USOO804888OB2

(12) United States Patent (10) Patent No.: US 8,048,880 B2 Trias et al. (45) Date of Patent: Nov. 1, 2011

(54) TREATMENT OF CARDIOVASCULAR 7,098,237 B1 8, 2006 Todo DISEASE AND DYSLIPIDEMIAUSING 2004/0248.898 A1 12/2004 Saiga et al. 2006/0094693 A1 5, 2006 Aziz et al. SECRETORY PHOSPHOLIPASE A, (SPLA) 2006/0223865 A1 10, 2006 Buch et al. INHIBITORS AND SPLA, INHIBITOR 2007/O135385 A1 6/2007 Chang et al. COMBINATION THERAPES FOREIGN PATENT DOCUMENTS EP 1378.246 A1 1, 2004 (75) Inventors: Joaquim Trias, Millbrae, CA (US); WO WO96,03376 2, 1996 Colin Hislop, Menlo Park, CA (US); WO WO 99,56752 11, 1999 Paul Truex, Pleasanton, CA (US); WO 00/28332 A1 5, 2000 Bernadine Fraser, Sunnyvale, CA (US); WO O1 (36420 A1 5, 2001 WO WOO1, 55.108 A2 8, 2001 Debra Odink, Oakland, CA (US); Scott WO 02/08189 A1 1, 2002 Chadwick, Redwood City, CA (US); WO 2004,103960 A2 12/2004 Kenneth Gould, Zionsville, IN (US); WO 2005/028653 A1 3, 2005 Marian Mosior, Indianapolis, IN (US); WO 2005/058310 A2 6, 2005 Patrick Eacho, Indianapolis, IN (US) WO WO 2007/056281 A2 5/2007 (73) Assignee: Anthera Pharmaceuticals, Inc., OTHER PUBLICATIONS Hayward, CA (US) Clarket al., “Potential therapeutic uses of phospholipase A2 inhibi tors'. Expert Opinion on Therapeutic Patents, vol. 14, No. 7, pp. (*) Notice: Subject to any disclaimer, the term of this 937-950 (2004).* patent is extended or adjusted under 35 Aviram, M. et al., “Phospholipase A2-Modified LDL Is Taken Up at Enhanced Rate by Macrophages.” Biochem Biophys Res Commun U.S.C. 154(b) by 390 days. 185:465-472 (1992). Boekholdt, S. M., et al., "Serum Levels of Type II Secretory (21) Appl. No.: 12/114,710 Phospholipase A2 and the Risk of Future in Apparently Healthy Men and Women: The EPIC-Norfolk Prospec (22) Filed: May 2, 2008 tive Population Study.” Arterioscler. Thromb. Vasc. Biol. 25:839-846 (2005). (65) Prior Publication Data Bostrom, M. A., et al., “Group V Secretory Phospholipase A2 Pro motes : Evidence From Genetically Altered Mice.” US 2009/O131396 A1 May 21, 2009 Arterioscler. Thromb. Vasc. Bio. 27:600-606 (2007). Boyanovsky, B., et al., “Group V Secretory Phospholipase A2-modi Related U.S. Application Data fied Low Density Lipoprotein Promotes Foam Cell Formation by a SR-A- and CD36- independent Process That Involves Cellular (60) Provisional application No. 61/190,044, filed on Oct. Proteoglycans.” J Biol Chem 280:32746-32752 (2005). 18, 2007, provisional application No. 60/969,591, filed on Aug. 31, 2007, provisional application No. (Continued) 60/915,910, filed on May 3, 2007. Primary Examiner — Kevin E Weddington (51) Int. Cl. (74) Attorney, Agent, or Firm — Perkins Coie LLP: Patrick A 6LX3/535 (2006.01) D. Morris A 6LX3/397 (2006.01) (57) ABSTRACT A6 IK3I/44 (2006.01) A6 IK3I/40 (2006.01) Administration of sPLA inhibitors has been found to (52) U.S. Cl...... 514/235.2: 514/210.02: 514/356; decrease cholesterol levels, atherosclerotic plaque formation 514/422:514/423 and aortic aneurysm in mice, and to decrease cholesterol and (58) Field of Classification Search ...... 514/235.2, triglyceride levels in humans. Interestingly, administration of 514/210.02, 356, 422, 423 sPLA inhibitors was found to decrease cholesterol levels See application file for complete search history. even when the inhibitors were administered only once per day. Therefore, provided herein are methods of treating dys (56) References Cited lipidemia, CVD, and conditions associated with CVD such as atherosclerosis and metabolic syndrome, by administering U.S. PATENT DOCUMENTS one or more SPLA inhibitors. Significantly, administration of 3,485,847 A * 12/1969 Bossert et al...... 546,321 SPLA inhibitors and various compounds used in the treat 5,273,995 A * 12/1993 Roth ...... 514,422 ment of CVD. Such as for example , resulted in greater 5,504,073. A 4, 1996 Homan decreases in LDL and LDL particle levels in a synergistic 5,578,634 A 11/1996 Bach et al. 5,641,800 A 6, 1997 Bach et al. manner. In addition, administration of SPLA inhibitors and 5,654,326 A * 8/1997 Bach et al...... 514,419 statins resulted in a synergistic decrease in plaque content. 5,684,034 A 11/1997 Bach et al. Therefore, also provided herein are compositions comprising 5,767,115 A * 6/1998 Rosenblum et al...... 514,210.02 one or more spIA inhibitors and one or more compounds 6,177.440 B1 1/2001 Bach et al. used in the treatment of CVD, such as for example statins, and 6,252,084 B1 6, 2001 Bach et al. methods of using these compositions to treat dyslipidemia, 6,274,578 B1 8/2001 Denney et al. 6,472.389 B1 10, 2002 Ohtani et al. CVD, and conditions associated with CVD such as athero 6,713,645 B1 3, 2004 Bach et al. Sclerosis and metabolic syndrome. 6,756,376 B1 6/2004 Fuji et al. 7,026,318 B2 4/2006 Ogawa et al. 30 Claims, 12 Drawing Sheets US 8,048.880 B2 Page 2

OTHER PUBLICATIONS Kovanen, P. T., et al., “Secretory Group II Phospholipase A2: A Camejo, G., et al., “Association of Apo B Lipoproteins with Arterial Newly Recognized Acute-Phase Reactant With a Role in Proteoglycans: Pathological Significance and Molecular Basis.” Atherogenesis.” Circ Res 86:610-612 (2000). Arterioscler. 139:205-222 (1998). Kugiyama, K., et al., “Circulating Levels of Secretory Type II Chait, A., et al., "Lipoprotein-Associated Inflammatory Proteins: Phospholipase A2 Predict Coronary Events in Patients with Coronary Markers or Mediators of Cardiovascular Disease?” J Lipid Res. Artery Disease.” Circulation 100: 1280-1284 (1999). 46:389-403 (2005). Leitinger, N., et al., “Role of Group II Secretory Phospholipase A2 in Daugherty, A., et al., “Angiotensin II Promotes Atherosclerotic Atherosclerosis : 2. Potential Involvement of Biologically Active Lesions and Aneurysms in Apollipoprotein E-Deficient Mice.” JClin. Oxidized Phospholipids.” Arterioscler Thromb Vasc Biol 19:1291 Invest. 105:1605-1612 (2000). 1298 (1999). De Beer, F. C., et al., “Secretory Non-Pancreatic Phospholipase A2: Levick, S. et al., “Antifibrotic Activity of an Inhibitor of Group IIA Influence on Lipoprotein Metabolism.” J Lipid Res. 38:2232-2239 Secretory Phospholipase A2 in Young Spontaneously Hypertensive (1997). Rats.” J Immunol 176:7000-7007 (2006). Divchev, D., et al., “The Secretory Phospholipase A2 Group IIA: A Liu, P.Y., et al., “Prognostic Value and the Changes of Plasma Levels Missing Link Between Inflammation, Activated Renin-Angiotensin of Secretory Type II Phospholipase A2 in Patients with Coronary System, and Atherogenesis.” Vasc Health Risk Manag 4:597–604 Artery Disease Undergoing Percutaneous Coronary Intervention.” (2008). Eur Heart J 24:1824-1832 (2003). Draheim, S.E., et al., “Indole Inhibitors of Human Nonpancreatic Mallat, Z. et al., “Circulating Secretory Phospholipase A2 Activity Secretory Phospholipase A2. 3. Indole-3-Glyoxamides,” J Med and Risk of Incident Coronary Events in Healthy Men and Women: Chem 39:5159-5175 (1996). The Epic-Norfolk Study.” Arterioslcer. Thromb. Vasc. Biol. 27:1177 Eckey, R., et al., “Increased Hepatic Cholesterol Accumulation in 1183 (2007). Transgenic Mice Overexpressing Human Secretory Phospholipase Mallat, Z. et al., “Circulating Secretory Phospholipase A2 Activity A Group IIA.” Inflammation 28:59-65 (2004). Predicts Recurrent Events in Patients with Severe Acute Coronary Elinder, L. S., et al., “Expression of Phospholipase A2 Isoforms in Syndromes.” JAm. Coll. Cardiol. 46:1249-1257 (2005). Human Normal and Atherosclerotic Arterial Wall.” Arterioscler. Menschikowski, M., et al., “Secretory Group II Phospholipase A2 in Human Atherosclerotic Plaques.” Atheroscler, 118:173-181 (1995). Thromb. Vasc. Biol. 17:2257-2263 (1997). Menschikowski, M., et al., “Secretory Phospholipase A2 of Group Flood, C., et al., “Molecular Mechanism for Changes in Proteoglycan IIA: Is Itan Offensive or a Defensive Player During Atherosclerosis Binding on Compositional Changes of the Core and the Surface of and Other Inflammatory Diseases?” Prostaglandins Other Lipid Low-Density Lipoprotein Containing Human Apollipoprotein Mediat 79:1-33 (2005). B100.” Arterioscler Thromb Vasc Biol 24:564-570 (2004). Murakami, M. & Kudo, I., “New phospholipase A2 isozymes with a Ghesquiere, S., et al., “Macrophage-Specific Overexpression of potential role in atherosclerosis.” Curr Opin Lipidol 14:431-436 Group IIa spLA2 Increases Atherosclerosis and Enhances Collagen (2003). Deposition.” J Lipid Res 46:20 1-210 (2005). Murakami, M., et al., “Diversity of Phospholipase A2 Enzymes: Hakala, J. 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Hartford, M., et al., "CRP Interleukin-6, Secretory Phospholipase A2 Oorni, K. et al., “PLA2-V: A Real Player in Atherogenesis.” Group IIA, and Intercellular Adhesion Molecule-1 During the Early Arterioscler Thromb Vasc Biol 27:445-447 (2007). Phase of Acute Coronary Syndromes and Long-Term Follow-Up.” J Paradis, M-E, et al., “Visceral Adipose Tissue Accumulation, Secre Cardiol. 108:55-62 (2006). tory Phospholipase A2-IIA and Atherogenecity of LDL.” Int. JObe Hurt-Camejo, E., et al., “Localization of Nonpancreatic Secretory sity 30:1615-1622 (2006). Phospholipase A2 in Normal and Atherosclerotic Arteries.” Piek, J., et al., “Type II Secretory Phospholipase A2: The Emerging Arterioscler Thromb Vasc Biol 17:300-309 (1997). Role of Biochemical Markers of Plaque Inflammation.” Eur Heart J Hurt-Camejo, E., et al., “Cellular Consequences of the Association of 24:1804-1806 (2003). ApoB Lipoproteins with Proteoglycans.” Arterioscler Thromb Vasc Pruzanski, W., et al., "Lipoproteins are Substrates for Human Secre Biol 17:1011-1017 (1997). tory Group IIA Phospholipase A2: Preferential Hydrolysis of Acute Hurt-Camejo, E., et al., “Phospholipase A2 in Vascular Disease.” Circ Phase HDL.” J Lipid Res. 39:2150-2160 (1998). Res 89:298-304 (2001). Ramoner, R., et al., “Dendritic-Cell Activation by Secretory Ivandic, B., et al., “Role of Group II Secretory Phospholipase A2 in Phospholipase A2.” Blood 105:3583-3587 (2005). Atherosclerosis: 1. Increased Atherogenesis and Altered Romano, M., et al., “Ultrastructural Localization of SecretoryType II Lipoproteins in Transgenic Mice Expressing Group IIa Phospholipase A2 in Atherosclerotic and Nonatherosclerotic Phospholipase A2.” Arterioscler. Thromb. Vasc. Biol. 19:1284-1290 Regions of Human Arteries.” Arterioscler Thromb Vasc Biol 18:519 (1999). 525 (1998). Jialal, I., “Evolving Lipoprotein Risk Factors: Lipoprotein(a) and Rosengren, B., et al., “Secretory Phospholipase A2 Group V: Lesion Oxidized Low-Density Lipoprotein.” Clin. Chem. 44:1827-1832 Distribution, Activation by Arterial Proteoglycans, and Induction in (1998). Aorta by a Western Diet.” Arterioscler. Thromb. Vasc. Biol. 26:1579 Karabina, S.A., et al., “Atherogenic Properties of LDL Particles 1585 (2006). Modified by Human Group X Secreted Phospholipase A2 on Human Saiga, A., et al., “Group X Secretory Phospholipase A2 Induces Endothelial Cell Function.” FASEB J 20:2547 (2006). Potent Productions of Various Lipid Mediators in Mouse Peritoneal Kimura-Matsumoto, M., et al., “Expression of Secretory Macrophages.” Biochim Biophys Acta 1530:67-76 (2001). Phospholipase A2s in Human Atherosclerosis Development.” Ath Sartipy, P. et al., “Binding of Human Phospholipase A2 Type II to erosclerosis, doi:10.1016/j.atherosclerosis.2006.08.062 (2007). Proteoglycans.” J Biol Chem 271:26307-26314 (1996). Kougias, P. et al., “LySophosphatidylcholine and Secretory Sartipy, P. et al., “Phospholipase A2 Modification of Low Density Phospholipase A2 in Vascular Disease: Mediators of Endothelial Lipoproteins Forms Small High Density Particles with Increased Dysfunction and Atherosclerosis.” Med Sci Monit 12:RA5-RA16 Affinity for Proteoglycans and Glycosaminoglycans,” J Biol. Chem. (2006). 274:25913-25920 (1999). US 8,048.880 B2 Page 3

Schiering, A., et al., “Analysis of secretory group II phospholipase A2 Labeque, R., et al., “Enzymatic Modification of Plasma Low Density expression in human aortic tissue in dependence on the degree of Lipoproteins in Rabbits: A Potential Treatment for atherosclerosis.” Atherosclerosis 144:73-78 (1999). Hypercholesterolemia.” Proc Natl Acad Sci USA 90:3476-3480 Scott, K.F., et al., “Secreted Phospholipase A2 Enzymes as Thera (1993). peutic Targets.” Expert Opin TherTargets 7:427-440 (2003). Melani, L., et al., “Efficacy and safety of coadministered Snyder, D. W., et al., “Pharmacology of LY315920/S-5920, 1 3 with in patients with primary hypercholesterolemia: a (Aminooxoacetyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4- prospective, randomized, double-blind trial.” Eur Heart J24:717-728 ylloxyacetate, A Potent and Selective Secretory Phospholipase A2 (2003). Inhibitor: A New Class of Anti-Inflammatory Drugs, SPI.” J Menschikowski, M., et al., “Expression of Human Secretory Group Pharmacol Exp. Ther 288: 1117 (1999). IIA Phospholipase A2 Is Associated with Reduced Concentrations of Sparrow, C. P. et al., “Enzymatic Modification of Low Density Plasma Cholesterol in Transgenic Mice.” Inflammation 24:227-237 Lipoprotein by Purified Lipoxygenase Plus Phospholipase APMim (2000). ics Cell-Mediated Oxidative Modification.” J Lipid Res 29:745-753 Menschikowski, M., et al., “Statins potentiate the IFN-y-induced (1988). upregulation of group IIA phospholipase A2 in human aortic Smooth Szmitko, P. E., et al., “Biomarkers of Vascular Disease Linking muscle cells and HepG2 hepatoma cells.” Biochim Biophys Acta Inflammation to Endothelial Activation: Part II. Circulation 1733:157-171 (2005). 108:2041-2048 (2003). Mohler, E. R. et al., “The Effect of Darapladib on Plasma Tietge, U.J. F., et al., “Overexpression of Secretory Phospholipase Lipoprotein-Associated Phospholipase A2 Activity and Cardiovas A2 Causes Rapid Catabolism and Altered Tissue Uptake of High cular Biomarkers in Patients with Stable Coronary Heart Disease or Density Lipoprotein Cholesteryl Ester and Apollipoprotein A-I.” J Coronary Heart Disease Risk Equivalent.” J Am Coll Cardiol Biol. Chem. 275: 10077-10084 (2000). 51: 1632-1641 (2008). Tietge, U., et al., “Macrophage-Specific Expression of Group IIA Pearson, T.A., et al., “Pooled analyses of effects on C-reactive protein SPLA2 Results in Accelerated Atherogenesis by Increasing Oxida and low density lipoprotein cholesterolin placebo-controlled trials of tive Stress,” J Lipid Res 46:1604-1614 (2005). eZetimibe monotherapy or eZetimibe added to baseline United States Patent and Trademark Office, Combined International therapy,” Biochim Biophys Acta 1636:108-118 (2004). Search Report and Written Opinion for PCT/US08/62577, dated Petry, C., et al., “Inhibition of Rho modulates cytokine-induced Aug. 4, 2008. Webb, N., et al., “Macrophage-Expressed Group IIA Secretory prostaglandin E2 formation in renal mesangial cells.” Biochim Phospholipase A2 Increases Atherosclerotic Lesion Formation in Biophys Acta 1636:108-118 (2004). LDL Receptor—Deficient Mice.” Arterioscler Thromb Vasc Biol Pirkova, A.A., et al., “Effect of Therapy with on the 23:263-268 (2003). Level of Secretory Phospholipase A2 Group IIA and Modification of Webb, N., “Secretory Phospholipase A2 Enzymes in Atherogenesis.” Low Density Lipoproteins in Patients with Ischemic Heart Disease.” Curr Opin Lipidology 16:341-344 (2005). Kardiologia 4:37-40 (2007). Wootton, P. et al., “Tagging SNP Haplotype Analysis of the Secre Porela, P. et al., “Level of circulating phospholipase A2 in prediction tory PLA2-V gene, PLA2G5, Shows Strong Association with LDL of the prognosis of patients with Suspected myocardial infarction.” and oxLDL Levels, Suggesting Functional Distinction from spLA2 Basic Res Cardiol 95:413-417 (2000). IIA: Results From the UDACS Study.” Hum Mol Genet 16:1437 Serruys, P. W., et al., “Effects of the Direct Lipoprotein-Associated 1444 (2007). Phospholipase A2 Inhibitor Darapladib on Human Coronary Athero Wooton-Kee, C. R. et al., “Group V spLA2 Hydrolysis of Low sclerotic Plaque.” Circulation 118: 1172-1182 (2008). Density Lipoprotein Results in Spontaneous Particle Aggregation Tietge, U.J.F., et al., “Human Secretory Phospholipase A2 Mediates and Promotes Macrophage Foam Cell Formation.” Arterioscler Decreased Plasma Levels of HDL Cholesterol and ApoA-I in Thromb Vasc Biol 24:762-767 (2004). Response to Inflammation in Human Apo A-I Transgenic Mice.” Ballantyne, C.M., et al., “Effect of eZetimibe coadministered with Arterioscler. Thromb Vasc Biol 22:1213-1218 (2002). atorvastatin in 628 patients with primary hypercholesterolemia: a Clark, J. D., et al., “Potential Therapeutic Uses of Phospholipase A2 prospective, randomized, double-blind trial.” Circulation 97:2409 Inhibitors.” Expert Opin. Ther. Patents 14 (7):937-950 (2004). 2415 (2003). Abraham, E., et al., “Efficacy and Safety of LY315920Na/S-5920, A Bays, H.E., et al., “Effects of hydrochloride on low Selective Inhibitor of 14-kDa Group IIA Secretory Phospholipase density lipoprotein cholesterol and high-sensitivity C-reactive pro A2, in Patients with Suspected Sepsis and Organ Failure.” Crit. Care. tein when added to statins in patients with hypercholesterolemia.” Med. 31:718-728 (2003). Am J Cardiol 97:1198-1205 (2006). Bowton, D. L., et al., “Impact of a Soluble Phospholipase A2 Inhibi Davidson, M.H., et al., “EZetimibe coadministered with tor on Inhaled Allergen Challenge in Subjects with Asthma.” Journal in patients with primary hypercholesterolemia.” JAm Coll Cardiol of Asthma 1:65-71 (2005). 40: 2125-2134 (2002). Bradley, J. D., et al., “A Randomized, Double-Blinded, Placebo Elsayed, R.K. and Evans, J.D., “Emerging lipid-lowering drugs: Controlled Clinical Trial ofLY333013, a Selective Inhibitor of Group squalene synthase inhibitors.” Expert Opin Emerging Drugs 13:309 II Secretory Phospholipase A2, in the Treatment of Rheumatoid 322 (2008). Arthritis.” J. Rheumatol. 32:417-23 (2005). Fichtlischerer, S., et al., “Elevated Secretory Non-Pancreatic Type II Pearson, T. A., et al., “Pooled Analyses of Effects on C-Reactive Phospholipase A2 Serum Activity is Associated with Impaired Protein and Low Density Lipoprotein Cholesterol in Placebo-Con Endothelial Vasodilator Function in Patients with Coronary Artery trolled Trials of Ezetimibe Monotherapy or Ezetimibe Added to Disease.” Clin Sci 106:51 1-517 (2004). Baseline Statin Therapy.” Am. J. Cardiol. 103:369-374 (2009). Hunninghake, D., et al., "Coadministration of colesevelam hydro Winkler, E., et al., “Decreased Serum Cholesterol Level After Snake chloride with atorvastatin lowers LDL cholesterol additively.” Ath Bite (Vipera Palaestinae) as a Marker of Severity of Envenomation.” erosclerosis 158:407-416 (2001). J. Lab. Clin. Med. 121(6):774-8 (1993). Jaross, W., et al., “Biological effects of secretory phospholipase A2 Zeiher, B. G., et al., “LY31592ONA/S-5920, A Selective Inhibitor of group IIA on lipoproteins and in atherogenesis.” Eur J. Clin Invest Group IIA Secretory Phospholipase A2, Fails to Improve Clinical 32:383-393 (2002). Outcome for Patients with Severe Sepsis.” Crit. Care Med. 33:1741 Kerzner, B., et al., “Efficacy and safety of eZetimibe coadministered 1748 (2005). with in primary hypercholesterolemia.” Am J Cardiol Chen, W.M., et al., “Control of Capillary Formation by Membrane 91:418-424 (2003). Anchored Extracellular Inhibitor of Phospholipase A.” FEBS Let Kugiyama, K. et al., “Prognostic Value of Plasma Levels of Secretory ters 522:113-118 (2002). Type II Phospholipase A2 in Patients with Unstable Angina Pectoris.” European Patent Office, Extended European Search Report dated Am J Cardiol 86:718–722 (2000). Apr. 1, 2011 for European Application No. 08747605.7. US 8,048.880 B2 Page 4

