CYTOTOXIC EFFECT AND PERMEABILITY ACTIVITIES OF CURCUMIN ANALOGUE; 2, 6-BIS (2, 5-DIMETHOXYBENZY-LIDENE) CYCLOHEXANONE (BDMC33) IN CACO-2 CELL MODEL
Abstract
Previously, curcumin analogue, 2, 6-bis (2, 5-dimethoxybenzylidene) cyclohexanone (BDMC33) with high anti-inflammatory activity was chemically synthesized in our laboratory to enhance the biological activity of curcumin. In this study, the toxicity and permeability activities of 2,6-bis(2,5-dimethoxybenzy-lidene)cyclohexanone (BDMC33) in Caco-2 cells was investigated. Toxicity effects using MTT assay and apparent permeability coefficient (Papp), uptake (UR) and efflux (ER) ratios, and mass balance of BDMC33 after permeation in Caco-2 cells for 180 min were evaluated in apical (A) to basolateral (B) and basolateral (B) to apical (A) directions. The similar analyses on 3-(2-fluoro-benzylidene)-5-(2-fluorocyclohexylmethylene)-piperidin-4-one; (EF-24) (check control) were also conducted. The 24 hr LC50 value for BDMC33 and EF-24 on Caco-2 cells were both 50 µM. The Papp value in A→B direction was 3.37 ± 0.47 cm/s (BDMC33) and 2.47 ± 0.15 cm/s (EF-24). Whereas in B→A direction, it was 1.9 ± 0.36 cm/s (BDMC33) and 1.8 ± 0.15 cm/s (EF-24) upon 120 min incubation. The UR and ER ratios calculated were 1.77% and 0.56%, respectively, and the mass balance calculated were 41-44% (BDMC33) and 31-34% (EF-24) in A→B and B→A direction. This study has suggested BDMC33 to be more absorbable than EF-24 in Caco-2 cells. Therefore, BDMC33 could be a leading feature, the anti-inflammatory agent, as it biological activities would be expected outside the intestine.
Keywords:
Apparent permeability coefficient, Caco-2 cells, Curcumin, Curcumin analogue, Efflux ratio, MTT assayDOI
https://doi.org/10.25004/IJPSDR.2015.070605References
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24. Yu H, Huang Q. Investigation of the absorption mechanism of solubilized curcumin using Caco-2 cell monolayers. Journal of Agriculture and Food Chemistry 2011; 59(17):9120-9126.
25. Artursson P, Karlsson J. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2). Biochemical and Biophysical Research Communications 1991; 175(3):880-885.
26. Hubatsch I, Ragnarsson EG, Artursson P. Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers. Nature protocols 2007; 2(9):2111-2119.
27. Shehzad A, Khan S, Shehzad O, Lee YS. Curcumin therapeutic promises and bioavailability in colorectal cancer. Drugs of Today 2010; 46(7): 523.
28. Yang KY, Lin LC, Tseng TY, Wang SC, Tsai TH. Oral bioavailability of curcumin in rat and the herbal analysis from Curcuma longa by LC-MS/MS. Journal of Chromatography B 2007; 853(1-2):183-189.
29. Marczylo TH, Verschoyle RD, Cooke DN, Morazzoni P, Steward WP, Gescher, AJ. Comparison of systemic availability of curcumin with that of curcumin formulated with phosphatidylcholine. Cancer Chemotherapy and Pharmacology 2007; 60(2):171-7.
30. Maubon N, Le Vee M, Fossati L, Audry M, Le Ferrec E, Bolze S, Fardel O. Analysis of drug transporter expression in human intestinal Caco‐2 cells by real‐time PCR. Fundamental and Clinical Pharmacology 2007; 21(6):659-663.
2. Goel A, Kunnumakkara AB, Aggarwal BB. Curcumin as “Curecumin’’: From kitchen to clinic. Biochemical Pharmacology 2008; 75:787-809.
3. Aggarwal BB, Harikumar BB. Potential therapeutic effects of curcumin, the anti-inflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. International Journal of Biochemistry and Cell Biology 2009; 41(1):40-59.
4. Yu H, Huang Q. Improving the oral bioavailability of curcumin using novel organogel-based nanoemulsions. Journal of Agriculture and Food Chemistry 2001; 260(21):5373-5379.
5. 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. Biochemical Pharmacology 2008; 76(11):1590-1611.
6. Anand P, Kunnumakkara AB, Newman RA. Aggarwal BB. Bioavailability of curcumin: Problems and promises. Molecular Pharmacology 2007; 4:807-818.
7. Cheng AL, Hsu CH, Lin JK, Hsu MM, Ho YF, Shen TS, Hsieh CY. Phase I clinical trial of curcumin, a chemopreventive agent, in patients with high-risk or pre-malignant lesions. Anticancer Research 2001; 21:2895-2900.
8. Wahlang B, Yogesh BP, Arvind KB. Identification of permeability-related hurdles in oral delivery of curcumin using the Caco-2 cell model. European Journal of Pharmaceutics and Biopharmaceutics 2011; 77(2):275-282.
9. Liang G, Li X, Chen L, Yang S, Wu X, Studer E, Zhou H. Synthesis and anti-inflammatory activities of mono-carbonyl analogue of curcumin. Bioorganic and Medical. Chemistry. Letters 2008; 18:1525-1529.
