure the instlltion of dibetes, we mesured blood glucose of rts fter 72 hours of lloxn injection, only rts with fsting glycemi greter thn 3 mg/dl were selected for the rest of the study [11]. Experimentl protocol nd niml tretment Rts were rndomly divided into 7 groups of 6 rts per cge (Tble 1): two control groups (helthy nd dibetics), 4 subgroups treted with 2 nd 4 mg of the two drugs nd group treted with 2 mg of the generic supplemented by n queous extrct of n ntidibetic plnt ccording to the locl trditionl medicine. Tretments were dministered dily to rts by single orl dose [12], for 32 dys. The rts were weighed nd their fsting blood glucose levels (FBG) were mesured using n Accu-check glucometer every three dys throughout the tretment period. Tble 1: Experimentl design HC DC 2mg-G 2mg-A 4mg-G 4mg-A G+Plt Chrcteristics Helthy controls received wter under the sme conditions s other groups. Dibetic controls received wter under the sme conditions s other groups. Dibetics treted with 2 mg of the generic drug in wter. Dibetics treted with 4 mg of the generic drug in wter. Dibetics treted with 2 mg of Amrel in wter. Dibetics treted with 4 mg of Amrel in wter. Dibetics treted with morning dose of 2 mg of the generic in wter nd 8 mg/kg b.w. of the queous extrct of the plnt. Biochemicl ssy At the end of experiment, overnight fsted rts were scrificed; blood smples were immeditely collected in lbeled tubes. After centrifugtion, the serum obtined will be used for ssys of serum glucose, totl cholesterol (TC), totl lipids (TL), triglycerides (TG), low density lipoprotein cholesterol (LDLc), high density lipoprotein cholesterol (HDLc) levels, the ctivity of serum minotrnsferses GOT nd GPT, mylse nd lipse, nd even the renl prmeters; Glycted Hb ws determined in whole blood. The ssys were performed ccording to the supplier s specifictions from the stndrd Kits. LDLc hve been clculted using the following formul: LDLc = TC (HDLc) (TG/5) [13]. Sttisticl nlysis of results The results were presented s men plus or minus stndrd devition (Men ± SEM). The sttisticl nlysis ws performed by one wy nlysis of vrince (ANOVA) followed by Dunnett s or Tukey multiple comprisons tests using MINITAB 16 pckge. Level of significnce ws: Significnt when P.5; highly significnt when P.1 nd very highly significnt when P.1. RESULTS AND DISCUSSION Qulity control of products The in vitro nlysis of the generic drug nd the brnded product Amrel re presented in Tble 2. Despite the complince with USP stndrds [8, 9]; results obtined from glimepiride ssy, disintegrtion time nd dissolution test for the generic drug re inferior to those of Amrel. Tble 2: Results of the qulity control of the two glimepiride drugs Anlysis USP limits Amrel Generic Glimepiride ssy (%) 9, - 11, 19,8 17, Disintegrtion time (min) Dissolution fter 15 min (%) 8 98,4 82,7 USP: United Sttes phrmcopei. 312

4 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): Chnges in body weights nd fsting glycemi before scrifice The following figures show the results of mesurements of body weights nd fsting blood glucose levels of nimls monitored during the whole period of tretment. Body weights Men body weights 28 (g) Helthy Control 26 Dibetic Control 24 2mg-Generic AA b b 4mg-Generic 2mg-Amrel 4mg-Amrel 2mg-Generic+plnt 14 D D4 D8 D12 D16 D2 D24 D28 D32 Dys Fig.1: Chnges in body weights of rts during the whole period of tretment Vs HC: P,1, P,1 ; Vs DC : AA P,1, P,1 ; 2mg-G Vs 4mg-G : P,1, b P,1. The results obtined (Fig.1) shows n increse in body weights of the helthy controls (HC) which is relted to the norml growth of nimls s well s fll in weights of the dibetic controls (DC) which would be linked to metbolic disorders due to dibetes. The lck of insulin ctivtes lipolysis in dipose tissue nd would cuse weight loss [14]; moreover, these nimls received no tretment. An increse in weights of the dibetic groups treted with the two doses of drugs hs been observed, but it remins lower thn tht observed in helthy control rts, HC. This