Ma, J., et al., “Are Evidence-Based Cardiovascular Prevention Thera Vervoordeldonk, M., et al., “Aspirin Inhibits Expression of the pies Being Used? A Review of Aspirin and Statin Therapies.” Pre Interleukin-1 f-Inducible Group II Phospholipase A.” FEBS Letters vention and Control 1:285-295 (2005). Robinson, J. G., et al., “Combination Therapy with Ezetimibe and 397: 108-112 (1996). Simvastatin to Achieve Aggressive LDL Reduction.” Expert Reu. Wiklund, O., et al., “Effects of Simvastatin and Atorvastatin on Cardiovasc. Ther. 4(4):461-476 (2006). Inflammation Markers in Plasma.” J. Int. Med. 251:338-347 (2002). Rodondi, N., et al., “Aspirin for the Primary Prevention of Cardio vascular Disease a Comprehensive Review.” Comp. Ther. 31(3): 186 193 (2005). * cited by examiner U.S. Patent Nov. 1, 2011 Sheet 1 of 12 US 8,048,880 B2

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-0.030 5.00 1OOO 15.OO 2O.OO 25.00 30.00 35.00 A.O.OO 45.00 50.00 55.0 SO.OO 55.00 7OOO Minutes US 8,048,880 B2 1. 2 TREATMENT OF CARDOVASCULAR be administered to the Subject twice or more per day, and in DISEASE AND DYSLPIDEMIAUSING certain of these embodiments the one or more spLA inhibi SECRETORY PHOSPHOLIPASE A, (SPLA) tors are administered twice per day. In other embodiments, INHIBITORS AND SPLA, INHIBITOR the one or more spLA inhibitors may be administered to the COMBINATION THERAPES Subject on a once a day basis. In certain embodiments, the one or more SPLA inhibitors may be administered at a dosage of CROSS-REFERENCE TO RELATED about 50 to about 500 mg. APPLICATIONS In certain embodiments, methods are provided for decreas ing cholesterol levels in a subject in need thereof by admin The subject application claims the benefit of U.S. Provi 10 istering a therapeutically effective amount of one or more sional Patent Application No. 61/190,044, filed Oct. 18, 2007, sPLA inhibitors. In certain embodiments, the one or more U.S. Provisional Patent Application No. 60/969,591, filed sPLA inhibitors are administered in a composition that fur Aug. 31, 2007, and U.S. Provisional Patent Application No. ther comprises one or more pharmaceutically acceptable car 60/915,910, filed May 3, 2007. The disclosures of each of riers. In certain embodiments, the one or more spLA inhibi these applications are incorporated herein by reference in 15 tors comprise A-001 or a pharmaceutically acceptable their entirety, including drawings. prodrug, salt, polymorph, co-crystal, or Solvate of A-001. In certain of these embodiments, the one or more spLA inhibi BACKGROUND tors comprise A-002, a prodrug of A-001. In certain embodi ments, administration of one or more SPLA inhibitors results In 2004, it was estimated that over 75 million Americans in a decrease in total cholesterol, non-HDL cholesterol, LDL, had one or more forms of cardiovascular disease (CVD). LDL particle, small LDL particle, oxidized LDL, and/or Coronary heart disease (CHD) and coronary artery disease ApoB levels. In certain embodiments, administration of one (CAD) are the most common types of CVD. CHD and CAD or more spLA inhibitors results in an improvement in HDL/ occur when coronary arteries that supply blood to the heart LDL ratio. In certain embodiments, the one or more spLA become hardened and narrowed due to atherosclerosis. A 25 inhibitors may be administered to the subject twice or more variety of therapeutic options are currently employed in the per day, and in certain of these embodiments the one or more treatment of CVD and conditions associated with CVD. sPLA inhibitors are administered twice per day. In other Many of these therapeutic options function by lowering cho embodiments, the one or more sRLA inhibitors may be lesterol levels, particularly LDL levels. Among the most administered to the Subject on a once a day basis. In certain popular and effective of these therapeutic options are statins, 30 embodiments, the one or more sRLA inhibitors may be a class of compounds that inhibit cholesterol biosynthesis and administered at a dosage of about 50 to about 500 mg. prevent the build-up of arterial plaque. Statin administration In certain embodiments, methods are provided for decreas has been shown to lower LDL and triglyceride levels and to ing triglyceride levels in a subject in need thereof by admin substantially reduce coronary events and death from CVD. istering a therapeutically effective amount of one or more However, statin therapy alone is insufficient to completely 35 sPLA inhibitors. In certain embodiments, the one or more treat CVD. Therefore, there is a need in the art for more sPLA inhibitors are administered in a composition that fur effective methods of treating CVD and conditions associated ther comprises one or more pharmaceutically acceptable car with CVD. riers. In certain of these embodiments, the one or more spLA inhibitors comprise A-001 or a pharmaceutically acceptable SUMMARY 40 prodrug, salt, polymorph, co-crystal, or Solvate of A-001. In certain of these embodiments, the one or more spLA inhibi In certain embodiments, methods are provided for treating tors comprise A-002, a prodrug of A-001. In certain embodi dyslipidemia in a subject in need thereof by administering a ments, the one or more spLA inhibitors may be administered therapeutically effective amount of one or more spLA to the Subject twice or more per day, and in certain of these inhibitors. In certain embodiments, the one or more spLA 45 embodiments the one or more spLA inhibitors are adminis inhibitors are administered in a composition that further com tered twice per day. In other embodiments, the one or more prises one or more pharmaceutically acceptable carriers. In sPLA inhibitors may be administered to the subjectona once certain embodiments, the one or more spIA inhibitors com a day basis. In certain embodiments, the one or more SPLA prise A-001 or a pharmaceutically acceptable prodrug, salt, inhibitors may be administered at a dosage of about 50 to polymorph, co-crystal, or solvate of A-001. In certain of these 50 about 500 mg. embodiments, the one or more spLA inhibitors comprise In certain embodiments, methods are provided for increas A-002, a prodrug of A-001. In certain embodiments, admin ing HDL levels in a subject in need thereof by administering istration of one or more spLA inhibitors results in a decrease a therapeutically effective amount of one or more spLA in cholesterol and/or triglyceride levels. In certain of these inhibitors. In certain embodiments, the one or more sRLA embodiments, administration of one or more spLA inhibi 55 inhibitors are administered in a composition that further com tors results in a decrease in total cholesterol, non-HDL cho prises one or more pharmaceutically acceptable carriers. In lesterol, LDL, LDL particle, small LDL particle, oxidized certain embodiments, the one or more spIA inhibitors com LDL, and/or ApoB levels. In certain embodiments, adminis prise A-001 or a pharmaceutically acceptable prodrug, salt, tration of one or more SPLA inhibitors results in an increase polymorph, co-crystal, or solvate of A-001. In certain of these in HDL levels and/or LDL particle size. In certain embodi 60 embodiments, the one or more spLA inhibitors comprise ments, administration of one or more SPLA inhibitors results A-002, a prodrug of A-001. In certain embodiments, admin in a decrease in levels of one or more inflammatory markers. istration of one or more spLA inhibitors results in an In certain of these embodiments, the inflammatory markers improvement in HDL/LDL ratio. In certain embodiments, may include, but are not limited to, spLA, CRP, and/or IL-6. improvement in HDL/LDL ratio is further accomplished by a In certain embodiments, administration of one or more spIA 65 reduction in LDL levels. In certain embodiments, the one or inhibitors results in an improvement in HDL/LDL ratio. In more spLA inhibitors may be administered to the subject certain embodiments, the one or more SPLA inhibitors may twice or more per day, and in certain of these embodiments US 8,048,880 B2 3 4 the one or more spLA inhibitors are administered twice per of A-001. In certain embodiments, the one or more com day. In other embodiments, the one or more spLA inhibitors pounds used in the treatment of CVD comprise one or more may be administered to the Subject on a once a day basis. In statins or statin combination drugs. In certain of these certain embodiments, the one or more SPLA inhibitors may embodiments, the one or more statins are selected from the be administered at a dosage of about 50 to about 500 mg. group consisting of atorvastatin, simvastatin, , In certain embodiments, methods are provided for treating lovastatin, pravastatin, , , , CVD or a condition associated with CVD in a subject in need , and various salts, Solvates, Stereoisomers, and thereof by administering a therapeutically effective amount prodrug derivatives thereof. In certain of these embodiments, of one or more spLA inhibitors. In certain embodiments, the the statin combination drugs are selected from the group one or more SPLA inhibitors are administered in a compo 10 consisting of atorvastatin plus eZetimibe, atorvastatin plus sition that further comprises one or more pharmaceutically amlodipine, atorvastatin plus CP-529414, atorvastatin plus acceptable carriers. In certain embodiments, the one or more APA-01, simvastatin plus eZetimibe, simvastatin plus sPLA inhibitors comprise A-001 or a pharmaceutically extended release , simvastatin plus MK-0524A, lovas acceptable prodrug, salt, polymorph, co-crystal, or Solvate of tatin plus extended release niacin, rosuvastatin plus fenofi A-001. In certain of these embodiments, the one or more 15 brate, pravastatin plus , and statin plus TAK-457. sPLA inhibitors comprise A-002, a prodrug of A-001. In In certain embodiments, the one or more compounds used in certain embodiments, the one or more SPLA inhibitors may the treatment of CVD comprise one or more non-statin com be administered to the Subject twice or more per day, and in pounds selected from the group consisting of bile acid certain of these embodiments the one or more spIA inhibi sequestrants, , niacin or niacin derivatives, cholesterol tors are administered twice per day. In other embodiments, absorption inhibitors, cholesteryl ester transfer protein the one or more spLA inhibitors may be administered to the (CETP) inhibitors, microsomal triglyceride transfer protein Subject on a once a day basis. In certain embodiments, the one (MTP) inhibitors, squalene synthase inhibitors, ACE inhibi or more spIA inhibitors may be administered at a dosage of tors, angiotensin II receptor antagonists, beta-adrenergic about 50 to about 500 mg. In certain embodiments, adminis blockers, calcium channel blockers, and antithrombotics. tration of one or more spLA inhibitors results in a decrease in 25 In certain embodiments, methods are provided for treating levels of one or more inflammatory markers. In certain of dyslipidemia in a subject in need thereof by administering a these embodiments, the inflammatory markers may include, therapeutically effective amount of one or more spLA but are not limited to, sli A, CRP, and/or IL-6. In certain inhibitors and a therapeutically effective amount of one or embodiments, CVD or a condition associated with CVD more compounds used in the treatment of CVD. In certain includes, but is not limited to, atherosclerosis, metabolic Syn 30 embodiments, the one or more spLA inhibitors comprise drome, coronary artery disease, coronary heart disease, cere A-001 or a pharmaceutically acceptable prodrug, salt, poly brovascular disease, peripheral vascular disease, and/or con morph, co-crystal, or solvate of A-001. In certain of these ditions associated with coronary artery disease, coronary embodiments, the one or more spLA inhibitors comprise heart disease, cerebrovascular disease, or peripheral vascular A-002, a prodrug of A-001. In certain embodiments, the one disease. 35 or more compounds used in the treatment of CVD comprise In certain embodiments, methods are provided for treating one or more statins or statin combination drugs. In certain of metabolic syndrome in a subject in need thereof by adminis these embodiments, the one or more statins are selected from tering a therapeutically effective amount of one or more the group consisting of atorvastatin, simvastatin, rosuvasta sPLA inhibitors. In certain embodiments, the one or more tin, lovastatin, pravastatin, cerivastatin, fluvastatin, mevasta sPLA inhibitors are administered in a composition that fur 40 tin, pitavastatin, and various salts, Solvates, Stereoisomers, ther comprises one or more pharmaceutically acceptable car and prodrug derivatives thereof. In certain of these embodi riers. In certain embodiments, the one or more spLA inhibi ments, the statin combination drugs are selected from the tors comprise A-001 or a pharmaceutically acceptable group consisting of atorvastatin plus eZetimibe, atorvastatin prodrug, salt, polymorph, co-crystal, or Solvate of A-001. In plus amlodipine, atorvastatin plus CP-529414, atorvastatin certain of these embodiments, the one or more spLA inhibi 45 plus APA-01, simvastatin plus eZetimibe, simvastatin plus tors comprise A-002, a prodrug of A-001. In certain embodi extended release niacin, simvastatin plus MK-0524A, lovas ments, the one or more spLA inhibitors may be administered tatin plus extended release niacin, rosuvastatin plus fenofi to the Subject twice or more per day, and in certain of these brate, pravastatin plus fenofibrate, and statin plus TAK-457. embodiments the one or more spLA inhibitors are adminis In certain embodiments, the one or more compounds used in tered twice per day. In other embodiments, the one or more 50 the treatment of CVD comprise one or more non-statin com sPLA inhibitors may be administered to the subject on a once pounds selected from the group consisting of bile acid a day basis. In certain embodiments, the one or more SPLA sequestrants, fibrates, niacin or niacin derivatives, cholesterol inhibitors may be administered at a dosage of about 50 to absorption inhibitors, cholesteryl ester transfer protein about 500 mg. In certain embodiments, administration of one (CETP) inhibitors, microsomal triglyceride transfer protein or more spLA inhibitors results in a decrease in levels of one 55 (MTP) inhibitors, squalene synthase inhibitors, ACE inhibi or more inflammatory markers. In certain of these embodi tors, angiotensin II receptor antagonists, beta-adrenergic ments, the inflammatory markers may include, but are not blockers, calcium channel blockers, and antithrombotics. In limited to, spLA, CRP, and/or IL-6. certain embodiments, administration of one or more SPLA In certain embodiments, compositions are provided com inhibitors and one or more compounds used in the treatment prising one or more SPLA inhibitors and one or more com 60 of CVD results in a decrease in cholesterol levels and/or pounds used in the treatment of CVD. In certain embodi triglyceride levels. In certain of these embodiments, admin ments, the composition further comprises one or more istration of one or more spLA inhibitors and one or more pharmaceutically acceptable carriers. In certain embodi compounds used in the treatment of CVD results in a decrease ments, the one or more spLA inhibitors comprise A-001 or a in total cholesterol, non-HDL cholesterol, LDL, LDL par pharmaceutically acceptable prodrug, salt, polymorph, co 65 ticle, small LDL particle, oxidized LDL, and/or ApoB levels. crystal, or solvate of A-001. In certain of these embodiments, In certain embodiments, administration of one or more SPLA the one or more spLA inhibitors comprise A-002, a prodrug inhibitors and one or more compounds used in the treatment US 8,048,880 B2 5 6 of CVD results in an increase in HDL levels and/or LDL administration of one or more SPLA inhibitors and one or particle size. In certain embodiments, administration of one more compounds used in the treatment of CVD results in an or more SPLA inhibitors and one or more compounds used in increase in LDL particle size. In certain embodiments, the treatment of CVD results in an improvement in HDL/LDL administration of one or more SPLA inhibitors and one or ratio. In certain embodiments, the one or more spLA inhibi 5 more compounds used in the treatment of CVD results in an tors and the one or more compounds used in the treatment of improvement in HDL/LDL ratio. In certain embodiments, the CVD may be administered simultaneously. In certain of these one or more spIA inhibitors and the one or more compounds embodiments, the one or more spLA inhibitors and the one used in the treatment of CVD may be administered simulta or more compounds used in the treatment of CVD may be neously. In certain of these embodiments, the one or more administered in a single formulation, while in other embodi 10 SPLA inhibitors and the one or more compounds used in the ments the compounds may be administered simultaneously in treatment of CVD may be administered in a single formula two or more formulations. In each of these embodiments, the tion, while in other embodiments the compounds may be formulation(s) may further comprise one or more pharmaceu administered simultaneously in two or more formulations. In tically acceptable carriers. In other embodiments, the one or each of these embodiments, the formulation(s) may further more SPLA inhibitors and the one or more compounds used 15 comprise one or more pharmaceutically acceptable carriers. in the treatment of CVD may be administered sequentially. In In other embodiments, the one or more spLA inhibitors and certain embodiments, the one or more SPLA inhibitors may the one or more compounds used in the treatment of CVD be administered to the Subject twice or more per day, and in may be administered sequentially. In certain embodiments, certain of these embodiments the one or more spIA inhibi the one or more spLA inhibitors may be administered to the tors are administered twice per day. In other embodiments, Subject twice or more per day, and in certain of these embodi the one or more spLA inhibitors may be administered to the ments the one or more sRLA inhibitors are administered Subject on a once a day basis. In certain embodiments, the one twice per day. In other embodiments, the one or more spLA or more spIA inhibitors may be administered at a dosage of inhibitors may be administered to the Subject on a once a day about 50 to about 500 mg. In certain embodiments, adminis basis. In certain embodiments, the one or more spLA inhibi tration of one or more spLA inhibitors and one or more 25 tors may be administered at a dosage of about 50 to about 500 compounds used in the treatment of CVD results in a decrease ng. in levels of one or more inflammatory markers. In certain of In certain embodiments, methods are provided for decreas these embodiments, the inflammatory markers may include, ing triglyceride levels in a subject in need thereof by admin but are not limited to, spLA, CRP, and/or IL-6. istering a therapeutically effective amount of one or more In certain embodiments, methods are provided for decreas 30 sPLA inhibitors and a therapeutically effective amount of ing cholesterol levels in a subject in need thereof by admin one or more compounds used in the treatment of CVD. In istering a therapeutically effective amount of one or more certain embodiments, the one or more spIA inhibitors com sPLA inhibitors and a therapeutically effective amount of prise A-001 or a pharmaceutically acceptable prodrug, salt, one or more compounds used in the treatment of CVD. In polymorph, co-crystal, or solvate of A-001. In certain of these certain embodiments, the one or more spIA inhibitors com 35 embodiments, the one or more spLA inhibitors comprise prise A-001 or a pharmaceutically acceptable prodrug, salt, A-002, a prodrug of A-001. In certain embodiments, the one polymorph, co-crystal, or solvate of A-001. In certain of these or more compounds used in the treatment of CVD comprise embodiments, the one or more spLA inhibitors comprise one or more statins or statin combination drugs. In certain of A-002, a prodrug of A-001. In certain embodiments, the one these embodiments, the one or more statins are selected from or more compounds used in the treatment of CVD comprise 40 the group consisting of atorvastatin, simvastatin, rosuvasta one or more statins or statin combination drugs. In certain of tin, lovastatin, pravastatin, cerivastatin, fluvastatin, mevasta these embodiments, the one or more statins are selected from tin, pitavastatin, and various salts, Solvates, Stereoisomers, the group consisting of atorvastatin, simvastatin, rosuvasta and prodrug derivatives thereof. In certain of these embodi tin, lovastatin, pravastatin, cerivastatin, fluvastatin, mevasta ments, the statin combination drugs are selected from the tin, pitavastatin, and various salts, Solvates, Stereoisomers, 45 group consisting of atorvastatin plus eZetimibe, atorvastatin and prodrug derivatives thereof. In certain of these embodi plus amlodipine, atorvastatin plus CP-529414, atorvastatin ments, the statin combination drugs are selected from the plus APA-01, simvastatin plus eZetimibe, simvastatin plus group consisting of atorvastatin plus eZetimibe, atorvastatin extended release niacin, simvastatin plus MK-0524A, lovas plus amlodipine, atorvastatin plus CP-529414, atorvastatin tatin plus extended release niacin, rosuvastatin plus fenofi plus APA-01, simvastatin plus eZetimibe, simvastatin plus 50 brate, pravastatin plus fenofibrate, and statin plus TAK-457. extended release niacin, simvastatin plus MK-0524A, lovas In certain embodiments, the one or more compounds used in tatin plus extended release niacin, rosuvastatin plus fenofi the treatment of CVD comprise one or more non-statin com brate, pravastatin plus fenofibrate, and statin plus TAK-457. pounds selected from the group consisting of bile acid In certain embodiments, the one or more compounds used in sequestrants, fibrates, niacin or niacin derivatives, cholesterol the treatment of CVD comprise one or more non-statin com 55 absorption inhibitors, cholesteryl ester transfer protein pounds selected from the group consisting of bile acid (CETP) inhibitors, microsomal triglyceride transfer protein sequestrants, fibrates, niacin or niacin derivatives, cholesterol (MTP) inhibitors, squalene synthase inhibitors, ACE inhibi absorption inhibitors, cholesteryl ester transfer protein tors, angiotensin II receptor antagonists, beta-adrenergic (CETP) inhibitors, microsomal triglyceride transfer protein blockers, calcium channel blockers, and antithrombotics. In (MTP) inhibitors, squalene synthase inhibitors, ACE inhibi 60 certain embodiments, the one or more spLA inhibitors and tors, angiotensin II receptor antagonists, beta-adrenergic the one or more compounds used in the treatment of CVD blockers, calcium channel blockers, and antithrombotics. In may be administered simultaneously. In certain of these certain embodiments, administration of one or more spIA embodiments, the one or more spLA inhibitors and the one inhibitors and one or more compounds used in the treatment or more compounds used in the treatment of CVD may be of CVD results in a decrease in total cholesterol, non-HDL 65 administered in a single formulation, while in other embodi cholesterol, LDL, LDL particle, small LDL particle levels, ments the compounds may be administered simultaneously in oxidized LDL, and/or ApoB levels. In certain embodiments, two or more formulations. In each of these embodiments, the US 8,048,880 B2 7 8 formulation(s) may further comprise one or more pharmaceu Subject on a once a day basis. In certain embodiments, the one tically acceptable carriers. In other embodiments, the one or or more spIA inhibitors may be administered at a dosage of more SPLA inhibitors and the one or more compounds used about 50 to about 500 mg. in the treatment of CVD may be administered sequentially. In In certain embodiments, methods are provided for treating certain embodiments, the one or more SPLA inhibitors may CVD or a condition associated with CVD in a subject in need be administered to the Subject twice or more per day, and in thereof by administering a therapeutically effective amount certain of these embodiments the one or more spLA inhibi of one or more SPLA inhibitors and one or more compounds tors are administered twice per day. In other embodiments, used in the treatment of CVD. In certain embodiments, the the one or more spLA inhibitors may be administered to the one or more spLA inhibitors comprise A-001 or a pharma Subject on a once a day basis. In certain embodiments, the one 10 ceutically acceptable prodrug, salt, polymorph, co-crystal, or or more SPLA inhibitors may be administered at a dosage of about 50 to about 500 mg. solvate of A-001. In certain of these embodiments, the one or In certain embodiments, methods are provided for increas more spIA inhibitors comprise A-002, a prodrug of A-001. ing HDL levels in a subject in need thereof by administering In certain embodiments, the one or more compounds used in a therapeutically effective amount of one or more spLA 15 the treatment of CVD comprise one or more statins or statin inhibitors and a therapeutically effective amount of one or combination drugs. In certain of these embodiments, the one more compounds used in the treatment of CVD. In certain or more statins are selected from the group consisting of embodiments, the one or more spLA inhibitors comprise atorvastatin, simvastatin, rosuvastatin, lovastatin, pravasta A-001 or a pharmaceutically acceptable prodrug, salt, poly tin, cerivastatin, fluvastatin, mevastatin, pitavastatin, and morph, co-crystal, or solvate of A-001. In certain of these various salts, Solvates, Stereoisomers, and prodrug derivatives embodiments, the one or more spLA inhibitors comprise thereof. In certain of these embodiments, the statin combina A-002, a prodrug of A-001. In certain of these embodiments, tion drugs are selected from the group consisting of atorvas the one or more compounds used in the treatment of CVD tatin plus eZetimibe, atorvastatin plus amlodipine, atorvasta comprise one or more statins or statin combination drugs. In tin plus CP-529414, atorvastatin plus APA-01, simvastatin certain of these embodiments, the one or more statins are 25 plus eZetimibe, simvastatin plus extended release niacin, sim selected from the group consisting of atorvastatin, simvasta vastatin plus MK-0524A, lovastatin plus extended release tin, rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvas niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofi tatin, mevastatin, pitavastatin, and various salts, Solvates, Ste brate, and statin plus TAK-457. In certain embodiments, the reoisomers, and prodrug derivatives thereof. In certain of one or more compounds used in the treatment of CVD com these embodiments, the statin combination drugs are selected 30 prise one or more non-statin compounds selected from the from the group consisting of atorvastatin plus eZetimibe, ator group consisting of bile acid sequestrants, fibrates, niacin or vastatin plus amlodipine, atorvastatin plus CP-529414, ator niacin derivatives, cholesterol absorption inhibitors, choles vastatin plus APA-01, simvastatin plus eZetimibe, simvastatin teryl ester transfer protein (CETP) inhibitors, microsomal plus extended release niacin, simvastatin plus MK-0524A. triglyceride transfer protein (MTP) inhibitors, squalene syn lovastatin plus extended release niacin, rosuvastatin plus 35 thase inhibitors, ACE inhibitors, angiotensin II receptor fenofibrate, pravastatin plus fenofibrate, and statin plus TAK antagonists, beta-adrenergic blockers, calcium channel 457. In certain embodiments, the one or more compounds blockers, and antithrombotics. In certain embodiments, used in the treatment of CVD comprise one or more non administration of one or more spLA inhibitors results in a statin compounds selected from the group consisting of bile decrease in levels of one or more inflammatory markers. In acid sequestrants, fibrates, niacin or niacin derivatives, cho 40 certain of these embodiments, the inflammatory markers may lesterol absorption inhibitors, cholesteryl ester transfer pro include, but are not limited to, spLA, CRP, and/or IL-6. In tein (CETP) inhibitors, microsomal triglyceride transfer pro certain embodiments, CVD or a condition associated with tein (MTP) inhibitors, squalene synthase inhibitors, ACE CVD includes, but is not limited to, atherosclerosis, meta inhibitors, angiotensin II receptor antagonists, beta-adrener bolic syndrome, coronary artery disease, coronary heart dis gic blockers, calcium channel blockers, and antithrombotics. 45 ease, cerebrovascular disease, peripheral vascular disease, In certain embodiments, administration of one or more spIA and/or conditions associated with coronary artery disease, inhibitors and one or more compounds used in the treatment coronary heart disease, cerebrovascular disease, or peripheral of CVD results in an improvement in HDL/LDL ratio. In vascular disease. In certain embodiments, the one or more certain embodiments, improvement in HDL/LDL ratio is fur SPLA inhibitors and the one or more compounds used in the ther accomplished by a reduction in LDL levels. In certain 50 treatment of CVD may be administered simultaneously. In embodiments, the one or more spLA inhibitors and the one certain of these embodiments, the one or more spLA inhibi or more compounds used in the treatment of CVD may be tors and the one or more compounds used in the treatment of administered simultaneously. In certain of these embodi CVD may be administered in a single formulation, while in ments, the one or more spIA inhibitors and the one or more other embodiments the compounds may be administered compounds used in the treatment of CVD may be adminis 55 simultaneously in two or more formulations. In each of these tered in a single formulation, while in other embodiments the embodiments, the formulation(s) may further comprise one compounds may be administered simultaneously in two or or more pharmaceutically acceptable carriers. In other more formulations. In each of these embodiments, the formu embodiments, the one or more spLA inhibitors and the one lation(s) may further comprise one or more pharmaceutically or more compounds used in the treatment of CVD may be acceptable carriers. In other embodiments, the one or more 60 administered sequentially. In certain embodiments, the one or SPLA inhibitors and the one or more compounds used in the more spLA inhibitors may be administered to the subject treatment of CVD may be administered sequentially. In cer twice or more per day, and in certain of these embodiments tain embodiments, the one or more spLA inhibitors may be the one or more spLA inhibitors are administered twice per administered to the Subject twice or more per day, and in day. In other embodiments, the one or more spLA inhibitors certain of these embodiments the one or more spIA inhibi 65 may be administered to the Subject on a once a day basis. In tors are administered twice per day. In other embodiments, certain embodiments, the one or more SPLA inhibitors may the one or more spLA inhibitors may be administered to the be administered at a dosage of about 50 to about 500 mg. US 8,048,880 B2 10 In certain embodiments, methods are provided for treating these embodiments, the statin may be selected from the group metabolic syndrome in a subject in need thereof by adminis consisting of atorvastatin, simvastatin, rosuvastatin, lovasta tering a therapeutically effective amount of one or more tin, pravastatin, cerivastatin, fluvastatin, mevastatin, pitavas SPLA inhibitors and one or more compounds used in the tatin, and various salts, Solvates, Stereoisomers, and prodrug treatment of CVD. In certain embodiments, the one or more derivatives thereof. In certain of these embodiments, the sta sPLA inhibitors comprise A-001 or a pharmaceutically tin combination drug may be selected from the group consist acceptable prodrug, salt, polymorph, co-crystal, or Solvate of ing of atorvastatin plus eZetimibe, atorvastatin plus amlo A-001. In certain of these embodiments, the one or more dipine, atorvastatin plus CP-529414, atorvastatin plus APA sPLA inhibitors comprise A-002, a prodrug of A-001. In 01, simvastatin plus eZetimibe, simvastatin plus extended certain embodiments, the one or more compounds used in the 10 release niacin, simvastatin plus MK-0524A, lovastatin plus treatment of CVD comprise one or more statins or statin extended release niacin, rosuvastatin plus fenofibrate, prav combination drugs. In certain of these embodiments, the one astatin plus fenofibrate, and statin plus TAK-457. In certain or more statins are selected from the group consisting of embodiments, the compound used in the treatment of CVD atorvastatin, simvastatin, rosuvastatin, lovastatin, pravasta may be a non-statin compound selected from the group con tin, cerivastatin, fluvastatin, mevastatin, pitavastatin, and 15 sisting of bile acid sequestrants, fibrates, niacin or niacin various salts, Solvates, Stereoisomers, and prodrug derivatives derivatives, cholesterol absorption inhibitors, cholesteryl thereof. In certain of these embodiments, the statin combina ester transfer protein (CETP) inhibitors, microsomal triglyc tion drugs are selected from the group consisting of atorvas eride transfer protein (MTP) inhibitors, squalene synthase tatin plus eZetimibe, atorvastatin plus amlodipine, atorvasta inhibitors, ACE inhibitors, angiotensin II receptor antago tin plus CP-529414, atorvastatin plus APA-01, simvastatin nists, beta-adrenergic blockers, calcium channel blockers, plus eZetimibe, simvastatin plus extended release niacin, sim and antithrombotics. In certain embodiments, the one or more vastatin plus MK-0524A, lovastatin plus extended release sPLA inhibitors may be administered simultaneously with niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofi the compound used in the treatment of CVD. In certain of brate, and statin plus TAK-457. In certain embodiments, the these embodiments, the one or more spLA inhibitors and the one or more compounds used in the treatment of CVD com 25 compound used in the treatment of CVD may be administered prise one or more non-statin compounds selected from the in a single formulation, while in other embodiments the com group consisting of bile acid sequestrants, fibrates, niacin or pounds may be administered simultaneously in two or niacin derivatives, cholesterol absorption inhibitors, choles more formulations. In each of these embodiments, the formu teryl ester transfer protein (CETP) inhibitors, microsomal lation(s) may further comprise one or more pharmaceutically triglyceride transfer protein (MTP) inhibitors, squalene syn 30 acceptable carriers. In other embodiments, the one or more thase inhibitors, ACE inhibitors, angiotensin II receptor sPLA inhibitors and the compound used in the treatment of antagonists, beta-adrenergic blockers, calcium channel CVD may be administered sequentially. In certain embodi blockers, and antithrombotics. In certain embodiments, ments, the one or more spLA inhibitors may be administered administration of one or more spLA inhibitors results in a to the Subject twice or more per day, and in certain of these decrease in levels of one or more inflammatory markers. In 35 embodiments the one or more spLA inhibitors are adminis certain of these embodiments, the inflammatory markers may tered twice per day. In other embodiments, the one or more include, but are not limited to, spLA, CRP, and/or IL-6. In sPLA inhibitors may be administered to the subject on a once certain embodiments, the one or more spLA inhibitors and a day basis. In certain embodiments, the one or more SPLA the one or more compounds used in the treatment of CVD inhibitors may be administered at a dosage of about 50 to may be administered simultaneously. In certain of these 40 about 500 mg. embodiments, the one or more spLA inhibitors and the one In certain embodiments, the use of one or more SPLA or more compounds used in the treatment of CVD may be inhibitors for preparation of a medicament for treating dys administered in a single formulation, while in other embodi lipidemia, treating CVD and conditions associated with ments the compounds may be administered simultaneously in CVD, lowering cholesterol levels, lowering triglyceride lev two or more formulations. In each of these embodiments, the 45 els, increasing HDL levels, and improving HDL/LDL ratios formulation(s) may further comprise one or more pharmaceu in a Subject is provided. In certain embodiments, the one or tically acceptable carriers. In other embodiments, the one or more spLA inhibitors comprise A-001 or a pharmaceutically more SPLA inhibitors and the one or more compounds used acceptable prodrug, salt, polymorph, co-crystal, or Solvate of in the treatment of CVD may be administered sequentially. In A-001. In certain of these embodiments, the one or more certain embodiments, the one or more SPLA inhibitors may 50 sPLA inhibitors comprise A-002, a prodrug of A-001. In be administered to the Subject twice or more per day, and in certain embodiments, the medicament further comprises one certain of these embodiments the one or more spIA inhibi or more pharmaceutically acceptable carriers. tors are administered twice per day. In other embodiments, In certain embodiments, the use of one or more SPLA the one or more spLA inhibitors may be administered to the inhibitors and one or more compounds used in the treatment Subject on a once a day basis. In certain embodiments, the one 55 of CVD for preparation of a medicament for treating dyslipi or more spIA inhibitors may be administered at a dosage of demia, treating CVD and conditions associated with CVD. about 50 to about 500 mg. lowering cholesterol levels, lowering triglyceride levels, In certain embodiments, methods are provided for increas increasing HDL levels, and/or improving HDL/LDL ratios in ing the effectiveness of a compound used in the treatment of a subject is provided. In certain embodiments, the one or more CVD in a subject by administering one or more spLA inhibi 60 sPLA inhibitors comprise A-001 or a pharmaceutically tors to said subject. In certain embodiments, the one or more acceptable prodrug, salt, polymorph, co-crystal, or Solvate of sPLA inhibitors comprise A-001 or a pharmaceutically A-001. In certain of these embodiments, the one or more acceptable prodrug, salt, polymorph, co-crystal, or Solvate of sPLA inhibitors comprise A-002, a prodrug of A-001. In A-001. In certain of these embodiments, the one or more certain embodiments, the one or more compounds used in the sPLA inhibitors comprise A-002, a prodrug of A-001. In 65 treatment of CVD comprise one or more statins or statin certain embodiments, the compound used in the treatment of combination drugs. In certain of these embodiments, the one CVD may be a statin or statin combination drug. In certain of or more statins are selected from the group consisting of US 8,048,880 B2 11 12 atorvastatin, simvastatin, rosuvastatin, lovastatin, pravasta weeks, and HDL levels were measured. A: low dose A-002; tin, cerivastatin, fluvastatin, mevastatin, pitavastatin, and B: high dose A-002; C: statin; D: low dose A-002 plus statin: various salts, Solvates, Stereoisomers, and prodrug derivatives E: high dose A-002 plus statin; and F: vehicle only. thereof. In certain of these embodiments, the statin combina FIG. 8: A-002/simvastatin combination tablet preparation tion drugs are selected from the group consisting of atorvas protocol. tatin plus eZetimibe, atorvastatin plus amlodipine, atorvasta FIG. 9: HPLC profiles. A. Simvastatin. B. A-002. C. tin plus CP-529414, atorvastatin plus APA-01, simvastatin A-002/simvastatin combination tablet #19. D. A-002/simv plus eZetimibe, simvastatin plus extended release niacin, sim astatin combination tablet #27. vastatin plus MK-0524A, lovastatin plus extended release niacin, rosuvastatin plus fenofibrate, pravastatin plus fenofi 10 DETAILED DESCRIPTION brate, and statin plus TAK-457. In certain embodiments, the one or more compounds used in the treatment of CVD com The following description of the invention is merely prise one or more non-statin compounds selected from the intended to illustrate various embodiments of the invention. group consisting of bile acid sequestrants, fibrates, niacin or As such, the specific modifications discussed are not to be niacin derivatives, cholesterol absorption inhibitors, choles 15 construed as limitations on the scope of the invention. It will teryl ester transfer protein (CETP) inhibitors, microsomal be apparent to one skilled in the art that various equivalents, triglyceride transfer protein (MTP) inhibitors, squalene syn changes, and modifications may be made without departing thase inhibitors, ACE inhibitors, angiotensin II receptor from the scope of the invention, and it is understood that Such antagonists, beta-adrenergic blockers, calcium channel equivalent embodiments are to be included herein. blockers, and antithrombotics. In certain embodiments, the medicament further comprises one or more pharmaceutically Abbreviations acceptable carriers. In addition to the exemplary embodiments described AA, arachidonic acid; ACE, angiotensin converting above, further embodiments and aspects will become appar enzyme; Ang, angiotensin; ApoB, apolipoprotein B; ARB, ent by reference to the drawings and by study of the following 25 Angiotensin Receptor Blocker; BID, twice daily: BMI, body descriptions. mass index; CAD, coronary artery disease; CETP, cholesteryl ester transfer protein; cfm, cubic feet per minute; CHD, coro BRIEF DESCRIPTION OF DRAWINGS nary heart disease; CRPC-reactive protein: CVD, cardiovas cular disease; ECG, electrocardiogram; ERN, extended FIG. 1: Effect of A-002 administration on body weight in 30 release niacin; HDL, high density lipoprotein; HMG-CoA, mice. ApoE-mice were administered vehicle only, 30 hydroxymethyl glutaryl coenzyme A: HPLC, high perfor mg/kg. A-002, or 90 mg/kg A-002 twice daily over 16 weeks. mance liquid chromatography; ICAM-1, intercellular adhe Body weight was measured once a week. sion molecule 1: IDL, intermediate density lipoprotein, IL, FIG. 2: Effect of A-002 administration on plasma total interleukin (e.g., IL-6, IL-8); ITT, intent to treat; LDL, low cholesterol levels in mice. ApoE mice were administered 35 density lipoprotein; LPA, lysophosphatidic acid: MCP-1, vehicle only, 30 mg/kg A-002, or 90 mg/kg A-002 twice daily monocyte chemotactic protein-1, MI, myocardial infarction; over 16 weeks. Total plasma cholesterol levels were measured MIP-1C, macrophage inflammatory protein 1 alpha; MTP, at 0, 4, 8, 12, and 16 weeks. microsomal triglyceride transfer protein: PAD, peripheral FIG. 3: Effect of A-002 administration on atherosclerotic artery disease; PAF, platelet activating factor; PLA2, phos plaque coverage in mice. ApoE mice were administered 40 pholipase A, QD, once daily; SPLA, secretory phospholi vehicle only, 30 mg/kg A-002, or 90 mg/kg A-002 twice daily pase A, TEAE, treatment-emergent adverse event, TG, trig over 16 weeks. At 16 weeks, aortic plaque coverage was lyceride; TIA, transient ischemic attack; TNFC., tumor measured. necrosis factor alpha; VCAM-1, vascular cell adhesion mol FIG. 4: Effect of A-002 administration on angiotensin II ecule 1: VLDL, very low density lipoprotein. mediated atherosclerotic plaque formation and aortic aneu 45 The terms “treat,” “treating,” or “treatment as used herein rysm. ApoE mice were administered saline plus water, with regards to a condition refers to preventing the condition, angiotensin II in water vehicle, angiotensin II in acacia slowing the onset or rate of development of the condition, vehicle, or angiotensin II in acacia Vehicle plus 30 mg/kg reducing the risk of developing the condition, preventing or A-002. At 4 weeks, aortic plaque coverage was measured. delaying the development of symptoms associated with the FIG. 5: En face lesions in A-002 and/or statin dosage 50 condition, reducing or ending symptoms associated with the groups. ApoE mice on a high fat diet were administered condition, generating a complete or partial regression of the various dosages of A-002, statin, or A-002 plus statin over condition, or some combination thereof. For example, with twelve weeks, and en face lesion content was measured using regard to atherosclerosis, “treatment may refer to a decrease digital imaging methods. Lesion size is expressed as percent in the likelihood of developing atherosclerotic plaque depos coverage over entire tissue sample. A: low dose A-002; B: 55 its, a decrease in the rate of development of deposits, a high dose A-002; C: statin; D: low dose A-002 plus statin: E: decrease in the number or size of existing deposits, or high dose A-002 plus statin; and F: vehicle only. improved plaque stability. Likewise, “treatment” with regard FIG. 6: Total plasma cholesterol levels in A-002 and/or to dyslipidemia may refer to a decrease in lipid levels, cho statin dosage groups. ApoE mice on a high fat diet were lesterol levels, and/or triglyceride (TG) levels. administered various dosages of A-002, statin, or A-002 plus 60 The term “subject' as used herein refers to any mammal, statin over twelve weeks, and total cholesterol levels were preferably a human. measured. A: low dose A-002; B: high dose A-002: C: statin: In certain embodiments, a “subject in need thereof refers D: low dose A-002 plus statin: E: high dose A-002 plus statin: to a subject diagnosed with CVD or exhibiting one or more and F: vehicle only. conditions associated with CVD, a subject who has been FIG. 7: HDL levels in A-002 and/or statin dosage groups. 