| Title of Invention | "QUINOLINONE-CARBOXAMIDE COMPOUNDS AS 5-HT4 RECEPTOR AGONISTS" |
|---|---|
| Abstract | The present invention relates to a compound of formula (I): wherein: R1 is hydrogen, halo, hydroxy, C1-4alkyl, or C1-4alkoxy; R is C3-4alkyl, or C3-6Cycloalkyl; R is hydrogen or C1-3alkyl; R4 is —S(O)2R6 or —C(O)R7; R5 is hydrogen, C1-3alkyl, C2-3alkyl substituted with -OH or C1-3alkoxy, or -CH2-pyridyl; R6 is C1-3alkyl; or, R5 and R6 taken together form C3-4alkylenyl; and R7 is hydrogen, C1-3alkyl, or pyridyl; or a pharmaceutically-acceptable salt or solvate or stereoisomer thereof. The present invention also relates to process for preparing the said compound. |
| Full Text | BACKGROUND OF THE INVENTION Field of the Invention The invention is directed to quinolinone-carboxamide compounds which are useful as 5-HT4 receptor agonists. The invention is also directed to pharmaceutical compositions comprising such compounds, methods of using such compounds for treating or preventing medical conditions mediated by 5-HT4 receptor activity, and processes and 20 intermediates useful for preparing such compounds. State of the Art Serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter that is widely distributed throughout the body, both in the central nervous system and in peripheral systems. At least seven subtypes of serotonin receptors have been identified and the interaction of serotonin with these different receptors is linked to a wide variety of physiological functions. There has been, therefore, substantial interest in developing therapeutic agents that target specific 5-HT receptor subtypes. In particular, characterization of 5-HT4 receptors and identification of pharmaceutical agents that interact with them has been the focus of significant recent activity. (See, for example, the review by Langlois and Fischmeister, J. Med. Chem. 2003, 46, 319-344.) 5-HT4 receptor agonists are useful for the treatment of disorders of reduced motility of the gastrointestinal tract. Such disorders include irritable bowel syndrome (DBS), chronic constipation, functional dyspepsia, delayed gastric emptying, gastroesophageal reflux disease (GERD), gastroparesis, post-operative ileus, intestinal pseudo-obstruction, and drug-induced delayed transit. In addition, it has been suggested that some 5-HT4 receptor agonist compounds may be used in the treatment of central nervous system disorders including cognitive disorders, behavioral disorders, mood disorders, and disorders of control of autonomic function. Despite the broad utility of pharmaceutical agents modulating 5-HT4 receptor activity, few 5-HT4 receptor agonist compounds are in clinical use at present. One agent, cisapride, that was utilized extensively for treatment of motility disorders of the gastrointestinal tract was withdrawn from the market, reportedly due to cardiac side effects. Late stage clinical trials of another agent, prucalopride, have been suspended. Accordingly, there is a need for new 5-HT4 receptor agonists that achieve their desired effects with minimal side effects. Preferred agents may possess, among other properties, improved selectivity, potency, pharmacokinetic properties, and/or duration of action. SUMMARY OF THE INVENTION The invention provides novel compounds that possess 5-HT4 receptor agonist activity. Among other properties, compounds of the invention have been found to be potent and selective 5-HT4 receptor agonists. In addition, compounds of the invention have been found to exhibit favorable pharmacokinetic properties which are predictive of 20 good bioavailability upon oral administration. Accordingly, the invention provides a compound of formula (I): CO wherein: R1 is hydrogen, halo, hydroxy, Chalky!, or R2 is Cs^alkyl, or R3 is hydrogen or R4 is —S(0)2R6 or —C(O)R7; 5 ^ R is hydrogen, Ci.3alkyl, Ca^alkyl substituted with -OH or Ci.3alkoxy, or — CHa — pyridyl; R6 is Ci_3alkyl; or, R5 and R6 taken together form Ca^alkylenyl; and R7 is hydrogen, Chalky!, or pyridyl; or a pharmaceutically-acceptable salt or solvate or stereoisomer thereof. The invention also provides a pharmaceutical composition comprising a compound of the invention and a pharmaceutically-acceptable carrier. The invention also provides a method of treating a disease or condition associated with 5-HT4 receptor activity, e.g. a disorder of reduced motility of the gastrointestinal tract, the method comprising administering to the mammal, a therapeutically effective amount of a compound of the invention. Further, the invention provides a method of treating a disease or condition associated with 5-HT4 receptor activity in a mammal, the method