THE PHARMACOKENETICS OF LEVOFLOXACIN IN HEALTHY RABBIT TREATED WITH ANTACID Aniek Setiya Budiatin, Weni Kristanti, Suharjono

  The Pharmacokenetics of Levofloxacin in Healthy Rabbit Treated with Antacid (Aniek Setiya Budiatin, Weni Kristanti, Suharjono) THE PHARMACOKENETICS OF LEVOFLOXACIN IN HEALTHY RABBIT TREATED WITH ANTACID Aniek Setiya Budiatin, Weni Kristanti, Suharjono Departement of Clinical Pharmacy.

  Airlangga University School of Pharmacy. Surabaya.

  ABSTRACT

The effect of antacid (Antasida DOEN) 14 ml/ kg BW which containing polyvalent cation Mg (OH) and Al(OH) on the

  2

   3

absorption of levofloxacin were examined in healthy rabbit. Twelve subjects were administered 23 mg/ml kg BW

levofloxacin alone. On day 8 the six subjects were administered antacid and levofloxacin concomitant and the other

were administered antacid two hours after levofloxacin. There were statistically significant different in pharmacokinetic

parameter produced by administration with antacid concomitant but the other one were not significant different. There

were C and AUC decrease (p<0.05) but t increase was not significant different (p>0.05) for administration

max max

concomitant and but the other one were C , AUC and t smaller than control but not significant different (p>0.05).

max max

  

The different had apparent effect on the plasma concentrations of levofloxacin and was determined with

spectrofluorometric.Antacid may reduce the absorption of levofloxacin, the extent of this interaction appears to

increase as time between administration of two drugs decreases.

  Keyword: levofloxacin, Mg (OH) and Al(OH) antacid, spetrofluorometric

  2 3, Correspondence: Aniek Setiya Budiatin, Departement of Clinical Pharmacy, Airlangga University

  School of Pharmacy, Jl Dharmawangsa Dalam, Surabaya, phone. 62-31-5033710

  INTRODUCTION

  Levofloxacin (fig. 1) is fluoroquinolone antibiotic and is the 1 ofloxacin isomer, more active 8 – 128 times against positive and negative microorganism than d- ofloxacin, and twice more potent than rasemat form (Fish and Chow, 1997). Levofloxacin were used for treatment disease : GIT, respiratory tract up and down; urinary tract, skin infection, which inhibit DNA synthesis as a potential antibacterial. Plasma concentrations in healthy volunteers reach a mean peak

  Figure 2. Ciprofloxacin drug plasma concentrations (C ) of approximately 2.8

  max

  and 5.2 mg/L within 1 to 2 hours after oral In separate study, the effect of antacid, sucralfat; food, administration of levofloxacin 250 and 500 mg tablets, H receptor antagonist on fluoroquinolon interaction

  2

  respectively. The oral absorption is very rapid and was examined (Radanst et al. 1992). The interactions complete, the bioavailability of oral levofloxacin with aluminum and magnesium containing antacid with approaches 100% and it little affected by administration fluoroquinolones (siprofloxacin, Figure 2), resulting in with food (Fish and Chow, 1997). significantly decrease absorption when administered concurrently (Fish and Chow 1997). A measure such as t does not adequately describe situations in which

  max

  double peaking may occur or which prolonged absorption is observed. Antacid (Mg (OH) and Al(OH)

  2

  ) is widely used for a variety of indications (duodenal

  3 ulcer disease, dyspepsia, saur stomach) (McEvoy 2002).

  The probably that levofloxacin will be given to patients receiving long term therapy with antacid/H blocker is

  2 Figure1. Levofloxacin

  high. Since antacid affect oral absorption of variety of Folia Medica Indonesiana Vol. 44 No. 4 October – December 2008 : 264-268

  agents, the effect of administration of antacid on the absorption of ciprofloxacin was studied (Radanst et al, 1992), Therefore studies were perform to determined the effect of antacid administration on the oral absorption of levofloxacin in healthy rabbits.

  ) and earliest time at which C

  max

  The mean plasma levofloxacin concentration-time curves for the administered antacid and levofloxacin concomitant groups were significant different, C max , AUC and t

  RESULTS

  ) were estimated directly from the experimental data. The intensity under the plasma concentration-time curve from time zero to the last time (t) plasma levofloxacin was measurable (AUC 0-300 ) was estimated by the linier trapezoidal approximation (Shargel 2005). Student’s t tests with 95% confidence limits were used to examine pair-wise differences between the groups. Value of p<0.05 were considered to be statistically significant (Santosa et al 2005).

  max

  occurred (t

  max

  max

  MATERIAL AND METHODS Levofloxacin was a kind gift of PT Kimia Farma.

