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Research articleFood Biotechnology (FB) | Bioactive compounds and Physiological properties
Two flavonoid-based compounds from Murraya paniculata as novel human carbonic anhydrase isozyme II inhibitors detected by a resazurin yeast-based assayAnyaporn Sangkaew 1, Nawara Samritsakulchai 2, Kamonpan Sanachai 3, Thanyada Rungrotmongkol 3, 4, Warinthorn Chavasiri 2 and Chulee Yompakdee *
1Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, 2Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, 3Structural and Computational Biology Research Unit, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok10330, Thailand, 4Structural and Computational Biology Research Unit, Department of Biochemistry and Program in Bioinformatics and Computational Biology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, 5Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, 6Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok 10330, ThailandReceived: October 17, 2019; Accepted: December 27, 2019
J. Microbiol. Biotechnol. 2020; 30(4): 552-560
Published April 28, 2020
Copyright © The Korean Society for Microbiology and Biotechnology.
AbstractHuman carbonic anhydrase isozyme II has been used as protein target for disorder treatment including glaucoma. Current clinically used sulfonamide-based CA inhibitors can induce side effects, and so alternatives are required. This study aimed to investigate a natural CA inhibitor from Murraya paniculata. The previously developed yeast-based assay was used to screen 14 compounds isolated from M. paniculata and identified by NMR analysis for anti-human CA isozyme II (hCAII) activity. Cytotoxicity of the compounds was also tested using the same yeast-based assay but in a different cultivation condition. Two flavonoid candidate compounds, 5, 6, 7, 8, 3, 4, 5-heptamethoxyflavone (4) and 3 ,5, 7, 8, 3, 4, 5-heptamethoxyflavone (9), showed potent inhibitory activity against hCAII with a minimal effective concentration of 10.8 and 21.5 µM, respectively, while they both exhibited no cytotoxic effect even at the highest concentration tested (170 µM). The results from an in vitro esterase assay of the two candidates confirmed their hCAII inhibitory activity with IC50 values of 24.0 and 34.3 µM, respectively. To investigate the potential inhibition mechanism of compound 4, in silico molecular docking was performed using the FlexX and Swissdock software. This revealed that compound 4 coordinated with the Zn2+ ion in the hCAII active site through its methoxy oxygen at a distance of 1.60 Å (FlexX) or 2.29 Å (Swissdock). The interaction energy of compound 4 with hCAII was -13.36 kcal/mol. Thus, compound 4 is a potent novel flavonoid-based hCAII inhibitor and may be useful for further anti-CAII design and development.
KeywordsHuman carbonic anhydrase isozyme II (hCAII), carbonic anhydrase inhibitor, glaucoma, murraya paniculata, yeast-based assay, flavonoid
- Song P, Wang J, Bucan K, Theodoratou E, Rudan I, Chan KY. 2017. National and subnational prevalence and burden of glaucoma in China: a systematic analysis. J. Glob. Health 7: 020705.
- Liang YB, Zhang Y, Musch DC, Congdon N. 2017. Proposing new indicators for glaucoma healthcare service. Eye Vis. (Lond) 4: 6.
- Varma R, Lee PP, Goldberg I, Kotak S. 2011. An assessment of the health and economic burdens of glaucoma. Am. J. Ophthalmol. 152:515-522.
- Goel M, Picciani RG, Lee RK, Bhattacharya SK. 2010. Aqueous humor dynamics: a review. Open Ophthalmol. J. 4: 52-59.
- Sly WS, Hu PY. 1995. Human carbonic anhydrases and carbonic anhydrase deficiencies. Annu. Rev. Biochem. 64: 375-401.
- Supuran CT. 2008. Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat. Rev. Drug Discov. 7: 168181.
- Supuran CT, Scozzafava A. 2000. Carbonic anhydrase inhibitors and their therapeutic potential. Expert Opin. Ther. Pat. 10: 575-600.
- Silver LH. 1998. Clinical efficacy and safety of brinzolamide (Azopt), a new topical carbonic anhydrase inhibitor for primary openangle glaucoma and ocular hypertension. Brinzolamide Primary Therapy Study Group. Am. J. Ophthalmol. 126: 400-408.
