The Inhibitory Activity of Plants from Central Argentina on p-Hydroxyphenylpyruvate Dioxygenase. Isolation and Mechanism of Inhibition of a Flavanone from Flourensia oolepis

Abstract

The enzyme 4-hydroxyphenylpyruvate dioxygenase catalyzes the second step in the tyrosine degradation pathway. In mammals, this enzyme is the molecular target of drugs used for the treatment of metabolic disorders associated with defects in the tyrosine catabolism, mainly the fatal hereditary disease tyrosinemia type 1. This study evaluated the inhibitory effect of 91 extracts on 4-hydroxyphenylpyruvate dioxygenase from mostly native plants from central Argentina. Flourensia oolepis ethanol extract showed itself to be the most effective, and bioguided fractionation yielded pinocembrin (1) as its active principle. This flavanone, with an IC₅₀ value of 73.1 µM and a K_I of 13.7 µM, behaved as a reversible inhibitor of the enzyme and as a noncompetitive inhibitor. Molecular modeling studies confirmed the inhibitory potency of 1 and explained its activity by means of in silico determination of its binding mode in comparison to inhibitors of known activity, cocrystallized with 4-hydroxyphenylpyruvate dioxygenase. The main structural determinants that confer its potency are discussed. Analysis of the binding mode of the flavanone 1 with 4-hydroxyphenylpyruvate dioxygenase revealed the basis of the noncompetitive reversible mechanism of inhibition at the molecular level, which seems to be a common feature in this ubiquitous family of natural compounds. The resulting information may establish the basis for obtaining novel 4-hydroxyphenylpyruvate dioxygenase inhibitors for the treatment of tyrosinemia type 1 and other disorders associated with tyrosinase catabolism.

Key words

Flourensia oolepis - Asteraceae - HPPD inhibitors - Argentinian native plants - pinocembrin - molecular docking

