Isolation and Identification of Urinary Metabolites of Berberine in Rats and Humans

Abstract

ABSTRACT:The urinary metabolites of berberine, an isoquinoline alkaloid isolated from several Chinese herbal medicines, were investigated in rats and humans. Using macroporous adsorption resin chromatography, open octadecyl silane column chromatography and preparative high-performance liquid chromatography, we isolated seven metabolites (HM1-HM7) from human urine and five metabolites (RM1-RM5) from rat urine after oral administration. Berberine is an isoquinoline alkaloid isolated from several Chinese herbal medicines such as rhizoma coptidis, cortex phellodendri, and caulis mahoniae. Its chemical structure is 5,8-dihydro-9,10-dimethoxy-6H-benzo(g)-1,3-benzodioxolo(5,6-a)quinolizine. Berberine exhibits a wide variety of bioactivities such as antidiarrheic (Sack and Froehlich, 1982;Taylor and Greenough, 1989), antimicrobial (Yan et al., 2008), hypolipidemic (Kong et al., 2004;Doggrell, 2005;Brusq et al., 2006;Cicero et al., 2007), hypoglycemic (Yin et al., 2002;Pan et al., 2003;Zhou et al., 2007;Turner et al., 2008;Yin et al., 2008), antiarrhythmic (Wang et al., 1994;Lau et al., 2001), anticancer (Inoue et al., 2005;Lanvers-Kaminsky et al., 2006;Lin et al., 2006Lin et al., , 2007Piyanuch et al., 2007;Serafim et al., 2008;Yu et al., 2007), anti-inflammatory (Kuo et al., 2004Lee et al., 2007), antiviral (Hayashi et al., 2007), antidepressant (Kulkarni and Dhir, 2007), and hepatoprotective (Zhang et al., 2008) effects. Its chloride salt has been used for several decades in clinical situations to treat gastroenteritis and secretory diarrhea in China. In recent years, berberine has also shown significant effects in treatment of diabetes mellitus (Ni, 1988), hyperlipemia (Kong et al., 2004), arrhythmia, and heart failure (Zeng and Zeng, 1999;Lau et al., 2001;Zeng et al., 2003). However, pharmacokinetic studies have indicated that berberine has poor oral bioavailability (Shen et al., 1993;Yu et al., 2000;Zuo et al., 2006), and a few of the metabolites have been identified in rats (Zuo et al., 2006) and in humans (Pan et al., 2002). To obtain more information about its metabolism to improve its clinical applications, we examined the biotransformation of berberine in rats and humans. In the present article we describe the isolation and identification of urinary metabolites of berberine in these two species. Materials and Methods Materials.Berberine chloride (purity Ͼ 99.5%) was supplied by the Northeast General Pharmaceutical Factory (Shenyang, China). Methanol was HPLC grade and water was double-distilled in our laboratory. All other reagents were of analytical grade and were purchased from Shenyang Chemical Company (Shenyang, China). Normal-phase and reverse-phase preparatory thin-layer chromatography was performed using products from Merck (Darmstadt, Germany). Macroporous resin D101 was purchased from the Chemical Plant of Nankai University (Tianjin, China) and Diaion ion exchange resin HP20 from Mitsubishi Chemical Corporation (Tokyo, Japan). Sephadex LH-20 and ODS were obtained from Pfizer (Freiburg, Germany). Spot...

Extensive Intestinal First-Pass Elimination and Predominant Hepatic Distribution of Berberine Explain Its Low Plasma Levels in Rats

“…Previous studies in vivo (Tsai and Tsai, 2004;Zuo et al, 2006;Qiu et al, 2008), including our more recent study in vitro (Liu et al, 2009), have well demonstrated that berberine is extensively metabolized in the body to generate M1 and M1 glucuronide via oxidative demethylation and subsequent glucuronidation and to form M2 and M2 glucuronide via oxidative demethylenation and subsequent glucuronidation. Therefore, UDP-glucuronosyltransferases have been identified as the major drug-metabolizing enzymes responsible for the formation of the phase II metabolites of berberine as shown by our more recent study in vitro (Liu et al, 2009) and this study.…”

“…In theory, an extremely low plasma concentration of berberine is associated mainly with the following PK causes: 1) extensive metabolism in the gut and/or liver (Tsai and Tsai, 2004;Zuo et al, 2006;Hao et al, 2007;Qiu et al, 2008); 2) marked excretion to intestinal lumen, bile (Tsai and Tsai, 2004), and urine (Qiu et al, 2008) (as a substrate of certain efflux transporters) (Xie, 2010), as well as enterohepatic circulation (Zuo et al, 2006;Xie, 2010); 3) poor absorption (due to some unique physicochemical properties); and 4) predominant tissue distribution.…”

“…Efforts have been made to explore the reason that berberine exhibits low bioavailability in the body after oral intake (Zuo et al, 2006;Qiu et al, 2008). For example, the metabolism of berberine was found to be catalyzed by several cytochromes P450 and UDP-glucuronosyltransferases in rat liver microsomes, whose major metabolic pathways were oxidative demethylenation and subsequent glucuronidation after intravenous administration (Liu et al, 2009).…”

Berberine induces cell death in human hepatoma cells in vitro by downregulating CD147

“…These metabolites are very polar and easily excreted in urine. (41) A recent study has shown that orally administered berberine is detectable in the lung tissues of mice and inhibits in vivo tumorigenesis and growth of xenografted lung cell tumors. This indicates that viable concentrations of berberine are achievable through oral administration.…”

Pharmacokinetics and Brain Distribution and Metabolite Identification of Coptisine, a Protoberberine Alkaloid with Therapeutic Potential for CNS Disorders, in Rats

“…By comparison with blank samples, full scanning in the m/z region of 50 to 700 allowed the assignment of several peaks to COP and its metabolites in the total ion chromatogram (TIC). Based on the drug metabolism rule and PBA (berberine, jatrorrhizine, and epiberberine) metabolite reports 15,16) (Table 4 and Fig. 6).…”