w wz 16«2y Kor. J. Clin. Pharm., Vol. 16, No. 2. 2006 k v p w w sƒ B Á z C Á½ k B Á B Á B Á x B Á x D Á x C Á ³ C Á E B w w w C w w D ƒm w w w E œw Bioequivalence Study on Two Talniflumate Preparations Commercially Available in Korean Uy Dong Sohn a, Ji Hoon Jeong b, Kyung Tae Kim a, Young Rae Cho a, Young Sil Min a, Hyun Ju Song a, Hyun Dong Je c, Sung Hyuk Yim b, Yong Kyoo Shin b, Seong Wan Cho d a College of Pharmacy and b College of Medicine, Chung Ang University, Seoul, 156-756, Republic of Korea c College of Pharmacy, Catholic University of Daegu, Gyeongbuk, 712-702, Korea d Department of Pharmaceutical Engineering, Konyang University, Nonsan-city, Chungnam, 320-711, Republic of Korea The aim of the present study was to evaluate the bioequivalence of two talniflumate preparations. We used Somalgen tablet (Kun Wha Pharmaceutical Co., Korea.) as a reference drug for bioequivalence of Crimain tablet (Samjini Pharmaceutical Ind. Co., Korea), and performed this whole study according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty four healthy male volunteers, 22.8±2.2 years in age and 64.6±5.3 kg in body weight, were divided into two groups and a randomized 2 2 cross-over study was employed. After one tablet containing 370 mg of talniflumate was orally administered, blood was taken at predetermined time intervals and the concentrations of talniflumate in plasma were determined using HPLC method with UV-detector. The analysis system was validated in specificity, accuracy, precision and linearity. These items of the analysis condition in this study conform to the guideline of KFDA. The pharmacokinetic parameters such as AUC t and C max were calculated using the analysis condition we established and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed AUCt and Cmax. Mean±SD of reference drug and test drug in AUC t and C max were 1.27±0.58 (µg/ml hr) and 0.27±0.13 (µg/ml) and 1.14±0.46 (µg/ml hr) and 0.26±0.10 (µg/ml) respectively. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to logg (1.25) for AUC t and C max, respectively. These results indicate that Samjin talniflumate tablet is bioequivalent to reference drug, Somalgen tablet. G Keywords Talniflumate, Bioequivalence, HPLC k v p l x ü p, (, m),, r, š ü v y (prodrug) xk 1-3).» v w» w š v w k v p š z w sƒ š 4). k v p Correspondence to : w w w 156-756 221 Tel: +82-2-820-5614, Fax: +82-2-826-8752 E-mail: udsohn@cau.ac.kr w p n z šx (T max ) 154 šx (C max ) 423 ng/ml» 121 š š 5). w sƒ w k v p wš j x w w sƒ w w sƒ wš w. w sƒw» w w q k C max x š w (AUC) sƒ w x t t z e z w x z w ü ³ w w. 101
102 Kor. J. Clin. Pharm., Vol. 16, No. 2, 2006 vx vx œš mw z w, vx, n, x, e wš, w xz» š l w. x w w,,, x x yw mw,» ƒ vx w, w x» ( t t š 2001-57y, 2001, 9, 5) e» w wš» w vx w. <vx» w > Á 19 ~55 Á y š ü (v w q,, { ü ) Á ƒ t p Á w x, x yw, vx ww š q Á vx y < w > ƒ x p w ( sw) w. Áx : Lymphocytes, Monocytes, Eosinophils, Hemoglobin, Hematocrit,WBC, RBC, Platelet, Neutrophil, Basophils Áx yw : Fasting glucose, BUN, Creatinine, Total Protein, Albumin, sgot, sgpt, Alkaline Phosphatase, Total Bilirubin, Total Cholesterol Á : Specific Gravity, Color, ph, Glucose, Albumin, Bilirubin, WBC, blood Á : ü (,, er,,», ), y z(x, ). vx, ü wš vx z w w. z x z w xƒ z w. vx n x x 6 x ƒ vx» k z wš 7 ¾ x w. vx heparin-locked catheter ewš blank x ƒƒ 10 ml xw. w x w» w x x vx» w. x x 1, x 2, 3, x 1 ƒ g. vx w n 8 l x ƒ ƒ 1 (k v p 370 mg) 240 ml wì n w. vx x š w 2 w. x x»ƒ k v p n 3.03 4.41 š m 3 15 w 6). vx w, w { x š w 1 {» w š, x z n n z 0.5, 1, 1.5, 2, 2.5, 3, 4, 6, 9, 12 15 12 w. x i.v. catheter û xq w» w 2ml x ü š vx y x» x» 10 ml x w. x z i.v. catheter w x š w» w xq w. x x 3000 rpm 10 w x w. x v p p»š p w 70 w deep freezer ¾ w.»» x y w š x z w x w k v p w. ü t œw v w k (Fisher Scientific., Fair Lawn, NJ, U.S.A), xq (, Korea) qt, Milli Q(Millipore Co., Milford, MA, U.S.A) 18 MΩ-cm m k w.»k p 1 w. w j m v(hplc/uv) x»». - HPLC: Younglin SP930D HPLC Pump Midas type 830 Autosampler Younglin UV730 Detector Autochro Data Module (Younglin autochro win - Chromatography Data System) - Balance: Sartorius analytic balance -»: Union 5KR, Hanil
k v p w 103 - Voltex mixer : CM-1000 Eyela x k v p v t t d wš ƒ 1000 µg/mlƒ 100% MeOH ƒw w k z w 0.1, 0.5, 1, 1.5, 3, 5 µg/ml t. 100 µl w» w k k.» œx 500 µl ƒwš 1 w 0.02, 0.1, 0.2, 0.3, 0.6, 1 µg/ml spiked blood sample w. spiked blood sample ü t k (Diluted 0.5 µg/ml with 100% MeOH) 1000 µl y ƒw z 1 vortexingw z 3,100 rpm 20 w 100 µl w HPLC w w j m v j w w. x w» w, w x 5 ww ü x wš w 5 x ww. x y w z w. vx l ƒ w -70 C w o x ew, w w z 100 µl HPLC w. w HPLC/UV (mobile phase) acetonitrile : 0.1M sodium acetate (adjusted ph6.4 with acetic acid) 37 : 63 yww w Column Luna C 18, 4.6 250 mm (Phenomenox, USA) w š Column 25 C o w. 1.2 ml/min w š» UV» q 288 nm w. x j m l ü t v j w k v p v j w w l x k v p w. 24, vx, ƒƒ 22.8±2.2, 174.2±4.1 cm, 64.6±5.3 kg x w vx» m w. p x w HPLC w j m Fig. 1, 2, 3, v v j 6.9, ü t v j 6.0 š, v ü t (I.S.)»k x (Fig. 1). v 0.02 ( w ), 0.1, 0.2, 0.3, 0.6, 1 µg/ml t ƒw x 14. 5w w HPLC w, x l w v ü y = 1.4104x + 0.0301(R 2 = 0.9992, y = 1.4313x + 0.0342(R 2 = 0.9997 yw ùkü š w w (Fig. 2). w sƒ w x» sƒw 4). x w sƒ w w e x - š l w x - š w (AUCt), š x (Cmax) w š x (Tmax) š w. Tmax w x s ƒw ye w x m v (K-BE Test 2002, ver. 1.2.1) w α( )= 0.05 w. š vx x vx w Fig. 1. Chromatograms of (A) blank human plasma, (B) blank human plasma spiked with taniflumate and internal standard. (C) human plasma taniflumate peak.
104 Kor. J. Clin. Pharm., Vol. 16, No. 2, 2006 š. Fig. 2. Mean (ÛS.D, n=24) plasma concentration-time curves of talniflumatel following oral administration of referrence (ù) and test drug (ø)., y ( w ), y, š w w» w inter-day, intra-day validation w. v 0.02, 0.1, 0.2, 0.3, 0.6, 1 µg/ml w validation Table 1, 2 ùkü. Inter-day validation C.V.(coefficient of variation) 2.14% 12.46%, accuracy 97.44% 105.46 %. Intra-day validation C.V.(coefficient of variation) 4.27% 10.10% š, accuracy w 97.60% 113.27%. j m y (S/N ratio) 5 w x w 0.02 µg/ml w. x talniflumate w HPLC 0.02 µg/ml 1 µg/ml w x w, p,, y q l ƒ vx AUC n z 15 ¾ ƒ vx x - š l Õ œ w w. Cmax ƒ vx x - š l ƒ x w, Tmax ƒ vx x - š l š x w w. ƒ vx q k w Table 3 vx -x š Fig. 2 ùkü. x m w x» sƒw 7). x w sƒ w w e x - š l w x - š w (AUCt), š x (Cmax) w š x (Tmax) š w. Tmax w x s ƒw ye w x m v (K-BE Test 2002, ver. 1.2.1) w α( )= 0.05 w. m yw AUC s³e 90% log 0.8309<δ<log 1.0282 log 0.8 log 1.25 ü w q» g (Table 4). sƒw AUC w w. m yw Cmax s³e 90% log 0.9102<δ<log 1.119 log 0.8 Table 1. Intra-day Accuracy of the HPLC analysis for talniflumate in