would suggest tht both generic drug nd Amrel hve exerted slight effect on insulin ctivity of the pncres, resulting slight lipogenesis, tht llowed the recovery of body weights of rts, this weight gin is lso due to norml growth of rts. Studies of the effect of glimepiride on body weight in dibetic mn show tht this mediction is not ssocited with weight gin [2, 15]. While the group treted with the mixture, generic nd plnt extrct (Zygophyllum cornutum), showed better result similr to tht of helthy controls. Fsting blood glucose Mesurements of fsting blood glucose performed in the tretment period re shown in Figure 2; the helthy control group shows no chnge in blood glucose concentrtions during 32 dys of experiment. However, in the dibetic control group, very highly significnt increse in blood glucose levels ws observed (29,74%) during this period, which would be linked to the cytotoxic effect of lloxn on the pncretic bet cells [16, 17], the lck of tretment of nimls [18]. Dibetic groups, treted with 4 mg per dy of generic nd origintor drugs show lower blood glucose levels of 57,52 % nd 6,53 % respectively, this reduction is higher thn tht obtined by the dose 2 mg per dy (54,2 % for the generic nd 54,66 % for Amrel). This reduction of the ntidibetic effect of the generic could be relted to its ctive ingredient content which is lower to tht of the origintor (Tble 2) or its excipient, different from tht of the originl, which could influence the relese rte of the principle ctive [19]. The combintion of the generic, t rte of 2 mg per dy, with the queous extrct of Zygophyllum cornutum Coss, t dose of 8 mg/kg, resulted in significnt decrese in blood glucose levels compred to the other used tretments nd very highly significnt compred to dibetic controls, DC (72,67%). 313

5 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): Glycemi (g/l) Helthy Control Dibetic Control 2mg-Generic 4mg-Generic 2mg-Amrel mg-Amrel 2mg-Generic+plnt. D D4 D8 D12 D16 D2 D24 D28 D32 Dys Fig.2: Chnges in fsting glycemi of rts during the tretment period Vs HC: P,1; Vs DC: P,1. Biochemicl prmeters fter scrifice Results of generic glimepiride effects, compred to Amrel, on some biochemicl prmeters, fter scrifice, (blood glucose, lipid profiles, enzymes ctivities nd renl prmeters) re presented in Figures 3, 4, 5, 6 nd Tbles 3, 4. Glycemi nd HbA1 C Glycemi (g/l) (A) CCC BB BB HbA1c (g/l) (B) 1 2 Fig.3: Evlution of glycemi (A) nd HbA1c levels (B) of rts fter scrifice Vs HC: P,1; Vs DC: P,1; 2 mg-a Vs 4 mg-a: BBB P,1; 4 mg-g Vs 4 mg-a: CCC P,1. The results obtined fter 32 dys of tretment (Fig.3 A) show high level of blood glucose in dibetic controls compred to helthy ones, which could be the result of dibetes [18]. In contrst, tretment with glimepiride exhibited decrese in blood glucose levels in ll treted groups [2]; this reduction remins reltively elevted compred to the helthy control group. We noted dose-dependent effect of Amrel; indeed very highly significnt difference ws observed between fsting glycemi in rts treted with 2 mg (1,76 g/l) compred to those treted with 4 mg (1,56 g/l). Wheres, for the two doses of generic, we noticed no significnt difference between the glucose levels (2,1 g/l for the 2 mg dose nd 2,8 g/l for the 4 mg dose). Only the mixture drug-plnt extrct reveled n hypoglycemic effect higher thn tht obtined in treted groups by both drugs; glucose levels observed in this group re similr to those of HC. This result ws lso obtined with glycemi before scrifice. The follow-up of HbA1 C in dibetics reflects the glycemi stbility over given period (three months in humn), llows to evlute the effectiveness of tretments nd is even used for predicting of dyslipidemi risks in the humn dibetes [21]. HbA1 C levels observed in treted groups re in the rnge of 5,62 to 6,28 % nd re in ccordnce with decreses in fsting glucose levels before nd fter scrifice of nimls (Fig.3 B) showing lso efficcy of these tretments. However, HbA1 C levels re very elevted in untreted dibetics DC hving very highly significnt differences compred to helthy controls nd treted groups; this increse is ssocited to high concentrtions of blood glucose in this group [22]. Animls treted with ll doses of the generic or the brnded drug reveled no significnt differences between their glycted Hb levels, but these vlues re slightly higher thn the HC ones. 314

