USOO8637524B2

(12) United States Patent (10) Patent No.: US 8,637,524 B2 Rao et al. (45) Date of Patent: Jan. 28, 2014

(54) PYRIMIDINONE INHIBITORS OF Tonn, Biological Mass Spectrometry vol. 22 Issue 11, pp. 633-642 LIPOPROTEIN-ASSOCATED (1993).* PHOSPHOLPASE A2 Hist Biomedical Spectrometry vol. 9 Issue 7, pp. 269-277 Wolen, Journal of Clinical Pharmacology 1986; 26: 419-424.* (75) Inventors: Tadimeti Rao, San Diego, CA (US); Browne, Journal of Clinical Pharmacology 1998; 38: 213-220.* Chengzhi Zhang, San Diego, CA (US) Baillie, Pharmacology Rev. 1981: 33:81-132.* Gouyette, Biomedical and Environmental Mass Spectrometry, vol. (73) Assignee: Auspex Pharmaceuticals, Inc, La Jolla, 15, 243-247 (1988).* CA (US) Cherrah, Biomedical and Environmental Mass Spectrometry vol. 14 Issue 11, pp. 653-657 (1987).* Pieniaszek, J. Clin Pharmacol. 1999; 39: 817-825.* (*) Notice: Subject to any disclaimer, the term of this Honma et al., Drug Metab Dispos 15 (4): 551 (1987).* patent is extended or adjusted under 35 Kushner, D. Jet al., Pharmacological uses and perspectives of heavy U.S.C. 154(b) by 58 days. water and deuterated compounds, Can. J. Physiol. Pharmacol. (1999), 77, 79-88. (21) Appl. No.: 12/840,725 Bauer et al., Influence of long-term infusions on lidocaine kinetics, Clin. Pharmacol. Ther. (1982), 31(4), 433-7. (22) Filed: Jul. 21, 2010 Borgstrom et al., Comparative Pharmacokinetics of Unlabeled and Deuterium-Labeled Terbutaline: Demonstration of a Small Isotope Effect, J Pharm. Sci., (1988), 77(11) 952-4. (65) Prior Publication Data Browne et al., Chapter 2. Isotope Effect: Implications for pharma US 2011/0306552 A1 Dec. 15, 2011 ceutical investigations, Stable isotopes in pharmaceutical research; s Elsevier; Amsterdam, (1997). Related U.S. Application Data Browne et al., Pharmacokinetic equivalence of stable-isotope-la beled and unlabeled drugs. Phenobarbital in man, JClin Pharmacol, (60) Provisional application No. 61/229,096, filed on Jul. (1982), 22, 309-15. 28, 2009. Burm et al., Pharmacokinetics of Lidocaine and Bupivacaine and stbel isotope labelled analogues: a study in healthy volunteers, (51) Int. Cl. Biopharma & Drug Disp, (1988), 9, 85-95. C07D 239/78 (2006.01) Elison et al., Effect of deuteration of N-CH3 Group on Potency and A 6LX3/57 (200 6. 01) enzymatic N-demethylation of morphine, Science, A61P II/06 (2006.015 (1961), 134(3485), 1078-9. A61P 29/00 (2006.01) (Continued) A61P 9/10 (2006.01) (52) U.S. Cl. Primary Examiner — Susanna Moore USPC ...... 514/258.1544/253 (74). Attorney, Agent, or Firm - Dennis A. Bennett; Mike (58) Field of Classification Search Sertic USPC ...... 544/253: 514/258.1 See application file for complete search history. (57) ABSTRACT The present invention relates to new pyrimidinone inhibitors (56) References Cited of lipoprotein-associated phospholipase A2 activity, pharma U.S. PATENT DOCUMENTS ceutical compositions thereof, and methods of use thereof

6,221,335 B1 * 4/2001 Foster ...... 424f1.81 6,440,710 B1* 8/2002 Keinan et al. ... 435,148 Formula I

6,603,008 B1* 8/2003 Ando et al...... 546,269.7 6,649,619 B1 * 1 1/2003 Hickey et al. 514,258.1 7,517,990 B2 * 4/2009 Ito et al...... 546, 184 2002fOO13372 A1 1/2002 Ekins et al. 2007/0082929 A1 4/2007 Gant et al...... 514,338 2007/0197695 A1* 8/2007 Potyen et al...... 524/110 2008, OO33011 A1 2/2008 Tung 2008/0287774 A1 11, 2008 Katz-Birull 2011/O136861 A1 6, 2011 Rao et al.

FOREIGN PATENT DOCUMENTS

WO 9526325 A2 10, 1995 WO O160805 A1 8, 2001 WO 02083144 A1 10, 2002 WO O3O16287 A2 2, 2003 WO 2006086978 A2 8, 2006 WO 2006086978 A3 8, 2006 WO 2007052274 A2 5/2007 WO 2007052274 A3 5/2007 OTHER PUBLICATIONS Dyck, Journal of Neurochemistry vol. 46 Issue 2, pp. 399-404 (1986).* 16 Claims, No Drawings US 8,637,524 B2 Page 2

(56) References Cited Haskins, N. J., The Application of Stable Isotopes in Biomedical Research, Biomedical Mass Spectrometry, (1982), 9(7), 269-277. OTHER PUBLICATIONS Honma et al., The Metabolism of Roxatidine Acetate Hydrochloride, Farmer et al., Synthesis, metabolism, and antitumor activity of Drug Metabolism and Disposition, (1987), 15(4), 551-559. deuterated analogues of 1-(2-chloroethyl)-3-cyclohexyl-1- Pieiaszek et al., Moricizine Bioavailability Via Simultaneous, Dual, nitrosourea, J Med Chem, (1978), 21(6), 514-520. Stable Isotope Administration: Bioecuivalence Implications, J. Clin. Fisher et al., The complexities inherent in attempts to decrease drug Pharmacol., (1999), 39, 817-825. clearance by blocking sites of CYP-mediated metabolism, Curr Opin Tonn et al., Simultaneous Analysis of Diphenylhydramine and a in Drug Disc & Develop, (2006), 9(1), 101-9. Stable Isotope Analog (2H10) Diphenylhydramine Using Capillary Foster et al., Deuterium isotope effects in studies of drug metabolism, Gas Chromatography With Mass Selective Detection in Biological Trends in Pharma Sci., (1984), 524-7. Fluids From Chronically Instrumented Pregnant Ewes, Biomedical Helfenbein et al., Isotopic Effect Study of Propofol Deuteration on Mass Spectrometry, (1993), 22, 633-642. the Metabolism, Activity, and Toxicity of the Anesthetic, J Med Wolen et al., The Application of Stable Isotopes to Studies of Drug Chem. (2002), 45, 5806-8. Lee et al., Deuterium magic angle spinning studies of Substrates Bioavailibility and Bioecuivalence, J. Clin. Pharmacol., (1986), 26. bound to cytochrome P450, Biochem. (1999), 38, 10808-13. 419-424. Mamada et al., Pharmacokinetic equivalence of deuterium-labeled Blackie et al., The Identification of Clinical Candidate SB-480848: A and unlabeled phenytoin, Drug Metab Disp., (1986), 14(4) 509-11. Potent Inhibitor of Lipoprotein-Associated Phospholipase A2. Nelson et al., The use of deuterium isotope effects to probe the active Bioorganic & Medicinal Chemistry Letters, (2003), 13, 1067-1070. site properties, mechanism, of cytochrome P450-catalyzed reactions, Buiet al., Darapladib, Expert Opin. Investig. Drugs, (2010), 19(1), and mechanisms of metabolically dependent toxicity, Drug Metab 161-168. Disp., (2003), 31(12), 1481-98. Magee et al... An Integrated Pharmacokinetic/Pharmacodynamic Nelson et al., Primary and B-Secondary Deuterium Isotope Effects in (PK/PD) Model for SB-480848 Inhibition of Plasma Lipoprotein N-Deethylation reactions, J Med Chem. (1975), 18(11) 1062-5. Associated Phospholipase A2 (LP-PLA2) Enzyme Activity in Pohl et al., Determination of Toxic Pathways of Metabolism by Human, Clin. Pharmacol. Ther. (2003), 73(2), P86, Abst PIII-87. Deuterium Substitution, Drug Metab Rev., (1984), 15(7), 1335-51. Glinski et al., Catalytic Ketonisation Over Oxide Catalysts, Part Rampe et al., Deuterated analogs of Verapamil and nifedipine. Syn VIII.a Synthesis of Cycloalkanones in Cycloketonisation of Various thesis and biological activity, Eur J Med Chem... (1993), 28, 259-63. Dialkyl Alkanodiates, Reaction Kinetics and Catalysis Letters, Baillie, Thomas, The Use of Stable Isotopes in Pharmaceutical (2004), 82(1), 157-163. Research, Pharmacological Reviews, (1981), 33(2), 81-132. Wilensky et al., Inhibition of lipoprotein-associated phospholipase Browne, Thomas, Stable Isotope Techniques in Early Drug Develop A2 reduces complex coronary atherosclerotic plaque development, ment: An Economic Evaluation, J. Clin. Pharmacol., (1998), 38. 213-220. Nature Medicine, (2008), 14(10), 1059-1066. Cherrahetal. Study of Deuterium Isotope Effects on Protein Binding Mohler et al. The Effect of Darapladib on Plasma Lipoprotein by Gas Chromatography/Mass Spectrometry. Caffeine and Deuter Associated Phospholipase A2 Activity and Cardiovascular Biomark ated Isomers, Biomedical and Environmental Mass Spectrometry, ers in Patients With Stable Coronary Heart Disease or Coronary Heart (1987), 14,653-657. Disease Risk Equivalent, J. Am. Coll. Cardiol. (2008), 51(17), 1632 Dyck et al., Effects of Deuterium Substitution on the Catabolism of 41. Beta-Phenethylamine: An In Vivo Study, J. Neurochem... (1986), Foster, A.B., Deuterium Isotope Effects in the Metabolism of Drugs 46(2), 399-404. and Xenobiotics: Implications for Drug Design, Adv. Drug Res., Gouyette, Alain, Use of Deuterium-Labelled Elliptinium and Its Use Academic Press, London, GB, vol. 14, Jan. 1, 1985, pp. 1-40. in Metabolic Studies, Biomedical and Environmental Mass Spec trometry, (1988), 15, 243-247. * cited by examiner US 8,637,524 B2 1. 2 PYRIMDNONE INHIBITORS OF hydrogen (C H) bond to either a carbon-oxygen (C–O) or LIPOPROTEIN-ASSOCATED a carbon-carbon (C C) L-bond. The resultant metabolites PHOSPHOLPASE A2 may be stable or unstable under physiological conditions, and can have Substantially different pharmacokinetic, pharmaco This application claims the benefit of priority of U.S. pro- 5 dynamic, and acute and long-term toxicity profiles relative to visional application No. 61/229,096, filed Jul. 28, 2009, the the parent compounds. For most drugs, such oxidations are disclosure of which is hereby incorporated by reference as if generally rapid and ultimately lead to administration of mul written herein in its entirety. tiple or high daily doses. Disclosed herein are new substituted pyrimidinone com The relationship between the activation energy and the rate pounds, pharmaceutical compositions made thereof, and 10 of reaction may be quantified by the Arrhenius equation, methods to inhibit lipoprotein-associated phospholipase A2 k=Ae'. The Arrhenius equation states that, at a given activity in a subject are also provided for, for the treatment of temperature, the rate of a chemical reaction depends expo disorders such as primary and secondary coronary disorders, nentially on the activation energy (E). acute coronary heart disorders, atherosclerosis, peripheral The transition state in a reaction is a short lived state along vascular atherosclerosis, cerebrovascular atherosclerosis, 15 the reaction pathway during which the original bonds have rheumatoid arthritis, diabetes, stroke, myocardial infarction, stretched to their limit. By definition, the activation energy reperfusion injury, acute and chronic inflammation, psoriasis, E. for a reaction is the energy required to reach the transition and asthma. state of that reaction. Once the transition state is reached, the Darapladib (SB-480848, CAS #356057-34-6), N-(2-di molecules can either revert to the original reactants, or form ethylamino-ethyl)-2-[2-(4-fluoro-benzylsulfanyl)-4-oxo-4, 2O new bonds giving rise to reaction products. A catalyst facili 5,6,7-tetrahydro-cyclopentapyrimidin-1-yl)-N-(4-trifluo tates a reaction process by lowering the activation energy romethyl-biphenyl-4-ylmethyl)-acetamide, is a lipoprotein leading to a transition State. Enzymes are examples of bio associated phospholipase A2 inhibitor. Darapladib is logical catalysts. currently under investigation for the treatment of primary and Carbon-hydrogen bond strength is directly proportional to secondary coronary disorders, acute coronary heart disorders, 25 the absolute value of the ground-state vibrational energy of atherosclerosis, peripheral vascularatherosclerosis, and cere the bond. This vibrational energy depends on the mass of the brovascularatherosclerosis (Wilensky et al., Nature Medicine atoms that form the bond, and increases as the mass of one or 2008, 14(10), 1059-66; WO 03/016287 A2; and both of the atoms making the bond increases. Since deuterium WO/O160805 A1). Darapladib has also shown promise in (D) has twice the mass of protium (H), a C-D bond is stron treating rheumatoid arthritis, diabetes, stroke, myocardial 30 ger than the corresponding C–H bond. If a C-H bond is infarction, reperfusion injury, acute and chronic inflamma broken during a rate-determining step in a chemical reaction tion, psoriasis, and asthma (WO/O160805 A1; Blackie et al., (i.e. the step with the highest transition state energy), then Bioorg Med. Chem. Lett. 2003, 13, 1067-70; and Mohler et Substituting a deuterium for that protium will cause a al., J. Am. Coll. Cardiol 2008, 51 (17), decrease in the reaction rate. This phenomenon is known as 35 the Deuterium Kinetic Isotope Effect (DKIE). The magnitude of the DKIE can be expressed as the ratio between the rates of O a given reaction in which a C H bond is broken, and the same reaction where deuterium is substituted for protium. The DKIE can range from about 1 (no isotope effect) to very 40 large numbers, such as 50 or more. Substitution of tritium for hydrogen results in yet a stronger bond than deuterium and S l N CF gives numerically larger isotope effects.