65 diagnosed with or exhibited one or more conditions associ ApoE mice on a high fat diet were administered various ated with CVD in the past, or a subject who has been deemed dosages of A-002, statin, or A-002 plus statin over twelve at risk of developing CVD or one or more conditions associ US 8,048,880 B2 13 14 ated with CVD in the future due to hereditary or environmen levels, or some combination thereof. An increase in LDL tal factors. “Cardiovascular disease' or "CVD'as used herein particle size as used herein refers to an increase in mean includes, for example, atherosclerosis, including coronary particle size. artery atherosclerosis and carotid artery atherosclerosis, The term "elevated cholesterol level” as used herein refers coronary artery disease (CAD), coronary heart disease to a cholesterol level that is above an accepted normal thresh (CHD), conditions associated with CAD and CHD, cere old level, such as those promulgated by the National Heart brovascular disease and conditions associated with cere Lung and Blood Institute (NHLBI) National Cholesterol brovascular disease, peripheral vascular disease and condi Education Program (NCEP). The accepted normal threshold tions associated with peripheral vascular disease, aneurysm, cholesterol level may vary from subject to subject based on vasculitis, venous thrombosis, diabetes mellitus, and meta 10 various risk factors, such as for example a prior history of bolic syndrome. “Conditions associated with CAD and CVD. In certain embodiments, a subject exhibiting elevated CHD’ as used herein include, for example, angina and myo cholesterol levels may have a blood LDL level greater than or cardial infarction (MI; heart attack). “Conditions associated equal to 70 mg/dl. In certain of these embodiments, a subject with cerebrovascular disease' as used herein include, for exhibiting elevated cholesterol levels may have a blood LDL example, transient ischemic attack (TIA) and stroke. “Con 15 greater than or equal to 100 mg/dl., in other embodiments ditions associated with peripheral vascular disease' as used greater than or equal to 130 mg/dl., in other embodiments herein include, for example, claudication. “Conditions asso greater than or equal to 160 mg/dl., and in still other embodi ciated with CVD as used herein include, for example, dys ments greater than or equal to 190 mg/dl. In certain embodi lipidemia, Such as for example hyperlipidemia (elevated lipid ments, a Subject exhibiting elevated cholesterol levels may levels), hypercholesterolemia (elevated cholesterol levels), have a blood total cholesterol level greater than or equal to and hypertriglyceridemia (elevated TG levels), elevated glu 200 mg/dl. In certain of these embodiments, a subject exhib cose levels, low HDL/LDL ratio, and hypertension. There iting elevated cholesterol levels may have a blood total cho fore, in certain embodiments, a Subject in need thereofmay be lesterol greater than or equal to 240 mg/dl. a subject exhibiting dyslipidemia or a Subject that has exhib The term “triglyceride level” as used herein refers to blood ited dyslipidemia in the past or has been deemed at risk for 25 triglyceride level, serum triglyceride level, plasma triglycer developing dyslipidemia in the future. In certain of these ide level, or triglyceride level from another biological fluid. embodiments, the subject may exhibit elevated cholesterol The term “elevated triglyceride level” as used herein refers to levels, or may have exhibited elevated cholesterol levels in the a triglyceride level that is above an accepted normal threshold past or been deemed at risk for developing elevated choles level. The accepted normal threshold triglyceride level may terol levels in the future. Likewise, incertain of these embodi 30 vary from Subject to Subject based on various risk factors, ments, the subject may exhibit elevated triglyceride levels, or such as for example a prior history of CVD. In certain may have exhibited elevated triglyceride levels in the past or embodiments, a subject exhibiting elevated triglyceride lev been deemed at risk for developing elevated triglyceride lev els may have a blood triglyceride level greater than or equal to els in the future. In certain embodiments, a Subject in need 150 mg/dl. In certain of these embodiments, a subject exhib thereof may have a condition associated with inflammation, 35 iting elevated triglyceride levels may have a blood triglycer may have been diagnosed with Such a condition in the past, or ide level greater than or equal to 200 mg/dl., in other embodi may have been deemed at risk for developing Sucha condition ments greater than or equal to 300 mg/dl., in other in the future. In addition to atherosclerosis and certain other embodiments greater than or equal to 400 mg/dl., and in still forms of CVD, conditions associated with inflammation other embodiments greater than or equal to 500 mg/dl. include, for example, multiple Sclerosis (Cunningham 2006), 40 The term “statin” as used herein refers to any compound Alzheimer's disease (Moses 2006), sickle cell (Styles 1996), that inhibits HMG-CoA reductase, an enzyme that catalyzes rheumatoid arthritis, and osteoarthritis (Jamal 1998). In these the conversion of HMG-CoA to mevalonate. embodiments, a subject in need thereof may exhibit elevated The term “sPLA inhibitor as used herein refers to any sPLA levels, may have exhibited elevated sPLA levels in compound or prodrug thereof that inhibits the activity of the past, or may have been deemed at risk for developing 45 sPLA. elevated spLA levels. In other embodiments, a subject in A “therapeutically effective amount of a composition as need thereofmay exhibitsPLA levels falling within a normal used herein is an amount of a composition that produces a range. In certain embodiments, a Subject in need thereof may desired therapeutic effect in a subject, Such as treating a target exhibit elevated levels of one or more additional markers condition. The precise therapeutically effective amount is an associated with inflammation, including but not limited to 50 amount of the composition that will yield the most effective CRP, IL-6, MCP-1, TNFO, IL-8, ICAM-1, VCAM-1, and results in terms of therapeutic efficacy in a given subject. This MIP-1 O. amount will vary depending upon a variety of factors, includ The term “cholesterol level” as used herein refers to blood ing but not limited to the characteristics of the therapeutic cholesterol level, serum cholesterol level, plasma cholesterol composition (including, e.g., activity, pharmacokinetics, level, or cholesterol level from another biological fluid. A 55 pharmacodynamics, and bioavailability), the physiological decrease in cholesterol levels as used herein may refer to a condition of the Subject (including, e.g., age, body weight, decrease in total cholesterol levels or a decrease in one or sex, disease type and stage, medical history, general physical more of total cholesterol, non-HDL cholesterol, LDL, VLDL, condition, responsiveness to a given dosage, and other present and/or IDL levels. A decrease in LDL as used herein may ), the nature of the pharmaceutically acceptable refer to a decrease in total LDL, a decrease in LDL particles, 60 carrier or carriers in the composition, and the route of admin a decrease in Small LDL particles, a decrease in oxidized LDL istration. One skilled in the clinical and pharmacological arts levels, and/or a decrease in ApoB levels. A decrease in VLDL will be able to determine a therapeutically effective amount as used herein may refer to a decrease in total VLDL or to a through routine experimentation, namely by monitoring a decrease in the level of one or more of VLDL subparticles V1 Subject's response to administration of a composition and to V6. An improvement in HDL/LDL ratio as used herein 65 adjusting the dosage accordingly. For additional guidance, refers to any increase in the ratio of HDL to LDL, and may be see, e.g., Remington: The Science and Practice of Pharmacy, accomplished by decreasing LDL levels, increasing HDL 21 Edition, Univ. of Sciences in Philadelphia (USIP), Lip US 8,048,880 B2 15 16 pincott Williams & Wilkins, Philadelphia, Pa., 2005, and metabolic syndrome are at increased risk of developing CAD, Goodman & Gilman's The Pharmacological Basis of Thera CHD, conditions associated with CAD and CHD, and type 2 peutics, 11th Edition, McGraw-Hill, New York, N.Y., 2006, diabetes. the entire disclosures of which are incorporated by reference Over time, hypercholesterolemia and hypertriglyceri herein. demia can lead to the development of atheromatous plaques A “pharmaceutically acceptable carrier as used herein on the inner arterial linings via the process of atherogenesis, refers to a pharmaceutically acceptable material, composi which in turn results in atherosclerosis. Atherosclerosis leads tion, or vehicle that is involved in carrying or transporting a to significantly reduced blood flow through the arteries, compound of interest from one tissue, organ, orportion of the which in turn leads to the development of CAD, CHD, and 10 conditions associated with CAD and CHD. body to another tissue, organ, or portion of the body. Such a Reduction of cholesterol levels, particularly LDL levels, carrier may comprise, for example, a liquid, Solid, or semi and/or reduction of TG levels have been shown to delay the Solid filler, solvent, Surfactant, diluent, excipient, adjuvant, onset and decrease the progression ofatherosclerosis, thereby binder, buffer, dissolution aid, solvent, encapsulating mate reducing the risk of developing CAD and CHD. In certain rial, sequestering agent, dispersing agent, preservative, lubri 15 cases, adequate cholesterol and/or TG reduction may be cant, disintegrant, thickener, emulsifier, antimicrobial agent, accomplished through diet adjustment. In other cases, how antioxidant, stabilizing agent, coloring agent, or some com ever, adequate reduction requires the administration of one or bination thereof. Each component of the carrier must be more compounds used in the treatment of CVD. Compounds “pharmaceutically acceptable' in that it must be compatible used in the treatment of CVD include compounds for lower with the other ingredients of the composition and must be ing cholesterol levels and/or increasing HDL levels, such as Suitable for contact with any tissue, organ, or portion of the for example statins, bile acid sequestrants such as body that it may encounter, meaning that it must not carry a cholestyramine resin (Questran?R), PrevaliteR), risk of toxicity, irritation, allergic response, immunogenicity, hydrochloride (ColestidR), or colesevelam hydrochloride or any other complication that excessively outweighs its (WelChol(R), Cholestagel(R), fibrates such as therapeutic benefits. Examples of pharmaceutically accept 25 (Bezalip(R), (ModalimR), , able carriers for use in the presently disclosed pharmaceutical (LopidR), or fenofibrate (Antara R, TriCorr), ABT-335), nia compositions include, but are not limited to, diluents such as cin or niacin derivatives such as Xanthinol niacinate, niacin microcrystalline cellulose or lactose (e.g., anhydrous lactose, immediate-release (Niacor(R), extended release forms of nia lactose fast flo), binders such as gelatin, polyethylene glycol, cin (ERN) (Niaspan R, Niaspan MF, or Niaspan CF), or wax, microcrystalline cellulose, synthetic gums such as poly 30 extended release niacin combinations (e.g., extended release vinylpyrrolidone, or cellulosic polymers such as hydroxypro niacin plus the DP-1 antagonist (MK-0524), com bination known as MK-0524A and marketed as Cordap pyl cellulose (e.g., hydroxypropyl methylcellulose (HPMC)), tive(R), cholesterol absorption inhibitors such as eZetimibe lubricants such as magnesium Stearate, calcium Stearate, (ZetiaR), AVE 5530, or MD-0727, cholesteryl ester transfer Stearic acid, or microcrystalline cellulose, disintegrants such 35 protein (CETP) inhibitors such as JTT-705/RO4607381 as starches, cross-linked polymers, or celluloses (e.g., cros (R1658), CP-529414 ((R), or MK-0859, microso carmellose sodium (CCNa), fillers such as silicon dioxide, mal triglyceride transfer protein (MTP) inhibitors such as titanium dioxide, microcrystalline cellulose, or powdered AEGR-733 and AEGR-733 combinations (e.g., AEGR-733 cellulose, Surfactants or emulsifiers such as polysorbates plus eZetimibe), squalene synthase inhibitors such as (e.g., Polysorbate 20, 40, 60, or 80: Span20, 40, 60, 65, or 80), 40 acetate (TAK-475) and lapaquistat acetate com antioxidant agents such as butylated hydroxyanisole (BHA), binations (e.g., TAK-475 plus one or more statins), and other butylated hydroxytoluene (BHT), propyl gallate, or ascorbic miscellaneous compounds such as , ISIS acid (either free acid or salt forms thereof), buffers such as 301012, cardioprotectants such as MC-1 antibody, glycopro phosphate or citrate buffers, sequestering agents such as eth tein IIb/IIIa inhibitors such as tirofiban hydrochloride (Aggr ylenediaminetetraacetic acid (EDTA), ethylene glycol tet 45 astat(R), TG100-115, AEGR 773, AEGR 427, stanols, or ste raacetic acid (EGTA), or edetate disodium, dispersing agents rols. In addition to compounds that lower cholesterol levels or Such as Sodium carboxymethylcelluose, hydroxypropyl increase HDL levels, compounds used in the treatment of methylcellulose, povidone, or polyvinylpyrrolidone, dissolu CVD include, for example, ACE inhibitors such as lisinopril, tion aids such as calcium carbonate, and excipients such as captopril, enalapril, nitrosated ACE inhibitors, or ACE inhibi water, Saline, dextrose, glycerol, or ethanol, citric acid, cal 50 tor combinations (e.g., lisinopril plus MC-1 antibody, com cium metabisulfite, lactic acid, malic acid, Succinic acid, or bination referred to as MC-4232), angiotensin II receptor tartaric acid. antagonists, nitrosated angiotensin II receptor antagonists, or Metabolic syndrome is a disorder characterized by a group angiotensin II receptor antagonist combinations (e.g., Angio of metabolic risk factors. These factors include, for example, tensin Receptor Blocker (ARB) plus MC-1, combination dyslipidemia, abdominal obesity, elevated blood pressure 55 referred to as MC-4262), beta-adrenergic blockers or nitro (hypertension), insulin resistance orglucose intolerance, pro sated beta-adrenergic blockers, calcium channel blockers, or thrombotic State, and proinflammatory state. Subjects are antithrombotics such as aspirin or nitrosated aspirin. generally classified as having metabolic syndrome if they Among the most well-known and commonly used com meet three of the five following criteria: 1) abdominal obesity pounds used in the treatment of CVD are statins. Statins are (waist circumference >35 inches in women, >40 inches in 60 compounds that inhibit HMG-CoA reductase from cata men); 2) low HDL levels (<50 mg/dL in women, <40 mg/dL lyzing the conversion of HMG-CoA to mevalonate, a rate in men); 3) high blood pressure (21130/85 mm Hg) or cur limiting step in the cholesterol biosynthetic pathway. As such, rent treatment with antihypertensive ; 4) hyper statins function as potent lipid lowering agent. Statin admin triglyceridemia (TG levels 2150 mg/dL); and 5) impaired istration significantly decreases blood LDL levels, and mod fasting glucose (blood glucose levels of 2110 mg/dL). Meta 65 erately decreases blood TG levels. In addition, it has been bolic syndrome is associated with elevated levels of various proposed that statins may prevent CVD by improving endot inflammatory markers, such as CRP or IL-6. Subjects with helial function, modulating inflammatory responses, main US 8,048,880 B2 17 18 taining plaque stability, and preventing thrombus formation. expression of SPLA type V has been shown to increase arte Examples of statins that may be used in conjunction with the riosclerosis in mice, while a deficiency of spLA type V has compositions and methods disclosed herein include, but are been shown to reduce arteriosclerosis (Rosengren 2006: Bos not limited to, atorvastatin oratorvastatin calcium (marketed trom 2007). as Lipitor R or Torvast(R); see, e.g., U.S. Pat. Nos. 4,681,893 or SPLA expression has also been correlated with an 5,273.995) and atorvastatin combinations (e.g., atorvastatin increased risk of development of CAD. Higher circulating plus amlodipine (marketed as Norvasc(R), combination mar levels of sRLA, and of spLA type IIA specifically, have keted as Caduet(R), see, e.g., U.S. Pat. No. 6,455,574; atorv been observed in patients with documented CAD than in astatin plus CP-529414 (marketed as TorcetrapibR); atorvas control patients (Kugiyama 1999; Liu 2003; Boekholdt 2005; tatin plus APA-01; atorvastatin plus eZetimibe), cerivastatin 10 Chait 2005; Hartford 2006). In addition, higher circulating (marketed as Lipobay(R) or BaycolR), fluvastatin (marketed levels of SPLA were found to provide an accurate prognostic as LescolR); U.S. Pat. No. 4,739,073), lovastatin (marketed as indicator for development of CAD in healthy individuals Mevacor R or Altocor R; see, e.g., U.S. Pat. No. 4,231.938), (Mallat 2007). Measurement of spLA activity has been lovastatin combinations (e.g., lovastatin plus Niaspan R, shown to be an independent predictor of death and new or combination marketed as Advicor R), mevastatin, pitavastatin 15 recurrent myocardial infarction in Subjects with acute coro (marketed as LivaloR or PitavaR), pravastatin (marketed as nary syndrome, and provides greater prognostic accuracy Pravachol(R), MevalotinR), Selektine(R), or Lipostat(R); see, than measuring type IIA concentration only (Mallat 2005). It e.g., U.S. Pat. No. 4.346.227), pravastatin combinations (e.g., has also been proposed that SPLA may have detrimental pravastatin plus fenofibrate), rosuvastatin (marketed as Cre effects in the setting of ischemic events. This is based largely storR), rosuvastatin combinations (e.g., rosuvastatin plus Tri on the finding of SPLA depositions in the necrotic center of Corr), simvastatin (marketed as Zocor(R) or LipexR; see, infarcted human myocardium (Nijmeijer 2002). e.g., U.S. Pat. Nos. 4,444,784; 4.916,239; and 4,820,850), As disclosed herein, administration of the spLA inhibitor and simvastatin combinations (e.g., simvastatin plus A-002, which is a prodrug form of A-001, decreased plasma eZetimibe, combination marketed as VytorinR), see, e.g., U.S. total cholesterol levels, increased plasma HDL levels, and Pat. No. 7,229.982; simvastatin plus Niaspan R, combination 25 decreased atherosclerotic plaque formation and aortic aneu marketed as Simcor R; simvastatin plus MK-0524A, combi rysm in mice. In humans with CVD, twice daily administra nation referred to as MK-0524B), as well as various pharma tion of A-002 decreased serum LDL, LDL particle, small ceutically acceptable salts, Solvates, salts, Stereoisomers, pro LDL particle, total cholesterol, and TG levels. Decreases in drugs derivatives, or nitroderivatives of the compounds listed LDL were observed in a diabetic subpopulation, as well as in above. In some cases, such as for example with simvastatin, 30 a subpopulation exhibiting elevated baseline LDL levels, the active form of the statin is a metabolite formed in the body while decreases in TG levels were observed in a metabolic of a subject following administration. In other cases, statins syndrome subpopulation. Additionally, administration of are administered in their active form. A-002 was found to decrease serum levels of the inflamma Phospholipases A (PLAs) are a family of enzymes that tory markers spLA, CRP, and IL-6. Surprisingly, similar catalyze hydrolysis of the sn-2 fatty acyl ester bond of phos 35 results were obtained with once a day administration of pholipids to produce free fatty acids and lysophospholipids A-002, which caused a decrease in LDL, non-HDL choles Such as arachidonic acid (AA) and lysophosphatidylcholine. terol, total cholesterol, small LDL particle, oxidized LDL, AA can then be converted into eicosanoids such as prostag triglyceride, and ApoB levels and an increase in LDL particle landins, leukotrienes, thromboxanes, and lipoxins, while size. In addition, once a day dosing with A-002 prevented the lysophosphatidylcholine can be metabolized to lysophospha 40 large increase in CRP levels observed in the placebo group. tidic acid (LPA) or platelet-activating factor (PAF). PLAs Based on the experimental results disclosed herein showing have been classified into several groups based on factors such that administration of spLA inhibitors decreases total cho as cellular localization, amino acid sequence, molecular lesterol levels, atherosclerotic plaque formation, and aortic mass, and calcium requirement for enzymatic activity (Ra aneurysm in mice, decreases total cholesterol, non-HDL cho moner 2005). 45 lesterol, LDL, LDL particle, small LDL particle, oxidized Secretory phospholipase A (SPLA) is an extracellular or LDL particle, TG, and ApoB levels in humans, and increases secreted subgroup of PLA that plays a role in inducing LDL particle size in humans, administration of a therapeuti inflammation. Elevated levels of spLA types IIA, IID, IIE, cally effective amount of one or more spLA inhibitors may IIF, III, V, and X have been observed in all stages of athero be used to treat CVD and conditions associated with CVD, Sclerosis development and have been implicated in athero 50 Such as for example dyslipidemia, atherosclerosis, metabolic genesis based on their ability to degrade phospholipid syndrome, CAD, CHD, and conditions associated with CAD (Kimura-Matsumoto 2007). SPLA type IIA has been found and CHD. to be expressed at vascular Smooth muscle cells and foam Provided herein in certain embodiments are methods of cells in human arteriosclerosis lesions, and this expression treating CVD and conditions associated with CVD in a sub has been recognized to have a correlation with the develop 55 ject in need thereof by administering a therapeutically effec ment of arteriosclerosis (Menschikowski 1995; Elinder tive amount of one or more spLA inhibitors. In certain of 1997). Transgenic mice that express high levels of human these embodiments, administration of one or more SPLA type IIA spLA have increased LDL levels, decreased HDL inhibitors may result in, among other effects, a decrease in levels, and arteriosclerotic lesions (Ivandic 1999; Tietge cholesterol levels, a decrease in TG levels, an increase in LDL 2000), and develop arteriosclerosis at a higher rate compared 60 particle size, and/or an improvement in HDL/LDL ratio. to normal mice when given a high fat diet (Ivandic 1999). Therefore, also provided herein are methods of treating dys Treatment with sPLA modifies LDL lipoproteins such that lipidemia in a subject in need thereof. In certain embodi they have higher affinity for extracellular matrix proteins ments, one or more spLA inhibitors for use in these methods (Cameo 1998; Sartipy 1999; Hakala 2001), resulting in an comprise A-001 or a pharmaceutically acceptable prodrug, increased retention of LDL particles in the arterial wall. In 65 salt, polymorph, co-crystal, or solvate of A-001. In certain of addition, there is some evidence that spLA remodels HDL, these embodiments, the pharmaceutically acceptable prodrug resulting in HDL catabolism (Pruzanski 1998). Increased is A-002. In certain embodiments, administration of one or US 8,048,880 B2 19 20 more SPLA inhibitors may resultina decrease in one or more hereinfortreating CVD and conditions associated with CVD. markers associated with inflammation. Such markers include, including dyslipidemia and atherosclerosis, in a Subject in but are not limited to, spLA, CRP, and IL-6. Also provided need thereof by administering a therapeutically effective herein are compositions comprising one or more SPLA amount of one or more spIA inhibitors and a therapeutically inhibitors for use in the methods disclosed herein, such as for effective amount of one or more compounds used in the example in the treatment of CVD, conditions associated with treatment of CVD, such as for example statins. Also provided CVD, and/or dyslipidemia or to lower total cholesterol, LDL, herein are methods of increasing the effectiveness of a com non-HDL cholesterol, LDL particle, small LDL particle, oxi pound used in the treatment of CVD, such as for example a dized LDL, ApoB, and/or triglyceride levels or to raise HDL statin, by administering one or more spIA inhibitors in levels. In certain embodiments, these compositions further 10 conjunction with the compound used in the treatment of comprise one or more pharmaceutically acceptable carriers. CVD. Compositions comprising one or more spIA inhibitors Provided herein in certain embodiments are methods of for use in the methods disclosed herein may be administered treating dyslipidemia in a Subject in need thereof by admin to a Subject on a one-time basis or in multiple administrations. istering a therapeutically effective amount of one or more In those embodiments wherein these compositions are given 15 sPLA inhibitors and a therapeutically effective amount of in multiple administrations, the compositions may be admin one or more compounds used in the treatment of CVD. In istered at set intervals over a particular time period deter certain embodiments, the one or more spIA inhibitors com mined in advance, or they may be administered indefinitely or prise A-001 or a pharmaceutically acceptable prodrug, salt, until a particular therapeutic benchmark is reached. Such as polymorph, co-crystal, or Solvate of A-001, and in certain of for example until a subject exhibits cholesterol levels below a these embodiments the pharmaceutically acceptable prodrug specified threshold. In certain embodiments, the one or more is A-002. In certain embodiments, the one or more com sPLA inhibitors may be administered from one or more pounds used in the treatment of CVD comprise one or more times per day to once every week, once every month, or once statins or statin combination drugs. In certain of these every several months. In certain embodiments, the one or embodiments, the one or more statins are selected from the more SPLA inhibitors may be administered twice a day, and 25 group consisting of atorvastatin, simvastatin, rosuvastatin, in other embodiments the one or more spLA inhibitors may lovastatin, pravastatin, cerivastatin, fluvastatin, mevastatin, be administered once a day. As disclosed herein, both twice a and pitavastatin, and the statin combination drugs are selected day and once a day administration of A-002 resulted in sig from the group consisting of atorvastatin plus eZetimibe, ator nificant decreases in serum lipid levels. vastatin plus amlodipine, atorvastatin plus CP-529414, ator In certain embodiments, kits are provided that comprise 30 vastatin plus APA-01, simvastatin plus eZetimibe, simvastatin one or more spLA inhibitors. In certain embodiments, the plus extended release niacin, simvastatin plus MK-0524A. one or more spILA inhibitors comprise A-001 or a pharma lovastatin plus extended release niacin, rosuvastatin plus ceutically acceptable prodrug, salt, polymorph, co-crystal, or fenofibrate, pravastatin plus fenofibrate, and statin plus TAK solvate of A-001, and in certain of these embodiments, the 457. In certain embodiments, the one or more compounds pharmaceutically acceptable prodrug is A-002. In certain 35 used in the treatment of CVD comprise one or more non embodiments, the kit provides instructions for usage, Such as statin compounds selected from the group consisting of bile dosage or administration instructions. acid sequestrants, fibrates, niacin or niacin derivatives, cho As disclosed herein, twice daily administration of A-002 lesterol absorption inhibitors, cholesteryl ester transfer pro was shown to further decrease total cholesterol levels in mice tein (CETP) inhibitors, microsomal triglyceride transfer pro and LDL, LDL particle, and small LDL particle levels in 40 tein (MTP) inhibitors, squalene synthase inhibitors, ACE humans that were receiving statin treatment. Likewise, A-002 inhibitors, angiotensin II receptor antagonists, beta-adrener was shown to further decrease serum LDL levels in humans gic blockers, calcium channel blockers, and antithrombotics. that were receiving eZetimibe treatment. Similar results were In certain of these embodiments, the one or more compounds obtained when A-002 was administered once a day in com used in the treatment of CVD comprise eZetimibe. In certain bination with statins. These results indicate that administra 45 embodiments, administration of one or more spLA inhibi tion of an SPLA inhibitor unexpectedly causes an additional tors and one or more statins results in a decrease in cholesterol decrease in cholesterol levels in subjects that are already levels, and in certain of these embodiments the decrease in being treated with another compound used in the treatment of cholesterol levels is greater than that observed following CVD, such as a statin. As further disclosed herein, adminis administration of one or more spLA inhibitors or one or tration of A-002 in conjunction with statins resulted in an 50 more statins alone. In certain of these embodiments, the unexpected Synergistic decrease in total cholesterol levels decrease in cholesterol levels is synergistic, meaning that the and aorticlesion formation in mice and in LDL and Small decrease is greater than the expected additive effect of one or LDL particle levels in humans. The synergistic decrease in more SPLA inhibitors and one or more statins. In certain of LDL following co-administration of A-002 and statins was these embodiments, administration of one or more spIA observed in the statin Subpopulation as a whole. In addition, 55 inhibitors and one or more statins results in an improved synergism appeared to occur between A-002 and each of the HDL/LDL ratio, either by decreasing LDL levels, increasing individual statins within the general statin Subpopulation, HDL levels, or both. although statistical analysis of this effect was complicated by Provided herein in certain embodiments are methods of the limited number of test and placebo subjects for each treating CVD or conditions associated with CVD in a subject statin. These results indicate that the synergistic decrease in 60 in need thereof by administering a therapeutically effective LDL levels generated by co-administration of A-002 and amount of one or more spIA inhibitors and a therapeutically statins is not limited to one particular statin, but rather occurs effective amount of one or more compounds used in the across the entire range of statins. Administration of A-002 in treatment of CVD. In certain embodiments, the one or more conjunction with eZetimibe led to a similar synergistic sPLA inhibitors comprise A-001 or a pharmaceutically decrease in LDL levels, indicating that the synergism 65 acceptable prodrug, salt, polymorph, co-crystal, or Solvate of between A-002 and compounds used in the treatment of CVD A-001, and in certain of these embodiments, the pharmaceu is not limited to statins. Therefore, methods are provided tically acceptable prodrug is A-002. In certain embodiments, US 8,048,880 B2 21 22 the one or more compounds used in the treatment of CVD acid sequestrants, fibrates, niacin or niacin derivatives, cho comprise one or more statins or statin combination drugs. In lesterol absorption inhibitors, cholesteryl ester transfer pro certain of these embodiments, the one or more statins are tein (CETP) inhibitors, microsomal triglyceride transfer pro selected from the group consisting of atorvastatin, simvasta tein (MTP) inhibitors, squalene synthase inhibitors, ACE tin, rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvas inhibitors, angiotensin II receptor antagonists, beta-adrener tatin, mevastatin, and pitavastatin, and the statin combination gic blockers, calcium channel blockers, and antithrombotics. drugs are selected from the group consisting of atorvastatin In certain of these embodiments, the one or more compounds plus eZetimibe, atorvastatin plus amlodipine, atorvastatin used in the treatment of CVD comprise eZetimibe. An plus CP-529414, atorvastatin plus APA-01, simvastatin plus increase in effectiveness of a compound used in the treatment eZetimibe, simvastatin plus extended release niacin, simvas 10 of CVD as used herein refers to an increase the therapeutic tatin plus MK-0524A, lovastatin plus extended release niacin, effect of the compound, a decrease in the dosage of the com rosuvastatin plus fenofibrate, pravastatin plus fenofibrate, and pound required to obtain a particular level of therapeutic statin plus TAK-457. In certain embodiments, the one or more effect, or some combination thereof. For example, an increase compounds used in the treatment of CVD comprise one or in effectiveness of a statinas used herein may refer to a greater more non-statin compounds selected from the group consist 15 decrease in LDL levels following administration of a particu ing of bile acid sequestrants, fibrates, niacin or niacin deriva lar dosage of a statin, a decrease in the dosage of Statin tives, cholesterol absorption inhibitors, cholesteryl ester required to bring about a particular decrease in LDL levels, or transfer protein (CETP) inhibitors, microsomal triglyceride Some combination thereof. transfer protein (MTP) inhibitors, squalene synthase inhibi Likewise, in certain embodiments methods are provided tors, ACE inhibitors, angiotensin II receptor antagonists, for increasing the effectiveness of an spIA inhibitor by beta-adrenergic blockers, calcium channel blockers, and anti administering one or more compounds used in the treatment thrombotics. In certain of these embodiments, the one or of CVD. In certain embodiments, the one or more spLA more compounds used in the treatment of CVD comprise inhibitors comprise A-001 or a pharmaceutically acceptable eZetimibe. In certain embodiments, treatment of CVD or a prodrug, salt, polymorph, co-crystal, or Solvate of A-001, and condition associated with CVD may be associated with a 25 in certain of these embodiments, the pharmaceutically decrease in cholesterol levels. In other embodiments, how acceptable prodrug is A-002. In certain embodiments, the one ever, treatment of certain forms of CVD may not be associ or more compounds used in the treatment of CVD comprise ated with a concomitant decrease in cholesterol levels. For one or more statins or statin combination drugs. In certain of example, SPLA inhibitor and statin administration may these embodiments, the one or more statins are selected from result in a decrease in atherosclerotic plaque formation with 30 the group consisting of atorvastatin, simvastatin, rosuvasta out a measurable decrease in cholesterol levels via a mecha tin, lovastatin, pravastatin, cerivastatin, fluvastatin, mevasta nism such as inhibition of inflammation. In these embodi tin, and pitavastatin, and the statin combination drugs are ments, treatment of CVD may be associated with a decrease selected from the group consisting of atorvastatin plus in levels of one or more inflammatory markers, such as for eZetimibe, atorvastatin plus amlodipine, atorvastatin plus example spLA, CRP, and/or IL-6. In certain embodiments, 35 CP-529414, atorvastatin plus APA-01, simvastatin plus administration of a combination of one or more spLA inhibi eZetimibe, simvastatin plus extended release niacin, simvas tors and one or more statins results in a greater degree of tatin plus MK-0524A, lovastatin plus extended release niacin, treatment than that observed following administration of one rosuvastatin plus fenofibrate, pravastatin plus fenofibrate, and or more spLA inhibitors or statins alone. In certain of these statin plus TAK-457. In certain embodiments, the one or more embodiments, the degree of treatment is synergistic, meaning 40 compounds used in the treatment of CVD comprise one or that the degree of treatment is greater than the expected addi more non-statin compounds selected from the group consist tive effect of one or more spLA inhibitors and one or more ing of bile acid sequestrants, fibrates, niacin or niacin deriva statins. tives, cholesterol absorption inhibitors, cholesteryl ester Provided herein in certain embodiments are methods for transfer protein (CETP) inhibitors, microsomal triglyceride increasing the effectiveness of a compound used in the treat 45 transfer protein (MTP) inhibitors, squalene synthase inhibi ment of CVD by administering one or more spIA inhibitors tors, ACE inhibitors, angiotensin II receptor antagonists, in conjunction with the compound used in the treatment of beta-adrenergic blockers, calcium channel blockers, and anti CVD. In certain embodiments, the one or more spLA inhibi thrombotics. In certain of these embodiments, the one or tors comprise A-001 or a pharmaceutically acceptable pro more compounds used in the treatment of CVD comprise drug, salt, polymorph, co-crystal, or Solvate of A-001, and in 50 eZetimibe. certain of these embodiments, the pharmaceutically accept In certain embodiments, one or more spIA inhibitors and able prodrug is A-002. In certain embodiments, the one or one or more compounds used in the treatment of CVD, such more compounds used in the treatment of CVD comprise one as for example one or more statins, may be administered to a or more statins or statin combination drugs. In certain of these Subject separately, i.e., as separate compositions. In these embodiments, the one or more statins are selected from the 55 embodiments, the one or more spLA inhibitors and the one group consisting of atorvastatin, simvastatin, rosuvastatin, or more compounds used in the treatment of CVD may be lovastatin, pravastatin, cerivastatin, fluvastatin, mevastatin, administered simultaneously or sequentially. Further, the one and pitavastatin, and the statin combination drugs are selected or more SPLA inhibitors and the one or more compounds from the group consisting of atorvastatin plus eZetimibe, ator used in the treatment of CVD may be administered at differ vastatin plus amlodipine, atorvastatin plus CP-529414, ator 60 ent times, and one compound may be administered more vastatin plus APA-01, simvastatin plus eZetimibe, simvastatin frequently than another. In certain embodiments wherein the plus extended release niacin, simvastatin plus MK-0524A. one or more compounds used in the treatment of CVD are lovastatin plus extended release niacin, rosuvastatin plus statins and wherein the spLA inhibitors and/or statins are fenofibrate, pravastatin plus fenofibrate, and statin plus TAK given in multiple administrations, one or both may be admin 457. In certain embodiments, the one or more compounds 65 istered anywhere from once or more times per day to once used in the treatment of CVD comprise one or more non every week, once every month, or once every several months. statin compounds selected from the group consisting of bile In certain preferred embodiments, the one or more spLA US 8,048,880 B2 23 24 inhibitors and/or statins may be administered twice a day, and ing of bile acid sequestrants, fibrates, niacin or niacin deriva in other preferred embodiments the one or more spLA tives, cholesterol absorption inhibitors, cholesteryl ester inhibitors and/or statins may be administered once a day. transfer protein (CETP) inhibitors, microsomal triglyceride In certain embodiments, kits are provided that comprise transfer protein (MTP) inhibitors, squalene synthase inhibi one or more SPLA inhibitors and one or more compounds tors, ACE inhibitors, angiotensin II receptor antagonists, used in the treatment of CVD. In certain embodiments, the beta-adrenergic blockers, calcium channel blockers, and anti one or more sRLA inhibitors comprise A-001 or a pharma thrombotics. In certain of these embodiments, the one or ceutically acceptable prodrug, salt, polymorph, co-crystal, or more compounds used in the treatment of CVD comprise solvate of A-001, and in certain of these embodiments, the eZetimibe. pharmaceutically acceptable prodrug is A-002. In certain 10 Pharmaceutical compositions comprising one or more embodiments, the one or more compounds used in the treat SPLA inhibitors and one or more compounds used in the ment of CVD comprise one or more statins or statin combi treatment of CVD, such as for example one or more statins, nation drugs. In certain of these embodiments, the one or may be administered to a subject on a one-time basis or in more statins are selected from the group consisting of atorV multiple administrations. In those embodiments wherein the astatin, simvastatin, rosuvastatin, lovastatin, pravastatin, 15 compositions are given in multiple administrations, they may cerivastatin, fluvastatin, mevastatin, and pitavastatin, and the be administered at set intervals over a particular time period statin combination drugs are selected from the group consist determined in advance, or they may be administered indefi ing of atorvastatin plus eZetimibe, atorvastatin plus amlo nitely or until a particular therapeutic benchmark is reached, dipine, atorvastatin plus CP-529414, atorvastatin plus APA such as for example until a subject exhibits cholesterol levels, 01, simvastatin plus eZetimibe, simvastatin plus extended triglyceride levels, or inflammatory marker levels that are release niacin, simvastatin plus MK-0524A, lovastatin plus below a specified threshold. In certain embodiments, the extended release niacin, rosuvastatin plus fenofibrate, prav compositions may be administered from once or more times astatin plus fenofibrate, and statin plus TAK-457. In certain per day to once every month or once every several months. In embodiments, the one or more compounds used in the treat certain of these embodiments, the compositions are adminis ment of CVD comprise one or more non-statin compounds 25 tered once or twice per day. selected from the group consisting of bile acid sequestrants, Pharmaceutical compositions comprising one or more fibrates, niacin or niacin derivatives, cholesterol absorption SPLA inhibitors, one or more compounds used in the treat inhibitors, cholesteryl ester transfer protein (CETP) inhibi ment of CVD, or one or more spLA inhibitors plus one or tors, microsomal triglyceride transfer protein (MTP) inhibi more compounds used in the treatment of CVD as disclosed tors, squalene synthase inhibitors, ACE inhibitors, angio 30 herein may be delivered to a subject by any administration tensin II receptor antagonists, beta-adrenergic blockers, pathway known in the art, including but not limited to oral, calcium channel blockers, and antithrombotics. Within the aerosol, enteral, nasal, ophthalmic, parenteral, or transdermal kit, the one or more spIA inhibitors and one or more com (e.g., topical cream or ointment, patch). "Parenteral refers to pounds used in the treatment of CVD may be divided into a route of administration that is generally associated with separate compartments. For example, the kit may comprise 35 injection, including infraorbital, infusion, intraarterial, intra multiple bottles or packets, wherein each bottle or packet capsular, intracardiac, intradermal, intramuscular, intraperi contains either one or more spLA inhibitors or one or more toneal, intrapulmonary, intraspinal, intrasternal, intrathecal, compounds used in the treatment of CVD. In other embodi intrauterine, intravenous, Subarachnoid, Subcapsular, Subcu ments, the one or more SPLA inhibitors and one or more taneous, transmucosal, or transtracheal. SPLA inhibitor, sta compounds used in the treatment of CVD may be found in a 40 tin, or combined sPLA inhibitor and statin pharmaceutical single, undivided container. In certain embodiments, the kit compositions as described herein may be administered in any provides instructions for usage. Such as dosage or adminis pharmaceutically acceptable form, including for example in tration instructions. the form of a solid, liquid Solution, Suspension, emulsion, Provided herein in certain embodiments are pharmaceuti dispersion, micelle, or liposome. Preparations for injection cal compositions comprising a therapeutically effective 45 may include sterile solutions ready for injection, sterile dry amount of one or more SPLA inhibitors and a therapeutically soluble products, such as lyophilized powders, ready to be effective amount of one or more compounds used in the combined with a solvent just prior to use, including hypoder treatment of CVD. In certain embodiments, the one or more mic tablets, sterile Suspensions ready for injection, Sterile dry sPLA inhibitors comprise A-001 or a pharmaceutically insoluble products ready to be combined with a vehicle just acceptable prodrug, salt, polymorph, co-crystal, or Solvate of 50 prior to use, and sterile emulsions. The Solutions may be A-001, and in certain of these embodiments, the pharmaceu either aqueous or nonaqueous. In certain embodiments, the tically acceptable prodrug is A-002. In certain embodiments, compositions may comprise one or more pharmaceutically the one or more compounds used in the treatment of CVD acceptable carriers or may be administered in conjunction comprise one or more statins or statin combination drugs. In with one or more pharmaceutically acceptable carriers. certain of these embodiments, the one or more statins are 55 In certain embodiments, pharmaceutical compositions selected from the group consisting of atorvastatin, simvasta comprising one or more spIA inhibitors, one or more com tin, rosuvastatin, lovastatin, pravastatin, cerivastatin, fluvas pounds used in the treatment of CVD, or one or more spLA tatin, mevastatin, and pitavastatin, and the statin combination inhibitors plus one or more compounds used in the treatment drugs are selected from the group consisting of atorvastatin of CVD as described herein may be formed into oral dosage plus eZetimibe, atorvastatin plus amlodipine, atorvastatin 60 units, such as for example tablets, pills, or capsules. Such an plus CP-529414, atorvastatin plus APA-01, simvastatin plus oral dosage unit may comprise the active ingredients (e.g., eZetimibe, simvastatin plus extended release niacin, simvas A-002 and one or more statins) and one or more pharmaceu tatin plus MK-0524A, lovastatin plus extended release niacin, tically acceptable carriers. As disclosed herein, the feasibility rosuvastatin plus fenofibrate, pravastatin plus fenofibrate, and of such an oral dosage unit was demonstrated by producing a statin plus TAK-457. In certain embodiments, the one or more 65 batch of tablets comprising A-002 and simvastatin as the compounds used in the treatment of CVD comprise one or active ingredients. Each tablet was formulated with 250mg of more non-statin compounds selected from the group consist A-002 and 40 mg of simvastatin. Pharmaceutically accept US 8,048,880 B2 25 26 able carriers utilized informulating the tablets included anhy methods well known in the art. In certain embodiments, a drous lactose, lactose fast flo, and microcrystalline cellulose therapeutically effective amount of ansPLA inhibitor foruse as diluents, hydroxypropyl cellulose as a binder, croScarmel alone or in combination with one or more compounds used in lose Sodium as a disintegrants, butylated hydroxyanisole as the treatment of CVD may be from about 5 to about 10,000 an antioxidant, magnesium Stearate as a lubricant, and mg/dose. In certain of these embodiments, a therapeutically polysorbate 80 as a surfactant. Water was used as a solvent effective amount of ansPLA inhibitor may be from about 25 when formulating the tablet. The final tablets contained high to about 5,000 mg/dose, and in certain of these embodiments concentrations of both active ingredients as determined by a therapeutically effective amount may be from about 50 to HPLC analysis, indicating that ansPLA inhibitor and one or about 500 mg/dose. more compounds used in the treatment of CVD, such as for 10 example one or more statins, may be formulated into a single In certain embodiments, an spLA inhibitor for use in the oral dosage unit. The specific tablet formulation disclosed compositions and methods disclosed herein may be an herein is provided as an example only. One of ordinary skill in indole-based sPLA inhibitor, meaning that the compound the art will recognize that the therapeutically acceptable car contains an indole nucleus having the structure: riers utilized in the formulation may be varied, and that such 15 variations are routine in the art. Likewise, the active ingredi ents may vary. As disclosed herein, administration of A-002 in combination with a variety of statins causes a synergistic decrease in serum lipid levels. Therefore, in certain embodi ments, A-002 may be combined with any statin known in the N art, including but not limited to atorvastatin oratorvastatin H calcium, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and/or simvastatin. In A variety of indole-based sPLA inhibitors are known in certain embodiments, an oral dosage unit may comprise a the art. For example, indole-based sPLA inhibitors that may coating that Surrounds the active ingredients and pharmaceu 25 be used in conjunction with the present invention include but tically acceptable carrier(s). are not limited to those set forth in U.S. Pat. Nos. 5,654,326 In certain embodiments, pharmaceutical compositions (Bach); 5,733,923 (Bach); 5,919,810 (Bach); 5,919,943 comprising one or more spIA inhibitors, one or more com (Bach); 6,175,021 (Bach): 6,177,440 (Bach); 6.274,578 pounds used in the treatment of CVD, or one or more spLA (Denney); and 6,433,001 (Bach), the entire disclosures of inhibitors plus one or more compounds used in the treatment 30 which are incorporated by reference herein. Methods of mak of CVD as disclosed herein may be administered via a time ing indole-based sPLA inhibitors are set forth in, for release delivery vehicle, such as for example a time release example, U.S. Pat. Nos. 5,986,106 (Khau); 6,265,591 oral dosage unit. A “time release vehicle' as used herein (Anderson); and 6.380,397 (Anderson), the entire disclosures refers to any delivery vehicle that releases active agent (e.g., of which are incorporated by reference herein. SPLA inhibi A-002 and one or more statins) at Some time after adminis 35 tration or over a period of time following administration tors for use in the present invention may be generated using rather than immediately upon administration. Time release these synthesis methods, or using any other synthesis method may be obtained by a coating on the vehicle that dissolves known in the art. In certain embodiments, spLA inhibitors over a set timeframe following administration. In certain for use in the present invention may be spIA type IIA, type embodiments, the time release vehicle may comprise mul 40 V, and/or type Xinhibitors. Various examples of indole-based tiple layers of coating alternated with multiple layers of active sPLA inhibitors are set forth below. These examples are ingredients, such that each layer of coating releases a certain merely provided as illustrations of the types of inhibitors that volume of active ingredients as it dissolves. In other embodi may be used in conjunction with the present invention, and as ments, spLA inhibitor, statin, or combined sPLA inhibitor such are not meant to be limiting. One of ordinary skill in the and Statin pharmaceutical compositions may be administered 45 art will recognize that a variety of other indole-based sPLA via an immediate release delivery vehicle. inhibitors may be used. Atherapeutically effective amount ofansPLA inhibitor or In certain embodiments, spLA inhibitors for use in the a compound used in the treatment of CVD for use in the current invention are 1H-indole-3-glyoxylamide compounds methods or compositions disclosed herein may be determined having the structure: for each compound individually. For example, statins or statin 50