comprising administering to the mammal, a therapeutically effective amount of a pharmaceutical composition of the invention. The compounds of the invention can also be used as research tools, i.e. to study biological systems or samples, or for studying the activity of other chemical compounds. Accordingly, in another of its method aspects, the invention provides a method of using a compound of formula (I), or a phannaceutically acceptable salt or solvate or stereoisomer thereof, as a research tool for studying a biological system or sample or for discovering new 5-HT4 receptor agonists, the method comprising contacting a biological system or sample with a compound of the invention and determining the effects caused by the compound on the biological system or sample. In separate and distinct aspects, the invention also provides synthetic processes and intermediates described herein, which are useful for preparing compounds of the invention. The invention also provides a compound of the invention as described herein for use in medical therapy, as well as the use of a compound of the invention in the manufacture of a formulation or medicament for treating a disease or condition associated with 5-HT4 receptor activity, e.g. a disorder of reduced motility of the gastrointestinal tract, in a mammal. ** DETAILED DESCRIPTION OF THE INVENTION The invention provides novel quinolinone-carboxamide 5-HT4 receptor agonists of formula (I), or pharmaceutically-acceptable salts or solvates or stereoisomers thereof. The following substituents and values are intended to provide representative examples of various aspects of this invention. These representative values are intended to further define such aspects and are not intended to exclude other values or limit the scope of the invention. Li a specific aspect of the invention, R1 is hydrogen, halo, Ci^alkyl, or Ci^alkoxy. In other specific aspects, R1 is hydrogen, halo, or Ci^alkyl; or R1 is hydrogen or halo; or R1 is fluoro; or R1 is bromo. In yet another specific aspect, R1 is hydrogen. In a specific aspect, R2 is Cs^alkyl or Cs-gcycloalkyl. In another specific aspect, R2 is Ca^alkyl. Representative R2 groups include i n-propyl, isopropyl, «-butyl, sec-butyl, and tert-bntyl. In another specific aspect, R2 is isopropyl. In yet other specific aspects R2 is Ca^alkyl or C^cycloalkyl; or R2 is isopropyl or In a specific aspect, R3 is hydrogen or Ci-salkyl. In other specific aspects, R3 is hydrogen, or R3 is methyl. In a specific aspect, R4 is — S(O)2R6 wherein R6 is Ci-3alkyl. In another specific aspect, R4 is — S(O)2CH3. In a specific aspect, R4 is — C(O)R7 wherein R7 is hydrogen, Ci-salkyl or pyridyl. In other specific aspects, R4 is — C(O)R7 wherein Jl7 is hydrogen or Chalky!; or R4 is — C(O)R7 wherein R7 is hydrogen or methyl; or R4 is — C(O)R7 wherein R7 is 25 hydrogen; or R4 is — C(O)R7 wherein R7 is methyl. In yet another specific aspect, R4 is — C(O)R7 wherein R7 is 3-pyridyl or 4-pyridyl. In a specific aspect, R5 is hydrogen; Chalky!; C2-3alkyl substituted with -OH or Cijjalkoxy; or — CHa — pyridyl. In other specific aspects R5 is hydrogen, Ci-salkyl, or — CHa — pyridyl; or R5 is 3 0 hydrogen or C i .3 alkyl. In yet other specific aspects, R5 is — CBb — 3-pyridyl; or R5 is hydrogen or methyl; or R5 is hydrogen; or R5 is methyl. t» In yet other specific aspects, R5 and R6 taken together form — (CH2)3 — or — (CH2)4; or R5 and R6 taken together form — (CH2)3— . hi one aspect, the invention provides a compound of formula (I) wherein R3 is hydrogen. 5 hi another aspect, the invention provides a compound of formula (I) wherein R4 is — S(0)2R6. hi another aspect, the invention provides a compound of formula (I) wherein R4 is — C(O)R7 The invention further provides a compound of formula (I) wherein R1 is hydrogen 10 or halo; R2 is isopropyl or C^scycloalkyl; and R3, R4, R5, R6, and R7 are defined as in formula (I). hi yet another aspect, the invention provides a compound of formula (I) wherein: R1 is hydrogen; R2 is Cs^alkyl or C^scycloalkyl; 15 R3 is hydrogen; R4 is — S(0)2R6 or — C(O)R7; R5 is hydrogen or Ci-salkyl; R6 is Ci_3a]kyl; and R7 is hydrogen or Ci-salkyl. 