Hepatobiliary excretion and enterohepatic circulation of colchicine in rats

doi:10.1016/j.ijpharm.2007.08.052

International Journal of Pharmaceutics

Volume 350, Issues 1–2, 28 February 2008, Pages 230-239

https://doi.org/10.1016/j.ijpharm.2007.08.052 Get rights and content

Abstract

This study investigated the pharmacokinetics of unbound colchicine in rat blood, liver and bile, and its interaction with cyclosporin A (CsA; P-glycoprotein inhibitor) and proadifen (non-specific cytochrome P450 inhibitor) by using a microdialysis and liquid chromatographic system. The pharmacokinetics of colchicine in rat blood showed elimination in a nonlinear manner within the dosage ranges of 1–10 mg/kg. Twenty minutes after administration, colchicine reached maximum concentration in the liver and bile. The liver-to-blood distribution ratios (AUC_liver/AUC_blood) were 1.8 ± 0.6, 1.0 ± 0.2 and 0.8 ± 0.1, and the bile-to-blood distribution ratios (AUC_bile/AUC_blood) were 121.6 ± 24.7, 102.2 ± 13.4 and 116.5 ± 18.4 at dosages of 1, 3 and 10 mg/kg, respectively. The high hepatobiliary excretion of colchicine may lead to increased toxicity in normal tissues and indicates that colchicine undergoes hepatobiliary excretion against the concentration gradient from bile-to-blood. The area under the curse (AUC) of colchicine in the liver increased in the proadifen-treated groups, suggesting that metabolism of colchicine may involve cytochrome P450. CsA pretreatment caused an increase in the AUC of colchicine in the blood, a decreased AUC in the bile, and a profound decline in the bile-to-blood distribution ratio. Furthermore, the acute diarrhea and body weight loss caused by colchicine were delayed by pretreatment with CsA. These results indicate that the hepatobiliary excretion of colchicine was regulated by P-glycoprotein (P-gp) and the related acute diarrhea could be modulated by CsA. By using a paired rats model, the enterohepatic circulation of colchicine was also observed.

Introduction

Colchicine is an alkaloid derived from the plant Colchicum autumnale which possesses anti-inflammatory and antimitotic characteristics and is most often used to treat symptoms of gout with rheumatic and nonrheumatic conditions. The anti-inflammatory action mechanism of colchicine may inhibit neutrophil chemotaxis, thereby decreasing the inflammatory process (Ben-Chetrit and Levy, 1998). Colchicine inhibits the function of polymorphonuclear leukocytes, and dermatoses with a strong presence of these cells may benefit the most from the administration of this medication (Sullivan et al., 1998).

The use of colchicine in the treatment of liver cirrhosis and primary biliary cirrhosis was reported (Sabouraud et al., 1992). We previously demonstrated that colchicine enhances the radiosensitivity of human hepatoma HA22T/VGH cells (Liu et al., 2005). However, detailed colchicine levels in the liver tissue and bile were not measured. The influence of hepatic diseases on colchicine disposition should be investigated in order to define the most appropriate therapeutic dosing. Pharmacokinetic studies have been relatively limited and their results somewhat contradictory, with mean terminal elimination half-lives of 19 min to 9 h being reported (Levy et al., 1991). Some of these differences may be attributed to assay difficulties. Radioimmunoassay has been used to study post-mortem tissue concentrations and the toxicokinetics of colchicine in cases of acute human poisoning (Rochdi et al., 1992). [Ring C-methoxy-³H]colchicine was used to study the pharmacokinetics of protein unbound colchicine with a limit of detection of 0.15 ng/ml (Desrayaud et al., 1997). In contrast to radioisotope labeling, liquid chromatography has a limit of detection of 1 ng/ml for colchicine in serum (Ko et al., 1990).

P-gp is normally located on the apical surface of enterocytes, the biliary canalicular membrane of hepatocytes, and the apical surface of endothelial cells in brain capillaries to protect the organism against xenobiotics by excreting them into the body (Thiebaut et al., 1987). Colchicine is a substrate for the multidrug resistance transporter, P-gp (Riordan and Ling, 1979). Co-administration of SDZ PSC 833 (P-gp inhibitor) increased the brain penetration of colchicine (Desrayaud et al., 1997).

Hepatic transformation, biliary excretion and enterohepatic circulation may have significant effects on the pharmacokinetics of a number of drugs. According to classical pharmacokinetics principles, several factors, including chemical properties and membrane-penetration capacities, can influence distribution or excretion. In addition, hepatic blood flow, protein binding and hepatic intrinsic clearance also play important role. The present study develops a microdialysis coupled to liquid chromatographic system to investigate the mechanism of hepatobiliary excretion and enterohepatic circulation of colchicine which may be regulated by P-gp. Besides, severe diarrhea of colchicine is delayed by pretreatment with P-gp inhibitor.

Section snippets

Animal experimentation

All experimental protocols involving animals were reviewed and approved by the institutional animal experimentation committee of the National Yang-Ming University and Mackay Memorial Hospital. All animal care and husbandry were conducted in accordance with the Guide for the Care and Use of Laboratory Animal in a facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. Male specific pathogen-free Sprague–Dawley rats weighing 250–300 g were obtained from

Method validation

Typical chromatograms of colchicine in rat blood, liver and bile dialysis are shown in Fig. 1, Fig. 2, Fig. 3 with a retention time of 3.8 min. Fig. 1A shows a standard injection of colchicine (1.0 μg/ml), and Fig. 1B shows the chromatogram of a blank blood dialysate from pre-drug administration. Fig. 1C shows the chromatogram of a blood dialysate sample containing colchicine (0.47 μg/ml) collected 10 min after colchicine administration (3 mg/kg).

Fig. 2A shows the chromatogram of a standard

Discussion

From literature review, the protein binding of colchicine was weak. The protein-binding rate was ranged from 23 to 38.9% (Lannoy et al., 1994, Sabouraud et al., 1994). Most colchicine was unbound and active form in vivo. The liquid chromatographic method described here has been applied to the determination of unbound colchicine in rat blood, liver and bile with a quantification limit of 10 ng/ml. However, it may not be sufficiently sensitive to detect unbound colchicine in brain extracellular

Acknowledgements

This study was supported by the following research grants: NSC96-2113-M-010-003-MY3, NSC96-2628-B-010-006-MY3 and NSC95-2314-B-195-014 from the National Science Council, MMH-9637 from Mackay Memorial Hospital, 95004-62-170 from Taipei City Hospital, Taipei, Taiwan.

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Hepatobiliary excretion and enterohepatic circulation of colchicine in rats

Abstract

Introduction

Section snippets

Animal experimentation

Method validation

Discussion

Acknowledgements

Med. Hypotheses

J. Chromatogr. B

Adv. Drug Del. Rev.

Life Sci.

Forensic Sci. Int.

J. Chromatogr. A

Adv. Drug Del. Rev.

Biochem. Pharmacol.

J. Biol. Chem.

J. Am. Acad. Dermatol.

Biochem. Pharmacol.

J. Chromatogr. B

Multicentre randomized placebo-controlled trial of ursodeoxycholic acid with or without colchicine in symptomatic primary biliary cirrhosis

Aliment. Pharmacol. Ther.

Aspects of bile secretion in the rabbit

J. Physiol.

Biliary excretion of colchicine

J. Pharmacol. Exp. Ther.

Modified irinotecan hydrochloride (CPT-11) administration schedule improves induction of delayed-onset diarrhea in rats

Cancer Chemother. Pharmacol.