New 2-aryloxy-3-phenyl-propanoic acids as peroxisome proliferator-activated receptors α/γ dual agonists with improved potency and reduced adverse effects on skeletal muscle function

Journal of Medicinal Chemistry

Volumn 52, Issue 20, 2009, Pages 6382-6393

  Fracchiolla, Giuseppe ^a   Laghezza, Antonio ^a   Piemontese, Luca ^a   Tortorella, Paolo ^a   Mazza, Fernando ^b,c   Montanari, Roberta ^c   Pochetti, Giorgio ^c   Lavecchia, Antonio ^d   Novellino, Ettore ^d   Pierno, Sabata ^a   Camerino, Diana Conte ^a   Loiodice, Fulvio ^a  

^a UNIVERSITY OF BARI   (Italy)

^b UNIVERSITY OF L'AQUILA   (Italy)

^c CNR   (Italy)

^d UNIVERSITY OF NAPLES FEDERICO II   (Italy)

Author keywords

[No Author keywords available]

Indexed keywords

2 (4 BENZYLAMINO PHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 BENZYLOXY PHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 BENZYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 BENZYLTHIO PHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 PHENETHYL PHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 PHENETHYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 PHENOXY PHENOXY) 3 PHENYL PROPANOIC ACID; 2 (4 PHENOXYMETHYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 PHENYLAMINOMETHYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 PHENYLETHYNYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 PHENYLTHIOMETHYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 (4 STYRYLPHENOXY) 3 PHENYLPROPANOIC ACID; 2 [4 (3 PHENYLPROPYL)PHENOXY] 3 PHENYLPROPANOIC ACID; PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR ALPHA AGONIST; PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR GAMMA AGONIST; PROPIONIC ACID DERIVATIVE; UNCLASSIFIED DRUG;

ARTICLE; CONTROLLED STUDY; DRUG ACTIVITY; DRUG BINDING; DRUG IDENTIFICATION; DRUG POTENCY; DRUG STRUCTURE; DRUG SYNTHESIS; ENZYME ACTIVITY; HUMAN; HUMAN CELL; MUSCLE FUNCTION; SKELETAL MUSCLE; X RAY CRYSTALLOGRAPHY;

ANIMALS; CELL LINE, TUMOR; CHLORIDE CHANNELS; CHLORIDES; CLOFIBRIC ACID; CRYSTALLOGRAPHY, X-RAY; DRUG DISCOVERY; ELECTRIC CONDUCTIVITY; HUMANS; HYDROXYMETHYLGLUTARYL-COA REDUCTASE INHIBITORS; LIGANDS; MALE; MODELS, MOLECULAR; MOLECULAR CONFORMATION; MUSCLE, SKELETAL; MUSCULAR DISEASES; PPAR ALPHA; PPAR GAMMA; PROPIONIC ACIDS; RATS; RATS, WISTAR; REST; STEREOISOMERISM; SUBSTRATE SPECIFICITY;

EID: 70350041556     PISSN: 00222623     EISSN: None     Source Type: Journal    
DOI: 10.1021/jm900941b     Document Type: Article

References (65)