O Deuterium (HorD) is a stable and non-radioactive isotope F of hydrogen which has approximately twice the mass of pro 45 tium ("H), the most common isotope of hydrogen. Deuterium N oxide (DO or “heavy water) looks and tastes like H2O, but has different physical properties. When pure DO is given to rodents, it is readily absorbed. -- The quantity of deuterium required to induce toxicity is extremely high. When about 0-15% of the body water has N 50 been replaced by DO, animals are healthy but are unable to gain weight as fast as the control (untreated) group. When Darapladib about 15-20% of the body water has been replaced with D.O. the animals become excitable. When about 20-25% of the Adverse effects associated with darapladibadministration 55 body water has been replaced with DO, the animals become includes unpleasant odor in feces?urine, diarrhea, and so excitable that they go into frequent convulsions when changes in taste. stimulated. Skin lesions, ulcers on the paws and muzzles, and necrosis of the tails appear. The animals also become very DEUTERIUM KINETIC ISOTOPE EFFECT aggressive. When about 30% of the body water has been 60 replaced with DO, the animals refuse to eat and become In order to eliminate foreign Substances such as therapeutic comatose. Their body weight drops sharply and their meta agents, the animal body expresses various enzymes, such as bolic rates drop far below normal, with death occurring at the cytochrome Paso enzymes (CYPs), esterases, proteases, about 30 to about 35% replacement with D.O.The effects are reductases, dehydrogenases, and monoamine oxidases, to reversible unless more than thirty percent of the previous react with and convert these foreign Substances to more polar 65 body weight has been lost due to D.O. Studies have also intermediates or metabolites for renal excretion. Such meta shown that the use of DO can delay the growth of cancer cells bolic reactions frequently involve the oxidation of a carbon and enhance the cytotoxicity of certain antineoplastic agents. US 8,637,524 B2 3 Deuteration of pharmaceuticals to improve pharmacoki netics (PK), pharmacodynamics (PD), and toxicity profiles (I) has been demonstrated previously with some classes of drugs. For example, the DKIE was used to decrease the hepatotox icity of halothane, presumably by limiting the production of 5 reactive species such as trifluoroacetyl chloride. However, this method may not be applicable to all drug classes. For example, deuterium incorporation can lead to metabolic Switching. Metabolic Switching occurs when Xenogens, sequestered by Phase I enzymes, bind transiently and re-bind 10 in a variety of conformations prior to the chemical reaction (e.g., oxidation). Metabolic switching is enabled by the rela tively vast size of binding pockets in many Phase I enzymes and the promiscuous nature of many metabolic reactions. Metabolic switching can lead to different proportions of 15 known metabolites as well as altogether new metabolites. This new metabolic profile may impart more or less toxicity. Such pitfalls are non-obvious and are not predictable a priori for any drug class. Darapladib is a lipoprotein-associated phospholipase A2 or a pharmaceutically acceptable salt thereof, wherein: inhibitor. The carbon-hydrogenbonds of darapladib contain a R-Rs are independently selected from the group consist naturally occurring distribution of hydrogenisotopes, namely ing of hydrogen and deuterium; and "H or protium (about 99.984.4%), H or deuterium (about at least one of R-Rs is deuterium. 0.0156%), and H or tritium (in the range between about 0.5 Certain compounds disclosed herein may possess useful and 67 tritium atoms per 10' protium atoms). Increased 25 lipoprotein-associated phospholipase A2 inhibiting activity, levels of deuterium incorporation may produce a detectable and may be used in the treatment or prophylaxis of a disorder Deuterium Kinetic Isotope Effect (DKIE) that could effect in which lipoprotein-associated phospholipase A2 plays an the pharmacokinetic, pharmacologic and/or toxicologic pro active role. Thus, certain embodiments also provide pharma files of darapladib in comparison with darapladib having ceutical compositions comprising one or more compounds naturally occurring levels of deuterium. 30 disclosed herein together with a pharmaceutically acceptable Based on discoveries made in our laboratory, as well as carrier, as well as methods of making and using the com considering the literature, darapladib likely to be metabolized pounds and compositions. Certain embodiments provide in humans at the N-ethyl groups, the N-methylene groups, the methods for inhibiting lipoprotein-associated phospholipase cyclopentyl methylene groups, and the S-methylene group. A2 activity. Other embodiments provide methods for treating The current approach has the potential to prevent metabolism 35 a lipoprotein-associated phospholipase A2-mediated disor at these sites. Other sites on the molecule may also undergo der in a patient in need of such treatment, comprising admin transformations leading to metabolites with as-yet-unknown istering to said patient a therapeutically effective amount of a pharmacology/toxicology. Limiting the production of these compound or composition according to the present invention. metabolites has the potential to decrease the danger of the Also provided is the use of certain compounds disclosed administration of Such drugs and may even allow increased 40 herein for use in the manufacture of a medicament for the dosage and/or increased efficacy. All of these transformations prevention or treatment of a disorder ameliorated by inhibit can occur through polymorphically-expressed enzymes, ing lipoprotein-associated phospholipase A2. exacerbating interpatient variability. Further, some disorders The compounds as disclosed herein may also contain less are best treated when the subject is medicated around the prevalent isotopes for other elements, including, but not lim clock or for an extended period of time. For all of the forego 45 ited to, C or C for carbon, S, S, or S for sulfur, 'N ing reasons, a medicine with a longer half-life may result in for nitrogen, and 'O or "O for oxygen. greater efficacy and cost savings. Various deuteration patterns In certain embodiments, the compound disclosed herein can be used to (a) reduce or eliminate unwanted metabolites, may expose apatient to a maximum of about 0.000005% DO (b) increase the half-life of the parent drug, (c) decrease the or about 0.00001% DHO, assuming that all of the C-D bonds number of doses needed to achieve a desired effect, (d) 50 in the compound as disclosed herein are metabolized and decrease the amount of a dose needed to achieve a desired released as DO or DHO. In certain embodiments, the levels effect, (e) increase the formation of active metabolites, if any of DO shown to cause toxicity in animals is much greater are formed, (f) decrease the production of deleterious than even the maximum limit of exposure caused by admin metabolites in specific tissues, and/or (g) create a more effec istration of the deuterium enriched compound as disclosed tive drug and/or a safer drug for polypharmacy, whether the 55 herein. Thus, in certain embodiments, the deuterium-en polypharmacy be intentional or not. The deuteration riched compound disclosed herein should not cause any addi approach has the strong potential to slow the metabolism of tional toxicity due to the formation of DO or DHO upon drug darapladib and attenuate interpatient variability. metabolism. Novel compounds and pharmaceutical compositions, cer In certain embodiments, the deuterated compounds dis tain of which have been found to inhibit lipoprotein-associ 60 closed herein maintain the beneficial aspects of the corre ated phospholipase A2 activity have been discovered, sponding non-isotopically enriched molecules while Substan together with methods of synthesizing and using the com tially increasing the maximum tolerated dose, decreasing pounds, including methods for the treatment of lipoprotein toxicity, increasing the half-life (T), lowering the maxi associated phospholipase A2-mediated disorders in a patient mum plasma concentration (C) of the minimum effica by administering the compounds as disclosed herein. 65 cious dose (MED), lowering the efficacious dose and thus In certain embodiments of the present invention, com decreasing the non-mechanism-related toxicity, and/or low pounds have structural Formula I: ering the probability of drug-drug interactions. US 8,637,524 B2 5 6 All publications and references cited herein are expressly tiomeric products followed by separation Such as conversion incorporated herein by reference in their entirety. However, to a mixture of diastereomers followed by separation or with respect to any similar or identical terms found in both the recrystallization, chromatographic techniques, direct separa incorporated publications or references and those explicitly tion of enantiomers on chiral chromatographic columns, or put forth or defined in this document, then those terms defi any other appropriate method known in the art. Starting com nitions or meanings explicitly put forth in this document shall pounds of particular Stereochemistry are either commercially control in all respects. available or can be made and resolved by techniques known in As used herein, the terms below have the meanings indi the art. Additionally, the compounds disclosed herein may cated. exist as geometric isomers. The present invention includes all The singular forms “a”, “an', and “the may refer to plural 10 cis, trans, syn, anti, entgegen (E), and Zusammen (Z) isomers articles unless specifically stated otherwise. as well as the appropriate mixtures thereof. Additionally, The term “about’, as used herein, is intended to qualify the compounds may exist as tautomers; all tautomeric isomers numerical values which it modifies, denoting Such a value as are provided by this invention. Additionally, the compounds variable within a margin of error. When no particular margin disclosed herein can exist in unsolvated as well as Solvated of error, such as a standard deviation to a mean value given in 15 forms with pharmaceutically acceptable solvents such as a chart or table of data, is recited, the term “about should be water, ethanol, and the like. In general, the Solvated forms are understood to mean that range which would encompass the considered equivalent to the unsolvated forms. recited value and the range which would be included by The term “bond refers to a covalent linkage between two rounding up or downto that figure as well, taking into account atoms, or two moieties when the atoms joined by the bond are significant figures. considered to be part of larger substructure. A bond may be When ranges of values are disclosed, and the notation single, double, or triple unless otherwise specified. A dashed "from n ... to n' or “n-n' is used, where n and n are the line between two atoms in a drawing of a molecule indicates numbers, then unless otherwise specified, this notation is that an additional bond may be present or absent at that intended to include the numbers themselves and the range position. between them. This range may be integral or continuous 25 The term “disorder as used herein is intended to be gen between and including the end values. erally synonymous, and is used interchangeably with, the The term “deuterium enrichment” refers to the percentage terms “disease”, “syndrome', and “condition' (as in medical of incorporation of deuterium at a given position in a mol condition), in that all reflect an abnormal condition of the ecule in the place of hydrogen. For example, deuterium human or animal body or of one of its parts that impairs enrichment of 1% at a given position means that 1% of mol 30 normal functioning, is typically manifested by distinguishing ecules in a given sample contain deuterium at the specified signs and Symptoms. position. Because the naturally occurring distribution of deu The terms “treat”, “treating, and “treatment” are meant to terium is about 0.0156%, deuterium enrichment at any posi include alleviating or abrogating a disorder or one or more of tion in a compound synthesized using non-enriched starting the symptoms associated with a disorder, or alleviating or materials is about 0.0156%. The deuterium enrichment can 35 eradicating the cause(s) of the disorder itself. As used herein, be determined using conventional analytical methods known reference to “treatment of a disorder is intended to include to one of ordinary skill in the art, including mass spectrometry prevention. The terms “prevent”, “preventing, and “preven and nuclear magnetic resonance spectroscopy. tion” refer to a method of delaying or precluding the onset of The term “is/are deuterium, when used to describe a given a disorder, and/or its attendant symptoms, barring a subject position in a molecule such as R-Rs or the symbol “D’. 