6 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): Supplementtion of the generic by the plnt extrct seems to hve the most potent effect on blood glucose nd HbA1 C levels. This result is similr to those found by other uthors [23, 24], which hve shown tht the combintion of low dose of certin nti-dibetic gents combined with dose of herbl extrcts could ssure good glycemic control. Lipid profile Tble 3: Results of lipidic prmeters ssessment Prmeters HC TG (g/l) 1,49 ±,14 LT (g/l) 1,91 ±,25 CT (g/l) 1,22 ±,9 LDLc (g/l),81 ±,16 HDLc (g/l) 1,36 ±,28 DC 2mg-G 2mg-A 4mg-G 4mg-A G+Plt cc b A A A b A BB A CC AA 3,97 ±,3 2,32 ±,27 1,86 ±,23 1,98 ±,21 1,76 ±,15 1,52 ±,8 8,43 ±,58 4,11 ±,47 4,16 ±,63 4,35 ±,24 4,52 ±,79 4,47 ±,9 2,36 ±,37 1,68 ±,46 1,42 ±,41 1,46 ±,21 1,42 ±,35 1,4 ±,2 1,41 ±,62 1,26 ±,47,92 ±,54,82 ±,31,73 ±,33,87 ±,25,9 ±,18 1,27 ±,32 1,33 ±,35 1,61 ±,14 1,95 ±,14 1,83 ±,14 Vs HC: P,5, P,1, P,1; Vs DC: A P,5, AA P,1, P,1; 2mg-G Vs 4mg-G : b P,5; 2mg-P Vs 4mg-P: BB P,1; 2mg-G Vs 2mg-P: cc P,1; 4mg-P Vs 4mg-P: CC P,1. According to the results, there ws very highly significnt increse in the concentrtion of TG in the DC group compred to HC (Tble 3), which is linked to the insulin-deficiency responsible for lipolysis in dipose tissue [18]. In ll treted groups, the rte of TG is reduced, indicting hypolipemic effect of Glimepiride [25]. We lso observed concentrtion of TG in the group treted with the mixture of the generic- plnt extrct similr to tht of helthy control one, this result hs been obtined previously with glucose nd HbA1 C. Very high levels of totl cholestero (TC), LDLc nd totl lipids (TL) in dibetic control rts (DC) were obtined, compred to helthy controls. In contrst, ll treted groups hve lower rtes thn DC group, with very highly significnt differences. The mesurement of HDLc, reveled lower serum level in the DC group compred to HC, with highly significnt difference. This concentrtion in the group treted with 2 mg of Amrel (2mg-A) is similr to tht of helthy control one. Compred to helthy nd dibetic controls, 4mg-A nd G+plt groups exhibited highh levels of HDLc, with very highly significnt differences. These results re in greement with those reported by Arki et l. [26]; studying the effect of Glimepiride on incresing rtes of HDLc. Totl proteins Totl proteins (g/dl) BBB Fig. 4: Evlution of totl proteins in rts t the end of the experiment Vs HC: P,1; Vs DC: P,1; 2mg-G Vs 4mg-G: BBB P,1; 2mg-A Vs 4mg-A: P,1. The results presented in Figure 4 indicte rte of totl serum proteins in untreted dibetic rts lower thn tht of helthy controls, with very highly significnt difference. This could be due to losss in the urine (proteinuri) 315

7 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): relted to long lsting hyperglycemi [27], to the toxic ction of lloxn on the kidneys [28] or the result of incresed protein ctbolism ssocited with insulin deficiency [29]. Totl proteins of treted groups with 4 mg of both drugs re similr to ech other but remin lower thn those of HC, wheres proteins of the group treted by mixing drug-extrct re similr to those of the HC. Enzymtic ctivities Serum minotrnsferses GOT (U/L) (A) AA b c C GPT (U/L) 7 6 (B) c CC Fig. 5: Evlution of serum trnsminses GOT (A) nd GPT (B) in rts fter the tretment period Vs HC : P,1; Vs DC: AA P,1, P,1; 2mg-G Vs 4mg-G : P,1, b P,1; 2mg-G Vs 2mg-A : c P,5; 4mg-G Vs 4mg-A: C P,5, CC P,1. A very highly significnt increse in the ctivity of serum minotrnsferses in dibetic control rts ws observed compred to helthy controls. The high enzymtic ctivity of GOT nd GPT cn be explined by the heptotoxic effect of lloxn [28, 