1. Elisaf, M. Effects of fibrates on serum metabolic parameters. Curr. Med. Res. Opin. 2002, 18, 269-276.
2. Staels, B.; Dallongeville, J.; Auwerx, J.; Schoonjans, K.; Leitersdorf, E.; Fruchart, J.-C. Mechanism of action of fibrates on lipid and lipoprotein metabolism. Circulation 1998, 98, 2088-2093. (Pubitemid 28512236)
3. Campbell, I. W. The Clinical Significance of PPAR Gamma Agonism. Curr. Mol. Med. 2005, 5, 349-363.
4. Sprecher, D. L. Lipids, lipoproteins, and peroxisome proliferator activated receptor-delta. Am. J. Cardiol. 2007, 100, n20-n24.
5. Sprecher, D. L.; Massien, C.; Pearce, G.; Billin, A. N.; Perlstein, I.; Willson, T. M.; Hassall, D. G.; Ancellin, N.; Patterson, S. D.; Lobe, D. C.; Johnson, T. G. Triglyceride: high-density lipoprotein cholesterol effects in healthy subjects administered a peroxisome proliferator activated receptor delta agonist. Arterioscler. Thromb. Vasc. Biol. 2007, 27, 359-365.
6. Reilly, S. M.; Lee, C. H. PPAR delta as a therapeutic target in metabolic disease. FEBS Lett. 2008, 582, 26-31. (Pubitemid 350297247)
7. Madrazo, J. A.; Kelly, D. P. The PPAR trio: regulators of myocardial energy metabolism in health and disease. J. Mol. Cell. Cardiol. 2008, 44, 968-975.
8. Schmuth, M.; Jiang, Y. J.; Dubrac, S.; Elias, P. M.; Feingold, K. R. Thematic review series: skin lipids. Peroxisome proliferator-activated receptors and liver X receptors in epidermal biology. J. Lipid Res. 2008, 49, 499-509.
9. Iwashita, A.; Muramatsu, Y.; Yamazaki, T.; Muramoto, M.; Kita, Y.; Yamazaki, S.; Mihara, K.; Moriguchi, A.; Matsuoka, N. Neuroprotective efficacy of the peroxisome proliferator-activated receptor delta-selective agonists in vitro and in vivo. J. Pharmacol. Exp. Ther. 2007, 320, 1087-1096.
10. Peters, J. M.; Hollingshead, H. E.; Gonzalez, F. J. Role of peroxisome- proliferator-activated receptor beta/delta (PPARbeta/delta) in gastrointestinal tract function and disease. Clin. Sci. (London) 2008, 115, 107-127.
11. Reifel Miller, A. Today's challenges and tomorrow's opportunities: ligands to Peroxisome Proliferator-Activated Receptors as Therapies for Type 2 Diabetes and Metabolic Syndrome. Drug. Dev. Res. 2006, 67, 574-578.
12. Willson, T. M.; Brown, P. J.; Sternbach, D. D.; Henke, B. R. The PPARs: from orphan receptors to drug discovery. J. Med. Chem. 2000, 43, 527-550.
13. Henke, B. R. Peroxisome proliferator-activated receptor alpha/ gamma dual agonists for the treatment of type 2 diabetes. J. Med. Chem. 2004, 47, 4118-4127.
14. Lehmann, J. M.; Moore, L. B.; Smith-Oliver, T. A.; Wilkison, W. O.; Willson, T. M.; Kliewer, S. A. An antidiabetic thiazolidi-nedione is a high affinity ligand for peroxisome proliferator-activated receptor gamma (PPAR gamma). J. Biol. Chem. 1995, 270, 12953-12956.
15. Willson, T. M.; Cobb, J. E.; Cowan, D. J.; Wiethe, R. W.; Correa, I. D.; Prakash, S. R.; Beck, K. D.; Moore, L. B.; Kliewer, S. A.; Lehmann, J. M. The structure-activity relationship between peroxisome proliferator-activated receptor gamma agonism and the antihyperglycemic activity of thiazolidinediones. J. Med. Chem. 1996, 39, 665-668.
