Investigation of Mammal Arginase Inhibitory Properties of Natural Ubiquitous Polyphenols by Using an Optimized Colorimetric Microplate Assay

Abstract

Polyphenols are plant secondary metabolites which possess many positive effects on human health. Although these beneficial effects could be mediated through an increase in nitric oxide synthase activity, little is known regarding the inhibitory effect of polyphenols on mammal arginase, an enzyme which competes with nitric oxide synthase for their common substrate, L-arginine. The aim of the present study was to determine the potential of a series of polyphenols as mammalian arginase inhibitors and to identify some structure-activity relationships. For this purpose, we first developed a simple and cost-effective in vitro colorimetric microplate method using commercially-available mammal bovine liver arginase (b-ARG 1). Among the ten tested polyphenolic compounds [chlorogenic acid, piceatannol, resveratrol, (−)-epicatechin, taxifolin, quercetin, fisetin, caffeic acid, quinic acid, and kaempferol], cholorogenic acid and piceatannol exhibited the highest inhibitory activities (IC₅₀ = 10.6 and 12.1 µM, respectively) but were however less active as (S)-(2-Boronoethyl)-L-cysteine (IC₅₀ = 3.3 µM), used as reference compound. Enzyme kinetic studies showed that both chlorogenic acid and piceatannol are competitive arginase inhibitors. Structural data identified the importance of the caffeoyl (3,4-dihydroxycinnamoyl)-part and of the catechol function in the inhibitory activity of the tested compounds. These results identified chlorogenic acid and piceatannol as two potential core structures for the design of new arginase inhibitors.

Key words

phenolic compounds - arginase inhibitors - in vitro microplate assay

^* Simon Bordage and Thanh-Nhat Pham contributed equally to this work.

