Investigation of the Intestinal Permeability of Ciclosporin Using the in situ Technique in Rats and the Relevance of P-Glycoprotein

Abstract

The primary endpoint of this study was to determine the intestinal permeability of Ciclosporin (cyclosporine A, CsA, CAS 59865-13-3) using the single-pass intestinal perfusion technique (SPIP) and a range of concentrations in rats. The second objective was to assess the quantitative contribution of P-glycoprotein (P-gp) -mediated efflux in limiting the oral bioavailability of CsA using erythromycin (Ery, CAS 114-07-8) as an inhibitor of P-gp efflux transporter. A solution containing CsA and phenol red either in the presence or in the absence of Ery as a P-gp inhibitor was perfused through a cannulated jejunal segment in rats. Outlet samples were collected every 10 min in micro tubes up to 90 min. Samples were analyzed using a modified reverse phase HPLC method. The mean effective permeability coefficients (P_eff ) of CsA in concentrations of 5,10,15 and 20 µmol/L in the perfusion solution were found to be 2.21 (± 0.26) × 10^–4 cm/s, 3.34 (± 1.29) × l0^–4 cm/s, 3.12 (± 0.23) x 10^–4 cm/s and 2.73 (± 0.28) × 10^–4 cm/s, respectively. The corresponding values in the presence of Ery were found to be 3.96 (± 1.04) × 10^–4 cm/s, 5.34 (± 1.29) × 10^–4 cm/s, 3.72 (± 0.21) × 10^–4 cm/s and 4.41 (± 0.89) × 10^–4 cm/s, respectively.

The two-tailed Student’s t-test showed that the intestinal permeability of CsA was significantly increased by Ery in all four CsA concentrations used (P < 0.05). However, there was no significant difference between the P_eff values of CsA in different concentrations, indicating that the CsA permeation was independent of the concentration. Therefore it is concluded that at least some part of the observed clinical interaction between Ery and CsA is due to the interaction in absorption level.

Key words

CAS 59865-13-3 - Ciclosporin, intestinal permeability, rat - Immunosuppressants - P-glycoprotein

• References

• 1 Cyclosporine Injection, USE Available: http://www.paddocklabs.com/forms/pi/20659%95Cyclosporine.pdf.
• 2 Chiu YY, Higaki K, Neudeck BL, Barnett JL, Welage LS, Amidon GL. Human jejunal permeability of cyclosporine A: Influence of surfactants on P-gp efflux in Caco-2 cells. Pharm Res. 2005; 20 (5) 749-756
• 3 Arimori K, Nakano M. Drug exsorption from blood into the gastrointestinal tract. Pharm Res. 1998; 15: 371-376
• 4 Hunter I, Hirst BH. Intestinal secretion of drugs. The role of P-gp and related drug efflux systems in limiting oral drug absorption. Adv Drug Del Rev. 1997; 25: 129-157
• 5 Terao T, Hisanaga E, Sai Y, Tamai I, Tsuji A. Active secretion of drugs from the small intestinal epithelium in rats by P-gp functioning as an absorption barrier. J Pharm Pharmacol. 1996; 48: 1083-1089
• 6 Wacher VI, Silverman JA, Zhang Y, Benet LZ. Role of P-gp and cytochrome P450 3A in limiting oral absorption of peptides and peptidomimetics. J Pharm Sci. 1998; 87: 1322-1330
• 7 Drewe J, Beglinger C, Kissei T. The absorption site of cyclosporine in the human gastrointestinal tract. Br J Clin Pharmacol. 1992; 33: 39-3
• 8 Ohba M, Ohnishi N, Romada F, Iwakawa S, Okumura K. Effect of clarithromycin on the bioavailability of cyclosporine in rats. Biol Pharm Bull. 1996; 19 (5) 733-737
• 9 Hughes CM, Swanton IG, Collier PS. Cyclosporine A and erythromycin: A study of a drug interaction in the in situ rat liver model. Biopharm Drug Dispos. 1993; 14: 615-625
• 10 Gupta SK, Bakran A, Johnson RWG, Rowland M. Cyclosporin-Erythromycine interaction in renal transplant patients. Br J Clin Pharmacol. 1989; 27: 475-481
• 11 Swenson ES, William B, Malison B. Curatolo W Intestinal permeability enhancement: Efficacy Acute local toxicity and reversibility. Pharm Res. 1994; 11 (8) 1132-2
• 12 Valizadeh H, Zakeri-Milani P, Islambolchilar Z, Tajerzadeh H. A simple and rapid HPLC method for determining Furosemide, Hydrochlorothiazide and Phenol red: Applicability to Intestinal permeability studies. J AOAC Int. 2006; 89 (1) 1-6
• 13 Zakeri-Milani P, Barzegar-Jalali M, Tajerzadeh H, Azarmi Y, Valizadeh H. Simultaneous determination of naproxen, ke-toprofen and phenol red in samples from rat intestinal permeability studies: HPLC method development and validation. J Pharm Biomed Anal. 2005; 39: 624-630
• 14 Ernest D, Alfert ED, Brenda M, Cross BM, McWilliam AA eds CCAC, Guide to the care and use of experimental animals, Canadian Council on Animal Care. 1993; 1: 1-150
• 15 Komiya I, Park JY, Kamani A, Ho NFH, Higuchi WI. Quantitative mechanistic studies in simultaneous fluid flow and intestinal absorption using steroids as model solutes. Int J Pharm. 1980; 4: 249-262
• 16 Levitt MD, Kneip IM, Levitt DG. Use of laminar flow and unstirred layer models to predict intestinal absorption in the rat. J Clin Invest. 1988; 81: 1365-1369
• 17 Varma MVS, Kapoor N, Srkar M, Panchagnula R. Simultaneous determination of digoxin and permeability markers in rat in situ intestinal perfusion samples by RP-HPLC. J Chrom. B. 2004; 813: 347-352
• 18 Zakeri-Milani P, Valizadeh H, Tajerzadeh H, Azarmi Y, Islambolchilar Z, Barzegar S et al. Predicting human intestinal permeability using single-pass intestinal perfusion in rat. J Pharm Pharmaceut Sci. 2007; 10 (3) 368-379