• References

• 1 Moran GR. 4-Hydroxyphenylpyruvate dioxygenase. Arch Biochem Biophys 2005; 433: 117-128
• 2 Moran GR. 4-Hydroxyphenylpyruvate dioxygenase and hydroxymandelate synthase: exemplars of the α-keto acid dependent oxygenases. Arch Biochem Biophys 2014; 544: 58-68
• 3 Imtiaz F, Rashed MS, Al-Mubarak B, Allam R, El-Karaksy H, Al-Hassnan Z, Al-Owain M, Al-Zaidan H, Rahbeeni Z, Qari A, Meyer BF, Al-Sayed M. Identification of mutations causing hereditary tyrosinemia type I in patients of Middle Eastern origin. Mol Genet Metab 2011; 104: 688-690
• 4 Neve S, Aarenstrup L, Tornehave D, Rahbek-Nielsen H, Corydon TJ, Roepstorff P, Kristiansen K. Tissue distribution, intracellular localization and proteolytic processing of rat 4-hydroxyphenylpyruvate dioxygenase. Cell Biol Int 2003; 27: 611-624
• 5 Raspail C, Graindorge M, Moreau Y, Crouzy S, Lefèbvre B, Robin AY, Dumas R, Matringe M. 4-hydroxyphenylpyruvate dioxygenase catalysis: identification of catalytic residues and production of a hydroxylated intermediate shared with a structurally unrelated enzyme. J Biol Chem 2011; 286: 26061-26070
• 6 Dayan FE, Duke SO, Sauldubois A, Singh N, McCurdy C, Cantrell C. p-Hydroxyphenylpyruvate dioxygenase is a herbicidal target site for β-triketones from Leptospermum scoparium . Phytochemistry 2007; 68: 2004-2014
• 7 Dayan FE, Singh N, McCurdy CR, Godfrey CA, Larsen L, Weavers RT, Van Klink JW, Perry NB. β-Triketone inhibitors of plant p-hydroxyphenylpyruvate dioxygenase: modeling and comparative molecular field analysis of their interactions. J Agric Food Chem 2009; 57: 5194-5200
• 8 Hüter O. Use of natural products in the crop protection industry. Phytochem Rev 2011; 10: 185-194
• 9 Meazza G, Scheffler BE, Tellez MR, Rimando AM, Romagni JG, Duke SO, Nanayakkara D, Khan IA, Abourashed EA, Dayan FE. The inhibitory activity of natural products on plant p-hydroxyphenylpyruvate dioxygenase. Phytochemistry 2002; 60: 281-288
• 10 Balunas MJ, Kinghorn AD. Drug discovery from medicinal plants. Life Sci 2005; 78: 431-441
• 11 Carpinella MC, Andrione DG, Ruiz G, Palacios SM. Screening for acetylcholinesterase inhibitory activity in plant extracts from Argentina. Phytother Res 2010; 24: 259-263
• 12 Chiari ME, Joray MB, Ruiz G, Palacios SM, Carpinella MC. Tyrosinase inhibitory activity of native plants from central Argentina: Isolation of an active principle from Lithrea molleoides . Food Chem 2010; 120: 10-14
• 13 Chiari ME, Vera DMA, Palacios SM, Carpinella MC. Tyrosinase inhibitory activity of a 6-isoprenoid-substituted flavanone isolated from Dalea elegans . Bioorg Med Chem 2011; 19: 3474-3482
• 14 Diaz Napal GN, Carpinella MC, Palacios SM. Antifeedant activity of ethanolic extract from Flourensia oolepis and isolation of pinocembrin as its active principle compound. Bioresour Technol 2009; 100: 3669-3673
• 15 Brownlee JM, Johnson-Winters K, Harrison DHT, Moran GR. Structure of the ferrous form of (4-hydroxyphenyl)pyruvate dioxygenase from Streptomyces avermitilis in complex with the therapeutic herbicide, NTBC. Biochemistry (Mosc) 2004; 43: 6370-6377
• 16 Ellis MK, Whitfield AC, Gowans LA, Auton TR, Provan WM, Lock EA, Lee DL, Smith LL. Characterization of the interaction of 2-[2-nitro-4-(trifluoromethyl)benzoyl]-4,4,6,6-tetramethylcyclohexane-1,3,5-trione with rat hepatic 4-hydroxyphenylpyruvate dioxygenase. Chem Res Toxicol 1996; 9: 24-27
• 17 Lee DL, Knudsen CG, Michaely WJ, Chin HL, Nguyen NH, Carter CG, Cromartie TH, Lake BH, Shribbs JM, Fraser T. The structure-activity relationships of the triketone class of HPPD herbicides. Pestic Sci 1998; 54: 377-384
• 18 Lin SW, Lin YL, Lin TC, Yang DY. Discovery of a potent, non-triketone type inhibitor of 4-hydroxyphenylpyruvate dioxygenase. Bioorg Med Chem Lett 2000; 10: 1297-1298
• 19 Lin YL, Wu CS, Lin SW, Yang DY. SAR studies of 2-o-substituted-benzoyl- and 2-alkanoyl-cyclohexane-1,3-diones as inhibitors of 4-hydroxyphenylpyruvate dioxygenase. Bioorg Med Chem Lett 2000; 10: 843-845
• 20 Wu CS, Huang JL, Sun YS, Yang DY. Mode of action of 4-hydroxyphenylpyruvate dioxygenase inhibition by triketone-type inhibitors. J Med Chem 2002; 45: 2222-2228
• 21 Yang C, Pflugrath JW, Camper DL, Foster ML, Pernich DJ, Walsh TA. Structural basis for herbicidal inhibitor selectivity revealed by comparison of crystal structures of plant and mammalian 4-hydroxyphenylpyruvate dioxygenases. Biochemistry (Mosc) 2004; 43: 10414-10423
• 22 Rasul A, Millimouno FM, Ali Eltayb W, Ali M, Li J, Li X. Pinocembrin: a novel natural compound with versatile pharmacological and biological activities. Biomed Res Int 2013; 2013: 379850
• 23 Liu R, Li JZ, Song JK, Sun JL, Li YJ, Zhou SB, Zhang TT, Du GH. Pinocembrin protects human brain microvascular endothelial cells against fibrillar amyloid-β(1 − 40)injury by suppressing the MAPK/NF-κB inflammatory pathways. Biomed Res Int 2014; 2014: 470393