combination drugs may be administered or included in a pharmaceutical composition at a dosage that is well known in the art to decrease cholesterol levels. One of skill in the art will recognize that in those embodiments wherein one or more compounds used in the treatment of CVD are combined 55 with one or more spIA inhibitors in a single composition, the amount of the compound used in the treatment of CVD that constitutes a therapeutically effective amount may be different than the amount of the compound that constitutes a therapeutically effective amount of the compound when 60 administered alone due to, for example, interactions between the compound and the one or more spLA inhibitors. For wherein: example, the effective dosage of a statin for use in combina each X is independently oxygen or Sulfur, tion therapy may be lower than the standard dosage for the R is selected from the group consisting of (a), (b), and (c), statin when administered alone. In this situation, one of skill 65 wherein: in the art will be able to readily determine a therapeutically (a) is C7-Co alkyl, C7-Coalkenyl, C7-Co alkynyl, car effective amount for the combination composition using bocyclic radicals, or heterocyclic radicals; US 8,048,880 B2 27 28 (b) is a member of (a) substituted with one or more inde hydroxyamino, iodo, nitro, phosphono, -SOH, thioacetal, pendently selected non-interfering Substituents; and thiocarbonyl, and C-C carbonyl, where n is from 1 to 8: (c) is the group -(L)-Rs, where, -(L)- is a divalent linking and pharmaceutically acceptable salts, Solvates, prodrug group of 1 to 12 atoms selected from carbon, hydrogen, derivatives, racemates, tautomers, or optical isomers thereof. oxygen, nitrogen, and Sulfur, wherein the combination of 5 In certain of these embodiments, -(L)- has the formula: atoms in -(L)- are selected from the group consisting of (i) carbon and hydrogen only, (ii) Sulfur only, (iii) oxygen only, (iv) nitrogen and hydrogen only, (v) carbon, hydrogen, and Sulfur only, and (vi) carbon, hydrogen, and oxygen only; and where Rso is a group selected from (a) or (b); 10 R is hydrogen, halo, C-C alkyl, C-C cycloalkyl, C-C, cycloalkenyl, -O-(C-C alkyl). —S-(C-C alkyl), or a non-interfering Substituent having a total of 1 to 3 atoms other than hydrogen; wherein Rs and Rs are eachindependently selected from the Ra and Rs are independently selected from the group consist 15 group consisting of hydrogen, C-Co alkyl, carboxy, car ing of hydrogen, a non-interfering substituent, and -(L)- balkoxy, and halo; p is a number from 1 to 5; and Z is selected (acidic group), wherein -(L)- is an acid linker having an acid from the group consisting of a bond, —(CH2)— —O—, linker length of 1 to 4: provided that at least one of Ra and —N(C-Co alkyl)-, -NH-, and —S—. Rs must be -(L)-(acidic group); In certain of these embodiments wherein Rais-(L)-(acidic Re and R7 are each independently selected from hydrogen, group), the acid linker -(L)- has the formula: non-interfering Substituents, carbocyclic radicals, carbocy clic radicals Substituted with non-interfering Substituents, heterocyclic radicals, and heterocyclic radicals substituted with non-interfering Substituents; provided that for any of the groups R. R. and R-7, the car 25 bocyclic radical is selected from the group consisting of o cycloalkyl, cycloalkenyl, phenyl, naphthyl, norbornanyl. R84 bicycloheptadienyl, toluoyl, Xylenyl, indenyl, stilbenzyl, ter phenylyl, diphenylethylenyl, phenyl-cyclohexenly, acenaph 30 wherein Q is selected from the group consisting of thylenyl, and anthracenyl, biphenyl, bibenzylyl and related (CH2)— —O— —NH-, and —S—; and Rs and Rs are bibenzylyl homologues represented by the formula (bb), each independently selected from the group consisting of hydrogen, C-Co alkyl, aryl, C-Co alkaryl, C-Co aralkyl, hydroxy, and halo. (bb) 35 In certain of these embodiments wherein Rsis-(L)-(acidic KX- (CH2)n -() group), the acid linker -(L)- has the formula: R85 where n is a number from 1 to 8: provided, that for any of the 40 groups R. R. and R-7, the heterocyclic radical is selected Q--C --- phenylene from the group consisting of pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, phenylimidazolyl, triazolyl, isox Rs6 azolyl, oxazolyl, thiazolyl, thiadiazolyl, indolyl, carbazolyl, norharmanyl, azaindolyl, benzofuranyl, dibenzofuranyl. 45 whereinris a number from 2 to 7; S is 0 or 1: Q is selected from thianaphtheneyl, dibenzothiophenyl, indazolyl, imidazo(1.2- the group consisting of —(CH2)— —O— —NH-, and A)pyridinyl, benzotriazolyl, anthranillyl, 1,2-benzisoxazolyl, —S—; and Rss and Rs are each independently selected from benzoxazolyl, benzotriazolyl, purinyl, pryidinyl, dipyridylyl, the group consisting of hydrogen, C-Co alkyl, aryl, C-Co phenylpyridinyl, benzylpyridinyl, pyrimidinyl, phenylpyrim alkaryl, C-C aralkyl, carboxy, carbalkoxy, and halo. idinyl, pyrazinyl, 1,3,5-triazinyl, quinolinyl, phthalazinyl, 50 In certain embodiments, a 1H-indole-3-glyoxylamide quinazolinyl, and quinoxalinyl; and compound for use in the present invention is selected from the provided that for the groups R. R. R. Rs. R. and R, the group consisting of ((3-(2-Amino-1,2-dioxoethyl)-2-ethyl non-interfering Substituent is selected from the group consist 1-(phenylmethyl)-1H-indol-4-yl)oxy)acetic acid; 3-(2- ing of C-C alkyl, C-C alkenyl, C-C alkynyl, C7-C Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-in aralkyl, C7-C alkaryl, C-C cycloalkyl, C-C cycloalk 55 dol-4-yloxyacetic acid methyl ester; (3-(2-Amino-1,2- enyl, phenyl, toluoyl, Xylenyl, biphenyl, C-C alkoxy, C-C, dioxoethyl)-2-methyl-1-(phenylmethyl)-1H-indol-4-yl)oxy) alkenyloxy, C-C alkynyloxy, C-C alkoxyalkyl, C-C2 acetic acid; d1-2-((3-(2-Amino-1,2-dioxoethyl)-2-methyl-1- alkoxyalkyloxy, C-C alkylcarbonyl, C-C alkylcarbony (phenylmethyl)-1H-indol-4-yl) oxy)propanoic acid; (3-(2- lamino, C-C alkoxyamino, C-C alkoxyaminocarbonyl, Amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2- C-C alkylamino, C-C alkylthio, C-C alkylthiocarbo 60 methyl-1H-indol-4-yl)oxy)acetic acid; (3-(2-Amino-1,2- nyl, C-C alkylsulfinyl, C-C alkylsulfonyl, C-C, dioxoethyl)-1-((1,1'-biphenyl)-3-ylmethyl)-2-methyl-1H haloalkoxy, C-Chaloalkylsulfonyl, C-Chaloalkyl, C-C, indol-4-yl)oxy)acetic acid; (3-(2-Amino-1,2-dioxoethyl)-1- hydroxyalkyl, -C(O)C(C-C alkyl). —(CH), O—(C- ((1,1'-biphenyl)-4-ylmethyl)-2-methyl-1H-indol-4-yl)oxy) Calkyl), benzyloxy, phenoxy, phenylthio. —(CONHSOR), acetic acid; ((3-(2-Amino-1,2-dioxoethyl)-1-((2,6- —CHO, amino, amidino, bromo, carbamyl, carboxyl, car 65 dichlorophenyl)methyl)-2-methyl-1H-indol-4-yl)oxy)acetic balkoxy, —(CH2), COH, chloro, cyano, cyanoguanidinyl, acid; ((3-(2-Amino-1,2-dioxoethyl)-1-(4(-fluorophenyl)me fluoro, guanidino, hydrazide, hydrazino, hydrazido, hydroxy, thyl)-2-methyl-1H-indol-4-yl)oxy)acetic acid; ((3-(2- US 8,048,880 B2 29 30 Amino-1,2-dioxoethyl)-2-methyl-1-((1-naphthalenyl)me -continued thyl)-1H-indol-4-yl)oxy)acetic acid; ((3-(2-Amino-1,2- dioxoethyl)-1-((3-chlorophenyl)methyl)-2-ethyl-1H-indol 4-yl)oxy)acetic acid; (3-(2-Amino-1,2-dioxoethyl)-1-((1,1'- biphenyl)-2-ylmethyl)-2-ethyl-1H-indol-4-yl)oxy)acetic acid; ((3-(2-amino-1,2-dioxoethyl)-1-((1,1'-biphenyl)-2-yl methyl)-2-propyl-1H-indol-4-yl)oxy)acetic acid; ((3-(2- Amino-1,2-dioxoethyl)-2-cyclopropyl-1-(phenylmethyl)- 1H-indol-4-yl)oxy)acetic acid; ((3-(2-Amino-1,2- dioxoethyl)-1-((1,1'-biphenyl)-2-ylmethyl)-2-cyclopropyl 10 No 1H-indol-4-yl)oxy)acetic acid; and 4-((3-(2-Amino-1,2- dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-5-yl)oxy) provided that the acid linker -(L)- for Rs is selected from the butanoic acid, or pharmaceutically acceptable salts, Solvates, group consisting of: prodrug derivatives, racemates, tautomers, or optical isomers 15 thereof. In certain embodiments, spLA inhibitors for use in the s o s current invention are 1H-indole-3-glyoxylamide compounds having the structure: O-C \ A R85

