EFFECTS OF CURCUMIN ANALOGUE, 2, 6-BIS (2, 5-DIMETHOXYBENZYLIDENE) CYCLOHEXANONE (BDMC33) ON THE ACTIVITIES OF DRUG-METABOLIZING ENZYMES IN CULTURED CACO-2 CELL MODEL
Abstract
Poor systemic delivery of curcumin outside the gut due to its rapid metabolism has severely limited its application to many chronic diseases. Previously, our research group synthesized curcumin analogues 2, 6-bis (2, 5-dimethoxybenzylidene) cyclohexanone (BDMC33) that has potent anti-inflammatory activities. Therefore, the aim of this study is to evaluate the effects of curcumin analog (BDMC33) on the activities of drug metabolizing enzymes in Caco-2 cells, which was compared with that of curcumin and 3-(2-Fluoro-benzylidene)-5-(2-fluorocyclohexylmethylene)-piperidin-4-one (EF-24). BDMC-33 was synthesized through the appropriate reaction of the aromatic aldehyde with cyclohexanone, under base catalyzed aldol condensation, at the ratio of ketone: aldehyde (1:2). Activity of drug metabolizing enzymes such as NADPH-cytochrome p450 reductase (CPR), UDP-glucuronosyltransferase (UGT), glutathione-S-transferase (GST) and Sulfotransferase (SULT) in Caco-2 cells were evaluated upon exposure to 50µM of BDMC33, curcumin, and EF-24, separately, for 4 hours. The BDMC33, EF-24, and curcumin treatments did not affect the activities of UGT, GST, SULT, and CPR in respect to their controls (29.45, 27.18, 23.64 and 2.08µmol/mg), respectively, at all periods of incubation. Hence, BDMC33 was able to maintain the activities of both phases I and II drug metabolizing enzymes, and therefore it could be a potential lead, anti-inflammatory agents.
Keywords:
Caco-2 cells, curcumin, drug metabolizing enzymes, glutathione-S-transferase, NADPH-cytochrome c reductase, sulfotransferase, UDP-glucuronosyltransferaseDOI
https://doi.org/10.25004/IJPSDR.2016.080109References
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28. Yu H, Huang Q Investigation of the absorption mechanism of solubilized curcumin using Caco-2 cell monolayers. J. Agric. Food Chem 2011; 59 (17):9120-9126.
29. Volak LP, Ghirmai S, Cashman JR Curcuminoids inhibit multiple human cytochromes P450, UDP-glucuronosyltransferase, and sulfotransferase enzymes, whereas piperine is a relatively selective CYP3A4 inhibitor. Drug Metab. Disp., 2008; 36(8):1594-1605.
30. Coleman MD. editor Human Drug Metabolism: An Introduction. England: John Wiley and Sons Ltd. 2005, pp. 24-26.
31. Odenthal J, van Heumen BW, Roelofs HM, te Morsche RH, Marian B, Nagengast FM, Peters WH. The influence of curcumin, quercetin, and eicosapentaenoic acid on the expression of phase II detoxification enzymes in the intestinal cell lines HT-29, Caco-2, HuTu 80, and LT97. Nutr. Cancer, 2012; 64(6):856-863.
32. Tukey RH, Strassburg CP. Human UDPglucuronosyltransferases: metabolism, expression, and disease. Annual Rev. Pharmacol. Toxicol. 2000; 40:581-616.
33. Talalay P. Chemoprotection against cancer by induction of phase 2 enzymes. Biofactors 2000; 12:5-11.
34. Van der Logt EMJ, Bergevoet SM, Roelofs HMJ, Van Hooijdonk Z, Te Morsche RHM, Wobbes T, Peters WHM. Genetic polymorphisms in UDPglucuronosyltransferases and glutathione Anticarcinogens and detoxification in intestinal tumor cell lines S-transferases and colorectal cancer risk. Carcinogenesis, 2004; 25:2407-2415.
35. Berkhout M, Roelofs HMJ, te Morsche RHM, den Dekker E., Van Krieken JHJM, Nagengast FM, Peters WHM. Detoxification enzyme polymorphisms are not involved in duodenal adenomatosis in familial adenomatous polyposis. Brazilian J Surg. 2008; 95:499-505.