16. Lohray, B. B.; Lohray,V. B.; Bajji, A. C.; Kalchar, S.; Poondra, R. R.; Padakanti, S.; Chakrabarti, R.; Vikramadithyan, R. K.; Misra, P.; Juluri, S.; Mamidi, N. V.; Rajagopalan, R. (-)3-[4-[2-(Phenoxazin-10- yl)ethoxy]phenyl]-2- ethoxypropanoic acid [(-)DRF 2725]: a dual PPAR agonist with potent antihyperglycemic and lipid modulating activity. J. Med. Chem. 2001, 44, 2675-2678.
17. Sauerberg, P.; Pettersson, I.; Jeppesen, L.; Bury, P. S.; Mogensen, J. P.; Wassermann, K.; Brand, C. L.; Sturis, J.; Woldike, H. F.; Fleckner, J.; Andersen, A. S.; Mortensen, S. B.; Svensson, L. A.; Rasmussen, H. B.; Lehmann, S. V.; Polivka, Z.; Sindelar, K.; Panajotova, V.; Ynddal, L.; Wulff, E. M. Novel tricyclic-alpha-alkyloxyphenylpropionic acids: dual PPARalpha/gamma agonists with hypolipidemic and antidiabetic activity. J. Med. Chem. 2002, 45, 789-804.
18. Ebdrup, S.; Pettersson, I.; Rasmussen, H. B.; Deussen, H. J.; Frost Jensen, A.; Mortensen, S. B.; Fleckner, J.; Pridal, L.; Nygaard, L.; Sauerberg, P. Synthesis and biological and structural characterization of the dual-acting peroxisome proliferator-activated receptor alpha/gamma agonist ragaglitazar. J. Med. Chem. 2003, 46, 1306-1317.
19. Devasthale, P. V.; Chen, S.; Jeon, Y.;Qu, F.; Shao, C.;Wang, W.; Zhang, H.; Cap, M.; Farrelly, D.; Golla, R.; Grover, G.; Harrity, T.;Ma, Z.;Moore, L.; Ren, J.; Seethala, R.; Cheng, L.; Sleph, P.; Sun, W.; Tieman, A.; Wetterau, J. R.; Doweyko, A.; Chandrasena, G.; Chang, S. Y.; Humphreys, W. G.; Sasseville, V. G.; Biller, S. A.; Ryono, D. E.; Selan, F.; Hariharan, N.; Cheng, P. T. Design and synthesis of N-[(4-methoxyphenoxy)carbonyl]-N-[[4-[2-(5- methyl-2-phenyl-4-oxazolyl)ethoxy]phenyl]methyl]glycine [Muraglitazar/ BMS-298585], a novel peroxisome proliferator-activated receptor alpha/gamma dual agonist with efficacious glucose and lipid-lowering activities. J. Med. Chem. 2005, 48, 2248-2250.
20. Koyama, H.; Miller, D. J.; Boueres, J. K.; Desai, R. C.; Jones, A. B.; Berger, J. P.; MacNaul, K. L.; Kelly, L. J.; Doebber, T. W.; Wu, M. S.; Zhou, G.; Wang, P. R.; Ippolito, M. C.; Chao, Y. S.; Agrawal, A. K.; Franklin, R.; Heck, J. V.; Wright, S. D.; Moller, D. E.; Sahoo, S. P. (2R)-2-ethylchromane-2- carboxylic acids: discovery of novel PPARalpha/gamma dual agonists as antihyperglycemic and hypolipidemic agents. J. Med. Chem. 2004, 47, 3255-3263.
21. Shi, G. Q.; Dropinski, J. F.; McKeever, B. M.; Xu, S.; Becker, J. W.; Berger, J. P.; MacNaul, K. L.; Elbrecht, A.; Zhou, G.; Doebber, T. W.; Wang, P.; Chao, Y. S.; Forrest, M.; Heck, J. V.; Moller, D. E.; Jones, A. B. Design and synthesis of alpha-aryloxyphenylacetic acid derivatives: a novel class of PPARalpha/gamma dual agonists with potent antihyperglycemic and lipid modulating activity. J. Med. Chem. 2005, 48, 4457-4468.
22. Ljung, B.; Bamberg, K.; Dahllof, B.; Kjellstedt, A.; Oakes, N. D.; Ostling, J.; Svensson, L.; Camejo, G. AZ 242, a novel PPARalpha/ gamma agonist with beneficial effects on insulin resistance and carbohydrate and lipid metabolism in ob/ob mice and obese Zucker rats. J. Lipid Res. 2002, 43, 1855-1863. (Pubitemid 35315803)
23. Grey, A. Skeletal consequences of thiazolidinedione therapy. Osteoporosis Int. 2008, 19, 129-137.
24. Rubenstrunk, A.; Hanf, R.; Hum, D. W.; Fruchart, J.-C.; Staels, B. Safety issues and prospects for future generations of PPAR modulators. Biochim. Biophys. Acta 2007, 1771, 1065-1081.