• References

• 1 Marquardt KC, Watson RR. Chapter 2 – Polyphenols and public Health. Polyphenols in human Health and Disease. San Diego: Academic Press; 2014: 9-15
• 2 Caldwell RB, Toque HA, Narayanan SP, Caldwell RW. Arginase: an old enzyme with new tricks. Trends Pharmacol Sci 2015; 36: 395-405
• 3 Jenkinson CP, Grody WW, Cederbaum SD. Comparative properties of arginases. Comp Biochem Physiol B Biochem Mol Biol 1996; 114: 107-132
• 4 Di Costanzo L, Sabio G, Mora A, Rodriguez PC, Ochoa AC, Centeno F, Christianson DW. Crystal structure of human arginase I at 1.29-Å resolution and exploration of inhibition in the immune response. Proc Natl Acad Sci U S A 2005; 102: 13058-13063
• 5 Pernow J, Jung C. Arginase as a potential target in the treatment of cardiovascular disease: reversal of arginine steal?. Cardiovasc Res 2013; 98: 334-343
• 6 Munder M. Role of arginase in asthma: potential clinical applications. Expert Rev Clin Pharmacol 2010; 3: 17-23
• 7 Munder M. Arginase: an emerging key player in the mammalian immune system. Br J Pharmacol 2009; 158: 638-651
• 8 Hegde ND, Kumari S, Hegde MN, Shetty S. Role of nitric oxide and arginase in the pathogenesis of oral cancer. ResearchGate 2012; 2: 14-17
• 9 Holowatz LA, Kenney WL. Up-regulation of arginase activity contributes to attenuated reflex cutaneous vasodilatation in hypertensive humans. J Physiol 2007; 581: 863-872
• 10 Shemyakin A, Kövamees O, Rafnsson A, Böhm F, Svenarud P, Settergren M, Jung C, Pernow J. Arginase inhibition improves endothelial function in patients with coronary artery disease and type 2 diabetes mellitus. Circulation 2012; 126: 2943-2950
• 11 Ivanenkov YA, Chufarova NV. Small-molecule arginase inhibitors. Pharm Pat Anal 2013; 3: 65-85
• 12 Da Silva MFL, Floeter-Winter LM. Arginase in Leishmania . In: Santos ALS, Branquinha MH, dʼAvila-Levy CM, Kneipp LF, Sodré CL. eds. Proteins and Proteomics of Leishmania and Trypanosoma . Dordrecht: Springer Netherlands; 2014: 103-117
• 13 Hai Y, Kerkhoven EJ, Barrett MP, Christianson DW. Crystal structure of an arginase-like protein from Trypanosoma brucei that evolved without a binuclear manganese cluster. Biochemistry (Mosc) 2015; 54: 458-471
• 14 Gobert AP, McGee DJ, Akhtar M, Mendz GL, Newton JC, Cheng Y, Mobley HLT, Wilson KT. Helicobacter pylori arginase inhibits nitric oxide production by eukaryotic cells: a strategy for bacterial survival. Proc Natl Acad Sci U S A 2001; 98: 13844-13849
• 15 Golebiowski A, Beckett RP, Van Zandt M, Ji MK, Whitehouse D, Ryder TR, Jagdmann E, Andreoli M, Mazur A, Padmanilayam M, Cousido-Siah A, Mitschler A, Ruiz FX, Podjarny A, Schroeter H. 2-Substituted-2-amino-6-boronohexanoic acids as arginase inhibitors. Bioorg Med Chem Lett 2013; 23: 2027-2030
• 16 Van Zandt MC, Whitehouse DL, Golebiowski A, Ji MK, Zhang M, Beckett RP, Jagdmann GE, Ryder TR, Sheeler R, Andreoli M, Conway B, Mahboubi K, DʼAngelo G, Mitschler A, Cousido-Siah A, Ruiz FX, Howard EI, Podjarny AD, Schroeter H. Discovery of (R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid and congeners as highly potent inhibitors of human arginases I and II for treatment of myocardial reperfusion injury. J Med Chem 2013; 56: 2568-2580
• 17 Hai Y, Christianson DW. Crystal structures of Leishmania mexicana arginase complexed with α,α-disubstituted boronic amino-acid inhibitors. Acta Crystallogr Sect F Struct Biol Commun 2016; 72: 300-306
• 18 Girard-Thernier C, Pham T-N, Demougeot C. The promise of plant-derived substances as inhibitors of arginase. Mini Rev Med Chem 2015; 15: 798-808
• 19 Ferreira C, Soares DC, do Nascimento MTC, Pinto-da-Silva LH, Sarzedas CG, Tinoco LW, Saraiva EM. Resveratrol is active against Leishmania amazonensis: in vitro effect of its association with amphotericin B. Antimicrob Agents Chemother 2014; 58: 6197-6208
• 20 Schnorr O, Brossette T, Momma TY, Kleinbongard P, Keen CL, Schroeter H, Sies H. Cocoa flavanols lower vascular arginase activity in human endothelial cells in vitro and in erythrocytes in vivo . Arch Biochem Biophys 2008; 476: 211-215
• 21 Kim SW, Cuong TD, Hung TM, Ryoo S, Lee JH, Min BS. Arginase II inhibitory activity of flavonoid compounds from Scutellaria indica . Arch Pharm Res 2013; 36: 922-926
• 22 Woo A, Min B, Ryoo S. Piceatannol-3′-O-β-D-glucopyranoside as an active component of rhubarb activates endothelial nitric oxide synthase through inhibition of arginase activity. Exp Mol Med 2010; 42: 524-532
• 23 Joe Y, Zheng M, Kim HJ, Kim S, Uddin MJ, Park C, Ryu DG, Kang SS, Ryoo S, Ryter SW, Chang KC, Chung HT. Salvianolic acid B exerts vasoprotective effects through the modulation of heme oxygenase-1 and arginase activities. J Pharmacol Exp Ther 2012; 341: 850-858
• 24 Corraliza IM, Campo ML, Soler G, Modolell M. Determination of arginase activity in macrophages: a micromethod. J Immunol Methods 1994; 174: 231-235
• 25 Iyamu EW, Asakura T, Woods GM. A colorimetric microplate assay method for high throughput analysis of arginase activity in vitro . Anal Biochem 2008; 383: 332-334
• 26 Hunter A, Downs CE. The inhibition of arginase by amino acids. J Biol Chem 1945; 157: 427-446
• 27 Venkatakrishnan G, Shankar V, Reddy SRR. Microheterogeneity of molecular forms of arginase in mammalian tissues. Indian J Biochem Biophys 2003; 40: 400-408
• 28 Jiang W, Sun B, Song X, Zheng Y, Wang L, Wang T, Liu S. Arginase inhibition protects against hypoxia-induced pulmonary arterial hypertension. Mol Med Rep 2015; 12: 4743-4749
• 29 You H, Gao T, Cooper TK, Morris SM, Awad AS. Arginase inhibition: a new treatment for preventing progression of established diabetic nephropathy. Am J Physiol Ren Physiol 2015; 309: F447-F455
• 30 Kim NN, Cox JD, Baggio RF, Emig FA, Mistry SK, Harper SL, Speicher DW, Morris SM, Ash DE, Traish A, Christianson DW. Probing erectile function: S-(2-Boronoethyl)-l-Cysteine binds to arginase as a transition state analogue and enhances smooth muscle relaxation in human penile corpus cavernosum . Biochemistry (Mosc) 2001; 40: 2678-2688
• 31 Colleluori DM, Ash DE. Classical and slow-binding inhibitors of human type II arginase. Biochemistry (Mosc) 2001; 40: 9356-9362
• 32 Ash DE. Structure and function of arginases. J Nutr 2004; 134: 2760S-2764S
• 33 Clifford MN. Chlorogenic acids and other cinnamates – nature, occurrence, dietary burden, absorption and metabolism. J Sci Food Agric 2000; 80: 1033-1043
• 34 White RE. High-throughput screening in drug metabolism and pharmacokinetic support of drug discovery. Annu Rev Pharmacol Toxicol 2000; 40: 133-157
• 35 Marques V, Farah A. Chlorogenic acids and related compounds in medicinal plants and infusions. Food Chem 2009; 113: 1370-1376