36. Aggarwal BB, Harikumar BB. Potential therapeutic effects of curcumin, the anti-inflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int. J. Biochem. Cell Biol 2009; 41 (1):40-5920.
37. Wang Z, Hasegawa J, Wang X, Lin LI, Ho YS, Hsieh CY, Lin JK. Protective effects of ginger against aspirin-induced gastric ulcers in rats. Yonago acta medica. 2010; 54(1):11.
38. Jiwapornkupt N, Niwattisaiwong N, Phivthong-ngam L, Piyachaturawat P, Suksamran A, Apipalakul K, Lawanprasert S. Effects of Curcuma comosa extracts on phase II drug-metabolizing enzymes in rat livers. J Health Res, 2010; 24(3):113-116.
39. Shao J, Sheng H, Inoue H, Morrow JD, DuBois RN. Regulation of constitutive cyclooxygenase-2 expression in colon carcinoma cells. J Biol. Chem. 2000; 275:33951-6.
40. Jurek D, Fleckl E, Marian B. Bile acid induced gene expression in LT97 colonic adenoma cells. Food Chem. Toxicol. 2005; 43:87-93.
41. Maier TJ, Schilling K, Schmidt R, Geisslinger G, Grösch S. Cyclooxygenase-2 (COX- 2)-dependent and –independent anticarcinogenic effects of celecoxib in human colon carcinoma cells. Biochem. Pharmacol. 2004; 67:1469-78.
2. Goel A, Kunnumakkara AB, and Aggarwal BB. Curcumin as “Curecumin’’: From kitchen to clinic. Biochem. Pharmacol 2008; 75: 787-809.
3. Anand P, Thomas SG, Kunnumakkara AB, Sundaram C, Harikumar KB, Sung B, Aggarwal BB. Biological activities of curcumin and its analogues (Congeners) made by man and Mother Nature. Biochem. Pharmacol. 2008; 76 (11):1590-1611.
4. Wang YJ, Pan MH, Cheng AL, Lin LI, Ho YS, Hsieh CY, Lin JK. Stability of curcumin in buffer solutions and characterization ot its degradation products. J Pharmaceut. Biomed. Anal. 1997; 15:1867-1876.
5. Pan MH, Huang TM, Lin JK Biotransformation of curcumin through reduction and glucuronidation in mice. Drug Metab. Disp. 1999; 27 (4):486-94.
6. Wahlang B, Yogesh BP, Arvind KB. Identification of permeability-related hurdles in oral delivery of curcumin using the Caco-2 cell model. Eur. J. Pharm. Biopharm. 2011; 77 (2):275-282.
7. Dempe JS, Scheerle RK, Pfeiffer E, Metzler M. Metabolism and permeability of curcumin in cultured Caco-2 cells. Mol. Nutr. Food Res 2012; 57:1543-1549.
8. Thomas SL, Zhong D, Zhou W Malik S, Liotta D, Snyder JP, Giannakakou P. EF24, a novel curcumin analog, disrupts the microtubule cytoskeleton and inhibits HIF-1. Cell Cycle 2008; 7 (15):2409-17.
9. Steward WP, Gescher AJ. Curcumin in cancer management: Recent results of Analogue design and clinical studies and desirable future research. Molecular Nutrition and Food Res. 2008; 52:1005-1009.
10. Aggarwal R, Goel SK, Behari JR. Detoxification and antioxidant effects of curcumin in rats experimentally exposed to mercury. J App Toxicol. 2010; 30(5):457-468.
11. Metzler M, Pfeiffer E, Schulz SI, Dempe JS. Curcumin uptake and metabolism. Biofactors. 2013; 39(1):14-20.
12. Lee KH, Abd Aziz FH, Syahida A, Abas F, Shaari K, Israf DA, Lajis NH. Synthesis and biological evaluation of curcumin-like diarylpentanoid analogues for anti-inflammatory, antioxidant and anti-tyrosinase activities. Eur. J. Med. Chem 2009; 44:3195–3200.