- Scozzafava A, Supuran CT. 2014. Glaucoma and the applications of carbonic anhydrase inhibitors. Subcell Biochem. 75: 349-359.
- Schmidl D, Schmetterer L, Garhofer G, Popa-Cherecheanu A. 2015. Pharmacotherapy of glaucoma. J. Ocul. Pharmacol. Ther. 31: 6377.
- Kelly TE, Hackett PH. 2010. Acetazolamide and sulfonamide allergy: a not so simple story. High Alt. Med. Biol. 11: 319-323.
- Guedes GB, Karan A, Mayer HR, Shields MB. 2013. Evaluation of adverse events in self-reported sulfa-allergic patients using topical carbonic anhydrase inhibitors. J. Ocul. Pharmacol. Ther. 29: 456-461.
- Macy E, Poon KYT. 2009. Self-reported antibiotic allergy incidence and prevalence: age and sex effects. Am. J. Med. 122: 778 e771777.
- Lomelino C, Supuran C, McKenna R. 2016. Non-Classical Inhibition of Carbonic Anhydrase. Int. J. Mol. Sci. 17: 1150.
- Bilsland E, Sparkes A, Williams K, Moss HJ, de Clare M, Pir P, et al. 2013. Yeast-based automated high-throughput screens to identify anti-parasitic lead compounds. Open Biol. 3: 120158.
- Barberis A, Gunde T, Berset C, Audetat S, Luthi U. 2005. Yeast as a screening tool. Drug Discov. Today Technol. 2: 187-192.
- Ann Bjornsti M. 2002. Cancer therapeutics in yeast. Cancer Cell 2: 267-273.
- Sangkaew A, Krungkrai J, Yompakdee C. 2018. Development of a high throughput yeast-based screening assay for human carbonic anhydrase isozyme II inhibitors. AMB Express. 8: 124.
- Li R. 2016. Natural Product-Based Drug Discovery. Med. Res. Rev. 36: 3.
- J. Mabberley D. 2016. The typification of Murraya, M. exotica, and M. paniculata (Rutaceae): its significance for the world citrus industry. Taxon. 65: 366-371.
- Ito C, Furukawa H. 1987. Constituents of Murraya exotica L. Structure Elucidation of New Coumarins. Chem. Pharm. Bull (Tokyo). 35: 4277-4285.
- Barik BR, Dey AK, Das PC, Chatterjee A, Shoolery JN. 1983. Coumarins of Murraya exotica—absolute configuration of auraptenol. Phytochemistry 22: 792-794.
- Shan J, Wang XZ, Ma YD, Yang RJ, Li XW, Jin YR. 2010. Studies on flavonoids from leaves of Murraya panaculata L. (I). Chin Pharm. J. 45: 1910-1912.
- Zhang Y, Li J, Zhou SX, Tu PF. 2010. Polymethoxylated flavonoids from the leaves of Murraya paniculata. Chin. Pharm. J. 45: 11391141.
- Chen C-H, Chan H-C, Chu Y-T, Ho H-Y, Chen P-Y, Lee T-H, et al. 2009. Antioxidant Activity of Some Plant Extracts Towards Xanthine Oxidase, Lipoxygenase and Tyrosinase. Molecules 14: 2947-2958.
- Gautam M, Gangwar M, Singh A, Rao C, Goel R. 2012. In-vitro Antioxidant properties of Murraya paniculata (L.) leaves extract. Inventi Rapid: Ethnopharmacology 2012: 1-3.
- Saeed S, Shah S, Mehmood R, Malik A. 2011. Paniculacin, a new coumarin derivative from Murraya paniculata. J. Asian Nat. Prod Res. 13: 724-727.
- Gautam M, Singh A, Rao C, Goel R. 2012. Toxicological evaluation of murraya paniculata (L.) leaves extract on rodents. AJPT. 7: 6267.
- Gautam M, Gangwar M, Nath G, Rao C, K Goel R. 2012. In-vitro antibacterial activity on human pathogens and total phenolic, flavonoid contents of Murraya paniculata Linn. leaves. Asian Pac. J. Trop Biomed. 2: S1660–S1663.