25. Shearer, B. G.; Billin, A. N. The next generation of PPAR drugs: do we have the tools to find them? Biochim. Biophys. Acta 2007, 1771, 1082-1093.
26. Montanari, R.; Saccoccia, F.; Scotti, E.; Crestani, M.; Godio, C.; Gilardi, F.; Loiodice, F.; Fracchiolla, G.; Laghezza, A.; Tortorella, P.; Lavecchia, A.; Novellino, E.; Mazza, F.; Aschi, M.; Pochetti, G. Crystal structure of the peroxisome proliferator-activated receptor gamma (PPARgamma) ligand binding domain complexed with a novel partial agonist: a new region of the hydrophobic pocket could be exploited for drug design. J. Med. Chem. 2008, 51, 7768-7776.
27. Feller, D. R.; Kamanna, V. S.; Newman, H. A.; Romstedt, K. J.; Witiak, D. T.; Bettoni, G.; Bryant, S. H.; Conte-Camerino, D.; Loiodice, F.; Tortorella, V. Dissociation of hypolipidemic and antiplatelet actions from adverse myotonic effects of clofibric acid related enantiomers. J. Med. Chem. 1987, 30, 1265-1267.
28. Bettoni, G.; Loiodice, F.; Tortorella, V.; Conte-Camerino, D.; Mambrini, M.; Ferrannini, E.; Bryant, S. H. Stereospecificity of the chloride ion channel: the action of chiral clofibric acid analogues. J. Med. Chem. 1987, 30, 1267-1270.
29. Hodel, C. Myopathy and rhabdomyolysis with lipid-lowering drugs. Toxicol. Lett. 2002, 128, 159-168.
30. Pierno, S.; Didonna, M. P.; Cippone, V.; De Luca, A.; Pisoni, M.; Frigeri, A.; Nicchia, G. P.; Svelto, M.; Chiesa, G.; Sirtori, C.; Scanziani, E.; Rizzo, C.; De Vito, D.; Conte Camerino, D. Effects of chronic treatment with statins and fenofibrate on rat skeletal muscle: a biochemical, histological and electrophysiological study. Br. J. Pharmacol. 2006, 149, 909-919.
31. Liang, Y.; Xie, Y. X.; Li, J. H. Modified palladium-catalyzed Sonogashira cross-coupling reactions under copper-, amine-, and solvent-free conditions. J. Org. Chem. 2006, 71, 379-381.
32. Bhanushali, M. J.; Nandurkar, N. S.; Bhor, M. D.; Bhanage, B. M. Direct reductive amination of carbonyl compounds using bis(triphenylphosphine)copper(I) tetrahydroborate. Tetrahedron Lett. 2007, 48, 1273-1276.
33. Tarzia, G.; Duranti, A.; Tontini, A.; Piersanti, G.; Mor, M.; Rivara, S.; Plazzi, P. V.; Park, C.; Kathuria, S.; Piomelli, D. Design, synthesis, and structure-activity relationships of alkylcarbamic acid aryl esters, a new class of fatty acid amide hydrolase inhibitors. J. Med. Chem. 2003, 46, 2352-2360.
34. Pinelli, A.; Godio, C.; Laghezza, A.; Mitro, N.; Fracchiolla, G.; Tortorella, V.; Lavecchia, A.; Novellino, E.; Fruchart, J. C.; Staels, B.; Crestani, M.; Loiodice, F. Synthesis, biological evaluation, and molecular modeling investigation of new chiral fibrates with PPARalpha and PPARgamma agonist activity. J. Med. Chem. 2005, 48, 5509-5519.
35. Kleywegt, G. J.; Zou, J. Y.; Kjeldgaard, M.; Jones, T. A. Around O. In International Tables for Crystallography, Vol.F. Crystallography of Biological Macromolecules; Rossmann, M. G., Arnold, E., Eds.; Kluwer Academic Publishers: Dordrecht, The Netherlands, 2001, pp 353-356, 366-367.
36. Meyer, E. A.; Castellano, R. K.; Diederich, F. Interactions with aromatic rings in chemical and biological recognition. Angew. Chem., Int. Ed. 2003, 42, 1210-1250. (Pubitemid 36410287)