o s S-C \ A 25 R85 -lR84 o s \ A 30 R85 wherein: both X are oxygen; 35 R is selected from the group consisting of: R84 R84 o–-ch s s--ch s 40 R85 R85 - (-CH2)2 \ / and R84 R84 -" H H2 (H3C4 N--ch * and C --ch R85 R85 ( )-cis-K) 45 wherein Rs and Rss are eachindependently selected from the wherein Rio is a radical independently selected from halo. group consisting of hydrogen, C1-Cio alkyl, aryl, C-Co C-C alkoxy, —S—(C-Co alkyl), and C-Co haloalkyl, alkaryl, C-C aralkyl, carboxy, carbalkoxy, and halo: pro and t is a number from 0 to 5; 50 vided that at least one of R and Rs must be -(L)-(acidic R is selected from the group consisting of halo, cyclopropyl. group), and (acidic group) on -(L)-(acidic group) of R or Rs methyl, ethyl, and propyl; is selected from —COH, -SOH, or—P(O)(OH): Re and R, are each independently selected from the group Ra and Rs are independently selected from the group consist consisting of hydrogen and non-interfering Substituents, with ing of hydrogen, a non-interfering substituent, and -(L)- 55 the non-interfering Substituents being selected from the group (acidic group), wherein -(L)- is an acid linker; provided that consisting of C-C alkyl, C-C alkenyl, C-C alkynyl, the acid linker -(L)- for R is selected from the group con C7-C aralkyl, C7-C alkaryl, C-Cs cycloalkyl, C-Cs sisting of: cycloalkenyl, phenyl, toluoyl, Xylenyl, biphenyl, C-C, alkoxy, C-C alkenyloxy, C-C alkynyloxy, C-C alkoxy 60 alkyl, C-C alkoxyalkyloxy, C-C alkylcarbonyl, C-C2 --O-CH2--, --S-CH2--, --N-CH-H, alkylcarbonylamino, C-C alkoxyamino, C-C, alkoxyaminocarbonyl, C-C alkylamino, C-C alkylthio. CH3 C-C alkylthiocarbonyl, C-C alkylsulfinyl, C-C alkyl Sulfonyl, C-C haloalkoxy, C-C haloalkylsulfonyl, C-C, O 65 haloalkyl, C-C hydroxyalkyl, —C(O)O(C-C alkyl). --HC-CH-H, , and —(CH), O—(C-C alkyl), benzyloxy, phenoxy, phe nylthio. —(CONHSOR), —CHO, amino, amidino, bromo,