- Menezes IR, Santana TI, Varela VJ, Saraiva RA, Matias EF, Boligon AA, et al. 2015. Chemical composition and evaluation of acute toxicological, antimicrobial and modulatory resistance of the extract of Murraya paniculata. Pharm. Biol. 53: 185-191.
- Rodanant P, Khetkam P, Suksamrarn A, Kuvatanasuchati J. 2015. Coumarins and flavonoid from Murraya paniculata (L.) Jack:antibacterial and anti-inflammation activity. Pak J. Pharm. Sci. 28: 1947-1951.
- Lv H-N, Wang S, Zeng K-W, Li J, Guo X-Y, Ferreira D, et al. 2015. Anti-inflammatory coumarin and benzocoumarin derivatives from Murraya alata. J. Nat. Prod. 78: 279-285.
- Davis RA, Vullo D, Maresca A, Supuran CT, Poulsen SA. 2013. Natural product coumarins that inhibit human carbonic anhydrases. Bioorgan. Med. Chem. 21: 1539-1543.
- Karatas MO, Alici B, Cakir U, Cetinkaya E, Demir D, Ergun A, et al. 2014. New coumarin derivatives as carbonic anhydrase inhibitors. Artif. Cells Nanomed. Biotechnol. 42: 192-198.
- Pustenko A, Stepanovs D, Zalubovskis R, Vullo D, Kazaks A, Leitans J, et al. 2017. 3H-1,2-benzoxathiepine 2,2-dioxides: a new class of isoform-selective carbonic anhydrase inhibitors. J. Enzyme. Inhib. Med. Chem. 32: 767-775.
- Karatas MO, Uslu H, Sari S, Alagoz MA, Karakurt A, Alici B, et al. 2016. Coumarin or benzoxazinone based novel carbonic anhydrase inhibitors: synthesis, molecular docking and anticonvulsant studies. J. Enzyme. Inhib. Med. Chem. 31: 760-772.
- Huyut Z, Beydemir Ş, Gülçin İ. 2017. Inhibition properties of some flavonoids on carbonic anhydrase I and II isoenzymes purified from human erythrocytes. J. Biochem. Mol. Toxicol. 31: e21930.
- Ekinci D, Karagoz L, Ekinci D, Senturk M, Supuran CT. 2013. Carbonic anhydrase inhibitors: in vitro inhibition of alpha isoforms (hCA I, hCA II, bCA III, hCA IV) by flavonoids. J. Enzyme Inhib. Med. Chem.. 28: 283-288.
- Rarey M, Kramer B, Lengauer T, Klebe G. 1996. A fast flexible docking method using an incremental construction algorithm. J. Mol. Biol. 261: 470-489.
- Grosdidier A, Zoete V, Michielin O. 2011. SwissDock, a protein-small molecule docking web service based on EADock DSS. Nucleic Acids Res. 39: W270-277.
- Petersson GA, Malick DK, Wilson WG, Ochterski JW, Montgomery JA, Frisch MJ. 1998. Calibration and comparison of the Gaussian-2, complete basis set, and density functional methods for computational thermochemistry. J. Chem. Phys. 109: 1057010579.
- Grosdidier A, Zoete V, Michielin O. 2007. EADock: docking of small molecules into protein active sites with a multiobjective evolutionary optimization. Proteins 67: 1010-1025.
- Barken FM, Gasteiger EL. 1980. Excitability of a penicillin-induced cortical epileptic focus. Exp. Neurol. 70: 539-547.
- Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, et al. 2004. UCSF chimera - A visualization system for exploratory research and analysis. J. Comput. Chem. 25: 1605-1612.
- Zhong HA. 2013. ADME and toxicity in early drug discovery. Curr. Top Med. Chem. 13: 1255-1256.
- Aguilera J, Van Dijken JP, De Winde JH, Pronk JT. 2005. Carbonic anhydrase (Nce103p): an essential biosynthetic enzyme for growth of Saccharomyces cerevisiae at atmospheric carbon dioxide pressure. Biochem. J. 391: 311-316.
- Polat MF, Nalbantolu B. 2002. In vitro esterase activity of carbonic anhydrase on total esterase activity level in serum. Turk J. Med. Sci. 32: 299-302.