13. Lee KH, Abas F, Alitheen NB, Shaari K, Lajis NH, Ahmad S. Curcumin Derivative, 2, 6-Bis(2, 5-dimethoxybenzylidene)- cyclohexanone (BDMC33) Attenuates Prostaglandin E2 Synthesis via Selective Suppression of Cyclooxygenase-2 in IFN-/LPS Stimulated Macrophages. Mol. 2011; 16:9728-9738.
14. Lee KH, Chow YL, Sharmili V, Abas F, Alitheen NBM, Shaari K, Syahida A. BDMC33, a curcumin derivative suppresses inflammatory responses in macrophage-like cellular system: Role of inhibition in NF-κB and MAPK signaling pathways. Int. J. Mol. Sci. 2012; 13:2985–3008.
15. Yee S. In vitro permeability across Caco-2 cells (colonic) can predict in vivo (small intestinal) absorption in man–factor myth. Pharm. Res 1997; 14:763–766.
16. Hung MW, Zhang ZJ, Li S, Lei B, Yuan S, Cui GZ, Lee SMY. From omics to drug metabolism and high content screen of natural product in zebrafish: a new model for discovery of neuroactive compound. Evidence-Based Complementary and Alternative Medicine, 2012; 1-20
17. Meunier V, Bourrie M, Berger, Fabre G. The human intestinal epithelial cell line Caco-2; pharmacological and pharmacokinetic applications. Cell Biol. Toxicol. 1995; 11 (3-4):187 -194.
18. Press B, Di Grandi D. Permeability for intestinal absorption: Caco-2 assay and related issues. Curr. Drug. Metab 2008; 9:893-900.
19. Bradford MM. A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976; 72:248-254.
20. Bock KW, Brian B, Geoffrey JD, Hänninen O, Mulder GJ, Owens IS, Tephly TR. UDP-glucuronosyltransferase activities: guidelines for consistent interim terminology and assay conditions. Biochem. Pharmacol. 1983; 32 (6):953-955.
21. Luquita MG, Sánchez Pozzi EJ, Catania VA, Mottino AD. Analysis of p-nitrophenol glucuronidation in hepatic microsomes from lactating rats. Biochem. Pharmacol. 1994; 47:1179-1185.
22. Habig WH Pabst MJ, Jakoby WN. Glutathione-s transferase: The first enzyme step in Mercapturic acid formation. J Bio. Chem. 1974; 249: 7130-7139.
23. Frame LT, Ozawa S, Nowell SA, Chou HC, DeLongchamp RR, Doerge DR, Kadlubar FF. A simple colorimetric assay for phenotyping the major human thermostable phenol sulfotransferase (SULT1A1) using platelet cytosols. Drug Metab. Disp. 2000; 28:1063-8.
24. Iwata H, Fujita KL, Kushida H, et al. High catalytic activity of human cytochrome P450 co-expressed with human NADPH-cytochrome P450 reductase in Escherichia coli. Biochem. Pharmacol. 1998; 55(8):1315-1325.
25. Yamazaki H, Ueng YF, Shimada T, Guengerich FP. Roles of divalent metal ions in oxidations catalyzed by recombinant cytochrome P450 3A4 and replacement of NADPH-cytochrome P450 reductase with other flavoproteins, ferredoxin, and oxygen surrogates. Biochem. 1995; 34(26): 8380-8389.
26. Gibson GG, Skett P, editors. Introduction to drug metabolism. 3rd edn. New York: Chapman. 2001, pp. 30-35.
27. Kittichanun C, Phivthong-ngam L, Niwattisaiwong N, Piyachaturawat P, Suksamran A, Apipalakul K, Sotharak W. Effects of curcuma comosa extract on hepatic cytochrome P450 activities in rats. J Health Res. 2010; 24(1):1-6.
28. Yu H, Huang Q Investigation of the absorption mechanism of solubilized curcumin using Caco-2 cell monolayers. J. Agric. Food Chem 2011; 59 (17):9120-9126.