20 hi yet another aspect, the invention provides a group of compounds of formula (H): (n) wherein R1 is hydrogen, R2 is isopropyl, and R3, R4, R5, and R6, or R3, R4, R5, and R7 take 25 the values shown in Table I and Table n, respectively. Table I R4 = — S(O)2R6 Example No. R3 R5 R6 1 H CH3 CH3 2 CH3 CH3 CH3 3 CH3 — CH2-3-pyridyl CH3 4 H H CH3 5 H — CH2-3-pyridyl CH3 6 H CH3 CH3 7 H CH3 CH3 21 H C2H5 CH3 22 H H CH3 23 H — (CH2)3— Table H R4 = — C(0)R7 Example No. R3 R5 R7 8 H CH3 4-pyridyl 9 H H 4-pyridyl 10 CH3 CH3 CH3 11 CH3 CH3 4-pyridyl 12 CH3 — CH2-3-pyridyl CH3 13 H H CH3 14 H CH3 CH3 15 H CH3 H 16 H H H 17 H CH3 CH3 18 H CH3 H 5 The chemical naming conventions used herein are illustrated for the compound of Example 1: which is designated l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lJS',3JR,5JR)-8-[2-hydroxy-3-(methaiiesulfonyl-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide, according to the AutoNom software, provided by MDL Information Systems, GmbH (Frankfurt, Germany). The designation (IS,3R,5R) describes 5 the relative orientation of the bonds associated with the bicyclic ring system that are depicted as solid and dashed wedges. The compound is alternatively denoted as 7V-[(3-endo)-8-[2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl]-l-(l-methylethyl)-2-oxo-l,2-dihydro-3-quinolinecarboxamide. In all of the compounds of the invention depicted above, the quinolinone-carboxamide is endo to the 10 azabicyclooctane group. Particular mention may be made of the following compounds l-isopropyl-2-oxo-l ,2-dihydroquinoline-3-carboxylic acid {(15,3^?,5^)-8-[2-hydroxy-3-(methanesulfonyl-methyl-ainino)propyl]-8-azabicyclo[3.2.1]oct-3-yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(ljS,3^?,57?)-8-[2-hydroxy-3-(methanesulfonylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(15.,3jf?55/?)-8-[(J?)-2- hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl} amide; 1 -isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(liS',3.K,5JR!)-8-[(jS)-2- hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(l^SJJjS^-S-fS- (acetyl-methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(l^S^.S^-S-fS-(formyl-methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylicacid {(15,3J2,5^)- (acetyl-methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl} amide; l-isopropyl-2-oxo-l32-dihydroquinoline-3-carboxylicacid {(l^S^S^-(formyl-methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; and l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylicacid {(liSr,3^,57?)- hydroxy-3-(methanesulfonylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl} amide. As exemplified by particular compounds listed above, the compounds of the invention may contain a chiral center, specifically, at the carbon atom in formulas (I) or (IT) bearing the substituent —OR3. Accordingly, the invention includes racemic mixtures, pure stereoisomers, and stereoisomer-enriched mixtures of such isomers, unless otherwise indicated. When a particular stereoisomer is shown, it will be understood by those skilled in the art, that minor amounts of other stereoisomers maybe present in the compositions of the invention unless otherwise indicated, provided that any utility of the composition as a whole is not eliminated by the presence of such other isomers. 5 Definitions When describing the compounds, compositions and methods of the invention, the following terms have the following meanings, unless otherwise indicated. The term "alkyl" means a monovalent saturated hydrocarbon group which may be 10 linear or branched or combinations thereof. Unless otherwise defined, such alkyl groups typically contain from 1 to 10 carbon atoms. Representative alkyl groups include, by way of example, methyl, ethyl, ;;-propyl (n-Pr), isopropyl (z"-Pr), n-butyl (w-Bu), sec-butyl, isobutyl, tert-butyl, w-pentyl, n-hexyl, n-heptyl, «-octyl, n-nonyl, n-decyl and the like. The term "alkylenyl" means a divalent saturated hydrocarbon group which may be 15 linear or branched or combinations thereof. Unless otherwise defined, such alkylenyl groups typically contain from 1 to 10 carbon atoms. Representative alkylenyl groups include, by way of example, methylene, ethylene, n-propylene, w-butylene, propane —1,2-diyl (1-methylethylene), 2-methylpropane-l,2-diyl (1,1-dimethylethylene) and the like. The term "alkoxy" means a monovalent group -O-alkyl, where alkyl is defined as 20 above. Representative alkoxy groups include, by way of example, methoxy, ethoxy, propoxy, butoxy, and the like. The term "cycloalkyl" means a monovalent saturated carbocyclic group which maybe monocyclic or multicyclic. Unless otherwise defined, such cycloalkyl groups typically contain from 3 to 10 carbon atoms. Representative cycloalkyl groups include, by 25 way of example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. The term "halo" means fiuoro, chloro, bromo or iodo. The term "compound" means a compound that was synthetically prepared or prepared in any other way, such as by metabolism. The term "therapeutically effective amount" means an amount sufficient to effect treatment when administered to a patient in need of treatment. The