40 from acquiring a disorder or reducing a subject’s risk of when used to represent a given position in a drawing of a acquiring a disorder. molecular structure, means that the specified position is The term “therapeutically effective amount” refers to the enriched with deuterium above the naturally occurring distri amount of a compound that, when administered, is sufficient bution of deuterium. In one embodiment deuterium enrich to prevent development of oralleviate to some extent, one or ment is no less than about 1%, in another no less than about 45 more of the symptoms of the disorder being treated. The term 5%, in another no less than about 10%, in another no less than “therapeutically effective amount” also refers to the amount about 20%, in another no less than about 50%, in another no of a compound that is sufficient to elicit the biological or less than about 70%, in another no less than about 80%, in medical response of a cell, tissue, system, animal, or human another no less than about 90%, or in another no less than that is being sought by a researcher, Veterinarian, medical about 98% of deuterium at the specified position. 50 doctor, or clinician. The term “isotopic enrichment” refers to the percentage of The term “subject” refers to an animal, including, but not incorporation of a less prevalent isotope of an element at a limited to, a primate (e.g., human, monkey, chimpanzee, given position in a molecule in the place of the more prevalent gorilla, and the like), rodents (e.g., rats, mice, gerbils, ham isotope of the element. sters, ferrets, and the like), lagomorphs, Swine (e.g., pig, The term “non-isotopically enriched refers to a molecule 55 miniature pig), equine, canine, feline, and the like. The terms in which the percentages of the various isotopes are Substan “subject' and “patient” are used interchangeably herein in tially the same as the naturally occurring percentages. reference, for example, to a mammalian Subject, such as a Asymmetric centers exist in the compounds disclosed human patient. herein. These centers are designated by the symbols “R” or The term “combination therapy’ means the administration 'S', depending on the configuration of Substituents around 60 of two or more therapeutic agents to treat a therapeutic dis the chiral carbonatom. It should be understood that the inven order described in the present disclosure. Such administration tion encompasses all Stereochemical isomeric forms, includ encompasses co-administration of these therapeutic agents in ing diastereomeric, enantiomeric, and epimeric forms, as a Substantially simultaneous manner, Such as in a single cap well as D-isomers and L-isomers, and mixtures thereof. Indi Sule having a fixed ratio of active ingredients or in multiple, vidual stereoisomers of compounds can be prepared syntheti 65 separate capsules for each active ingredient. In addition, Such cally from commercially available starting materials which administration also encompasses use of each type of thera contain chiral centers or by preparation of mixtures of enan peutic agent in a sequential manner. In either case, the treat US 8,637,524 B2 7 8 ment regimen will provide beneficial effects of the drug com response, immunogenecity, are commensurate with a reason bination in treating the disorders described herein. able benefit/risk ratio, and are effective for their intended use. The term “lipoprotein-associated phospholipase A2 The term “pharmaceutically acceptable carrier”, “pharma refers to a member of the phospholipase A2 enzyme family, ceutically acceptable excipient”, “physiologically acceptable and is primarily found in blood and atherosclerotic plaque. carrier, or “physiologically acceptable excipient” refers to a Lipoprotein-associated phospholipase A2 is a potent pro pharmaceutically-acceptable material, composition, or inflammatory mediator involved in the vascular inflammation vehicle. Such as a liquid or Solid filler, diluent, excipient, process. It has been suggested that elevated lipoprotein-asso Solvent, or encapsulating material. Each component must be ciated phospholipase A2 activity is partly responsible for the “pharmaceutically acceptable' in the sense of being compat development and progression of atherosclerosis and cardio 10 ible with the other ingredients of a pharmaceutical formula vascular disorders. Thus, inhibition of lipoprotein-associated tion. It must also be suitable for use in contact with the tissue phospholipase A2 is expected to be useful in the treatment of or organ of humans and animals without excessive toxicity, these disorders. Mechanistically, lipoprotein-associated irritation, allergic response, immunogenecity, or other prob phospholipase A2 enzyme preferentially cleaves LDL cho lems or complications, commensurate with a reasonable ben lesterol to form lysophosphatidylcholine and oxidized fatty 15 efit/risk ratio. See, Remington. The Science and Practice of acids. These molecules lead to several atherogenic actions, Pharmacy, 21st Edition; Lippincott Williams & Wilkins: including the expression of adhesion molecules and the Philadelphia, Pa., 2005, Handbook of Pharmaceutical attraction of macrophages and lymphocytes to the atheroscle Excipients, 5th Edition; Rowe et al., Eds. The Pharmaceuti roticlesion. These events lead to the buildup of plaque within cal Press and the American Pharmaceutical Association: the arterial wall. The rupture of this unstable atherosclerotic 2005; and Handbook of Pharmaceutical Additives, 3rd Edi plaque is the primary cause of most heart attacks and strokes. tion; Ash and Ash Eds. Gower Publishing Company: 2007; Thus, it is hypothesized that inhibiting lipoprotein-associated Pharmaceutical Preformulation and Formulation, Gibson phospholipase A2 activity will prevent and/or reverse the Ed., CRC Press LLC: Boca Raton, Fla., 2004). build up of these macrophage enriched lesions. The terms “active ingredient”, “active compound', and The term "lipoprotein-associated phospholipase A2-medi 25 “active substance” refer to a compound, which is adminis ated disorder', refers to a disorder that is characterized by tered, alone or in combination with one or more pharmaceu abnormal lipoprotein-associated phospholipase A2 activity, tically acceptable excipients or carriers, to a subject for treat or normal lipoprotein-associated phospholipase A2 activity ing, preventing, or ameliorating one or more symptoms of a that when modulated leads to amelioration of other abnormal disorder. biochemical processes. A lipoprotein-associated phospholi 30 The terms “drug”, “therapeutic agent, and “chemothera pase A2-mediated disorder may be completely or partially peutic agent” refer to a compound, or a pharmaceutical com mediated by modulating lipoprotein-associated phospholi position thereof, which is administered to a subject for treat pase A2 activity. In particular, a lipoprotein-associated phos ing, preventing, or ameliorating one or more symptoms of a pholipase A2-mediated disorder is one in which inhibiting disorder. lipoprotein-associated phospholipase A2 activity results in 35 The term “release controlling excipient” refers to an Some effect on the underlying disorder e.g., administration of excipient whose primary function is to modify the duration or a lipoprotein-associated phospholipase A2 inhibitor results in place of release of the active Substance from a dosage form as Some improvement in at least some of the patients being compared with a conventional immediate release dosage treated. form. The term “lipoprotein-associated phospholipase A2 inhibi 40 The term “nonrelease controlling excipient” refers to an tor, refers to the ability of a compound disclosed herein to excipient whose primary function do not include modifying alter the function of lipoprotein-associated phospholipase the duration or place of release of the active substance from a A2. Alipoprotein-associated phospholipase A2 inhibitor may dosage form as compared with a conventional immediate block or reduce the activity of lipoprotein-associated phos release dosage form. pholipase A2 by forming a reversible or irreversible covalent 45 The term “prodrug” refers to a compound functional bond between the inhibitor and lipoprotein-associated phos derivative of the compound as disclosed herein and is readily pholipase A2 or through formation of a noncovalently bound convertible into the parent compound in vivo. Prodrugs are complex. Such inhibition may be manifest only in particular often useful because, in Some situations, they may be easier to cell types or may be contingent on a particular biological administer than the parent compound. They may, for instance, event. The term "lipoprotein-associated phospholipase A2 50 be bioavailable by oral administration whereas the parent inhibitor, also refers to altering the function of lipoprotein compound is not. The prodrug may also have enhanced solu associated phospholipase A2 by decreasing the probability bility in pharmaceutical compositions over the parent com that a complex forms between lipoprotein-associated phos pound. A prodrug may be converted into the parent drug by pholipase A2 and a natural Substrate. In some embodiments, various mechanisms, including enzymatic processes and inhibiting lipoprotein-associated phospholipase A2 activity 55 metabolic hydrolysis. See Harper, Progress in Drug Research may be assessed using the methods described in Mohleret al., 1962, 4, 221–294; Morozowich et al. in “Design of Biophar J. Am. Coll. Cardiol 2008, 51 (17), 1632–41; Wilensky et al., maceutical Properties through Prodrugs and Analogs. Roche Nature Medicine 2008, 14(10), 1059-66; and WO 01/60805. Ed., APHA Acad. Pharm. Sci. 1977: “Bioreversible Carriers The term “inhibiting lipoprotein-associated phospholipase in Drug in Drug Design, Theory and Application. Roche Ed., A2 activity” or “inhibition of lipoprotein-associated phos 60 APHA Acad. Pharm. Sci. 1987: “Design of Prodrugs.” Bund pholipase A2 activity” refers to altering the function of lipo gaard, Elsevier, 1985; Wang et al., Curr: Pharm. Design 1999, protein-associated phospholipase A2 activity by administer 5,265-287: Pauletti et al., Adv. Drug. Delivery Rev. 1997,27, ing a lipoprotein-associated phospholipase A2 inhibitor. 235-256; Mizen et al., Pharm. Biotech. 1998, 11, 345-365; The term “therapeutically acceptable' refers to those com Gaignault et al., Pract. Med. Chem. 1996, 671-696; pounds (or salts, prodrugs, tautomers, Zwitterionic forms, 65 Asgharnejad in “Transport Processes in Pharmaceutical Sys etc.) which are suitable for use in contact with the tissues of tems. Amidon et al., Ed., Marcell Dekker, 185-218, 2000; patients without excessive toxicity, irritation, allergic Balant et al., Eur: J. Drug Metab. Pharmacokinet. 