• 24 Yang N, Qin S, Wang M, Chen B, Yuan N, Fang Y, Yao S, Jiao P, Yu Y, Zhang Y, Wang J. Pinocembrin, a major flavonoid in propolis, improves the biological functions of EPCs derived from rat bone marrow through the PI3 K-eNOS-NO signaling pathway. Cytotechnology 2013; 65: 541-551
• 25 Meng F, Wang Y, Liu R, Gao M, Du G. Pinocembrin alleviates memory impairment in transient global cerebral ischemic rats. Exp Ther Med 2014; 8: 1285-1290
• 26 Barton AF, Dell B, Knight AR. Herbicidal activity of cineole derivatives. J Agric Food Chem 2010; 58: 10147-10155
• 27 Lee DL, Prisbylla MP, Cromartie TH, Dagarin DP, Howard SW, Mc Lean Povan W, Ellis MK, Fraser T, Mutter LC. The discovery and structural requirements of inhibitors of p-hydroxyphenylpyruvate dioxygenase. Weed Sci 1997; 45: 601-609
• 28 Ling TS, Shiu S, Yang DY. Design and synthesis of 3-fluoro-2-oxo-3-phenylpropionic acid derivatives as potent inhibitors of 4-hydroxyphenylpyruvate dioxygenase from pig liver. Bioorg Med Chem 1999; 7: 1459-1465
• 29 Barrero AF, Herrador MM, Arteaga P, Rodríguez-García I, García-Moreno M. Resorcinol derivatives and flavonoids of Ononis natrix subspecies ramosissima . J Nat Prod 1997; 60: 65-68
• 30 Rüetschi U, Dellsén A, Sahlin P, Stenman G, Rymo L, Lindstedt S. Human 4-hydroxyphenylpyruvate dioxygenase. Eur J Biochem 1993; 213: 1081-1089
• 31 Taniguchi K, Armstrong MD. The enzymatic formation of o-hydroxyphenylacetic acid. J Biol Chem 1963; 238: 4091-4097
• 32 Roche PA, Moorehead TJ, Hamilton GA. Purification and properties of hog liver 4-hydroxyphenylpyruvate dioxygenase. Arch Biochem Biophys 1982; 216: 62-73
• 33 Schulz A, Ort O, Beyer P, Kleinig H. SC-0051, a 2-benzoyl-cyclohexane-1, 3-dione bleaching herbicide, is a potent inhibitor of the enzyme p-hydroxyphenylpyruvate dioxygenase. FEBS Lett 1993; 318: 162-166
• 34 Knox WE, Pitt BM. Enzymic catalysis of the keto-enol tautomerization of phenylpyruvic acids. J Biol Chem 1957; 225: 675-688
• 35 Neidig ML, Kavana M, Moran GR, Solomon EI. CD and MCD Studies of the non-heme ferrous active site in (4-hydroxyphenyl)pyruvate dioxygenase: correlation between oxygen activation in the extradiol and α-KG-dependent dioxygenases. J Am Chem Soc 2004; 126: 4486-4487
• 36 Yanai T, Tew DP, Handy NC. A new hybrid exchange–correlation functional using the Coulomb-attenuating method (CAM-B3LYP). Chem Phys Lett 2004; 393: 51-57
• 37 Bayly CI, Cieplak P, Cornell W, Kollman PA. A well-behaved electrostatic potential based method using charge restraints for deriving atomic charges: the RESP model. J Phys Chem 1993; 97: 10269-10280
• 38 Morris GM, Goodsell DS, Halliday RS, Huey R, Hart WE, Belew RK, Olson AJ. Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comput Chem 1998; 19: 1639-1662
• 39 Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheeseman JR, Scalmani G, Barone V, Mennucci B, Petersson GA, Nakatsuji H, Caricato M, Li X, Hratchian HP, Izmaylov AF, Bloino J, Zheng G, Sonnenberg JL, Hada M, Ehara M, Toyota K, Fukuda R, Hasegawa J, Ishida M, Nakajima T, Honda Y, Kitao O, Nakai H, Vreven T, Montgomery jr. JA, Peralta JE, Ogliaro F, Bearpark M, Heyd JJ, Brothers E, Kudin KN, Staroverov VN, Kobayashi R, Normand J, Raghavachari K, Rendell A, Burant JC, Iyengar SS, Tomasi J, Cossi M, Rega N, Millam JM, Klene M, Knox JE, Cross JB, Bakken V, Adamo C, Jaramillo J, Gomperts R, Stratmann RE, Yazyev O, Austin AJ, Cammi R, Pomelli C, Ochterski JW, Martin RL, Morokuma K, Zakrzewski VG, Voth GA, Salvador P, Dannenberg JJ, Dapprich S, Daniels AD, Farkas O, Foresman JB, Ortiz JV, Cioslowski J, Fox DJ. Gaussian 09, Revision A.01. Wallingford CT: Gaussian, Inc.; 2009
• 40 ACD labs. Predict accurate acid/base dissociation constants from structure – the industry standard.. Available at http://www.acdlabs.com/products/percepta/predictors/pka/ Accessed 17 August 2015.
• 41 Liao C, Nicklaus MC. Comparison of nine programs predicting pKa values of pharmaceutical substances. J Chem Inf Model 2009; 49: 2801-2812
• 42 Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, Olson AJ. AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. J Comput Chem 2009; 16: 2785-2791
• 43 Humphrey W, Dalke A, Schulten K. VMD: visual molecular dynamics. J Mol Graph 1996; 14: 33-38
• 44 Cancès E, Mennucci B. Comment on “Reaction field treatment of charge penetration” [J Chem Phys 112, 5558 (2000)]. J Chem Phys 2001; 114: 4744-4745
• 45 Chipman DM. Reaction field treatment of charge penetration. J Chem Phys 2000; 112: 5558-5565
• 46 Tomasi J, Mennucci B, Cances E. The IEF version of the PCM solvation method: an overview of a new method addressed to study molecular solutes at the QM ab initio level. J Mol Struc: THEOCHEM 1999; 464: 211-226