Evaluation of Clopidogrel Conjugation Metabolism: PK Studies in Man and Mice of Clopidogrel Acyl Glucuronide

Savu, Simona Nicoleta; Silvestro, Luigi; Surmeian, Mariana; Remis, Lina; Rasit, Yuksel; Savu, Simona Rizea; Mircioiu, Constantin

doi:10.1124/dmd.116.071092

Cited by 8 publications

(12 citation statements)

References 21 publications

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“…Despite its widespread use in clinical settings as an antiplatelet drug (Xie et al, 2011;Saeed et al, 2017), clopidogrel continues to draw attention worldwide. Previous studies demonstrated that clopidogrel undergoes extensive metabolism in the liver (Kazui et al, 2010;Xie et al, 2011;Zhu et al, 2013;Savu et al, 2016;Tai et al, 2016). In the human body, ;85% of ingested clopidogrel is rapidly hydrolyzed to clopidogrel carboxylate (CLP-C, an inactive carboxylic acid form), an immediate metabolite, by hepatic carboxylesterase 1 (Zhu et al, 2013); the remaining 15% is metabolized to clopidogrel active metabolite (CAM) by multiple cytochrome P450-mediated two-step oxidative pathways in the liver (Savi et al, 1992(Savi et al, , 2000Kazui et al, 2010;Xie et al, 2011Xie et al, , 2017.…”

Section: Introductionmentioning

confidence: 99%

Mrp3 Transports Clopidogrel Acyl Glucuronide from the Hepatocytes into Blood

Tai

Huang

et al. 2017

Drug Metab Dispos

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Clopidogrel acyl glucuronide (CLP-G) is a major phase II metabolite of clopidogrel generated in the liver for further excretion into urine; however, it is unclear whether CLP-G transports from hepatocytes into blood. Because multidrug resistance-associated protein 3 (MRP3) is predominantly expressed in the sinusoidal side of hepatocytes and preferentially transports glucuronide conjugates of drug metabolites from hepatocytes into bloodstream, we hypothesized that MRP3 could be such an efflux transporter for CLP-G. In this study, we compared the liver-to-plasma ratios of clopidogrel and its metabolites (including CLP-G) between (ATP-binding cassette, subfamily C, member 3) knockout (KO) and wild-type (WT) mice. We also evaluated the ATP-dependent uptake of clopidogrel and CLP-G as well as estradiol-17-d-glucuronide into human recombinant MRP3 inside-out membrane vesicles in the presence or absence of ATP. The results indicated that the liver-to-plasma ratio of CLP-G was 11-fold higher in KO mice than in WT mice, and that uptake of CLP-G (1 or 10 M each) into the membrane vesicles was 11.8- and 3.8-fold higher in the presence of ATP than in the presence of AMP, respectively. We conclude that Mrp3 transports CLP-G from the hepatocytes into blood in an ATP-dependent manner.

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Section: Introductionmentioning

confidence: 99%

Mrp3 Transports Clopidogrel Acyl Glucuronide from the Hepatocytes into Blood

Tai

Huang

et al. 2017

Drug Metab Dispos

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“…Similarly, a mean CAG plasma concentration was 336‐fold higher than a mean clopidogrel plasma concentration (470 vs. 1.4 ng/mL) in patients receiving this drug (Silvestro et al, ). More recently, a study indicated that CAG is confirmed as one of the major terminal metabolite of clopidogrel (Savu et al, ). These data indicated that the glucuronidation of CCA is the major phase II metabolic pathway for the elimination of clopidogrel from the body.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, UGT2B7 exhibits a lower apparent K m value and 3-to The most proportion of the ingested clopidogrel (85%) is hydrolyzed to CCA, and subsequently CCA is glucuronidated to CAG by UGTs (Kazui et al, 2010;Xie et al, 2011). In patients, an observed CCA concentration in plasma was 2736 ng/mL, 3000-fold higher than clopidogrel plasma concentration after taking a single oral dose of 75 mg clopidogrel (Savu et al, 2016), and 10400 ng/mL, also 3000-fold higher than plasma clopidogrel concentration after taking an oral dose of 300 mg clopidogrel (Kim et al, 2016). Similarly, a mean CAG plasma concentration was 336-fold higher than a mean clopidogrel plasma concentration (470 vs. 1.4 ng/mL) in patients receiving this drug (Silvestro et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…In addition to aspirin, clopidogrel has been added into the list of the World Health Organization essential medicine as a second antiplatelet drug (Patel, Vidula, Kishore, Vedanthan, & Huffman, ). Previous studies have been demonstrated that clopidogrel undergoes extensive metabolism in the human liver (Kazui et al, ; Xie et al, ,), where clopidogrel is predominantly hydrolyzed to clopidogrel carboxylic acid (CCA, or called clopidogrel carboxylate) by carboxylesterase‐1 (Tang et al, ; Zhu et al, ), and subsequently CCA is conjugated with the glucuronide moiety from UDP‐glucuronic acid (UDPGA) by the UDP‐glucuronosyltransferase (UGT) to form clopidogrel acyl glucuronide (CAG) (Silvestro et al, ; Savu et al, ). However, the UGT enzyme(s) involved have not been identified to date.…”