1990, 15, US 8,637,524 B2 9 10 143-53; Balimane and Sinko, Adv. Drug Delivery Rev. 1999, glucamine, hydrabamine, 1H-imidazole, L-lysine, morpho 39, 183-209; Browne, Clin. Neuropharmacol. 1997, 20, 1-12; line, 4-(2-hydroxyethyl)-morpholine, methylamine, piperi Bundgaard, Arch. Pharm. Chem. 1979, 86, 1-39: Bundgaard, dine, piperazine, propylamine, pyrrolidine, 1-(2- Controlled Drug Delivery 1987, 17, 179–96: Bundgaard, Adv. hydroxyethyl)-pyrrolidine, pyridine, quinuclidine, quinoline, Drug Delivery Rev. 1992, 8, 1-38; Fleisher et al., Adv. Drug isoquinoline, secondary amines, triethanolamine, trimethy Delivery Rev. 1996, 19, 115-130; Fleisher et al., Methods lamine, triethylamine, N-methyl-D-glucamine, 2-amino-2- Enzymol. 1985, 112,360-381: Farquhar et al., J. Pharm. Sci. (hydroxymethyl)-1,3-propanediol, and tromethamine. 1983, 72, 324-325; Freeman et al., J. Chem. Soc., Chem. While it may be possible for the compounds of the subject Commun. 1991, 875-877: Friis and Bundgaard, Eur: J. invention to be administered as the raw chemical, it is also Pharm. Sci. 1996, 4, 49-59; Gangwar et al., Des. Biopharm. 10 possible to present them as a pharmaceutical composition. Prop. Prodrugs Analogs, 1977, 409–421; Nathwani and Accordingly, provided herein are pharmaceutical composi Wood, Drugs 1993, 45,866-94: Sinhababu and Thakker, Adv. tions which comprise one or more of certain compounds Drug Delivery Rev. 1996, 19, 241-273; Stella et al., Drugs disclosed herein, or one or more pharmaceutically acceptable 1985, 29, 455-73; Tan et al., Adv. Drug Delivery Rev. 1999, salts, prodrugs, or Solvates thereof, together with one or more 39, 117-151; Taylor, Adv. Drug Delivery Rev. 1996, 19, 131 15 pharmaceutically acceptable carriers thereof and optionally 148; Valentino and Borchardt, Drug Discovery Today 1997.2, one or more other therapeutic ingredients. Properformulation 148-155; Wiebe and Knaus, Adv. Drug Delivery Rev. 1999, is dependent upon the route of administration chosen. Any of 39, 63-80; Waller et al., Br. J. Clin. Pharmac. 1989, 28, the well-known techniques, carriers, and excipients may be 497-507. used as Suitable and as understood in the art; e.g., in Reming The compounds disclosed herein can exist as therapeuti ton’s Pharmaceutical Sciences. The pharmaceutical compo cally acceptable salts. The term “pharmaceutically accept sitions disclosed herein may be manufactured in any manner able salt”, as used herein, represents salts or Zwitterionic known in the art, e.g., by means of conventional mixing, forms of the compounds disclosed herein which are therapeu dissolving, granulating, dragee-making, levigating, emulsi tically acceptable as defined herein. The salts can be prepared fying, encapsulating, entrapping or compression processes. during the final isolation and purification of the compounds or 25 The pharmaceutical compositions may also be formulated as separately by reacting the appropriate compound with a Suit a modified release dosage form, including delayed-, able acid or base. Therapeutically acceptable salts include extended-, prolonged-, Sustained-, pulsatile-, controlled acid and basic addition salts. For a more complete discussion accelerated- and fast-, targeted-, programmed-release, and of the preparation and selection of salts, refer to “Handbook gastric retention dosage forms. These dosage forms can be of Pharmaceutical Salts, Properties, and Use.” Stah and Wer 30 prepared according to conventional methods and techniques muth, Ed., (Wiley-VCH and VHCA, Zurich, 2002) and Berge known to those skilled in the art (see, Remington. The Science et al., J. Pharm. Sci. 1977, 66, 1-19. and Practice of Pharmacy, supra; Modified-Release Drug Suitable acids for use in the preparation of pharmaceuti Deliver Technology, Rathbone et al., Eds. Drugs and the cally acceptable salts include, but are not limited to, acetic Pharmaceutical Science, Marcel Dekker, Inc.: New York, acid, 2,2-dichloroacetic acid, acylated amino acids, adipic 35 N.Y., 2002; Vol. 126). acid, alginic acid, ascorbic acid, L-aspartic acid, benzene The compositions include those suitable for oral, Sulfonic acid, benzoic acid, 4-acetamidobenzoic acid, boric parenteral (including Subcutaneous, intradermal, intramuscu acid, (+)-camphoric acid, camphorsulfonic acid, (+)-(1S)- lar, intravenous, intraarticular, and intramedullary), intraperi camphor-10-Sulfonic acid, capric acid, caproic acid, caprylic toneal, transmucosal, transdermal, rectal and topical (includ acid, cinnamic acid, citric acid, cyclamic acid, cyclohexane 40 ing dermal, buccal, Sublingual and intraocular) Sulfamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic administration although the most Suitable route may depend acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, upon for example the condition and disorder of the recipient. formic acid, fumaric acid, galactaric acid, gentisic acid, glu The compositions may conveniently be presented in unit dos coheptonic acid, D-gluconic acid, D-glucuronic acid, age form and may be prepared by any of the methods well L-glutamic acid, C-OXO-glutaric acid, glycolic acid, hippuric 45 known in the art of pharmacy. Typically, these methods acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, include the step of bringing into association a compound of (+)-L-lactic acid, (t)-DL-lactic acid, lactobionic acid, lauric the Subject invention or a pharmaceutically salt, prodrug, or acid, maleic acid, (-)-L-malic acid, malonic acid, (+)-DL solvate thereof (“active ingredient') with the carrier which mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic constitutes one or more accessory ingredients. In general, the acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naph 50 compositions are prepared by uniformly and intimately thoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, bringing into association the active ingredient with liquid oxalic acid, palmitic acid, pamoic acid, perchloric acid, phos carriers or finely divided solid carriers or both and then, if phoric acid, L-pyroglutamic acid, Saccharic acid, salicylic necessary, shaping the product into the desired formulation. acid, 4-amino-salicylic acid, sebacic acid, Stearic acid, suc Formulations of the compounds disclosed herein suitable cinic acid, Sulfuric acid, tannic acid, (+)-L-tartaric acid, thio 55 for oral administration may be presented as discrete units cyanic acid, p-toluenesulfonic acid, undecylenic acid, and Such as capsules, cachets or tablets each containing a prede Valeric acid. termined amount of the active ingredient, as a powder or Suitable bases for use in the preparation of pharmaceuti granules; as a solution or a suspension in an aqueous liquid or cally acceptable salts, including, but not limited to, inorganic a non-aqueous liquid; or as an oil-in-water liquid emulsion or bases, such as magnesium hydroxide, calcium hydroxide, 60 a water-in-oil liquid emulsion. The active ingredient may also potassium hydroxide, Zinc hydroxide, or sodium hydroxide; be presented as a bolus, electuary or paste. and organic bases, such as primary, secondary, tertiary, and Pharmaceutical preparations which can be used orally quaternary, aliphatic and aromatic amines, including L-argi include tablets, push-fit capsules made of gelatin, as well as nine, benethamine, benzathine, choline, deanol, diethanola soft, sealed capsules made of gelatin and a plasticizer, Such as mine, diethylamine, dimethylamine, dipropylamine, diiso 65 glycerol or sorbitol. Tablets may be made by compression or propylamine, 2-(diethylamino)-ethanol, ethanolamine, molding, optionally with one or more accessory ingredients. ethylamine, ethylenediamine, isopropylamine, N-methyl Compressed tablets may be prepared by compressing in a US 8,637,524 B2 11 12 Suitable machine the active ingredient in a free-flowing form formulated in conventional manner. Such compositions may Such as a powder or granules, optionally mixed with binders, comprise the active ingredient in a flavored basis such as inert diluents, or lubricating, Surface active or dispersing Sucrose and acacia or tragacanth. agents. Molded tablets may be made by molding in a Suitable The compounds may also be formulated in rectal compo machine a mixture of the powdered compound moistened 5 sitions such as Suppositories or retention enemas, e.g., con with an inert liquid diluent. The tablets may optionally be taining conventional Suppository bases such as cocoa butter, coated or scored and may beformulated so as to provide slow polyethylene glycol, or other glycerides. or controlled release of the active ingredient therein. All for Certain compounds disclosed herein may be administered mulations for oral administration should be in dosages Suit topically, that is by non-systemic administration. This able for Such administration. The push-fit capsules can con 10 tain the active ingredients in admixture with filler such as includes the application of a compound disclosed herein lactose, binders such as starches, and/or lubricants such as externally to the epidermis or the buccal cavity and the instil talc or magnesium Stearate and, optionally, stabilizers. In soft lation of such a compound into the ear, eye and nose, such that capsules, the active compounds may be dissolved or Sus the compound does not significantly enter the blood stream. pended in Suitable liquids, such as fatty oils, liquid paraffin, or 15 In contrast, Systemic administration refers to oral, intrave liquid polyethylene glycols. In addition, stabilizers may be nous, intraperitoneal and intramuscular administration. added. Dragee cores are provided with Suitable coatings. For Formulations suitable for topical administration include this purpose, concentrated Sugar Solutions may be used, liquid or semi-liquid preparations Suitable for penetration which may optionally contain gum arabic, talc, polyvinyl through the skin to the site of inflammation Such as gels, pyrrolidone, carbopol gel, polyethylene glycol, and/or tita liniments, lotions, creams, ointments or pastes, and drops nium dioxide, lacquer Solutions, and Suitable organic solvents Suitable for administration to the eye, ear or nose. or solvent mixtures. Dyestuffs or pigments may be added to For administration by inhalation, compounds may be the tablets or dragee coatings for identification or to charac delivered from an insufflator, nebulizer pressurized packs or terize different combinations of active compound doses. other convenient means of delivering an aerosol spray. Pres The compounds may be formulated for parenteral admin 25 Surized packs may comprise a suitable propellant Such as istration by injection, e.g., by bolus injection or continuous dichlorodifluoromethane, trichlorofluoromethane, dichlo infusion. Formulations for injection may be presented in unit rotetrafluoroethane, carbon dioxide or other suitable gas. In dosage form, e.g., in ampoules or in multi-dose containers, the case of a pressurized aerosol, the dosage unit may be with an added preservative. The compositions may take Such determined by providing a valve to deliver a metered amount. forms as Suspensions, Solutions or emulsions in oily or aque 30 Alternatively, for administration by inhalation or insufflation, ous vehicles, and may contain formulatory agents such as the compounds according to the invention may take the form suspending, stabilizing and/or dispersing agents. The formu of a dry powder composition, for example a powder mix of the lations may be presented in unit-dose or multi-dose contain compound and a suitable powder base such as lactose or ers, for example sealed ampoules and vials, and may be stored starch. The powder composition may be presented in unit in powder form or in a freeze-dried (lyophilized) condition 35 dosage form, in for example, capsules, cartridges, gelatin or requiring only the addition of the sterile liquid carrier, for blister packs from which the powder may be administered example, saline or sterile pyrogen-free water, immediately with the aid of an inhalator or insufflator. prior to use. Extemporaneous injection solutions and Suspen Preferred unit dosage formulations are those containing an sions may be prepared from Sterile powders, granules and effective dose, as herein below recited, or an appropriate tablets of the kind previously described. 