term "treatment" as used herein means the treatment of a disease, disorder, or medical condition in a patient, such as a mammal (particularly a human) which includes: ^ • (a) preventing the disease, disorder, or medical condition from occurring, i.e., prophylactic treatment of a patient; (b) ameliorating the disease, disorder, or medical condition, i.e., eliminating or causing regression of the disease, disorder, or medical condition in a patient; (c) suppressing the disease, disorder, or medical condition, i.e., slowing or arresting the development of the disease, disorder, or medical condition in a patient; or (d) alleviating the symptoms of the disease, disorder, or medical condition in a patient. The term "pharmaceutically-acceptable salt" means a salt prepared from an acid or base which is acceptable for administration to a patient, such as a mammal. Such salts can be derived from pharmaceutically-acceptable inorganic or organic acids and from pharmaceutically-acceptable bases. Typically, pharmaceutically-acceptable salts of compounds of the present invention are prepared from acids. Salts derived from pharmaceutically-acceptable acids include, but are not limited to, acetic, adipic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, xinafoic (l-hydroxy-2-naphthoic acid), naphthalene-l,5-disulfonic acid and the like. The term "solvate" means a complex or aggregate formed by one or more molecules of a solute, i.e. a compound of the invention or a pharmaceutically-acceptable salt thereof, and one or more molecules of a solvent. Such solvates are typically crystalline solids having a substantially fixed molar ratio of solute and solvent. Representative solvents include byway of example, water, methanol, ethanol, isopropanol, acetic acid, and the like. When the solvent is water, the solvate formed is a hydrate. It will be appreciated that the term "or a pharmaceutically-acceptable salt or solvate of stereoisomer thereof is intended to include all permutations of salts, solvates and stereoisomers, such as a solvate of a pharmaceutically-acceptable salt of a stereoisomer of a compound of formula (I). The term "ammo-protecting group" means a protecting group suitable for preventing undesired reactions at an ammo nitrogen. Representative amino-protecting groups include, but are not limited to, formyl; acyl groups, for example alkanoyl groups, such as acetyl; alkoxycarbonyl groups, such as terf-butoxycarbonyl (Boc); 5 arylmethoxycafbonyl groups, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl groups, such as benzyl (Bn), trityl (Tr), and l,l-di-(4'-methoxyphenyl)methyl; silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBDMS); and the like. General Synthetic Procedures Compounds of the invention can be prepared from readily available starting materials using the following general methods and procedures. Although a particular aspect of the present invention is illustrated in the schemes below, those skilled in the art will recognize that all aspects of the present invention can be prepared using the methods described herein or by using other methods, reagents and starting materials known to those skilled in the art. It will also be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary wife the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures. Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group, as well as suitable conditions for protection and deprotection, are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in T. W. Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, Third Edition, Wiley, New York, 1999, and references cited therein. In one method of synthesis, compounds of formula (I) are prepared as illustrated in Scheme A. (The substituents and variables shown in the following schemes have the definitions provided above unless otherwise indicated). Scheme A hi Scheme A, L represents a leaving group such as chloro, bronio, iodo, or ethoxy, or the reagent L—R4 is the carboxylic acid HO-C(O)R7, i.e. L formally represents hydroxy. Optimal reaction conditions for the reaction of Scheme A may vary depending on the chemical properties of the reagent L—R4, as is well known to those skilled in the art. For example, when L is a halo leaving group, such as chloro, the reaction is typically conducted by contacting intermediate (ID) with between about 1 and about 4 equivalents of a compound of formula L—R4 in an inert diluent, such as dichloromethane, in the presence of an excess of base, for example between about 3 and about 6 equivalents, of base, such as A^-diisopropylethylamine or l,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Suitable inert diluents