US 8,048,880 B2 33 34 wherein: -continued both X are oxygen; R84 R84 R is selected from the group consisting of: o–-ch s s--ch s R85 R85 R84 R84 - (-CH2)2 \ / and H H2 - " N--city * and C --ch (H3C4 R85 R85

( )-cis-( ) wherein Rs and Rss are eachindependently selected from the group consisting of hydrogen, C1-Cio alkyl, aryl, C-Co wherein Rio is a radical independently selected from halo. 15 alkaryl, C-C aralkyl, carboxy, carbalkoxy, and halo: pro C-C alkyl, C-C alkoxy, —S—(C-C alkyl), and vided that at least one of R and Rs must be -(L)-(acidic C-Chaloalkyl, and t is a number from 0 to 5: group), and (acidic group) on -(L)-(acidic group) of R or Rs R is selected from the group consisting of halo, cyclopropyl. is selected from —COH, -SOH, or - P(O)(OH): methyl, ethyl, and propyl; Re and R, are each independently selected from the group Ra and Rs are independently selected from the group consist consisting of hydrogen and non-interfering Substituents, with ing of hydrogen, a non-interfering Substituent, and -(L)- the non-interfering Substituents being selected from the group (acidic group), wherein -(L)- is an acid linker; provided that consisting of C-C alkyl, C-C alkenyl, C-C alkynyl, the acid linker -(L)- for R is selected from the group con C7-C aralkyl, C7-C alkaryl, C-Cs cycloalkyl, C-Cs sisting of: cycloalkenyl, phenyl, toluoyl, Xylenyl, biphenyl, C-C, 25 alkoxy, C-C alkenyloxy, C-C alkynyloxy, C-C alkoxy alkyl, C-C alkoxyalkyloxy, C-C alkylcarbonyl, C-C2 --O-CH-H - HS-CH2-H --N-CH2-H alkylcarbonylamino, C-C alkoxyamino, C-C, alkoxyaminocarbonyl, C-C alkylamino, C-C alkylthio. CH C-C alkylthiocarbonyl, C-C alkylsulfinyl, C-C alkyl 30 Sulfonyl, C-C haloalkoxy, C-C haloalkylsulfonyl, C-C, -- HC-CH2-H s O alld haloalkyl, C-C hydroxyalkyl, —C(O)O(C-C alkyl). —(CH), O—(C-C alkyl), benzyloxy, phenoxy, phe nylthio. —(CONHSOR), —CHO, amino, amidino, bromo, carbamyl, carboxyl, carbalkoxy, —(CH2), COH, chloro, 35 cyano, cyanoguanidinyl, fluoro, guanidino, hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo, nitro, phosphono. —SOH, thioacetal, thiocarbonyl, and C-C car bonyl; wherein n is from 1 to 8: so 40 and pharmaceutically acceptable salts, Solvates, prodrug derivatives, racemates, tautomers, or optical isomers thereof. In certain embodiments, spLA inhibitors for use in the provided that the acid linker -(L)- for Rs is selected from the current invention are (acyloxy) alkyl esterprodrug derivatives group consisting of of 1H-indole-3-glyoxylamide compounds having the struc 45 ture:

/=y-1 +- \ / /—y-1 HK)R85 /—y-1 wherein: 60 both X are oxygen; +- \ / R is selected from the group consisting of:

(C )- - yu R10), 8584 65 -cis-( ) and US 8,048,880 B2 35 36 -continued -continued R84 R84 (HCA \ H H CH2)0-2 N--city and C --city o 5 R85 wherein Rio is a radical independently selected from halo, wherein Rs and Rss are eachindependently selected from the C-C alkyl, C-C alkoxy, —S—(C-C alkyl), and group consisting of hydrogen, C-Co alkyl, aryl, C-Co C-Chaloalkyl, and t is a number from 0 to 5: 10 alkaryl, C-C aralkyl, carboxy, carbalkoxy, and halo, pro R is selected from the group consisting of halo, cyclopropyl. vided that at least one of Ra and Rs must be -(L)-(acidic methyl, ethyl, and propyl; group), and (acidic group) on -(L)-(acidic group) of R or Rs is selected from —COH, -SOH, or - P(O)(OH): Ra and Rs are independently selected from the group consist Re and R, are each independently selected from the group ing of hydrogen, a non-interfering Substituent, and -(L)- consisting of hydrogen and non-interfering Substituents, with (acidic group), wherein -(L)- is an acid linker; provided that 15 the non-interfering Substituents being selected from the group the acid linker -(L)- for R is selected from the group con consisting of C-C alkyl, C-C alkenyl, C-C alkynyl, sisting of: C7-C aralkyl, C7-C alkaryl, C-Cs cycloalkyl, C-Cs cycloalkenyl, phenyl, toluoyl, Xylenyl, biphenyl, C-C, alkoxy, C-C alkenyloxy, C-C alkynyloxy, C-C alkoxy alkyl, C-C alkoxyalkyloxy, C-C alkylcarbonyl, C-C, --O-CH-H - HS-CH2-H --N-CH2-H alkylcarbonylamino, C-C alkoxyamino, C-C2 alkoxyaminocarbonyl, C-C alkylamino, C-C alkylthio. CH3 C-C alkylthiocarbonyl, C-C alkylsulfinyl, C-C alkyl Sulfonyl, C-C haloalkoxy, C-C haloalkylsulfonyl, C-C, 25 haloalkyl, C-C hydroxyalkyl, —C(O)O(C-C alkyl). -- HC-CH2-H s O alld —(CH), O—(C-C alkyl), benzyloxy, phenoxy, phe nylthio. —(CONHSOR), —CHO, amino, amidino, bromo, carbamyl, carboxyl, carbalkoxy, —(CH2), COH, chloro, cyano, cyanoguanidinyl, fluoro, guanidino, hydrazide, 30 hydrazino, hydrazido, hydroxy, hydroxyamino, iodo, nitro, phosphono. —SOH, thioacetal, thiocarbonyl, and C-C car bonyl; wherein n is from 1 to 8: and pharmaceutically acceptable salts, Solvates, prodrug derivatives, racemates, tautomers, or optical isomers thereof. So 35 In certain embodiments, spLA inhibitors for use in the current invention are Substituted tricyclics having the struc ture: provided that the acid linker -(L)- for Rs is selected from the

group consisting of 40

45

wherein: 50 R is selected from the group consisting of —NHNH and —NH; R is selected from the group consisting of —OH and —O(CH2)Rs, wherein Rs is selected from the group con sisting of H. —CO.H. —CO(C-C alkyl). —SOH, -SO 55 (C-C alkyl), tetrazolyl, —CN, —NH —NHSORs, —CONHSORs, phenyl, phenyl substituted with —COH or —CO(C-C)alkyl, and

60 O -P(RR):

wherein R and R, are each independently selected from the o–-ch s s--ch s 65 group consisting of —OH, -O(C-C)alkyl, Ris is selected R85 R85 from the group consisting of-(C-C)alkyl and —CF; and m is 1-3:

US 8,048,880 B2 43 44 In certain other preferred embodiments, ansPLA inhibitor The following examples are provided to better illustrate the for use in the present invention is 3-(2-Amino-1,2-dioxoet claimed invention and are not to be interpreted as limiting the hyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxyacetic scope of the invention. To the extent that specific materials are acid methyl ester, also referred to herein as compound A-002. mentioned, it is merely for purposes of illustration and is not Compound A-002 has the structure: 5 intended to limit the invention. One skilled in the art may develop equivalent means or reactants without the exercise of inventive capacity and without departing from the scope of the invention. It will be understood that many variations can NH2 be made in the procedures herein described while still HC-O 10 remaining within the bounds of the present invention. It is the intention of the inventors that such variations are included within the scope of the invention. EXAMPLES 15 CH Example 1