40 fraction thereof, of the active ingredient. Formulations for parenteral administration include aque Compounds may be administered orally or via injection at ous and non-aqueous (oily) sterile injection solutions of the a dose of from 0.1 to 500 mg/kg per day. The dose range for active compounds which may contain antioxidants, buffers, adult humans is generally from 5 mg to 2 g/day. Tablets or bacteriostats and solutes which render the formulation iso other forms of presentation provided in discrete units may tonic with the blood of the intended recipient; and aqueous 45 conveniently contain an amount of one or more compounds and non-aqueous sterile Suspensions which may include Sus which is effective at Such dosage or as a multiple of the same, pending agents and thickening agents. Suitable lipophilic for instance, units containing 5 mg to 500 mg, usually around Solvents or vehicles include fatty oils such as sesame oil, or 10 mg to 200 mg. synthetic fatty acid esters, such as ethyl oleate or triglycer The amount of active ingredient that may be combined ides, or liposomes. Aqueous injection Suspensions may con 50 with the carrier materials to produce a single dosage form will tain Substances which increase the Viscosity of the Suspen vary depending upon the host treated and the particular mode Sion, such as Sodium carboxymethyl cellulose, Sorbitol, or of administration. dextran. Optionally, the Suspension may also contain Suitable The compounds can be administered in various modes, e.g. stabilizers or agents which increase the solubility of the com orally, topically, or by injection. The precise amount of com pounds to allow for the preparation of highly concentrated 55 pound administered to a patient will be the responsibility of Solutions. the attendant physician. The specific dose level for any par In addition to the formulations described previously, the ticular patient will depend upon a variety of factors including compounds may also be formulated as a depot preparation. the activity of the specific compound employed, the age, body Such long acting formulations may be administered by weight, general health, sex, diets, time of administration, implantation (for example Subcutaneously or intramuscu 60 route of administration, rate of excretion, drug combination, larly) or by intramuscular injection. Thus, for example, the the precise disorder being treated, and the severity of the compounds may be formulated with Suitable polymeric or disorder being treated. Also, the route of administration may hydrophobic materials (for example as an emulsion in an vary depending on the disorder and its severity. acceptable oil) or ion exchange resins, or as sparingly soluble In the case wherein the patient’s condition does not derivatives, for example, as a sparingly soluble salt. 65 improve, upon the doctor's discretion the administration of For buccal or Sublingual administration, the compositions the compounds may be administered chronically, that is, for may take the form of tablets, lozenges, pastilles, or gels an extended period of time, including throughout the duration US 8,637,524 B2 13 14 of the patient’s life in order to ameliorate or otherwise control described by Li et al. Rapid Communications in Mass Spec or limit the symptoms of the patient’s disorder. trometry 2005, 19, 1943-1950; and any references cited In the case wherein the patient’s status does improve, upon therein and any modifications made thereof. the doctor's discretion the administration of the compounds Examples of cytochrome Paso isoforms in a mammalian may be given continuously or temporarily Suspended for a subject include, but are not limited to, CYP1A1, CYP1A2, certain length of time (i.e., a "drug holiday). CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, Once improvement of the patient’s conditions has CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, occurred, a maintenance dose is administered if necessary. CYP2G1 CYP2J2, CYP2R1, CYP2S1, CYP3A4, CYP3A5, Subsequently, the dosage or the frequency of administration, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11, CYP4B1, or both, can be reduced, as a function of the symptoms, to a 10 level at which the improved disorder is retained. Patients can, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms. CYP17, CYP19, CYP21, CYP24, CYP26A1, CYP26B1, Disclosed herein are methods of treating a lipoprotein CYP27A1, CYP27B1, CYP39, CYP46, and CYP51. associated phospholipase A2-mediated disorder comprising 15 Examples of monoamine oxidase isoforms in a mamma administering to a subject having or Suspected of having Such lian subject include, but are not limited to, MAO, and a disorder, a therapeutically effective amount of a compound MAO. as disclosed herein or a pharmaceutically acceptable salt, The inhibition of the cytochrome Paso isoform is measured Solvate, or prodrug thereof. by the method of Ko et al., British Journal of Clinical Phar Lipoprotein-associated phospholipase A2-mediated disor macology 2000, 49, 343-351. The inhibition of the MAO ders, include, but are not limited to, coronary disorders, pri isoform is measured by the method of Weyler et al., J. Biol. mary and secondary coronary disorders, acute coronary dis Chem. 1985, 260, 13199-13207. The inhibition of the MAO orders, atherosclerosis, peripheral vascular atherosclerosis, isoform is measured by the method of Uebelhacket al., Phar cerebrovascular atherosclerosis, rheumatoid arthritis, diabe macopsychiatry 1998, 31, 187-192. tes, stroke, myocardial infarction, reperfusion injury, acute 25 Examples of polymorphically-expressed cytochrome Paso and chronic inflammation, psoriasis, and asthma, and/or any isoforms in a mammalian Subject include, but are not limited disorder which can lessened, alleviated, or prevented by to, CYP2C8, CYP2C9, CYP2C19, and CYP2D6. administering a lipoprotein-associated phospholipase A2 The metabolic activities of liver microsomes, cytochrome inhibitor. Paso isoforms, and monoamine oxidase isoforms are mea In certain embodiments, a method of treating a lipoprotein 30 sured by the methods described herein. associated phospholipase A2-mediated disorder comprises Examples of improved disorder-control and/or disorder administering to the subject a therapeutically effective eradication endpoints, or improved clinical effects include, amount of a compound as disclosed herein, or a pharmaceu but are not limited to, reduced heart attacks and/or strokes, tically acceptable salt, Solvate, or prodrug thereof. So as to reduced coronary lesion development, and Stabilization of affect: (1) decreased inter-individual variation in plasma lev 35 atherosclerotic plaques. els of the compound or a metabolite thereof; (2) increased Examples of diagnostic hepatobiliary function endpoints average plasma levels of the compound or decreased average include, but are not limited to, alanine aminotransferase plasma levels of at least one metabolite of the compound per (ALT), serum glutamic-pyruvic transaminase (“SGPT), dosage unit; (3) decreased inhibition of and/or metabolism aspartate aminotransferase (AST or “SGOT), ALT/AST by at least one cytochrome Paso or monoamine oxidase iso 40 ratios, serum aldolase, alkaline phosphatase (ALP), ammo form in the Subject; (4) decreased metabolism via at least one nia levels, bilirubin, gamma-glutamyl transpeptidase polymorphically-expressed cytochrome Paso isoform in the (“GGTP” “Y-GTP or “GGT), leucine aminopeptidase Subject; (5) at least one statistically-significantly improved (“LAP), liver biopsy, liver ultrasonography, liver nuclear disorder-control and/or disorder-eradication endpoint; (6) an scan, 5'-nucleotidase, and blood protein. Hepatobiliary end improved clinical effect during the treatment of the disorder, 45 points are compared to the stated normal levels as given in (7) prevention of recurrence, or delay of decline or appear “Diagnostic and Laboratory Test Reference', 4' edition, ance, of abnormal alimentary or hepatic parameters as the Mosby, 1999. These assays are run by accredited laboratories primary clinical benefit, or (8) reduction or elimination of according to standard protocol. deleterious changes in any diagnostic hepatobiliary function Besides being useful for human treatment, certain com endpoints, as compared to the corresponding non-isotopi 50 pounds and formulations disclosed herein may also be useful cally enriched compound. for veterinary treatment of companion animals, exotic ani In certain embodiments, inter-individual variation in mals and farm animals, including mammals, rodents, and the plasma levels of the compounds as disclosed herein, or like. More preferred animals include horses, dogs, and cats. metabolites thereof, is decreased; average plasma levels of Combination Therapy the compound as disclosed herein are increased; average 55 The compounds disclosed herein may also be combined or plasma levels of a metabolite of the compound as disclosed used in combination with other agents useful in the treatment herein are decreased; inhibition of a cytochrome Paso or of lipoprotein-associated phospholipase A2-mediated disor monoamine oxidase isoform by a compound as disclosed ders. Or, by way of example only, the therapeutic effective herein is decreased; or metabolism of the compound as dis ness of one of the compounds described herein may be closed herein by at least one polymorphically-expressed 60 enhanced by administration of an adjuvant (i.e., by itself the cytochrome Paso isoform is decreased; by greater than about adjuvant may only have minimal therapeutic benefit, but in 5%, greater than about 10%, greater than about 20%, greater combination with another therapeutic agent, the overall thera than about 30%, greater than about 40%, or by greater than peutic benefit to the patient is enhanced). about 50% as compared to the corresponding non-isotopi Such other agents, adjuvants, or drugs, may be adminis cally enriched compound. 