also include TV^V-dimethylformamide, trichloromethane, 1,1,2,2-tetrachloroethane, tetrahydrofuran, and the like. The reaction is typically conducted at a temperature in the range of about -100 °C to about 30 °C for about a quarter hour to about 2 hours, or until the reaction is substantially complete. Exemplary reagents L—R4 in which L is chloro include methanesulfonylchloride and acetylchloride. When the reagent L—R4 is a carboxylic acid, Scheme A represents an amide coupling reaction which is typically conducted by contacting intermediate (in) with between about 1 and about 4 equivalents of a compound of a carboxylic acid L—R in an inert diluent, for example, AyV-dimethylformamide, in the presence of a coupling agent such as benzotriazol-1-yloxytripyrrolidino-phosphonium hexafluorophosphate (PyBop). The reaction is typically conducted at ambient temperature, for about a quarter hour to about 2 hours, or until the reaction is substantially complete. Suitable alternative coupling agents include 1,3 dicyclohexylcarbodiimide (DCC), l-(3- dimethylaniinopropyl)-3-ethylcarbodiimide (BDC), and PyBop combined with 1-hydroxy-7-azabenzotriazole (HOAt). The amide coupling of intermediate (10) with the carboxylic acid L—R4 alternatively can be performed by converting L—R4 to an activated ester, such as an 11 ** .AT-hydroxy succmimide (NHS) ester or ap-nitrophenyl ester, or an acid imidazole, which is then reacted with intermediate (LET). Alternatively, when the reagent L—R4 is a liquid, for example ethyl formate, the reaction can be performed by dissolving (ID) in a large excess of the reagent L—R4, and 5 heating to a temperature of between about 50 °C and about 100 °C for about 12 to about 24 hours. The product of formula (I) is isolated and purified by conventional procedures. For example, the product can be concentrated to dryness under reduced pressure, taken up in an aqueous weak acid solution and purified by HPLC chromatography. 10 Alternatively, compounds of formula (I) can be prepared by JV-alkylating a compound of formula (I) in which R2 is hydrogen, which can be prepared according to Scheme A. The AT-alkylation reaction is typically conducted by contacting a compound of formula (I) in which R2 is hydrogen with between about 1 and about 4 equivalents of a compound of the formula L'—R2 in which L' is a leaving group such as iodo or brorno. 15 This reaction is typically conducted in a polar aprotic solvent such as dimethylfbrmamide in the presence of between about 2 and about 4 equivalents of strong base, such as potassium terf-butoxide. Typically, the reaction is performed at a temperature of between about 60 °C and about 100 °C for between about 6 and about 24 hours, or until the reaction is substantially complete. 20 In yet another alternative, compounds of formula (I) in which R1 is other than hydrogen are prepared by conventional processes from compounds of formula (I) in which R1 is hydrogen. Intermediates of formula (EOT) are prepared from readily available starting materials. For example, when the carbon bearing the substituent —OR3 is not chiral, an 25 intermediate of formula (ffi) is prepared by the procedure illustrated in Scheme B. 12 WO 2005/100350 PCT/US2005/011393 R = H,Ci.3alkyl Scheme B where L1 independently represents a halo leaving group such as bromo, chloro, or iodo. A negatively-charged counterion is also present associated with the positively-charged 5 intermediate (V) or (V1). First, an intermediate of formula (IV) is reacted with an oxirane compound, for example, 2-bromomethyloxirane (commonly, epibromohydrin) to form an azetidine salt of formula (V). This reaction is typically conducted by contacting (IV) with between about 2 and about 4 equivalents of 2-bromomethyloxirane in a polar diluent, such as 10 ethanol. The reaction is typically conducted at ambient temperature for between about 24 and about 48 hours or until the reaction is substantially complete. It will be understood that in the process of Scheme B and in other processes described below using intermediate (IV), intermediate (IV) can be supplied in the form of the freebase or in a salt form, with appropriate adjusment of reaction conditions, as 15 necessary, as known to those skilled in the art. * An intermediate of formula (V1), in which R3 is Ci-salkyl, can be prepared by contacting intermediate (V) with from slightly less than one equivalent to about one 13 WO 2005/100350 PCT/TJS2005/011393 %r equivalent of a compound of formula L'—R3, where R3 is Ci_3alkyl, in an inert diluent in the presence of between about 1 and about 3 equivalents of a strong base, such as potassium terf-butoxide or sodium