10. Wang J, Ao G, Chu X, Ji Y. Antioxidant Properties and PC12 Cell Protective Effects of a Novel Curcumin Analogue (2E,6E)-2,6-Bis(3,5 dimethoxybenzylidene) cyclohexanone (MCH). International Journal of Molecular Sciences 2014; 15: 3970-3988; doi: 10.3390/ijms15033970
11. Lee KH, Aziz FHA, 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. European Journal Medical Chemistry 2009; 44:3195-3200.
12. Lee KH, Abas F, Alitheen NBM, 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. Molecules 2011; 16:9728-9738.
13. 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. International Journal of Molecular Sciences 2012; 13:2985-3008.
14. 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.
15. Yee S. In vitro permeability across Caco-2 cells (colonic) can predict in vivo (small intestinal) absorption in man–factor myth. Pharmaceutical Research 1997; 14:763-766.
16. Press B, Di Grandi D. Permeability for intestinal absorption: Caco-2 assay and related issues. Current Drug Metabolism 2008; 9:893-900.
17. Dempe JS, Scheerle RK, Pfeiffer E, Metzler M. Metabolism and permeability of curcumin in cultured Caco-2 cells. Molecular Nutrition and Food Research 2013; 57:1543-1549, doi: 10.1002/mnfr.201200113.
18. Sumanasekera WK, Ivanova MM, Johnston BJ, Dougherty SM, Sumanasekera GU, Myers SR, Klinge CM. Rapid effects of diesel exhaust particulate extracts on intracellular signaling in human endothelial cells. Toxicological Letters 2007a; 174(1-3):61-73.
19. Sumanasekera WK, Sumanasekera GU, Mattingly KA, Dougherty SM, Keynton RS, Klinge CM. Estradiol and dihydrotestosterone regulate endothelial cell barrier function after hypergravity-induced alterations in MAPK activity. American Journal of Cell Physiology 2007b; 293(2):566-573.
20. Artursson P, Palm K, Luthman K. Caco-2 monolayers in experimental and theoretical predictions of drug transport. Advanced drug delivery Review 2001; 64:280-289.
21. Metzler M, Pfeiffer E, Schulz SI, Dempe JS. Curcumin uptake and metabolism. Biofactors 2013; 39(1):14-20.
22. Wang YJ, Pan MH, Cheng AL, Lin LI, Ho YS, Hsieh CY, Lin JK. Stability of curcumin in buffer solutions and characterization of its degradation products. Journal of Pharmaceutical and Biomedical Analysis 1997; 15:1867-1876.
23. Hou XL, Takahashi K, Tanaka K, Tougou K, Qiu F, Komatsu K, Azuma J. Curcuma drugs and curcumin regulate the expression and function of P-gp in Caco-2 cells in completely opposite ways. International Journal of Pharmacology 2008; 358(1-2):224-229.
24. Yu H, Huang Q. Investigation of the absorption mechanism of solubilized curcumin using Caco-2 cell monolayers. Journal of Agriculture and Food Chemistry 2011; 59(17):9120-9126.
25. Artursson P, Karlsson J. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2). Biochemical and Biophysical Research Communications 1991; 175(3):880-885.
26. Hubatsch I, Ragnarsson EG, Artursson P. Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers. Nature protocols 2007; 2(9):2111-2119.
27. Shehzad A, Khan S, Shehzad O, Lee YS. Curcumin therapeutic promises and bioavailability in colorectal cancer. Drugs of Today 2010; 46(7): 523.
28. Yang KY, Lin LC, Tseng TY, Wang SC, Tsai TH. Oral bioavailability of curcumin in rat and the herbal analysis from Curcuma longa by LC-MS/MS. Journal of Chromatography B 2007; 853(1-2):183-189.
29. Marczylo TH, Verschoyle RD, Cooke DN, Morazzoni P, Steward WP, Gescher, AJ. Comparison of systemic availability of curcumin with that of curcumin formulated with phosphatidylcholine. Cancer Chemotherapy and Pharmacology 2007; 60(2):171-7.
30. Maubon N, Le Vee M, Fossati L, Audry M, Le Ferrec E, Bolze S, Fardel O. Analysis of drug transporter expression in human intestinal Caco‐2 cells by real‐time PCR. Fundamental and Clinical Pharmacology 2007; 21(6):659-663.
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01-11-2015
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“CYTOTOXIC EFFECT AND PERMEABILITY ACTIVITIES OF CURCUMIN ANALOGUE; 2, 6-BIS (2, 5-DIMETHOXYBENZY-LIDENE) CYCLOHEXANONE (BDMC33) IN CACO-2 CELL MODEL”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 7, no. 6, Nov. 2015, pp. 465-73, https://doi.org/10.25004/IJPSDR.2015.070605.
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“CYTOTOXIC EFFECT AND PERMEABILITY ACTIVITIES OF CURCUMIN ANALOGUE; 2, 6-BIS (2, 5-DIMETHOXYBENZY-LIDENE) CYCLOHEXANONE (BDMC33) IN CACO-2 CELL MODEL”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 7, no. 6, Nov. 2015, pp. 465-73, https://doi.org/10.25004/IJPSDR.2015.070605.