  The pharmacokinetic analysis of plasma concentration of levofloxacin were determined using spectrofluorometric method. The assay had a dynamic range of 10 to 60 ng/ml (El-Kommos et al. 2003). . The maximum plasma concentration of levofloxacin (C

  μg /ml) was added and pH was adjusted to 3.5 using 1 ml of Teorell and Stenhagen buffer solution. The volume was completed with methanol. The relative fluorescence intensity of resulting solutions was measured against reagent blanks treated similarly at the excitation and emission maxima specified for levofloxacin. (El-Kommos et al. 2003) Five milliliter of plasma were deproteinized by the addition of 10 ml acetonitrile, centrifuged at 4000 rpm for 5 minutes. One milliliter of clear supernatant was spiked with 1 ml of drug stock solution. The mixture was then extracted with 2 portions; each of 5 ml chloroform. The chloroformic extract was collected, evaporated on a boiling water bath, then appropriate dilutions were made to obtain drug solutions containing 100, 300 and 500 ng/ ml, then general procedure was followed. The twelve rabbits were the same treatment as levofloxacin alone, and the day 8, six rabbits were administered antacid and levofloxacin concomitant and the other one were administered antacid 2 hours after levofloxacin (El-Kommos et. al., 2003).

  One milliliter of the ammonium molybdenum solution (7.5

  injection syringe 2.5 cc containing anticoagulant via ear of marginal venous. Samples were collecting in a vacuum test tube. Blood samples were obtained immediately before and 5, 10, 20, 30,45, 60, 75, 90, 120, 300 minutes after ingestion of the orally administered formulation.. Plasma was harvested and samples were stored at -20 C until assayed. Plasma concentration of levofloxacin was determined by spectrofluorometric. The assay was linier over the calibration range 100 to 600 ng/ml (El-Kommos et al, 2003) One milliliter aliquot volumes of standard or sample solutions were transferred into 10-ml calibrated flask.

  libitum 10 ml. Blood sample (2ml) were collected with

  Stock solutions containing 1 μg/ ml of each levofloxacin was prepared in methanol. Working standard solutions containing 100-600 ng/ml was prepared by suitable dilution of the stock solutions with methanol. The experimental animals were Australian Rabbit from Batu Malang, weight 2.5 kg. Treatment antacide 14 mg/ kg BW of rabbit, 70 mg/ml oral solution form and levofloxacin 23 mg/ml kg BW of rabbits (Paget 1964). The twelve rabbits were fast overnight before drug administration and 2 hours after the dose was administered. Water was allowed ad

  Heparin 500 unit/ml, Ammonium molybdat were purchased from Fluka AG, Citric acid was purchased from Riedel-de Häen, Xylol was obtained from Brataco Chemika, Acetonitril was purchased from Riedel-de Häen, Hitachi F-4000 Spectrofluorometer (Japan) was used for fluorometric measurements, Direct Reading Micro Balance LM -20 Libror, Ultrasonic Cleaner Sakura US-10E, Centrifuge Hettich, Protofix 32, Thermolyne Barnstead type 37600, glassware and injection syringe.

  Methanol (HPLC gradient) was purchased from JT Baker, Antacida DOEN was a kind gift of First Medipharma, Dipotassium hydrogenphosphate and Phosphoric acid 85% were purchased from Merck (Darmstadt, Germany), Double distilled Water was obtained from PT. Ikapharmindo Putramas (Indonesia).

  (Figure 3, Table 1: with treatment 6 rabbits number 1 until 6) and the administered antacid two hours after levofloxacin were not significant different (Figure 4, Table 2, with treatment 6 rabbits number 7 until 12). The Pharmacokenetics of Levofloxacin in Healthy Rabbit Treated with Antacid (Aniek Setiya Budiatin, Weni Kristanti, Suharjono)

  Table 1. Summary of pharmacokinetic parameter of the administered antacid and levofloksasin concomitant (treatment rabbits number 1 until 6)

  Rabbit Control Treatment number

  T max C max AUC0-300 t max C max AUC0-300 (minute) (µg/ ml) minute (minute) (µg/ ml) minute

  (µg.minute/ml) (µg.minute/ml)

  1

  90 4.21 754.10 120 1.19 225.47

  2

  90 3.29 484.48 180 1.24 216.77

  3

  90 2.27 494.00 120 1.34 276.64

  4

  60 4.59 985.17 180 2.10 491.13

  5

  60 3.96 497.83 180 0.90 156.51

  6

  60 2.08 415.08

  30 2.43 306.80 Mean

  75.00 3.40 605.11 135.00 1.53 278.89 ± SD

  16.43 1.04 219.62

  59.24 0.59 116.21

  Control 1 Treatment

  1 Time (minute)

  Figure 3. Curve concentration (µg/mL) vs time (minute), Treatment I = were administered levofloxacin and antacid concomitant Table 2. Summary of pharmacokinetic parameter of the administered antacid 2 hours after levofloxacin (treatment 6 rabbits number 7 until 12)