65 tered, by a route and in an amount commonly used therefor, Plasma levels of the compound as disclosed herein, or simultaneously or sequentially with a compound as disclosed metabolites thereof, may be measured using the methods herein. When a compound as disclosed herein is used con US 8,637,524 B2 15 16 temporaneously with one or more other drugs, a pharmaceu melagatran, Ximelagatran, bivalirudin, dabigatran etexilate, tical composition containing Such other drugs in addition to defibrotide, dermatan Sulfate, fondaparinux, and rivaroXaban. the compound disclosed herein may be utilized, but is not In certain embodiments, the compounds provided herein required. can be combined with one or more anti-platelet agents, In certain embodiments, the compounds disclosed herein including, but not limited to, abciximab, eptifibatide, can be combined with one or more alpha adrenergic receptor tirofiban, clopidogrel, praSugrel, ticlopidine, ticagrelor, bera antagonists, Angiotensin II receptor antagonists, angiotensin prost, prostacyclin, iloprost, treprostinil, acetylsalicylic acid, converting enzyme inhibitors, anti-arrhythmics, anti-throm aloxiprin, carbasalate calcium, indobufen, dipyridamole, botics, anti-platelet agents, beta adrenergic receptor antago picotamide, terutroban, cilostazol, dipyridamole, triflusal, nists, calcium channel blockers, fibrates, and HMG-CoA 10 cloricromen, and ditazole. reductase inhibitors. In certain embodiments, the compounds disclosed herein In certain embodiments, the compounds disclosed herein can be combined with one or more beta-adrenergic antago can be combined with one or more alpha adrenergic receptor nists, including, but not limited to, acebutolol, adaprolol. antagonists known in the art, including, but not limited to, adimolol, afurolol, alprenolol, alprenoXime, amoSulalol, abanoquil, adimolol, ajmalicine, alfuZosin, amoSulalol, aro 15 ancarolol, arnolol, arotinolol, atenolol, befunolol, betaxolol. tinolol, atiprosin, benoxathian, buflomedil, bunaZosin, bevantolol, bisoprolol, bopindolol, bormetolol, bornaprolol, carvedilol, CI-926, corynanthine, dapiprazole, DL-017, brefonalol, bucindolol, bucumolol, bufetolol, buftiralol, domesticine, doxazosin, eugenodilol, fenspiride, GYKI-12, bufuralol, bunitrolol, bunolol, bupranolol, burocrolol, butax 743, GYKI-16,084, indoramin, ketanserin, L-765,314, labe amine, butidrine, butofilolol, capsinolol, carazolol, carpin talol, mephendioxan, metazosin, monatepil, moxisylyte (thy dolol, carteolol, carvedilol, celiprolol, cetamolol, cicloprolol. moxamine), naftopidil, nantenine, neldaZosin, nicergoline, cinamolol, cloranolol, cyanopindolol, dalbraminol, dexpro niguldipine, pelanserin, phendioxan, phenoxybenzamine, pranolol, diacetolol, dichloroisoprenaline, dihydroalpre phentolamine, piperoxan, prazosin, quinaZosin, ritanserin, nolol, dilevalol, diprafenone, draquinolol, dropranolol, ecas RS-97.078, SGB-1,534, silodosin, SL-89.0591, spiperone, tolol, epanolol, ericolol, ersentilide, esatenolol, esmolol. talipexole, tamsulosin, teraZosin, tibalosin, tiodaZosin, tipen 25 esprolol, eugenodilol, exaprolol, fallintolol, flestolol, flusox tosin, tolazoline, trimaZosin, upidosin, urapidil. Zolertine, olol, hydroxycarteolol, hydroxytertatolol, ICI-118.551, idro 1-PP. adimolol, atipamezole, BRL-44408, buflomedil, cira pranolol, indenolol, indopanolol, iodocyanopindolol, ipro Zoline, efaroxan, esmirtazapine, fluparoxan, GYKI-12,743, crolol, isoxaprolol, isamoltane, labetalol, landiolol. GYKI-16,084, idazoxan, mianserin, mirtazapine, MK-912, levobetaxolol, levobunolol, levocicloprolol, levomoprolol, NAN-190, olanzapine, phentolamine, phenoxybenzamine, 30 medroxalol, mepindolol, metalol, metipranolol, metoprolol. piperoxan, piribedil, rauwolscine, rotigotine, SB-269,970, moprolol, nadolol, nadoxolol, nafetolol, nebivolol, nerami setiptiline, spiroXatrine, Sunepitron, tolazoline, and yohim nol, nifenalol, nipradillol, oberadillol, Oxprenolol, pacrinolol, bine. pafenolol, pamatolol, pargolol, parodilol, penbutolol. In certain embodiments, the compounds disclosed herein penirolol, PhOA-33, pindolol, pirepolol, practolol, primi can be combined with one or more Angiotensin II receptor 35 dolol, procinolol, pronethalol, propafenone, propranolol. antagonists (AIIRA), including, but not limited to, cande ridazolol, ronactolol, Soquinolol, Sotalol, spirendolol, SR Sartan, eprosartan, irbesartan, losartan, olmesartan, tasosar 59230A, sulfinalol, TA-2005, talinolol, tazolol, teoprolol, ter tan, telmisartan, and Valsartan. tatolol, terthianolol, tienoxolol, tilisolol, timolol, tiprenolol, In certain embodiments, the compounds disclosed herein tolamolol, toliprolol, tribendilol, trigevolol. xibenolol, and can be combined with one or more angiotensin-converting 40 Xipranolol. enzyme inhibitors (ACE inhibitors), including, but not lim In certain embodiments, the compounds disclosed herein ited to, captopril, enalapril, lisinopril, perindopril, ramipril, can be combined with one or more calcium channel blockers, quinapril, benazepril, cilaZapril, fosinopril, trandolapril, spi including, but not limited to amlodipine, felodipine, israd rapril, delapril, moexipril, temocapril. Zofenopril, and imi ipine, nicardipine, nifedipine, nimodipine, nisoldipine, dapril. 45 nitrendipine, lacidipine, nilvadipine, manidipine, barnid In certain embodiments, the compounds disclosed herein ipine, lercanidipine, cilnidipine, benidipine, mibefradil, Vera can be combined with one or more anti-arrhythmics, includ pamil, gallopamil, diltiazem, fendiline, bepridil, lidoflazine, ing, but not limited to, quinidine, procainamide, disopyra and perhexyline. mide, Sparteine, ajmaline, prajmaline, lorajmine, lidocaine, In certain embodiments, the compounds provided herein mexiletine, tocamide, aprindine, propafenone, flecamide, 50 can be combined with one or more fibrates, including, but not lorcainide, encainide, amiodarone, bretylium tosilate, bunaf limited to, clofibrate, bezafibrate, aluminium clofibrate, gem tine, dolfetilide, ibutilidem, tedisamil, moracizine, and ciben fibrozil, fenofibrate, simfibrate, ronifibrate, ciprofibrate, Zoline. etofibrate, and clofibride. In certain embodiments, the compounds provided herein In certain embodiments, the compounds disclosed herein can be combined with one or more anti-thrombotics, includ 55 can be combined with one or more HMG-CoA reductase ing, but not limited to, dicoumarol, phenindione, warfarin, inhibitors (statins), including, but not limited to, atorvastatin, phenprocoumon, acenocoumarol, ethyl biscoumacetate, clo cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, rindione, diphenadione, tioclomarol, heparin, antithrombin pravastatin, rosuvastatin, and simvastatin. III, dalteparin, enoxaparin, nadroparin, parnaparin, reviparin, The compounds disclosed herein can also be administered danaparoid, tinzaparin, Sulodexide, bemiparin, ditazole, clo 60 in combination with other classes of compounds, including, ricromen, picotamide, clopidogrel, ticlopidine, acetylsali but not limited to, norepinephrine reuptake inhibitors (NRIs) cylic acid, dipyridamole, carbasalate calcium, epoprostenol, Such as atomoxetine; dopamine reuptake inhibitors (DARIs), indobufen, iloprost, abciximab, aloxiprin, eptifibatide, Such as methylphenidate; serotonin-norepinephrine reuptake tirofiban, triflusal, beraprost, treprostinil, prasugrel, Strep inhibitors (SNRIs), such as milnacipran; sedatives, such as tokinase, alteplase, urokinase, fibrinolysin, brinase, 65 diazepham; norepinephrine-dopamine reuptake inhibitor reteplase, Saruplase, ancrod, drotrecogin alfa (activated), (NDRIs), Such as bupropion; serotonin-norepinephrine tenecteplase, protein C, desirudin, lepirudin, argatroban, dopamine-reuptake-inhibitors (SNDRIs), such as Venlafax US 8,637,524 B2 17 18 ine; monoamine oxidase inhibitors, such as selegiline; hypo agents, such as pacitaxel, docetaxel, and epothilones A-F. thalamic phospholipids; endothelin converting enzyme plant-derived products, such as Vinca alkaloids, epipodophyl (ECE) inhibitors, such as phosphoramidon; opioids. Such as lotoxins, and taxanes; topoisomerase inhibitors; prenyl-pro tramadol; thromboxane receptor antagonists, such as tein transferase inhibitors; cyclosporins; steroids, such as ifetroban; potassium channel openers; thrombin inhibitors, prednisone and dexamethasone; cytotoxic drugs, such as aza Such as hirudin; hypothalamic phospholipids; growth factor thiprine and cyclophosphamide; TNF-alpha inhibitors, such inhibitors, such as modulators of PDGF activity; platelet acti as tenidap; anti-TNF antibodies or soluble TNF receptor, such Vating factor (PAF) antagonists; anticoagulants, such as war as etanercept, rapamycin, and leflunimide; and cyclooxyge farin; low molecular- 0 weight heparins, such- 0 as enoxaparin;'''". 10 nase-2 (COX-2) inhibitors, such as celecoxib and rofecoxib: Factor VIIa Inhibitors and Factor Xa Inhibitors; renin inhibi- and miscellaneous agents such as, hydroxyurea, procarba tors; neutral endopeptidase (NEP) inhibitors; vasopepsidase Zine. mitotane. hexamethylmelamine gold compounds, plati inhibitors (dual NEP-ACE inhibitors), such as omapatrilat s r sy re. and gemopatrilat, squalene synthetase inhibitors;- - - - - bile acid num coordination complexes, such as cisplatin, satraplatin, sequestrants, such as questran; niacin; anti-atherosclerotic and carbop latin. agents, such as ACAT inhibitors; MTP Inhibitors; potassium Thus, in another aspect, certa1 embodiments provide channel activators; alpha-muscarinic agents; beta-muscarinic methods for treating lipoprotein-associated phospholipase or re A2-mediated disorders in a human or animal Subject in need agents. Such as carvedilol and metoprolol, diuretics, such as of such treatment comprising administering to said Subject an chlorothlazide, hydrochlorothiazide, flumethiazide, hydrof- 20 amount of a compound disclosed herein effective to reduce or lumethiazide, bendroflumethiazide, methylchlorothiazide, prevent said disorder in the subject, in combination with at trichloromethiazide, polythiazide, benzothlazide, ethacrynic least one additional agent for the treatment of said disorder acid, tricrynafen, chlorthalidone, furosenilde, musolimine, that is known in the art. In a related aspect, certain embodi bumetanide, triamterene, amiloride, and Spironolactone; ments provide therapeutic compositions comprising at least thrombolytic agents, such as tissue plasminogen activator 25 compound disclosed herein in combination with Ole O (tPA), recombinant tEPA, Streptokinase, urokinase, prouroki- more additional agents for the treatment of lipoprotein-asso nase, and anisoylated plasminogen Streptokinase activator in E"S.S.FFRReaponds complex (APSAC). anti-diabetic agents, Such as biguanides Isotopic hydrogen can be introduced into a compound as (e.g. metformin), glucosidase inhibitors (e.g., acarbose), so disclosed herein by synthetic techniques that employ deuter insulins, meglitinides (e.g., repaglinide), Sulfonylureas (e.g., ated reagents, whereby incorporation rates are pre-deter glimepiride, glyburide, and glipizide), thioZolidinediones mined; and/or by exchange techniques, wherein incorpora (e.g. troglitaZone, rosiglitaZone and pioglitaZone), and tion rates are determined by equilibrium conditions, and may PPAR-gamma agonists; mineralocorticoid receptor antago- be highly variable depending on the reaction conditions. Syn nists, such as Spironolactone and eplerenone; growth hor- 35 thetic techniques, where tritium or deuterium is directly and mone secretagogues; aP2 inhibitors; phosphodiesterase specifically inserted by tritiated or deuterated reagents of inhibitors, such as PDE III inhibitors (e.g., cilostazol) and known isotopic content, may yield high tritium O deuterium PDE V inhibitors (e.g., sildenafil, tadalafil. Vardenafil); pro- ENNA",R. thin'...list tein tyrosine kinase inhibitors; antiinflammatories: antipro- 40 tritium or deuterium incorporation, often with the isotope liferatives, such as methotrexate, FK506 (tacrolimus, Pro- being distributed over many sites on the molecule. graf), mycophenolate mofetil: chemotherapeutic agents; The compounds as disclosed herein can be prepared by immunosuppressants; anticancer agents and cytotoxic agents methods known to one of skill in the art and routine modifi (e.g., alkylating agents. Such as nitrogen mustards, alkyl Sul- cations thereof, and/or following procedures similar to those fonates, nitrosoureas, ethylenimines, and triazenes); antime- 45 described herein and routine modifications thereof, and/or tabolites, such as folate antagonists, purine analogues, and procedures found in Blackie et al., Bioorg. Med. Chem. Lett. pyrridine analogues; antibiotics, such as anthracyclines, bleo- 2003, 13, 1067-70, WO 03/016287; and WO 01/60805, mycins, mitomycin, dactinomycin, and plicamycin; which are hereby incorporated 1. their entirety, and refer enzymes, such as L-asparaginase; farnesyl-protein trans- ences cited therein and routine modifications thereof. Com ferase inhibitors; hormonal agents, such as glucocorticoids 50 runs its listlsE. (e.g., cortisone), estrogens/antiestrogens, androgens/antian- thereof. drogens, progestins, and luteinizing hormone-releasing hor- The following schemes can be used to practice the present O anatagonists, and octreotide acetate; microtubule-dis- invention. Any position shown as hydrogen may optionally be ruptor agents, such as ecteinascidins; microtubule-stablizing replaced with deuterium.