37. Kawatkar, S. P.; Kuntz, D. A.; Woods, R. J.; Rose, D. R.; Boons, G. J. Structural basis of the inhibition of Golgi alpha-mannosidase II by mannostatin A and the role of the thiomethyl moiety in ligand-protein interactions. J. Am. Chem. Soc. 2006, 128, 8310-8319.
38. Wang, W.; Devasthale, P.; Farrelly, D.; Gu, L.; Harrity, T.; Cap, M.; Chu, C.; Kunselman, L.; Morgan, N.; Ponticiello, R.; Zebo, R.; Zhang, L.; Locke, K.; Lippy, J.; O'Malley, K.; Hosagrahara, V.; Kadiyala, P.; Chang, C.; Muckelbauer, J.; Doweyko, A. M.; Zahler, R.; Ryono, D.; Hariharan, N.; Cheng, P. T. Discovery of azetidinone acids as conformationally-constrained dual PPARalpha/ gamma agonists. Bioorg. Med. Chem. Lett. 2008, 18, 1939-1944.
39. GOLD, version 4.0; CCDC Software Limited: Cambridge, U.K., 2008.
40. Jones, G.; Willett, P.; Glen, R. C.; Leach, A. R.; Taylor, R. Development and validation of a genetic algorithm for flexible docking. J. Mol. Biol. 1997, 267, 727-748.
41. Schulz-Gasch, T.; Stahl, M. Binding site characteristics in structure- based virtual screening: evaluation of current docking tools. J. Mol. Model. 2003, 9, 47-57.
42. Wang, R.; Lu, Y.; Wang, S. Comparative evaluation of 11 scoring functions for molecular docking. J. Med. Chem. 2003, 46, 2287-2303.
43. Kellenberger, E.; Rodrigo, J.; Muller, P.; Rognan, D. Comparative evaluation of eight docking tools for docking and virtual screening accuracy. Proteins 2004, 57, 225-242. (Pubitemid 39223729)
44. Warren, G. L.; Andrews, C. W.; Capelli, A. M.; Clarke, B.; LaLonde, J.; Lambert, M. H.; Lindvall, M.; Nevins, N.; Semus, S. F.; Senger, S.; Tedesco, G.; Wall, I. D.; Woolven, J. M.; Peishoff, C. E.; Head, M. S. A critical assessment of docking programs and scoring functions. J. Med. Chem. 2006, 49, 5912-5931.
45. Verdonk, M. L.; Cole, J. C.; Hartshorn, M. J.; Murray, C. W.; Taylor, R. D. Improved protein-ligand docking using GOLD. Proteins 2003, 52, 609-623.
46. Eldridge, M. D.; Murray, C. W.; Auton, T. R.; Paolini, G. V.; Mee, R. P. Empirical scoring functions: I. The development of a fast empirical scoring function to estimate the binding affinity of ligands in receptor complexes. J. Comput.-Aided Mol. Des. 1997, 11, 425-445.
47. Rimon, D.; Ludatscher, R.; Cohen, L. Clofibrate-induced muscular syndrome. Case report with ultrastructural findings and review of the literature. Isr. J. Med. Sci. 1984, 20, 1082-1086.
48. Pierce, L. R.; Wysowski, D. K.; Gross, T. P. Myopathy and rhabdomyolysis associated with lovastatin-gemfibrozil combination therapy. JAMA, J. Am. Med. Assoc. 1990, 264, 71-75.
49. Langer, T.; Levy, R. I. Acute muscular syndrome associated with administration of clofibrate. N. Engl. J. Med. 1968, 279, 856-858.
50. Reaven, P.; Witztum, J. L. Lovastatin, nicotinic acid, and rhabdomyolysis. Ann. Intern. Med. 1988, 109, 597-598.
51. Hodel, C. M.; Kruslin, E. Risk profile of lipid-lowering drugs. Arh. Hig. Rada Toksikol. 1997, 48, 67-86.
52. Kruslin, E.; Hodel, C. M. Myotoxicity in lipid-lowering therapy. Arh. Hig. Rada Toksikol. 1997, 48, 7-26.
53. Chauvin, M.; Zupan, M.; Brechenmacher, C. Sindrome musculaire atypique avec myolyse au cours d'un traitement chronique par fibrates (Atypical muscle syndrome characterized by myolysis during a chronic treatment with fibrates). Arch. Mal. Coeur. Vaiss. 1988, 81, 921-923.