  Rabbit Control Treatment number t max C max AUC0-300 t max C max AUC0-300

  (minute) (µg/ ml) minutes (minute) (µg/ ml) menit (µg.minute/ ml) (µg.minute/ ml)

  7

  90 1.79 336.59 120 1.75 356.38

  8

  60 4.05 630.54

  60 2.26 488.18

  9

  60 3.21 606.19

  60 3.43 518.38

  10

  60 2.31 447.16

  60 2.79 608.46

  11

  60 2.46 424.99

  60 1.85 352.72

  12

  60 0.67 143.71

  60 1.20 263.21 Mean 65..0 2.09 431.53

  70.00 2.21 431.22 ± SD 12..25 0.94 180.14

  24.49 0.80 128.25 Folia Medica Indonesiana Vol. 44 No. 4 October – December 2008 : 264-268

  Figure 4. Curve concentration (µg/mL) vs time (minute), Treatment 2 = were administered antacid 2 hours after levofloxacin

  max

  Figure 5. Levofloxacin and metal complex form the reaction similar with Ciprofloxacin (Nix et al., 1989; Brighty et al, 2000)

  The inhibition of absorption can be significant and potentially lead to the failure of the treatment, even when levofloxacin dose are separated from antacid dose by more than 2 hours (Radanst et al 1992).

  max increased 0.77 % but not significant different (p> 0.05).

  0.07 % (p> 0.05) respectively and t

  0-300

  5.74 % and AUC

  increased 180 % but not significant different (p>0.05) (Table 1 and figure 3). Administration of the antacid 2 hours after levofloxacin decrease but not significant different in C

  DISCUSSION

  max

  ) of levofloxacin 55.29 %, 53.91 % significant different (p<0.05) (Santosa et al, 2005) respectively and t

  0-300

  and the mean area under the concentration-vs-time curve (AUC

  max

  1989;) so the effect was same. Antacid was given with levofloxacin concomitant decreased the mean C

  To determine the effect of timing on absorption of antacid and levofloxacin administered of single 23 mg / kg BW dose of levofloxacin to 12 healthy rabbits in two-way crossover study, levofloxacin was given alone or combined with ether an magnesium/aluminum containing antacid 14 ml /kg BW; the timing of antacid and levofloxacin administration was varied (concomitant and antacid 2 hours after levofloxacin). The literature describing reduced absorption of fluoroquinolones due to antacid administration to patients and volunteers is most extensive for ciprofloxacin (Figure 5) where in the gastro intestinal region the complex chelate form of antacid (metal = Mg and Al) and ciprofloxacin couldn’t absorbed, the structure of levofloxacin (Figure 3) was similar with ciprofloxacin (Figure 4) (Brighty et al. 2000; Nix et al.

  Time (minute) Control 2 Treatment 2 Insoluble complex of levofloxacin-metal Oxo group Carboxil group The Pharmacokenetics of Levofloxacin in Healthy Rabbit Treated with Antacid (Aniek Setiya Budiatin, Weni Kristanti, Suharjono)

CONCLUSION Fish, DN, Chow, AT 1997, ‘The clinical

  pharmacokinetics of levofloxacin’, Pubmed, vol. 32, There were statistically significant different in no. 2, pp. 101-119. pharmacokinetic parameter produced by administration McEvoy, GK 2002, AHFS Drug Information, American with antacid concomitant but the other one were not Society Of Health System Pharmacists, pp. 788-790. significant different. There were C and AUC Nix, DE, Watson, WA, Lenes ME, Frost, RW, Krol, G,

  max

  decrease(p<0.05) but t increase was not significant Goldstein, H, Lettieri, J, Schentag, JJ 1989, ‘Effects

  max

  different (p>0.05) for administration concomitant and of aluminium and magnesium antacids and ranitidine but the other one were C , AUC and t smaller than on the absorption of ciprofloxacin’, Clin Pharmacol

  max max control but not significant different (p>0.05). It’s Ther , no. 46, pp. 700-705.

  suggested that levofloxacin dose are separated from Paget, GE. and Barnes, JE 1964, ‘Toxicity test’, in DR antacid dose by more than 2 hours. Laurence, AL Bacharach, Evaluation of Drugs

  Activities: Pharmacometrics , vol. I, Academic Press, London, pp. 161.

  

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