Scheme I O R32 R3. R27 Ras Rs. R32 R27 Rs R31.R.32 1no R30 to-X-X-so -e- N-X-X---> -- O R R29 O R29 R30 O R29 R30 ..'s 1 2

US 8,637,524 B2 21 22 Compound 1 is reacted with an appropriate alcohol. Such as ing deuterium Substitutions can be used. To introduce deute ethanol, in the presence an appropriate acid, such as Sulfuric rium at Rs. sodium borodeuteride can be used. acid, in an appropriate solvent. Such as toluene, to give com The following compounds can generally be made using the pound 2. Compound 2 is treated with an appropriate base, methods described above. It is expected that these com Such as potassium tert-butoxide, in an appropriate solvent, pounds when made will have activity similar to those Such as toluene, to give compound 3. Compound 3 is reacted described in the examples above. with compound 4 in the presence of an appropriate base, Such as sodium ethoxide, in the presence of an appropriate catalyst, 10 Such as diazabicyclo5.4.0]undec-7ene, in an appropriate Sol vent, such as acetonitrile, to give compound 5. Compound 5 is reacted with compound 6 in the presence of an appropriate 15 base. Such as Sodium ethoxide, in an appropriate solvent, Such as ethanol, to give compound 7. Compound 7 is reacted with compound 8 in the presence of an appropriate base. Such as N,N-diisopropylethylamine, in an appropriate solvent, Such as dichloromethane, to give compound 9. Compound 9 is treated with an appropriate acid, such as trifluoroacetic acid, in an appropriate solvent. Such as dichloromethane, to give compound 10. Compound 11 is reacted with compound 12 in 25 the presence of an appropriate catalyst, such as tetrakis(triph enylphosphine)palladium(0), in the presence of an appropri ate base, such as sodium carbonate, in an appropriate solvent, 30 Such as an appropriate mixture of water and dimethoxy ethane, to give compound 13. Compound 13 is reacted with compound 14 in the presence of an appropriate dehydrating CF agent, such as 4A molecular sieves, and an appropriate reduc 35 ing agent, such as Sodium borohydride, in an appropriate Solvent. Such as dichloromethane, to give compound 15. Compound 15 is reacted with compound 10 in the presence of an appropriate coupling agent, such as a combination of 1-(3- 40 dimethylaminopropyl)-3-ethylcarbodiimide and 1-hydroxy benzotriazole hydrate, in an appropriate solvent. Such as F dichloromethane, to give a compound 16 of Formula I. 45 Deuterium can be incorporated to different positions syn O thetically, according to the synthetic procedure as shown in Scheme I, by using appropriate deuterated intermediates. For example, to introduce deuterium at one or more positions of 50 -- R7-R compound 1 with the corresponding deuterium Sub O stitutions can be used. To introduce deuterium at one or more positions of R-Rs, compound 6 with the corresponding N 55 deuterium substitutions can be used. To introduce deuterium 1n 1\-1 at one or more positions of Rs-R compound 8 with the - D D corresponding deuterium Substitutions can be used. To intro duce deuterium at one or more positions of R-Ra, com 60 D D pound 11 with the corresponding deuterium Substitutions can D D be used. To introduce deuterium at one or more positions of Re-Ro compound 12 with the corresponding deuterium D D, 65 Substitutions can be used. To introduce deuterium at one or CF more positions of R-Ra, compound 14 with the correspond US 8,637,524 B2 23 24 -continued -continued