3]. In contrst, tretment of dibetic rts by Amrel nd its generic reveled decresed ctivities of these two enzymes, with very highly significnt differences compred to DC (Fig.5). These ctivities were higher in the group treted by 2 mg of the generic compred to the group treted with 4 mg, with highly significnt differencee (P.1). While, both doses of Amrel showed no significnt differences. The tretment resulted in reduction in the ctivity of these two enzymes nd therefore, exerted protective effect ginst liver dmge cused by lloxn; the best effect ws obtined with the 4 mg dose of Amrel nd the combintion of the generic with the plnt extrct. Serum mylse nd lipse ctivity Amylse (U/L) (A) AA AA BB Lipse (U/L) (B) A AA A Fig.6: Evlution of serum mylse (A) nd lipse (B) ctivities fter scrifice of rts fter tretment Vs HC: P,5, P,1, P,1; Vs DC: A P,5, AA P,1, P,1; 2mg-G Vs 4mg-G: b P,1; 2mg-A Vs 4mg-A: BB P,1. The determintion of mylse nd serum lipse llows the dignosis of pncretic dmge [31]. In dibetic controls, we noticed very highly significnt increse of serum mylse nd lipse ctivities, compred to the helthy control group, suggesting n ltertion of the pncres cused by lloxn [32]. The 4 mg dose of both generic nd even the combintion of the generic with the plnt extrct cused very highly significnt decreses in the ctivity of mylse (Fig.6 A), compred to dibetic controls (p.1). Reduced ctivities of serum mylse were lso observed in dibetic groups treted with 2 mg of the two medicments compred to DC group but remined elevted compred 316

8 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): to helthy controls (p.1 Vs HC). The lipse ctivity, in ll treted groups ws lower thn tht of DC; only the groups treted with 4 mg of both drugs nd the mixture generic-plnt showed decreses with highly significnt differences for the generic nd very highly significnt for Amrel nd the mixture, compred to DC (Fig.6 B). Renl prmeters Chronic hyperglycemi nd dyslipidemi re ssocited with metbolic disorders in humns nd nimls with dibetes [32], which cuses oxidtive stress, resulting high levels of ROS [33]. Oxidtive environments cn cuse dmge in heptic nd renl cells nd tissues [34]. High levels of ure nd cretinine (indictors of renl dysfunction) observed in untreted dibetic rts (Tble 4) would resulted in from the ction of ROS nd the toxic effect of lloxn on the kidneys [28]. Tble 4: Evlution of renl serum prmeters in rts Prmeters Uric cid Ure (mg/dl) (g/l) Cretinine (mmol/l) HC AA 2,58 ±,44,66 ±,8 63,64 ± 15,34 DC 3,37 ±,42 1,34 ±,36 111,68 ± 13,99 2mg-G A AA NS 2,84 ±,21,78 ±,18 9,88 ± 28,84 2mg-A A AA AA 2,8 ±,37,7 ±,19 75,3 ± 17,12 4mg-G NS AA 3,5 ±,4,8 ±,13 97,56 ± 14,26 4mg-A NS A 2,99 ±,38,74 ±,7 87,16 ± 21,23 G+Plt A 2,71 ±,57,67 ±,8 73,54 ± 14,1 Vs HC: P,1, P,1; Vs DC: A P,5, AA P,1, P,1; NS : Not Significnt. The tretment ws effective in improving these two prmeters. No significnt differences were observed between the different tretments for the three renl prmeters studied. CONCLUSION This study imed to evlute the phrmceuticl qulity of generic glimepiride, n orl ntidibetic gent, compred to the origintor product Amrel, nd to verify its effects, ccording to two selected doses nmely 2 nd 4 mg per dy, in rts with lloxn dibetes. Despite the complince of their physicochemicl chrcteristics with Americn stndrds, the results of the biologicl study reveled: recovery of body weights in rts treted with the mixture generic - plnt, followed by groups treted with 4 mg of Amrel nd the generic respectively. These results re in ccord with those obtined for glycemi declines following different tretments (Fig.2). After scrifice, the most importnt decreses of fsting glycemi nd HbA1c levels were obtined with the combintion generic-plnt extrct, nd with doses 4 nd 2 mg of Amrel. For the lipid profile, Amrel ws more effective thn the generic; nd the mixture still shows beneficil effect on these prmeters. Totl proteins nd lso enzymtic ctivities of serum trnsminses, mylse nd lipse were improved