29. Volak LP, Ghirmai S, Cashman JR Curcuminoids inhibit multiple human cytochromes P450, UDP-glucuronosyltransferase, and sulfotransferase enzymes, whereas piperine is a relatively selective CYP3A4 inhibitor. Drug Metab. Disp., 2008; 36(8):1594-1605.
30. Coleman MD. editor Human Drug Metabolism: An Introduction. England: John Wiley and Sons Ltd. 2005, pp. 24-26.
31. Odenthal J, van Heumen BW, Roelofs HM, te Morsche RH, Marian B, Nagengast FM, Peters WH. The influence of curcumin, quercetin, and eicosapentaenoic acid on the expression of phase II detoxification enzymes in the intestinal cell lines HT-29, Caco-2, HuTu 80, and LT97. Nutr. Cancer, 2012; 64(6):856-863.
32. Tukey RH, Strassburg CP. Human UDPglucuronosyltransferases: metabolism, expression, and disease. Annual Rev. Pharmacol. Toxicol. 2000; 40:581-616.
33. Talalay P. Chemoprotection against cancer by induction of phase 2 enzymes. Biofactors 2000; 12:5-11.
34. Van der Logt EMJ, Bergevoet SM, Roelofs HMJ, Van Hooijdonk Z, Te Morsche RHM, Wobbes T, Peters WHM. Genetic polymorphisms in UDPglucuronosyltransferases and glutathione Anticarcinogens and detoxification in intestinal tumor cell lines S-transferases and colorectal cancer risk. Carcinogenesis, 2004; 25:2407-2415.
35. Berkhout M, Roelofs HMJ, te Morsche RHM, den Dekker E., Van Krieken JHJM, Nagengast FM, Peters WHM. Detoxification enzyme polymorphisms are not involved in duodenal adenomatosis in familial adenomatous polyposis. Brazilian J Surg. 2008; 95:499-505.
36. Aggarwal BB, Harikumar BB. Potential therapeutic effects of curcumin, the anti-inflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int. J. Biochem. Cell Biol 2009; 41 (1):40-5920.
37. Wang Z, Hasegawa J, Wang X, Lin LI, Ho YS, Hsieh CY, Lin JK. Protective effects of ginger against aspirin-induced gastric ulcers in rats. Yonago acta medica. 2010; 54(1):11.
38. Jiwapornkupt N, Niwattisaiwong N, Phivthong-ngam L, Piyachaturawat P, Suksamran A, Apipalakul K, Lawanprasert S. Effects of Curcuma comosa extracts on phase II drug-metabolizing enzymes in rat livers. J Health Res, 2010; 24(3):113-116.
39. Shao J, Sheng H, Inoue H, Morrow JD, DuBois RN. Regulation of constitutive cyclooxygenase-2 expression in colon carcinoma cells. J Biol. Chem. 2000; 275:33951-6.
40. Jurek D, Fleckl E, Marian B. Bile acid induced gene expression in LT97 colonic adenoma cells. Food Chem. Toxicol. 2005; 43:87-93.
41. Maier TJ, Schilling K, Schmidt R, Geisslinger G, Grösch S. Cyclooxygenase-2 (COX- 2)-dependent and –independent anticarcinogenic effects of celecoxib in human colon carcinoma cells. Biochem. Pharmacol. 2004; 67:1469-78.
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01-01-2016
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“EFFECTS OF CURCUMIN ANALOGUE, 2, 6-BIS (2, 5-DIMETHOXYBENZYLIDENE) CYCLOHEXANONE (BDMC33) ON THE ACTIVITIES OF DRUG-METABOLIZING ENZYMES IN CULTURED CACO-2 CELL MODEL”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 8, no. 1, Jan. 2016, pp. 57-64, https://doi.org/10.25004/IJPSDR.2016.080109.
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How to Cite
“EFFECTS OF CURCUMIN ANALOGUE, 2, 6-BIS (2, 5-DIMETHOXYBENZYLIDENE) CYCLOHEXANONE (BDMC33) ON THE ACTIVITIES OF DRUG-METABOLIZING ENZYMES IN CULTURED CACO-2 CELL MODEL”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 8, no. 1, Jan. 2016, pp. 57-64, https://doi.org/10.25004/IJPSDR.2016.080109.