CF 3 25 CF

30

35

40

F F D D D D 45 O O D DN D DN l

S ulN 50 S1 N O O N 1n 1-1 N 1n 1a- 55 - D D

60 D D O D D, 65 CF CF US 8,637,524 B2 25 26 -continued -continued F

10

15 l

CF 25 CF

30

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50

55

60

CF 65 CF US 8,637,524 B2 27 28 -continued -continued F

D D

5 O

D DN

- N 10

15

25 CF

30

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45

50

55

60

65 CF CF US 8,637,524 B2 29 30 -continued -continued

CF 25 CF

30

35

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F F

45 O O -- -- 50 O O D D D D DC X N 1n-N DC X N 1n-N 55 DC ul-D D D D

D D

60 D D

D D,

65 CF CF US 8,637,524 B2 31 32 -continued -continued F

S 10 X---D D ul-d D 15 DC D

D D O D

CF 25 CF

30

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50

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60

65 CF CF US 8,637,524 B2 33 34 -continued -continued

10

15

25 CF

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65 CF CF US 8,637,524 B2 35 36 -continued -continued

10

15

CF 25 CF

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65 CF CF US 8,637,524 B2 37 38 -continued -continued

10

15

CF 25 CF

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60

65 CF CF US 8,637,524 B2 39 40 -continued -continued

10

15

25 CF

30

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50

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60

65 CF CF US 8,637,524 B2 41 42 -continued -continued

10

DC N 15

D

CF 25 CF

30

35

40

F

45 O

- 50

O D D D D

55

60 D D D D

D D, 65 CF CF US 8,637,524 B2 43 44 -continued -continued

30

35

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50

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60

65 CF CF US 8,637,524 B2 45 46 -continued -continued

CF 25 CF

30

35

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F F

45 O O

D S l N 50 D S l N D D O O

1n 1\-1 N 55 1n 1\-1 N - - Null

60 D D D D

s D D, 65 CF CF

US 8,637,524 B2 57 58 -continued -continued

CF 25 CF

30

35

40

F

45 O

D -- 50 D O D D

1n N N 55 - D D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 59 60 -continued -continued

CF 25 CF

30

35

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F F

D D 45 D D O O D p-NullDN D p-NullDN S N 50 S N D D O O D D D D N N ~- XXD D 55 ~XX- DDD D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 61 62 -continued -continued

30

35

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65 US 8,637,524 B2 63 64 -continued -continued

CF 25 CF

30

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F F

D D 45 D D O O

D DN D DN

D S us N 50 D S l N D D O O -X-X-D D D D DXX. D D D

D D

60 D D D D

s D D, 65 CF CF US 8,637,524 B2 65 66 -continued -continued

CF 25 CF

30

35

40

F F

45 O O

50 D-Null D-NullS N S N D D O O D D D D D D D D N X N DC N 55 DC N

ld D DC ul Di?e D DC D D

60 D D D D

D D, 65 CF CF

US 8,637,524 B2 69 70 -continued -continued

10

15

CF 25 CF

30

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F F

45 O O

-- 50 --

D O D O D D 1n 1a- N 55 1SN1a N - u D D

D D

s D D, 65 CF CF US 8,637,524 B2 71 72 -continued -continued

10

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CF 25 CF

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65 CF CF US 8,637,524 B2 73 74 -continued -continued

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65 US 8,637,524 B2 75 76 -continued -continued

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CF 25 CF

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45 D O ”, 50 -- D O D D N 55 N1\-1 ulp D D D

60 D D D D

D D, 65 CF CF

US 8,637,524 B2 81 82 -continued -continued

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CF 25 CF

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65 CF US 8,637,524 B2 83 84 -continued -continued

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65 CF CF US 8,637,524 B2 85 86

-continued -continued

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65 US 8,637,524 B2 87 88 -continued -continued

CF 25 CF

30

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F F

D 45 D D O O " D " us 50 l

55

D D

60 D D D D

s D D, 65 CF CF US 8,637,524 B2 89 90 -continued -continued

25 CF

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65

US 8,637,524 B2 93 94 -continued -continued

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65 CF CF US 8,637,524 B2 95 96 -continued -continued

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CF 25 CF

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F

D 45 O " -- 50 D D

D N 1SN1\- 55

60

65 CF CF US 8,637,524 B2 97 98 -continued -continued

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65

US 8,637,524 B2 101 102 -continued -continued

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US 8,637,524 B2 105 106 -continued -continued

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65 US 8,637,524 B2 107 108 -continued -continued

CF 25 CF

30

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F F

D D 45 D D O O

D DN D DN

D S l N 50 D S l N D D D D O O D DD D DD -X. N 55 ~XX N - D D DDD D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 109 110 -continued -continued

30

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65 US 8,637,524 B2 111 112 -continued -continued

CF 25 CF

30

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F F

D D 45 D D O O

D DN D DN

S N 50 S N

N 55

60

65 CF CF US 8,637,524 B2 113 114 -continued -continued

CF 25 CF

30

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F F

45 O O

50 D-Null D-NullS N S N D D D D O O D D D, DD D D D, DD

X N 55 X N DC N DC N

lo D DC ld DDD D DC D D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 115 116 -continued -continued

CF 25 CF

30

35

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F F

D D 45 D D O O

D DN D DN

p-Null 50 D ul

X N N DC N 55 DC ^x up D D DC

60 D

CF CF US 8,637,524 B2 117 118 -continued -continued

10

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65 CF CF US 8,637,524 B2 119 120 -continued -continued

CF 25 CF

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15

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65 US 8,637,524 B2 123 124 -continued -continued F F

D D

5 D D DN D, S l D 10 D O D D

15 ---D D D D

2O

CF 25 CF

30

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F F

D 45 D D O O "r D " -- 50 --

O O D D D D -X---- I 55 -X---- I

60 D

65 CF CF

US 8,637,524 B2 129 130 -continued -continued

10

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CF 25 CF

30

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S N 50 S N

D D 1NN N D, 55 ~ XX N D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 131 132 -continued -continued

D

10

15

2O

CF 3 25 CF

30

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F F

D 45 D D O O " D " -- 50 --

O O D D D D D D 1n N D, 55 ~ XX N D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 133 134 -continued -continued

30

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65 US 8,637,524 B2 135 136 -continued -continued

10

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25 CF

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65 CF CF US 8,637,524 B2 137 138 -continued -continued

10

DC N D 15

D CF 25 CF

30

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F F

45 O O

-- 50 --

O O D D D D D D D D D D N N DC N D, 55 DC N D

DC ld D DC ul Di?e D 3 D D

60 D D D D

D D, 65 CF CF

US 8,637,524 B2 145 146 -continued -continued

30

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65

US 8,637,524 B2 151 152 -continued -continued F D D

O 5 D DN

D-NullS D 10

D D

DC X---- 15 D DC1 VD

CF CF 25

30

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F

D 45 O " -- 50

O D D X 1n-1 P 55 D D1N3 D

60

65 CF CF US 8,637,524 B2 153 154 -continued -continued

CF 25 CF

30

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F F

45 O O

D-NullS N 50 D S l N D D O O D D D D D D 1n N N D, 55 1n N N D - D D - DDD D

60 D D D D

D D, 65 CF CF

US 8,637,524 B2 157 158

-continued -continued

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65

US 8,637,524 B2 161 162 -continued -continued

CF 25 CF

30

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F

45 O

50 D-NullS N D O D D D D D N 55 DC N D ul di?e D DC D

60 D D D D

D D, 65 CF CF

US 8,637,524 B2 165 166 -continued -continued

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25 CF

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65 CF CF US 8,637,524 B2 167 168 -continued -continued

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65 CF CF US 8,637,524 B2 169 170 -continued -continued

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65 US 8,637,524 B2 171 172 -continued -continued

C 10

15

CF 25 CF

30

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45 D O " 50 -- D O D N. P 55 N1\-1 D ulp D D D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 173 174 -continued -continued

30

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65

US 8,637,524 B2 177 178 -continued -continued

10

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CF 25 CF

30

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45 O O l, 50 -- D D O D DD D DD D N N D, 55 1NN D D u DDD

60 D D

D 65 CF CF US 8,637,524 B2 179 180 -continued -continued

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65 US 8,637,524 B2 181 182 -continued -continued

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US 8,637,524 B2 185 186 -continued -continued

D

CF 25 CF

30

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45 O O

-- 50 -- D D O O D D D, DD D D D, DD D X N D N DC N D, 55 DC N D ulpife D DC ldi D DC D D

60 D D D D

D D, 65 CF CF US 8,637,524 B2 187 188 -continued -continued

CF 25 CF 3

30

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F F

D D 45 D D O O

D DN D DN l 50 l

X N N DC N D, 55 DC N D

DC ldi D

60 D

65 CF CF US 8,637,524 B2 189 190 -continued -continued

30

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65 US 8,637,524 B2 191 192 -continued -continued

10

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CF 25 CF

30

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F

D 45 O " -- 50 D D

D N 1SN1\- 55

60

65 CF CF US 8,637,524 B2 193 194 -continued -continued

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65