Effect of A-002 Administration on Plasma Cholesterol Levels and Atherosclerotic Plaque Formation in Mice Compound A-002, which is sometimes referred to in the art as S-3013 or LY333013, is a prodrug form of A-001 that is Male ApoE mice were fed a high fat diet (21% fat, rapidly absorbed and hydrolyzed to A-001 following admin 0.15% cholesterol. 19.5% casein) ad libitum for two weeks in istration to a Subject. order to acclimate to the diet. Plasma cholesterol levels and In certain other preferred embodiments, ansPLA inhibitor 25 body weight were measured in order to obtain baseline levels, for use in the present invention is 9-(phenyl)methyl-5- and mice were randomized into three groups of 20 mice based carbamoylcarbazol-4-yl)oxyacetic acid, also referred to on these measurements. After the acclimation period, mice herein as compound A-003 or LY433771. Compound A-003 remained on the high fat diet and were administered A-002 has the structure: (30 mg/kg or 90 mg/kg) or vehicle only (5% acacia) twice a 30 day for 16 weeks by oral gavage. Plasma cholesterol levels and body weight were measured at 4, 8, 12, and 16 weeks after the start of A-002 administra tion. Comparisons of measurements between test and control mice at each time period were performed using a two-way 35 analysis of variance test (ANOVA) for repeated measures, followed by post-hoc Bonferroni test for significance. At the end of the 16th week, mice were sacrificed and e plasma samples, heart tissue, and aortas from the heart to approximately 3 mm distal to the iliac bifurcation were col 40 lected. Aortas were placed on microscope slides and stained with Oil Red O for Scanning and image analysis of athero Sclerotic lesion size. Vessel images were captured using a C Microtek Scanmaker 9600XL scanner (Microtek, Carson, Calif.) and Photoshop 6.0 software (Adobe Systems Inc., San In still other preferred embodiments, an spLA inhibitor 45 Jose, Calif.). Atherosclerotic plaque coverage was quantified for use in the present invention is (3-(2-amino-1,2-dioxoet for the entire length of the vessel including the arch using hyl)-2-methyl-1-(phenylmethyl)-1H-indol-4-yl)oxy)acetic Image-Pro Plus software. Vessels were also analyzed for acid N-morpholino ethyl ester, also referred to herein as com plaque coverage in the descending abdominal aorta, starting pound 421079. Compound 421079 has the structure: in the region of the first intercostal branches to the iliac 50 bifurcation (not including the arch). Plaque content was expressed as a percent of the aortic Surface. For assessment of significance in atherosclerotic plaque content between test and control mice, a one-way analysis of variance test fol lowed by Bonferroni post-hoc test was used. 55 All three groups of mice (control, 30 mg/kg A-002, and 90 mg/kg A-002) had similar baseline body weights (FIG. 1). Body weight increased over the 16 week test period by approximately 155%, 150%, and 14.1% for control, 30 mg/kg A-002, and 90 mg/kg A-002 mice, respectively (FIG. 1). 60 There was no statistically significant difference in body weight between the three groups. Baseline total plasma cholesterol levels were not signifi cantly different between the three groups (FIG. 2). At four weeks, mice receiving A-002 at either dosage exhibited a 65 significant decrease in total cholesterol. This effect remained consistent throughout the remainder of the 16 week test Like A-002, compound 421079 is a prodrug of A-001. period. At 16 weeks, the change from baseline in total cho US 8,048,880 B2 45 46 lesterol was +15%, -10%, and -12% in control, 30 mg/kg tion that each mouse weighed 22.5g and ate 4.5 g/day. Based A-002, and 90 mg/kg A-002 mice, respectively (FIG. 2). on body weight and food consumption, the diet was modified There was no apparent dose-response effect. over the 12 week course of treatment to adjust for a body Control mice exhibited approximately 13% plaque cover weight of 26 g and a food intake of 2.5 g/day. The dosages of age in aortic tissue at 16 weeks (FIG.3).30 mg/kg A-002 and A-002 were selected to cover a range of 100-fold based on 90 mg/kg A-002 mice exhibited 6.3% and 6.8% plaque cov pharmacokinetics in ApoE mice as well as toxicology and erage, respectively (FIG. 3). Therefore, A-002 treatment previously observed efficacy in this model. resulted in a significant decrease in plaque content. After twelve weeks, mice were sacrificed and plasma, heart 10 tissue, and aortic tissue were collected. En face lesion size Example 2 was determined via digital imaging analysis. Percent lesion coverage for each dosage group is Summarized in Table 2 and Effect of A-002 Administration on Angiotensin II-Mediated Atherosclerotic Plaque Formation and FIG. 5. Plaque content (as measured as en face lesion) was Aneurysm reduced in mice administered A-002 alone, statin alone, or 15 A-002 in combination with statin. The reduction in plaque A mouse model of accelerated atherosclerosis was utilized content in mice administered A-002 plus statin was substan to determine the effect of A-002 administration on atheroscle tially greater than the reduction in mice administered either rosis and aneurysm formation. ApoE mice acclimated to compound alone (Table 2, compare Group DVS. Groups. A the same high fat diet utilized in Example 1 were adminis and C: Group E vs. Groups B and C). Further, the reduction in tered angiotensin II with water, angiotensin with 5% acacia, plaque content in mice administered A-002 plus statin was or saline infusion and water twice daily for four weeks. Substantially greater than the sum of the reduction in plaque Angiotensin II has been shown to promote atherosclerosis content in mice administered A-002 alone and mice admin and aneurysm formation in apoE deficient mice (Daugherty istered statin alone. (Table 2, compare the change in mean 96 2000). At the end of the test period, plaque coverage was 25 enface lesion vs. control for Group D (i.e., -4.210%) with the assessed by three independent reviewers, and their assess Sum of the change in mean % en face lesion vs. control for ments were averaged to determine plaque coverage. Group A and Group C (i.e., the sum of -2.241% and Angiotensin II with water or with 5% acacia resulted in -0.813%, or -3.054%).) Thus, the administration of A-002 similar rates of plaque coverage (FIG. 4) and aneurysm (Table plus statin reduces plaque content in a synergistic manner. 1). Administration of A-002 at 30 mg/kg twice daily signifi 30 cantly decreased plaque content and aorticlevel aneurysms in TABLE 2 angiotensin II/acacia mice (FIG. 4). Effect of A-002 and/or statin administration on en face lesion size: TABLE 1. 35 Change in Effect of A-002 administration on aneurysm rate: mean 96 en Treatment group Aneurysm rate (%) Dosage Mean 96 en face lesion P-value vs. group # of mice face lesion vs. control control Angiotensin II + 5% acacia 22% (2/9) Angiotensin + water 25% (5/20) Low A-002 1 3.386 -2.241 0.0597 Saline + water 0% (0/25) 40 Angiotensin II + 5% acacia + 30 mg/kg 0% (0/16) (A) A-002 High A-002 2 1469 -4.158 O.OOO6 (B) Statin 2 4.814 -0.813 O.4797 (C) Example 3 45 Low A-002 O 1417 -4.210 O.OOO8 Effect of A-002 or A-002 Plus Statin on Lesion plus statin (D) Formation and Composition in Mice High A-002 O 0.774 -4.853 O.OOO1 plus statin Body weight, plasma total cholesterol, and plasma HDL 50 levels were measured for six groups of twelve ApoE' mice. (E) The mice were then placed on a high fat diet for twelve weeks Control 2 5.627 to allow for formation of significant fatty atherosclerotic (F) plaques. The high fat diet was a modified version of the Western diet (TD.88137, Harlan Teklad, Madison, Wis.) con 55 sisting of casein (195g/kg), DL-methionine (3 g/kg). Sucrose Plasma total cholesterol and HDL levels were measured. (341.44 g/kg), corn starch (150 g/kg), anhydrous milkfat (210 Mean levels of total cholesterol were decreased in mice g/kg), cholesterol (1.5 g/kg), cellulose (50 g/kg), mineral mix administered A-002 alone or in combination with statin AlN-76 (Teklad; 35 g/kg), calcium carbonate (4 g/kg), Vita (Table 3, FIG. 6). Administration of high dosage A-002 plus min mix (TekladiA0060; 10 g/kg), ethyoxyquin (antioxidant, 60 statin resulted in a greater decrease in total cholesterol than 0.04 g/kg), and either 0.0156 g/kg (“low dose”)A-002 (Group administration of high dosage A-002 or statin alone (Table 3, A), 1.56 g/kg (“high dose”) A-002 (Group B), 0.02 g/kg compare Group E vs. Groups B and C: FIG. 6). Mean levels pravastatin (Group C), low dose A-002 plus 0.02 g/kg prav of HDL were increased in mice administered A-002 at either astatin (Group D), high dose A-002 plus 0.02 g/kg pravastatin dosage (Table 4). This increase was greater in mice adminis (Group E), or vehicle only (Group F). Mice were adminis 65 tered high dose A-002 plus statin than in mice administered tered diets A-F in a blinded manner, and food was provided ad high dose A-002 or statin alone (Table 4 compare Group E vs. libitum. The diet was initially prepared based on the assump Groups B and C: FIG. 7). US 8,048,880 B2 47 48 TABLE 3 TABLE 5 Effect of A-002 and/or statin administration on plasma total Changes in serum LDL concentration in ITT population following cholesterol levels A-002 administration

Mean total A-OO2 Placebo cholesterol Dosage concentration P-value vs. Baseline # of 154 37 group # of mice (mg/dl) control Subjects Mean 76.8 mg/dl 77.7 mg/dl Low A-002 11 355.545 O.2296 LDL (A) 10 Week 8 # of 143 36 High A-002 12 301.750 O.O792 Subjects (B) observed Statin 12 40S.417 O.4647 Change in -8.2 mg/dl -1.5 mg/dl (C) (8. Low A-002 10 399.7OO O.4S21 LDL vs. plus statin 15 baseline (D) % change -10.68% -1.93% High A-002 10 219.300 O.O152 WS. plus statin baseline (E) p-value

TABLE 4 TABLE 6 Effect of A-002 and/or statin administration on plasma HDL levels 25 Changes in serum LDL particle concentration in ITT population following A-002 administration Mean HDL Dosage concentration P-value vs. A-002 Placebo group # of mice (mg/dl) control Baseline # of 158 38 Low A-002 (A) 11 21.000

Effect of A-002 or A-002 Plus Statin Administration 45 TABLE 7 on Serum Lipid Levels in Humans Changes in serum Small LDL particle concentration in ITT 204 human subjects from the United States with CVD, population following A-002 administration specifically stable coronary artery disease, were randomized A-002 Placebo to receive placebo or A-002 via oral administration twice a 50 Baseline # of 158 38 day over an eight week administration period at dosages of 50 Subjects mg, 100 mg, 250 mg. or 500 mg. Levels of various lipids and Mean 735.8 nmol/L. 661.7 nmol/L inflammatory markers were measured at the outset of the trial Small and at the end of weeks four and/or eight. For data sets with a LDL normal distribution, mean levels of lipids or inflammatory 55 particle Week 8 # of 129 33 markers were analyzed. For data sets with non-normal distri Subjects bution, median levels were analyzed. Administration of observed A-002 at all dosages tested resulted in a decrease in mean Change in -61.3 nmol/L +77.0 nmol/L levels of serum LDL (Table 5). LDL particles (Table 6), small (8. Small LDL particles (Table 7), total cholesterol (Table 8), and TG 60 LDL (Table 9), and median levels of sPLA (Table 10). In addition, particle administration of A-002 decreased median levels of CRP WS. (Table 11). The magnitude of the observed decreases in LDL baseline % change -8.33% +11.64% particle, small LDL particle, and CRP levels are particularly WS. noteworthy because subjects treated with placebo exhibited 65 baseline increases in these markers over the course of the trial (Tables 6, 7, and 11). US 8,048,880 B2 49 50 TABLE 8 TABLE 11 Changes in serum CRP concentration in ITT population Changes in serum total cholesterol concentration in ITT following A-002 administration population following A-002 administration Placebo

A-OO2 Placebo Baseline #o 155 38 subjects Baseline # of 158 39 Median 1.50 mg/L 2.00 mg/L Subjects ICRP) 10 Week 8 #o 143 36 Mean total 156.8 mg/dl 156.9 mg/dl subjects cholesterol observed Week 8 # of 146 37 Change in -0.20 mg/L +0.10 mg/L. median Subjects CRP vs. observed 15 baseline % change -13.33% +5.00% Change in -12.8 mg/dl -4.7 mg/dl WS. mean total baseline cholesterol p-value O.4478 WS. vs. baseline baseline % change -8.16% -2.99% vs. baseline p-value vs.

Baseline #o 159 41 Subiects Median 2.30 ng/ml 2.20 ng/ml TABLE 13 SPLA2 Week 8 #o 110 29 55 Changes in serum LDL concentration in diabetic Subiects subpopulation following A-002 administration observed Change in -2.00 ng/ml -0.30 ng/ml Placebo median SPLA2 Baseline # of 43 8 WS. Subjects baseline 60 Mean 63.7 mg/dl 75.1 mg/dl % change -86.96% -13.64% LDL WS. Week 8 # of 37 8 baseline Subjects p-value

TABLE 20 Summary of the synergistic effects of combined A-002 and statin administration versus administration of A-002 or statin alone Expected change from administration of A-002 plus Observed Observed Observed statin change change change (change for A- following following following 002 alone plus administration administration administration change for of A-002 plus of A-002 alone of statin alone statin alone) statin Mean serum -3.25% --13.26% --10.01% -9.00% Small LDL (Table 17) (Table 14) (-3.25% plus (Table 14) particle at +13.26%) week 8 Mean serum -6.93% -3.36% -10.29% -14.04% LDL at week 4 (Table 18) (Table 16) (-6.93% plus (Table 16) -3.36%) Mean serum -4.22% -2.15 -6.37% -12.48 LDL at week 8 (Table 18) (Table 16) (-4.22% plus (Table 16) -2.15% Mean serum -8.23% -4.40% -12.63% -16.27% LDL in 272 mg/dl (Table 19) (Table 15) (-8.23% plus (Table 15) Subpopulation -4.40%) at week 8 US 8,048,880 B2 55 56 Serum LDL data for subjects in the statin subpopulation TABLE 21-continued were subdivided based on the specific statin each subject was receiving. Statins with significant representation included Changes in serum LDL concentration in eZetimibe atorvastatin, rosuvastatin, simvastatin, lovastatin, pravasta subpopulation following A-002 administration tin, and fluvastatin, as well as the statin combination drugs simvastatin/ezetimibe (VytorinR), atorvastatin/ezetimibe, A-OO2 Placebo atorvastatin/amlodipine (Caduet(R), lovastatin/extended Change in -15.4 mg/dl -1.0 mg/dl release niacin (Advicor(R), rosuvastatin/TriCor(R), rosuvasta (8. tin/ABT-335, simvastatin/extended release niacin (Sim LDL vs. cor(R), simvastatin/MK-0524A (MK-0524B), pravastatin/ baseline fenofibrate, atorvastatin/APA-01, and TAK-457/statin. Statin 10 % change -17.8% -1.7% and Statin combination dosages varied within each individual WS. statin Subgroup. The number of test and placebo Subjects in baseline each individual statin Subgroup from the larger statin Sub TABLE 22

Summary of the synergistic effects of combined A-002 and ezetimibe administration versus administration of A-002 or ezetimibe alone

Expected change from administration Observed of A-002 plus Observed Observed change ezetimibe change change following (change for A- following following administration 002 alone plus administration administration of ezetimibe change for of A-002 plus of A-002 alone alone ezetimibe) ezetimibe

Mean serum -4.22% -1.7% -5.92% -17.8% LDL at week 8 (Table 18) (Table 21) (-4.22% plus (Table 21) -1.7%)

35 population was too low to allow for detailed Statistical analy Serum TG, CRP, and IL-6 levels were measured at eight sis. However, there appeared to be a trend towards the same weeks in 86 subjects from the ITT population diagnosed with synergistic decrease in LDL levels that was observed in the statin Subpopulation as a whole in several of the statin Sub metabolic syndrome. Administration of A-002 resulted in a groups, such as for example with certain dosages of atorvas decrease in serum levels of each of these markers (Tables tatin, rosuvastatin, and simvastatin. These results indicate that 40 23-25). Subjects administered placebo exhibited an increase the observed synergy between A-002 and statins is not limited in CRP and IL-6 levels (Tables 24-25). to a particular statin. The synergistic effect of A-002 and compounds used in the treatment of CVD on LDL levels was not limited to statins. TABLE 23 For example, 30 subjects from the ITT population were 45 receiving eZetimibe at a dosage of 10 mg during the course of Changes in serum TG concentration in metabolic syndrome the A-002 trial. Administration of A-002 resulted in a Subpopulation following A-002 administration decrease in mean serum LDL levels in the eZetimibe sub population after eight weeks (Table 21), indicating that administration of A-002 plus eZetimibe results in a greater A-OO2 Placebo decrease in LDL levels than administration of eZetimibe 50 alone (Table 21). As with the statin subpopulation, adminis Baseline #o 70 16 tration of A-002 in conjunction with eZetimibe resulted in a subjects synergistic decrease in mean LDL levels that was greater than Median 171.5 mg/dl 168.0 mg/dl the expected additive effect of A-002 and ezetimibe (Table TG 22). 55 Week 8 #o 63 13 subjects TABLE 21 observed Changes in serum LDL concentration in eZetimibe Change in -24.0 mg/dl -3.0 mg/dl Subpopulation following A-002 administration median 60 A-OO2 Placebo ITG) vs. baseline Baseline # of 28 12 Subjects % change -13.99% -1.79% Mean 81.0 mg/dl 79.6 mg/dl WS. LDL 65 baseline Week 8 # of 28 12 Subjects US 8,048,880 B2 57 58 TABLE 24 TABLE 26-continued Changes in serum CRP concentration in metabolic syndrome Generic A-002, statin formulation: subpopulation following A-002 administration Typical weight A-002 Placebo Component percentage Baseline # of 69 16 Statin Varies depending on Subjects desired dosage Median 2.40 mg/L. 2.60 mg/dl Anhydrous lactose 20-50% ICRP) Lactose monohydrate 20-50% Week 8 # of 62 13 10 Hydroxypropyl cellulose 2-6% Subjects Croscarmellose sodium O.S-5% observed Polysorbate 80 O. 1-3% Change in -0.3 mg/L. +0.1 mg/L. Microcrystalline cellulose 5-20% median Magnesium stearate O.25-3% CRP vs. baseline 15 % change -12.50% +3.84% To verify the feasibility of including both A-002 and a WS. statin in a single formulation, fixed dose tablets containing baseline 250 mg A-002 and 40 mg simvastatin (marketed as Zocor R) were generated. Unit and batch formulas for these tablets are set forth in Tables 27 and 28, respectively. All inactive ingre TABLE 25 dients were purchased from Spectrum Chemicals. Butylated hydroxyanisole was included in the formulation for consis Changes in serum IL-6 concentration in metabolic syndrome tency with the marketed form of simvastatin (Zocorr). subpopulation following A-002 administration

A-OO2 Placebo TABLE 27 25 Baseline # of 68 16 Unit formula for A-002, simvastatin tablets: Subjects Mean IL- 5.60 pg/ml 5.17 pg/ml Component Function mg tablet 6 (15.30) (2.60) Week 8 # of 44 10 A-002 Active ingredient 250 Subjects 30 Simvastatin Active ingredient 40 observed Anhydrous lactose Diluent 119.18 Change in -0.24 pg/ml +0.29 pg/ml Lactose fast flo Diluent 59.58 mean IL Hydroxypropyl cellulose Binder 13.5 (HPC) baseline Croscarmellose sodium Disintegrant 24.3 % change -4.29% +5.61% 35 (divided into two equal WS. portions for inner/extra baseline granular Polysorbate 80 Surfactant O.S4 Butylated hydroxyanisole Antioxidant O.01% (BHA) Microcrystalline cellulose Diluent 70.2O Example 5 40 200 (MCC) Magnesium stearate Lubricant 2.70 A-002 Plus Statin Combination Tablet Purified water Solvent Total tablet weight 580 mg Fixed dose tablets containing A-002 and one or more statins may be generated using methods known in the art. For 45 example, a fixed dose tablet containing a therapeutically effective amount of A-002 (e.g., 250 or 500 mg) and a thera TABLE 28 peutically effective amount of a statin (e.g., 10, 20, 40, or 80 mg), may be generated using a formula Such as that set forth Batch formula for A-002, simvastatin tablets: in Table 26. One of ordinary skill in the art will recognize that 50 Component Function gfbatch additional components may be added to this generic formu A-002 Active ingredient 259 lation. For example, a compound Such as calcium carbonate Simvastatin Active ingredient 41 may be added to the formulation to enhance dissolution and Anhydrous lactose Diluent 123 solubility. Likewise, one of skill in the art will recognize that Lactose fast flo Diluent 62 this formulation is just one example of a generic A-002/statin 55 Hydroxypropyl cellulose Binder 14 formulation, and that the identity and weight of the recited (HPC) Croscarmellose sodium Disintegrant 6.68.4 components within the formulation may be varied without (divided into two equal undue experimentation. portions for inner/extra granular TABLE 26 Polysorbate 80 Surfactant 1 60 Butylated hydroxanisole Antioxidant O.1 Generic A-002, statin formulation: (BHA) Microcrystalline cellulose Diluent 62 Typical weight 200 (MCC) Component percentage Magnesium stearate Lubricant 2.5 Purified water Solvent 93 A-OO2 Varies depending on 65 Batch size 673 g desired dosage US 8,048,880 B2 59 60 A flow chart detailing one method of preparing the tablet (around 58°C.), exhaust air temperature (around 45° C.), air above is set forth in FIG.8. BHA, polysorbate 80, and purified Volume inlet (around 1500 cfm), and negative pan pressure water were mixed to create granulation fluid, which was differential are verified. The tablets are pre-heated, and a stored overnight. Lactose anhydrous, lactose fast flo, and the sample number of tablets are weighed to determine the aver first portion of croScarmellose sodium were weighed and age core tablet weight. The coating solution is sprayed onto screened through a coarse mesh, then combined with A-002 the tablets while being gently mixed. At various intervals of and simvastatin (which can also be sieved if necessary) and around 5-15 minutes, the average tablet weight is recalcu dry blended for 1-2 minutes using a mixer at slow speed. In lated. Once target tablet weight is reached, the Supply air certain embodiments of this protocol, microcrystalline cellu temperature is reduced to around 45° C. and the pan is jogged lose may be included in the dry mix. Hydroxypropyl cellulose 10 at intervals for around five minutes to allow the tablets to dry. was combined with the granulation fluid, resulting in greatly The supply air temperature is reduced to around 30° C. and increased viscosity. The resultant solution was added slowly tablets are jogged manually for around ten more minutes. The to the mixer containing the A-002/simvastatin mix. During coated tablets are then discharged from the coating pan. addition, mixing speed was gradually increased. Granulation end-point was reached when fine granules formed without the 15 Example 6 mass becoming wet or Sticky. The granulation was screened through coarse mesh onto Once a Day Dosing of A-002 or A-002 Plus Statin foil lined trays and placed in an oven at 50° C. and ambient humidity for approximately three hours. After drying (and 135 human subjects over the age of 18 with CVD, specifi optional milling and/or sieving), granules were placed in a cally stable CAD, were randomized to receive either placebo low-density polyethylene (LDPE) bag. Approximately one or one of two dosages (250 mg or 500 mg) of A-002 via once fourth of the dried granules were pre-blended with microc a day oral administration over an eight week time period. 89 rystalline cellulose and magnesium Stearate in a small baggy, of the subjects received A-002, while 46 of the subjects and then the remainder of the granules were combined with received placebo. Subjects that were receiving statins or other the pre-blend in a mixer. The second portion of croscarmel 25 compounds used in the treatment of CVD at the outset of the lose sodium was added in, and the entire mixture was mixed trial continued to receive those therapeutics throughout the for several minutes. Tablets were prepared by weighing trial. 121 of the 135 subjects were on statins during the trial. approximately 580 mg of the final blend into a tablet press and There was little variation between dosage groups with respect compressing at 200-2500 psi (Carver press). The resultant to age, height, weight, or BMI. tablets were weighed individually, and had an average weight 30 Levels of various lipids and inflammatory markers were of approximately 0.6 g. Tablets appeared off-white to lighttan measured at the outset of the trial and at weeks two, four, in color. and/or eight weeks after the start of A-002 administration. In Two representative tablets (numbers 19 and 27) were ana addition, plasma A-002 levels were measured at various inter lyzed by reverse phase HPLC. Tablets were placed in a sta vals. Lipids that were measured included LDL, small LDL bility chamber at 25° C./60% RH. Representative HPLC 35 particle, oxidized LDL, non-HDL cholesterol, total choles results for two of the tablets are set forth in FIG. 9 and terol, ApoB, and triglycerides. In addition, levels of the summarized in Table 29. Tablets maintained high potency inflammatory marker CRP were measured. Lipid and inflam (i.e., high concentrations of A-002 and simvastatin), thereby matory marker levels at each timepoint were compared to the validating the feasibility of generating A-002/statin combi baseline measurements to determine the effect of A-002 nation tablets for the treatment of various conditions. 40 administration. For data sets with a normal distribution, mean levels of lipids or inflammatory markers were analyzed. For TABLE 29 data sets with non-normal distribution, median levels were analyzed. LDL particle size was also assessed. HPLC analysis of A-002/sinvastatin combination tablets: Subjects in the ITT population receiving A-002 at either 45 % A-002 % simvastatin dosage exhibited Substantial decreases at week eight in mean Tablet # Weight (RT: 28.5 minutes) (RT:44.5 minutes) serum LDL, non-HDL cholesterol, and total cholesterol lev els (Tables 30-32, respectively) and in median small LDL 19 0.64g 92% 89% particle, oxidized LDL, TG, and ApoB levels (Tables 33-36). 27 0.60 g 88% 86% Subjects receiving A-002 also exhibited an increase in mean 50 LDL particle size (Table 37). In addition, subjects receiving A film coating may be applied to combination A-002/statin A-002 did not show the same increase in median CRP levels tablets using methods well known in the art. In one Such that was observed in subjects receiving placebo (Table 38). method, the coating Suspension is prepared by adding a film coat mixture such as Opadry YS-1-18027-A to purified water TABLE 30 in a mixing vessel. The mixing speed is reduced to avoid 55 foaming and the Suspension is mixed for 60 or more minutes, Changes in serum LDL concentration in ITT population following until uniform. After mixing, the Suspension is allowed to A-002 administration: stand for 60 or more minutes to deaerate. A coater such as the A-OO2 Placebo Accela Coater is set up and the theoretical amount of coating Baseline # of 89 46 Suspension to be sprayed is calculated based on the number 60 Subjects and size of tablets being coated. Tablets are loaded onto the Mean 75.4 mg/dl 82.0 mg/dl coating pan, and the nozzle air pressure (around 80-100 psi), LDL Week 8 # of 81 43 atomizing air pressure (around 25-45 psi), patternair pressure Subjects (around 20-40 psi), delivery rate (around 300-400 g/minute), observed pan speed (around 4-10 rpm), and dew point setting (around 65 Change in -7.1 mg/dl -0.9 mg/dl 5-15°C.) are verified. The fan is turned on, and gun-to-bed (8. distance (around 6-10 inches), Supply air temperature US 8,048,880 B2 61 62 TABLE 30-continued TABLE 33-continued Changes in serum LDL concentration in ITT population following Changes in serum Small LDL particle concentration in ITT A-002 administration: population following A-002 administration:

A-OO2 Placebo 5 A-002 Placebo LDL vs. LDL baseline ic % change -8.3% -O.7% etic e WS. baseline 10 baseline % change -9.1% --0.6% WS. baseline TABLE 31 Changes in serum non-HDL cholesterol concentration in ITT population following A-002 administration: 15 TABLE 34 A-002 Placebo Changes in serum oxidized LDL concentration in ITT population Baseline # of 89 46 following A-002 administration: Subjects Mean non- 105.9 mg/dl 112.5 mg/dl 2O A-002 Placebo HDL cholesterol Baseline # of 89 46 Week 8 # of 81 43 Subjects Subjects Median 41.5 UFL 43.3 UFL observed oxidized Change in -10.5 mg/dl -2.1 mg/dl 25 LDL mean non- Week 8 # of 81 43 HDL Subjects cholesterol observed vs. baseline Change in -1.4UL +0.3 UFL % change -9.9% -O.7% median vs. baseline 30 oxidized LDL vs. baseline % change -3.6% +0.4%

TABLE 32 aseline Changes in serum total cholesterol concentration in ITT 35 population following A-002 administration: TABLE 35 A-OO2 Placebo Changes in serum TG concentration in ITT population following Baseline # of 89 46 A-002 administration: Subjects Mean total 154.9 mg/dl 159.9 mg/dl 40 A-OO2 Placebo cholesterol Week 8 # of 81 43 Baseline # of 88 45 Subjects Subjects observed Median 137.5 mg/dl 135.0 mg/dl Change in -11.5 mg/dl -3.1 mg/dl TG mean total 45 Week 8 # of 81 43 cholesterol Subjects vs. baseline observed % change -7.3% -1.7% Change in -13.0 mg/dl +3.0 mg/dl vs. baseline median TG) vs. 50 baseline % change -9.8% +2.6% TABLE 33 vs. baseline Changes in serum Small LDL particle concentration in ITT population following A-002 administration: 55 A-002 Placebo TABLE 36 Baseline # of 89 46 Changes in serum ApoB concentration in ITT population Subjects following A-002 administration: Median 759.0 nmol/L. 782.0 nmol/L. Small 60 A-OO2 Placebo LDL particle Baseline # of 89 46 Week 8 # of 81 43 Subjects Subjects Median 68.6 mg/dl 62.8 mg/dl observed ApoB Change in -74.0 nmol/L +2.0 nmol/L Week 8 # of 81 43 median 65 Subjects Small observed US 8,048,880 B2 63 64 TABLE 36-continued had baseline LDL levels greater than 70 mg/dl (Tables 48-55). The number of Subjects in the non-statin Subpopulation was Changes in serum ApoB concentration in ITT population too low to allow for a detailed statistical analysis of the following A-002 administration: combined effect of A-002 and statins versus the effect of A-OO2 Placebo either compound alone. However, once additional non-statin Change in -8.1 mg/dl +2.5 mg/dl controls are obtained, it is expected that a synergistic decrease median in LDL levels similar to that observed following twice daily ApoB vs. A-002 administration (Example 4) will be observed in sub baseline jects receiving A-002 once a day. % change -11.9% +3.5% 10 WS. baseline TABLE 39 Changes in serum LDL concentration in statin Subpopulation following A-002 administration:

TABLE 37 15 A-OO2 Placebo Changes in LDL particle size in ITT population following A-002 Baseline # of 81 40 administration: Subjects Mean 70.5 mg/dl 75.5 mg/dl A-002 Placebo LDL Week 8 # of 75 37 Baseline # of 89 46 Subjects Subjects observed Mean LDL 20.50 mm 20.50 mm Change in -6.7 mg/dl -2.7 mg/dl particle (8. size LDL vs. Week 8 # of 81 43 25 baseline Subjects % change -8.2% -1.8% observed WS. Change in +0.11 nm. 0.00 nm. baseline mean LDL particle size vs. baseline 30 TABLE 40 % change --0.6% O.0% WS. Changes in serum non-HDL cholesterol concentration in statin baseline subpopulation following A-002 administration: A-OO2 Placebo 35 Baseline #o 81 40 TABLE 38 subjects Mean non- 100.4 mg/dl 104.8 mg/dl Changes in serum CRP concentration in ITT population following A HDL 002 administration: cholesterol Week 8 #o 75 37 A-OO2 Placebo 40 subjects observed Baseline # of 89 46 Subjects Change in -10.1 mg/dl -3.2 mg/dl Median 1.10 mg/L. 1.30 mg/L mean non ICRP) cholesterol Week 8 # of 81 43 45 vs. baseline Subjects % change -9.9% -1.3% observed Change in +0.10 ng/ml +0.25 ng/ml vs. baseline median CRP vs. baseline % change +5.4% +25.4% 50 TABLE 41 WS. baseline Changes in serum total cholesterol concentration in statin subpopulation following A-002 administration: The decrease in mean serum LDL, non-HDL cholesterol, A-OO2 Placebo 55 and total cholesterol levels and median small LDL particle, Baseline #o 81 40 oxidized LDL, TG, and ApoB levels, the increase in mean subjects LDL particle size, and the lower than expected increase in Mean total 149.3 mg/dl 152.6 mg/dl median CRP levels observed in the ITT population were also cholesterol Week 8 #o 75 37 observed at eight weeks in the 121 subject subpopulation that subjects was receiving statins during the trial (Tables 39-47, respec 60 observed tively), indicating that administration of A-002 plus statin Change in -11.2 mg/dl -4.4 mg/dl results in a greater therapeutic effect on these markers than mean total cholesterol administration of statin alone. Likewise, a decrease in LDL, vs. baseline non-HDL cholesterol, total cholesterol, small LDL particle, % change -7.3% -2.3% oxidized LDL, and ApoB levels, an increase in LDL particle 65 vs. baseline size, and a lower than expected increase in CRP levels were observed in 51 subjects within the statin subpopulation that US 8,048,880 B2 65 66 TABLE 42 TABLE 45 Changes in serum Small LDL particle concentration in statin Subpopulation following A-002 administration: Changes in serum ApoB concentration in statin Subpopulation following A-002 administration: A-002 Placebo Placebo Baseline 81 40

743.0 nmol/L 773.5 nmol/L. Baseline #o 81 40 10 subjects Median 66.1 mg/dl 58.2 mg/dl Week 8 75 37 ApoB Week 8 75 37 subjects 15 -66.0 nmol/L -6.5 nmol/L observed Change in -7.9 mg/dl +2.5 mg/dl Small LDL median particle ApoB vs. WS. baseline baseline % change -11.9% +3.5% % change -8.2% -1.0% WS. WS. baseline baseline 25

TABLE 43 TABLE 46 Changes in serum oxidized LDL concentration in Changes in LDL particle size in statin Subpopulation statin Subpopulation following A-002 administration: 30 following A-002 administration: A-002 Placebo A-002 Placebo

Baseline # of 81 40 Baseline # of 81 40 Subjects Subjects Median 41.1 UFL 42.2UL 35 Mean LDL 20.50 mm 20.48 mm oxidized particle LDL size Week 8 # of 76 33 Week 8 # of 75 37 Subjects Subjects observed observed Change in -1.1 UFL +0.3 UFL 40 Change in +0.10 mm -0.01 mm median mean LDL oxidized particle LDL vs. size vs. baseline baseline % change -2.9% +0.4% % change --0.5% O.0% WS. WS. baseline 45 baseline

TABLE 44 TABLE 47 Changes in serum TG concentration in statin 50 Changes in serum CRP concentration in statin Subpopulation following A-002 administration: Subpopulation following A-002 administration: Placebo Placebo

Baseline 8O 39 Baseline #o 81 40 55 subjects 136.5 mg/dl 130.0 mg/dl Median 1.10 mg/L. 1.30 mg/L ICRP) Week 8 75 37 Week 8 #o 75 37 Subiects subjects observed observed Change in -13.0 mg/dl +3.0 mg/dl 60 Change in +0.10 mg/L. +0.30 mg/L median median ITG) vs. CRP vs. baseline baseline % change -10.9% +2.6% % change +8.7% +40.6% WS. WS. baseline baseline 65 US 8,048,880 B2 67 68 TABLE 48 TABLE 51 Changes in serum Small LDL particle concentration in statin Changes in serum LDL concentration in statin Subpopulation Subpopulation with baseline serum LDL levels greater than with baseline serum LDL levels greater than 70 mg/dl 5 20 mg/dl following A-002 administration: following A-002 administration: A-002 Placebo

A-OO2 Placebo Baseline # of 31 2O Subjects Baseline # of 31 2O 10 Median 964.0 nmol/L 919.5 nmol/L. Small Subjects LDL Mean 91.6 mg/dl 93.2 mg/dl particle LDL Week 8 # of 30 18 Subjects Week 8 # of 30 18 15 observed Subjects Change in -115.0 nmol/L -70.0 nmol/L observed median Change in 13.9 mg/dl 5.0 mg/dl LDLSmall le:8 particle LDL vs. 2O WS. baseline baseline % change -12.3% -7.6% % change -15.0% -3.5% WS. WS. baseline baseline 25 TABLE 52 TABLE 49 Changes in serum oxidized LDL concentration in statin Subpopulation with baseline serum LDL levels greater than 70 mg/dl following A-002 Changes in serum non-HDL cholesterol concentration in statin 30 administration: Subpopulation with baseline serum LDL levels greater than 70 mg/d following A-002 administration: A-002 Placebo A-OO2 Placebo Baseline # of 31 2O Subjects Baseline # of 31 2O Median 47.9 UFL S1.6 USL Subjects 35 oxidized Mean non- 123.3 mg/dl 126.6 mg/dl LDL HDL Week 8 # of 76 33 cholesterol Subjects Week 8 # of 30 18 observed Subjects Change in -2.8 UL +1.3 U.L. observed 40 median Change in -16.7 mg/dl –7.2 mg/dl oxidized mean non- LDL vs. HDL baseline cholesterol % change -5.6% +2.5% vs. baseline WS. % change - 13.7% -4.1% 45 baseline vs. baseline

TABLE 53 TABLE SO 50 Changes in serum ApoB concentration in statin Subpopulation Changes in serum total cholesterol concentration in statin with baseline serum LDL levels greater than 70 mg/dl following Subpopulation with baseline serum LDL levels greater A-002 administration: than 70 mg/dl following A-002 administration: A-OO2 Placebo A-OO2 Placebo 55 Baseline # of 31 2O Baseline # of 31 2O Subjects Subjects Median 84.6 mg/dl 71.9 mg/dl Mean total 172.5 mg/dl 176.6 mg/dl ApoB cholesterol Week 8 # of 30 18 Week 8 # of 30 18 Subjects Subjects 60 observed observed Change in -13.1 mg/dl +1.1 mg/dl Change in -18.0 mg/dl -8.6 mg/dl median mean total ApoB vs. cholesterol baseline vs. baseline % change -18.7% +2.1% % change -10.5% -4.2% WS. vs. baseline 65 baseline US 8,048,880 B2 69 70 TABLE 54 9. Ivandic et al. 1999. Arterioscler Thromb Vasc Biol 19:1284-129O. Changes in LDL particle size in statin Subpopulation 10. Jialal, I. 1998. Clin Chem 44:1827-1832. with baseline serum LDL levels greater than 70 mg/dl following A-002 administration: 11. Kimura-Matsumoto et al. Atherosclerosis Epub Mar. 10, 5 2007. A-002 Placebo 12. Kugiyama et al. 1999. Circulation 100: 1280-1284. 13. Liu et al. 2003. Eur Heart J 24:1824-1832. Baseline # of 31 2O Subjects 14. Mallat et al. 2007. Arterioscler Thromb Vasc Biol Mean LDL 20.24 mm 20.62mm 27:1177-1183. particle 10 15. Mallat et al. 2005. JAm Coll Cardiol 46:1249-1257. size 16. Menschikowski et al. 1995. Atherosclerosis 118:173-181. Week 8 # of 30 18 Subjects 17. Nijmeijer et al. 2002. Cardiovasc Res 53:138-146. observed 18. Pruzanski et al. 1998. J. Lipid Res 39:2150-2160. Change in +0.19 mm +0.02 nm. 19. Ramoner et al. 2005. Blood 105:3583. mean LDL particle 15 20. Rosengren, B., et al. 2006. Arterioscler Thromb Vasc Biol size vs. 26:1579-1585. baseline 21. Sartipy et al. 1999. J. Biol Chem 274:25913-25920. % change +1.0% +0.1% 22. SZmitko et al. 2003. Circulation 108:2041. WS. 23. Tietge et al. 2000. J. Biol Chem 275: 10077-10084. baseline What is claimed is: 1. A composition comprising ((3-(2-Amino-1,2-dioxoet hyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy)acetic TABLE 55 acid or a pharmaceutically acceptable salt, polymorph, co crystal, Solvate, or prodrug derivative thereof and a statin. Changes in serum CRP concentration in statin Subpopulation with baseline serum LDL levels greater than 70 mg/dl following 25 2. The composition of claim 1, wherein said prodrug A-002 administration: derivative is 3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phe nylmethyl)-1H-indol-4-yl)oxyacetic acid methyl ester. A-OO2 Placebo 3. The composition of claim 1, wherein said statin is Baseline # of 31 2O selected from the group consisting of atorvastatin, cerivasta Subjects 30 tin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravasta Median 1.10 mg/L. 1.30 mg/L tin, rosuvastatin, simvastatin, and pharmaceutically accept ICRP) Week 8 # of 30 18 able salts, solvates, stereoisomers, or prodrug derivatives Subjects thereof. observed 4. The composition of claim 1, further comprising one or Change in +0.10 mg/L. +0.45 mg/L 35 more compounds selected from the group consisting of median CRP vs. eZetimibe, amlodipine, CP-529414, APA-01, extended baseline release niacin, MK-0524A, fenofibrate, and TAK-457. % change --9.1% +26.0% 5. The composition of claim 1, wherein the combination of WS. ((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)- baseline 40 1H-indol-4-yl)oxy)acetic acid or a pharmaceutically accept able salt, polymorph, co-crystal, Solvate, or prodrug deriva As stated above, the foregoing is merely intended to illus tive thereof and a statin has a greater than additive effect in the trate various embodiments of the present invention. The spe treatment of a cardiovascular disease associated with elevated cific modifications discussed above are not to be construed as LDL levels and/or atherosclerosis when administered to a limitations on the scope of the invention. It will be apparent to 45 subject in need thereof. one skilled in the art that various equivalents, changes, and 6. The composition of claim 1, wherein the combination of modifications may be made without departing from the scope ((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)- of the invention, and it is understood that such equivalent 1H-indol-4-yl)oxy)acetic acid or a pharmaceutically accept embodiments are to be included herein. All references cited able salt, polymorph, co-crystal, Solvate, or prodrug deriva 50 tive thereof and a statin has a synergistic effect in the herein are incorporated by reference as if fully set forth treatment of a cardiovascular disease associated with elevated herein. LDL levels and/or atherosclerosis when administered to a subject in need thereof. REFERENCES 7. The composition of claim 1, wherein the combination of 55 ((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)- 1. Boekholdt et al. 2005. Arterioscler Thromb Vasc Biol 1H-indol-4-yl)oxy)acetic acid or a pharmaceutically accept 25:839-846. able salt, polymorph, co-crystal, Solvate, or prodrug deriva 2. Bostrom et al. 2007. Arterioscler Thromb Vasc Biol tive thereof and a statin has a greater than additive effect in the 27:600-606. reduction of LDL levels when administered to a subject in 3. Cameo et al. 1998. Atherosclerosis 139:205-222. 60 need thereof. 4. Chait, A., et al. 2005. J. Lipid Res 46:389-403. 8. The composition of claim 1, wherein the combination of 5. Daugherty et al. 2000. J. Clin Invest 105:1605-1612. ((3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)- 6. Elinder et al. 1997. Arterioscler Thromb Vasc Biol 1H-indol-4-yl)oxy)acetic acid or a pharmaceutically accept 17:2257-2263. able salt, polymorph, co-crystal, Solvate, or prodrug deriva 7. Hakala et al. 2001. Arterioscler Thromb VascBiol 21:1053 65 tive thereof and a statin has a synergistic effect in the 1058. reduction of LDL levels when administered to a subject in 8. Hartford et al. 2006. J Cardiol 108:55-62. need thereof. US 8,048,880 B2 71 72 9. A composition comprising ((3-(2-Amino-1,2-dioxoet 20. The pharmaceutical composition of claim 19, wherein hyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy)acetic said prodrug derivative is 3-(2-Amino-1,2-dioxoethyl)-2- acid or a pharmaceutically acceptable salt, polymorph, co ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxyacetic acid methyl ester. crystal, Solvate, or prodrug derivative for administration in 21. A pharmaceutical composition for administration in combination with a statin. combination with a statin comprising ((3-(2-Amino-1,2-di 10. The composition of claim 9, wherein said composition oxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy) is administered simultaneous with a statin. acetic acid or a pharmaceutically acceptable salt, polymorph, 11. The composition of claim 9, wherein said composition co-crystal, Solvate, or prodrug derivative thereof and one or is administered sequentially with a statin. more pharmaceutically acceptable carriers, wherein said 12. The composition of claim 9, wherein said prodrug 10 composition is administered simultaneous with or sequen derivative is 3-(2-Amino-1,2-dioxoethyl)-2-ethyl-1-(phe tially with a statin. nylmethyl)-1H-indol-4-yl)oxyacetic acid methyl ester. 22. The pharmaceutical composition of claim 21, wherein 13. The composition of claim 9, wherein administration of said prodrug derivative is 3-(2-Amino-1,2-dioxoethyl)-2- ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxyacetic acid said composition in combination with a statin to a subject in 15 need thereof results in a greater than additive effect in the methyl ester. treatment of a cardiovascular disease associated with elevated 23. A kit comprising ((3-(2-Amino-1,2-dioxoethyl)-2- LDL levels and/or atherosclerosis. ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy)acetic acid or a 14. The composition of claim 9, wherein administration of pharmaceutically acceptable salt, polymorph, co-crystal, Sol said composition in combination with a statin to a subject in Vate, or prodrug derivative thereof and a statin. 24. The kit of claim 23, wherein said ((3-(2-Amino-1,2- need thereof results in a synergistic effect in the treatment of dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy) a cardiovascular disease associated with elevated LDL levels acetic acid or a pharmaceutically acceptable salt, polymorph, and/or atherosclerosis. co-crystal, Solvate, or prodrug derivative thereof and said 15. The composition of claim 9, wherein administration of statin are divided into separate compartments. said composition in combination with a statin to a subject in 25 25. The kit of claim 23, wherein said ((3-(2-Amino-1,2- need thereof results in a greater than additive effect in the dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy) reduction of LDL levels. acetic acid or a pharmaceutically acceptable salt, polymorph, 16. The composition of claim 9, wherein administration of co-crystal, Solvate, or prodrug derivative thereof and said said composition in combination with a statin to a subject in statin are divided into separate compartments. need thereof results in a synergistic effect in the reduction of 30 LDL levels. 26. The kit of claim 23, wherein said ((3-(2-Amino-1,2- 17. A composition comprising ((3-(2-Amino-1,2-dioxoet dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy) hyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy)acetic acetic acid or a pharmaceutically acceptable salt, polymorph, acid or a pharmaceutically acceptable salt, polymorph, co co-crystal, Solvate, or prodrug derivative thereof and said statin are held within a single undivided container. crystal, Solvate, or prodrug derivative for administration in 35 27. The kit of claim 26, wherein said ((3-(2-Amino-1,2- combination with a statin to treat a cardiovascular disease dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxy) associated with elevated LDL levels and/or atherosclerosis. acetic acid or a pharmaceutically acceptable salt, polymorph, 18. A composition comprising 3-(2-Amino-1,2-dioxoet co-crystal, Solvate, or prodrug derivative thereof and said hyl)-2-ethyl-1-(phenylmethyl)-1H-indol-4-yl)oxyacetic statin are part of a single pharmaceutical composition. acid methyl ester for administration in combination with a 40 statin to treat a cardiovascular disease associated with 28. The kit of claim 23, wherein said kit further comprises elevated LDL levels and/or atherosclerosis. instructions for usage. 19. A pharmaceutical composition comprising ((3-(2- 29. The kit of claim 23 for use in lowering LDL and/or Amino-1,2-dioxoethyl)-2-ethyl-1-(phenylmethyl)-1H-in small LDL particle levels in a subject in need thereof. dol-4-yl)oxy)acetic acid or a pharmaceutically acceptable 45 30. The kit of claim 23 for use in treating a cardiovascular salt, polymorph, co-crystal, Solvate, or prodrug derivative disease associated with elevated LDL levels and/or athero thereof, a statin, and one or more pharmaceutically accept Sclerosis. able carriers. UNITED STATES PATENT AND TRADEMARK OFFICE CERTIFICATE OF CORRECTION PATENT NO. : 8,048,880 B2 Page 1 of 1 APPLICATIONNO. : 12/114710 DATED : November 1, 2011 INVENTOR(S) : Joaquim Trias et al. It is certified that error appears in the above-identified patent and that said Letters Patent is hereby corrected as shown below:

On the Face page, in Field (75) “INVENTORS, Column 1, Line 7, please delete “Kenneth Gould, Zionsville, IN (US):”. On the Face page, in Field (75) “INVENTORS, Column 1, Line 8, please delete “Marian Mosior, Indianapolis, IN (US);. On the Face page, in Field (75) “INVENTORS, Column 1, Line 9, please delete “Patrick Eacho, Indianapolis, IN (US).

Signed and Sealed this Fifth Day of June, 2012

David J. Kappos Director of the United States Patent and Trademark Office