Human plasma IntradayG(µg/ml) AG(Ratio) BG(Ratio) CG(Ratio) DG(Ratio) EG(Ratio) Mean SD C.V.G(%) AccuracyG(%) 0.02 ( w ) 0.06 0.06 0.07 0.06 0.06 0.06 0.01 9.41 111.09 0.1 0.19 0.19 0.18 0.20 0.18 0.19 0.01 4.27 113.27 0.2 0.32 0.34 0.27 0.28 0.33 0.31 0.03 10.10 97.76 0.3 0.47 0.48 0.40 0.42 0.44 0.44 0.03 7.22 97.70 0.6 0.90 0.89 0.80 0.90 0.78 0.86 0.06 6.70 97.60 1 1.51 1.51 1.29 1.48 1.47 1.45 0.09 6.28 101.02 Table 2. Inter-day Accuracy of the HPLC analysis for talniflumate in Human plasma InterdayG(µg/ml) 1G(Ratio) 2G(Ratio) 3G(Ratio) 4G(Ratio) IntradayGRatio Mean Mean SD C.V.G(%) AccuracyG(%) 0.02G( w ) 0.06 0.06 0.06 0.07 0.06 0.06 0.00 7.65 99.38 0.1 0.16 0.17 0.21 0.20 0.19 0.19 0.02 12.46 105.46 0.2 0.33 0.31 0.35 0.34 0.31 0.33 0.02 5.68 102.29 0.3 0.46 0.44 0.45 0.46 0.44 0.45 0.01 2.14 97.44 0.6 0.87 0.86 0.90 0.91 0.86 0.88 0.03 2.84 98.73 1 1.43 1.43 1.51 1.54 1.45 1.47 0.05 3.32 100.54
k v p w 105 Table 3. Bioavailability parameters s in normal and logarithmic scales for each volunteer obtained after oral administration of test and reference of talniflumate tablet AUC Cmax Tmax Reference drug test drug Reference drug test drug Reference drug test drug No. A-1 0.75-0.13 0.60-0.22 0.20-0.71 0.15-0.81 2.5 3 A-2 1.08 0.04 0.96-0.02 0.36-0.44 0.36-0.44 2.5 3 A-3 0.28-0.55 0.37-0.43 0.11-0.97 0.19-0.73 2.5 1.5 A-4 1.49 0.17 1.04 0.01 0.25-0.60 0.19-0.72 2 2 A-5 1.52 0.18 1.30 0.11 0.32-0.50 0.29-0.54 2 1.5 A-6 0.57-0.24 0.63-0.20 0.17-0.77 0.20-0.70 4 4 A-7 1.63 0.21 1.61 0.21 0.33-0.48 0.28-0.56 2.5 2 A-8 1.19 0.08 1.30 0.11 0.17-0.76 0.20-0.70 4 4 A-9 1.55 0.19 1.15 0.06 0.32-0.50 0.33-0.48 2.5 3 A-10 1.04 0.02 1.19 0.08 0.18-0.73 0.28-0.55 1 2 A-11 1.98 0.30 1.31 0.12 0.33-0.49 0.37-0.44 3 3 A-12 2.12 0.33 2.34 0.37 0.48-0.32 0.60-0.22 3 2 B-1 1.14 0.05 1.07 0.03 0.20-0.70 0.16-0.79 2.5 2.5 B-2 1.17 0.07 1.22 0.09 0.32-0.49 0.29-0.54 1.5 1.5 B-3 2.03 0.31 1.57 0.20 0.27-0.57 0.27-0.56 2 2 B-4 0.81-0.09 1.51 0.18 0.21-0.68 0.28-0.55 2 3 B-5 1.41 0.15 1.34 0.13 0.24-0.61 0.19-0.73 2 2 B-6 0.60-0.22 0.42-0.38 0.11-0.94 0.18-0.75 2.5 1.5 B-7 0.61-0.21 1.20 0.08 0.15-0.82 0.24-0.63 3 3 B-8 1.58 0.20 1.17 0.07 0.35-0.45 0.32-0.50 3 2 B-9 2.70 0.43 1.81 0.26 0.65-0.18 0.34-0.47 3 1.5 B-10 0.59-0.23 0.46-0.33 0.10-1.02 0.10-1.00 2 2.5 B-11 1.31 0.12 1.01 0.00 0.38-0.42 0.25-0.60 1.5 1.5 B-12 1.31 0.12 0.87-0.06 0.22-0.66 0.19-0.73 2.5 4 Average 1.27 0.05 1.14 0.02 0.27-0.62 0.26-0.61 2.46 2.42 S.D. 0.58 0.23 0.46 0.20 0.13 0.20 0.10 0.16 0.71 0.83 Table 4. Statistical results of bioequivalence evaluation in AUCt between two talniflumate tablets Anova F F table Group or Sequence 0.005 4.301 Subjects / Group 10.031 2.048 Period 0.003 4.301 Drug 1.611 4.301 least significant difference (%) 19.943 90% confidence interval lower limit 0.8309 upper limit 1.0282 Table 5. Statistical results of bioequivalence evaluation in Cmax between two talniflumate tablets Anova F F table Group or Sequence 0.516 4.301 Subjects / Group 7.408 2.048 Period 1.172 4.301 Drug 0.023 4.301 least significant difference (%)19.28 90% confidence interval lower limit 0.9102 upper limit 1.119 log1.25 ü w q» g. sƒw Cmax w w (Table 5). ww x j (k v p 370 mg) y q y (k v p 370 mg) w w x w ùkù w w š. w.
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