in groups treted with 4 mg of both drugs nd the mixture. The different tretments exerted similr effects on serum ure, cretinine nd uric cid. No toxic effects were observed in rts treted with the two drugs, Amrel nd its generic product, t doses of 2 nd 4 mg per dy, during the period of tretment. The nlyzed generic glimepiride exhibits lower therpeutic potency compred to the origintor drug Amrel, which requires dose djustment for tretments. According to this study, it is suggested to mnufcturers to crry out studies on their finished generic products, in order to djust the recommended doses to void overdosing or under dosing which could be dngerous for ptients. Acknowledgments This study ws prtly supported by the Ministry of Higher Eduction nd Scientific Reserch (MESRS) of Algeri (PNR, Helth project 211). Authors thnk ssistnce of Inph-Médis lbortories. REFERENCES [1] JC Mbny. Globl Dibetes Pln, Interntionl Dibetes Federtion, 21. Avilble t: Accessed: April 212. [2] SN Dvis. J. Dibetes Complictions, 24, 18,

9 Cherif Henchiri et l J. Chem. Phrm. Res., 213, 5(5): [3] F Cludot nd Y Juillière. Les génériques: spects juridiques, questions éthiques. Consensus Crdio pour le prticien, 27, 33, [4] C Abelli. Générique humnitires: intérêts et limites des cinétiques de dissolution dns le contrôle qulité des gélules. Appliction à l tétrcycline et à l indométcine. Phrm. thesis, Clermont-Fd, 1996, 114. Avilble t: Accessed: October 212. [5] MC Chemtob-Conce. Brevets phrmceutiques et snté publique des pys en développement. Med. & Droit, 25, Vol 25(7), [6] Ekette Akpbio, Clement Jckson, Clister Ugwu, Mfon Etim nd Mfon Udofi. Nigeri. J. Chem. Phrm. Res., 211, 3(3): [7] Committee for the Updte of the Guide for the Cre nd Use of Lbortory Animls, Ntionl Reserch Council. Guide for the Cre nd Use of Lbortory Animls, 8 th edition. ISBN: , 21, [8] Glimepiride Tblets. Phrmcopeil Forum, 33(3), 411. [9] Glimepiride Tblets: Performnce Tests/Dissolution/Test 1. USP 35 NF3, 212, [1] E Mtteucci nd O Gimpietro. J. Ethnophrmcol., 28, 115, [11] A Boudjell, C Henchiri, L Sircus, M Sri nd R Giuseppe. Fitoterpi, 212, 83, [12] J Rosenstock, E Smols, DB Muchmore nd J Schneider. Dibetes Cre, 1996, 19(11), [13] WT Friedewld, RI Levy, DS Fredrickson. Clin. Chem., 1972, 18, [14] PK Prbhkr nd M Doble. Curr. Dibetes Rev., 28, 4, [15] R Weitgsser, M Lechleitner, A Luger nd A Klingler. Dibetes Res. Clin. Prct., 23, 61, [16] P Kemsri, S Sngeeth nd P Venktlkshmi, J. Chem. Phrm. Res., 211, 3(5), [17] GO Alde1, OR Omobuwjo1, CA Adebjo nd EJ Verspohl. J. Chem. Phrm. Res., 211, 3(2), [18] PJ Guillusseu nd M Lloi-Michelin. Rev. Med. Interne, 23, 24, [19] H Bgheri. Génériques, équivlents thérpeutiques, copies, princeps: similitudes et différences. Rélités en Gynécologie-Obstétrique, 29, 14, 1-6. [2] E Dreger. Dibetes Res. Clin. Prct. Suppl., 1995, 28, S139-S146. [21] RV Mhto, P Gywli, P Psd. Rut, P Regmi, K Psd. Singh, DR Pndey nd P Gywli. Biomed. Res., 211, 22(3), [22] A Dnish, S Mnju, M Alok, WR Prmod nd K Viks. Altern. Med., 213, 13, 1. [23] SL Bdole nd SL Bodhnkr. Eur. J. Integr. Med., 29, 1, [24] M Ydv, A Lvni, R Tomr, G B K S Prsd, S Jin nd H Ydv. Appl. Biochem. Biotechnol., 21, 16, [25] Dn-yn Xu, Shui-Ping Zho, Qiu-xi Hung, Wei Du, Yu-hu Liu, Ling Liu nd Xio-mei Xie. Dibetes Res. Clin. Prct., 21, 88, [26] T Arki, M Emoto, T Konishi, Y Ikuno, E Lee, M Termur, K Motoym, H Yokoym, K Mori, H Koym, T Shoji nd Y Nishizw. Metb., 29, 58, [27] SM Muer, MW Steffes nd DM Brown. Am. J. Med., 1981, 7, 63. [28] S Lenzen. Dibetologi, 28, 51, [29] TP Almdl nd H Vilstrup. Dibetologic, 1988, 31, [3] M Rjn, V Kishor Kumr, P Stheesh Kumr, Kotm Reddy Swthi nd Sngm Hrith. J. Chem. Phrm. Res., 212, 4(6), [31] H Niecheiles. J. Ntl. Med. Assoc., 1961, 53(3), [32] K Hmden, F Aydi, K Jmoussi, H Msmoudi nd A Elfeki. Biofctors, 28, 33(3), [33] K Hmden, B Joudi, N Zri, T Rebi, S Crreu nd A Elfeki. J. Physiol. Biochem., 211, 67(1), [34] K Hmden, B Joudi, S Crreu, A Aouidet nd A Elfeki. Nt. Prod. Res., 211, 25(3),