- Balaydin HT, Durdagi S, Ekinci D, Senturk M, Goksu S, Menzek A. 2012. Inhibition of human carbonic anhydrase isozymes I, II and VI with a series of bisphenol, methoxy and bromophenol compounds. J. Enzyme Inhib. Med. Chem. 27: 467-475.
- Ghorab MM, Alsaid MS, Ceruso M, Nissan YM, Supuran CT. 2014. Carbonic anhydrase inhibitors: synthesis, molecular docking, cytotoxic and inhibition of the human carbonic anhydrase isoforms I, II, IX, XII with novel benzenesulfonamides incorporating pyrrole, pyrrolopyrimidine and fused pyrrolopyrimidine moieties. Bioorg. Med. Chem. 22: 3684-3695.
- Abuelizz H, El Dib R, Marzouk M, Anouar EH, A. Maklad Y, N. Attia H, et al. 2017. Molecular docking and anticonvulsant activity of newly synthesized quinazoline derivatives. Molecules. 22: 1094.
- Sethi KK, Verma SM, Tanc M, Purper G, Calafato G, Carta F, et al. 2014. Carbonic anhydrase inhibitors: synthesis and inhibition of the human carbonic anhydrase isoforms I, II, IX and XII with benzene sulfonamides incorporating 4- and 3-nitrophthalimide moieties. Bioorg. Med. Chem. 22: 1586-1595.
- Durdagi S, Korkmaz N, Isik S, Vullo D, Astley D, Ekinci D, et al. 2016. Kinetic and docking studies of cytosolic/tumor-associated carbonic anhydrase isozymes I, II and IX with some hydroxylic compounds. J. Enzyme Inhib. Med. Chem. 31: 1214-1220.
- Ekhteiari Salmas R, Mestanoglu M, Durdagi S, Senturk M, Kaya AA, Kaya EC. 2016. Kinetic and in silico studies of hydroxy-based inhibitors of carbonic anhydrase isoforms I and II. J. Enzyme Inhib. Med. Chem. 31: 31-37.
- Kinoshita T, Shimada M. 2002. Isolation and structure elucidation of a new prenylcoumarin from Murraya paniculata var. omphalocarpa (Rutaceae). Chem. Pharm. Bull (Tokyo). 50: 118-120.
- Xin-Jia Y, Wei L, Ying Z, Ning C, Ying X, Jian W, et al. 2016. A New biphenyl neolignan from Leaves of Patrinia villosa (Thunb.) Juss. Pharmacogn. Mag. 12: 1-3.
- Kinoshita T, Jin-Bin W, Feng-Chi H. 1996. The isolation of a prenylcoumarin of chemotaxonomic significance from Murraya paniculata var. omphalocarpa. Phytochemistry 43: 125-128.
- Nour AMM, Khalid SA, Kaiser M, Brun R, Abdalla WlE, Schmidt TJ. 2010. The antiprotozoal activity of methylated flavonoids from Ageratum conyzoides L. J. Ethnopharmacol. 129: 127-130.
- Tantishaiyakul V, Pummangura S, Chaichantipyuth C, Ma WW, McLaughlin JL. 1986. Phebalosin from the bark of Micromelum minutum. J. Nat. Prod. 49: 180-181.
- Ferracin RJ, das G.F. da Silva MF, Fernandes JB, Vieira PC. 1998. Flavonoids from the fruits of Murraya paniculata. Phytochemistry 47: 393-396.
- Longhuo Wu JL, Xiaohua Guo, Hao Huang, Haibo Hu, Rui Zhang. 2013. Chondroprotective evaluation of two natural coumarins:murrangatin and murracarpin. J. Intercult. Ethnopharmacol. 2: 91-98.
- Kinoshita T, Firman K. 1997. Myricetin 5,7,3’,4’,5’-pentamethyl ether and other methylated flavonoids from Murraya paniculata. Phytochemistry 45: 179-181.
- Karioti A, Ceruso M, Carta F, Bilia AR, Supuran CT. 2015. New natural product carbonic anhydrase inhibitors incorporating phenol moieties. Bioorg. Med. Chem. 23: 7219-7225.
- Wu T-S, Liou M-J, Kuoh C-S. 1989. Coumarins of the flowers of Murraya paniculata. Phytochemistry 28: 293-294.