Section: Introductionmentioning

confidence: 99%

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Human UGT2B7 is the major isoform responsible for the glucuronidation of clopidogrel carboxylate

Huang

et al. 2018

Biopharm & Drug Disp

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Clopidogrel is predominantly hydrolyzed to clopidogrel carboxylic acid (CCA) by carboxylesterase 1, and subsequently CCA is glucuronidated to clopidogrel acyl glucuronide (CAG) by uridine diphosphate-glucuronosyltransferases (UGTs); however, the UGT isoenzymes glucuronidating CCA remain unidentified to date. In this study, the glucuronidation of CCA was screened with pooled human liver microsomes (HLMs) and 7 human recombinant UGT (rUGT) isoforms. Results indicated that rUGT2B7 exhibited the highest catalytical activity for the CCA glucuronidation as measured with a mean V value of 120.9 pmol/min/mg protein, 3- to 12-fold higher than that of the other rUGT isoforms tested. According to relative activity factor approach, the relative contribution of rUGT2B7 to CCA glucuronidation was estimated to be 58.6%, with the minor contributions (3%) from rUGT1A9. Moreover, the glucuronidation of CCA followed Michaelis-Menten kinetics with a mean K value of 372.9 μM and 296.4 μM for pooled HLMs and rUGT2B7, respectively, showing similar affinity for both. The formation of CAG was significantly inhibited by azidothymidine and gemfibrozil (well-characterized UGT2B7 substrates) in a concentration-dependent manner, or by fluconazole (a typical UGT2B7-selective inhibitor) in a time-dependent manner, for both HLMs and rUGT2B7, respectively. In addition, CCA inhibited azidothymidine glucuronidation (catalyzed almost exclusively by UGT2B7) by HLMs and rUGT2B7 in a concentration-dependent manner, indicating that CCA is a substrate of UGT2B7. These results reveal that UGT2B7 is the major enzyme catalyzing clopidogrel glucuronidation in the human liver, and that there is the potential for drug-drug interactions between clopidogrel and the other substrate drugs of UGT2B7.

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“…Clopidogrel is prodrug and becomes active in the form of its thiol metabolite in the presence of cytochrome P450 enzymes in the liver while major fraction (approximately 85%) of the administered drug converts into a pharmacologically inactive metabolite, Clopidogrel carboxylic acid 2 . The thiol metabolite binds with P2Y12 receptor of platelets irreversibly and inhibits its aggregation 3 .…”

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confidence: 99%

Untitled

2019

IJPSR

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Clopidogrel, a poorly water-soluble drug, has been the mainstay of platelet management in acute coronary syndrome (ACS) or percutaneous coronary intervention (PCI) for over a decade. Since Clopidogrel is a prodrug and requires hepatic metabolism for activation, it exhibits a slower onset of action upon oral administration. This time-lag before the onset on the action is highly undesirable for the treatment of critical conditions such as coronary intervention, which is a medical emergency during which the rapid onset of antiplatelet action is of paramount importance. To achieve rapid onset, intravenous is the preferred route of administration; however, due to poor aqueous solubility, development of the IV formulation of Clopidogrel presents a considerable challenge. In the present investigation, the liposomal formulation of Clopidogrel was developed, which enables IV administration and may potentially provide a rapid onset of action. The formulation was prepared and optimized using 3 2 full factorial designs with Design Expert 11 software and evaluated for critical quality attributes (% entrapment, particle size, PDI, zeta potential and Morphology, pH/dilution induced stability and short term stability studies. The particle size of optimized formulation was 94.5 ± 2.8 nm, with PDI of 0.126 ± 0.012 and entrapment efficiency (EE) of 89.2 ± 2.1. The developed formulation was stable over a study period of 3 months at 2°-8 °C. This formulation has great potential as rapid-acting IV formulation to fulfill the unmet need in the management of cardiovascular emergency like (ACS) and PCI.

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Evaluation of Clopidogrel Conjugation Metabolism: PK Studies in Man and Mice of Clopidogrel Acyl Glucuronide

Cited by 8 publications

References 21 publications

Mrp3 Transports Clopidogrel Acyl Glucuronide from the Hepatocytes into Blood

Mrp3 Transports Clopidogrel Acyl Glucuronide from the Hepatocytes into Blood

Human UGT2B7 is the major isoform responsible for the glucuronidation of clopidogrel carboxylate

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