54. Bermingham, R. P.; Whitsitt, T. B.; Smart, M. L.; Nowak, D. P.; Scalley, R. D. Rhabdomyolysis in a patient receiving the combination of cerivastatin and gemfibrozil. Am. J. Health Syst. Pharm. 2000, 57, 461-464.
55. Pierno, S.; Camerino, G. M.; Cippone, V.; Rolland, J. F.; Desaphy, J. F.; De Luca, A.; Liantonio, A.; Bianco, G.; Kunic, J. D.; George, A. L., Jr.; Conte Camerino, D. Statins and fenofibrate affect skeletal muscle chloride conductance in rats by differently impairing ClC-1 channel regulation and expression. Br. J. Pharmacol. 2009, 156, 1206-1215.
56. Pochetti, G.; Godio, C.; Mitro, N.; Caruso, D.; Galmozzi, A.; Scurati, S.; Loiodice, F.; Fracchiolla, G.; Tortorella, P.; Laghezza, A.; Lavecchia, A.; Novellino, E.; Mazza, F.; Crestani, M. Insights into the mechanism of partial agonism: crystal structures of the peroxisome proliferator-activated receptor gamma ligand-binding domain in the complex with two enantiomeric ligands. J. Biol. Chem. 2007, 282, 17314-17324.
57. Leslie, A. G. W. Jt. CCP4 ESF-EACMB Newlett. Protein Crystallogr. 1992, 26.
58. Brunger, A. T.; Adams, P. D.; Clore, G. M.; DeLano, W. L.; Gros, P.; Grosse-Kunstleve, R. W.; Jiang, J. S.; Kuszewski, J.; Nilges, M.; Pannu, N. S.; Read, R. J.; Rice, L. M.; Simonson, T.; Warren, G. L. Crystallography NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr., Sect. D: Biol. Crystallogr. 1998, 54 (Part 5), 905-921.
59. Molecular Operating Environment (MOE), version 2005.06; Chemical Computing Group, Inc.: Montreal, Canada, 2005.
60. Huang, C. C.; Couch, G. S.; Pettersen, E. F.; Ferrin, T. E. Chimera: an extensible molecular modeling application constructed using standard components. Pac. Symp. Biocomput. 1996, 1, 724, http:// www.cgl.ucsf.edu/chimera
61. Case, D. A.; Darden, T. A.; Cheatham, T. E., III; Simmerling, C. L.; Wang, J.; Duke, R. E.; Luo, R.; Merz, K. M.; Pearlman, D. A.; Crowley, M.; Walker, R. C.; Zhang, W.; Wang, B.; Hayik, S.; Roitberg, A.; Seabra, G.; Wong, K. F.; Paesani, F.; Wu, X.; Brozell, S.; Tsui, V.; Gohlke, H.; Yang, L.; Tan, C.; Mongan, J.; Hornak, V.; Cui, G.; Beroza, P.; Mathews, D. H.; Schafmeister, C.; Ross, W. S.; Kollman, P. A. AMBER9; University of California, San Francisco, 2006.
62. Cornell, W. D.; Cieplak, P.; Bayly, C. I.; Gould, I. R.; Merz, K. M.; Ferguson, D. M.; Spellmeyer, D. C.; Fox, T.; Caldwell, J. W.; Kollman, P. A. A second generation force field for the simulation of proteins, nucleic acids, and organic molecules. J. Am. Chem. Soc. 1995, 117, 5179-5197.
63. Bernstein, F. C.; Koetzle, T. F.; Williams, G. J.; Meyer, E. F., Jr.; Brice, M. D.; Rodgers, J. R.; Kennard, O.; Shimanouchi, T.; Tasumi, M. The Protein Data Bank: a computer-based archival file for macromolecular structures. J. Mol. Biol. 1977, 112, 535-542.
64. Raspe, E.; Madsen, L.; Lefebvre, A. M.; Leitersdorf, I.; Gelman, L.; Peinado-Onsurbe, J.; Dallongeville, J.; Fruchart, J. C.; Berge, R.; Staels, B. Modulation of rat liver apolipoprotein gene expression and serum lipid levels by tetradecylthioacetic acid (TTA) via PPARalpha activation. J. Lipid Res. 1999, 40, 2099-2110. (Pubitemid 29523436)
65. Hollon, T.; Yoshimura, F. K. Variation in enzymatic transient gene expression assays. Anal. Biochem. 1989, 182, 411-418.