hydride. The reaction is typically conducted at ambient temperature for between about a quarter hour to an hour, or until the reaction is 5 substantially complete. Suitable inert diluents include dichloromethane, trichloromethane, 1,1,2,2-tetrachloroethane, and the like. Next, the azetidine intermediate (V) or (V1) is reacted with an amine of the formula EySIR5 to provide the intermediate (HI). Typically, the azetidine intermediate is dissolved in an inert diluent, such as ethanol, and contacted with between about 1 and 10 about 8 equivalents of the amine H2NR5. For example, when the amine EySTR5 is a volatile reagent, such as methylamine, preferably, between about 5 and about 7 equivalents of the amine are used. The reaction is typically conducted at a temperature of between about 50 °C and about 100 °C for between about 12 and about 24 hours or until the reaction is substantially complete. 15 An intermediate of formula (DT) in which R5 is hydrogen, can be prepared from the azetidine intermediate (V) or (V1) using ammonium formate in place of ammonia, i.e. in place of the reagent EkNR5 indicated in Scheme B. Alternatively, to prepare intermediate (HT) where R5 is hydrogen, the azetidine ring of (V) or (V) can be opened by reaction with an azide, such as sodium azide, which is then followed by a reduction 20 reaction to provide intermediate (HT), or the ring can be opened by reaction with ammonium hydroxide. As described in detail in Example 4a, when R3 and R5 are hydrogen and the carbon bearing the substituent —OR3 is not chiral, an intermediate of formula (HI) can be prepared by reacting intermediate (IV) with an oxiranyhnethyl compound having a 25 protected nitrogen atom and then deprotecting. One useful reagent is 2-oxiranylmethyl-isoindole-l,3-dione, commonly epoxypropylphthalimide, which is reacted with intermediate (TV) to form an intermediate in which aphthalimidyl-substituted 2-hydroxy propyl group: O 14 WO 2005/100350 PCT/US2005/011393 is joined to the nitrogen of the azabicylcooctane ring of formula (IV). The phthalimidyl group is then removed by refluxing in hydrazine to form an intermediate of formula (ID) in which R3 and R5 are hydrogen. An intermediate of formula (Tfl) in which R3 and R5 are hydrogen can also be prepared by reaction of the azetidine (V) with the anion of phthahmide and subsequent treatment with hydrazine. In an alternative method of synthesis, an intermediate of formula (HT) in which R3 is hydrogen, can be prepared by reaction of intermediate (TV) with a protected intermediate (VI): 10 OH R5 (VI) followed by a deprotection step. In formula (VT), P1 is an amino-protecting group, L' is a halo leaving group, and the asterisk denotes a chiral center. The process utilizing an intermediate of formula (VI) is useful for preparing forms of intermediate (IH) in which 1 5 the stereochemistry at the center marked by the aserisk is specifically (R) or (5) as well as for preparing non-chiral forms of intermediate (HI). Typically, intermediate (IV) is contacted with between about 1 and about 2 equivalents of intermediate (VI) in a polar diluent, such as methanol, in the presence of more than one equivalent of a base, such as JV^-diisopropylethylamine. The reaction is 20 typically conducted at a temperature of between about 60 °C and about 100 °C for between about 12 and about 24 hours, or until the reaction is substantially complete. The protecting group P1 is removed by standard procedures to provide an intermediate of formula (in). A useful protecting group P1 is Boc, which is typically removed by treatment with an acid, such as trifluoroacetic acid. 25 In yet another alternative process for the preparation of intermediate QJL), intermediate (VI) can first be converted to a cyclized form (VTT): d R5 (VH) before reaction with intermediate (TV) to provide intermediate (HI). Intermediate (VH) is 30 typically prepared by dissolving intermediate (VI) in an inert diluent, for example, tetrahydrofuran, in the presence of base, for example sodium hydroxide. The reaction of 15 AVO 2005/100350 PCT/US2005/011393 (VTI) with (TV) to provide intermediate (DT) is typically performed by contacting intermediate (TV) with between about 1 and about 4 equivalents of intermediate (VH) in a polar diluent, such as methanol. The reaction is typically conducted at a temperature of between about 60 °C to about 100 °C for between about 1 and about 4 hours, or until the 5 reaction is substantially complete. The protecting group P1 is removed by standard procedures to provide an intermediate of formula (IH). The protected intermediate (VI) can be prepared from an oxirane as illustrated in Scheme C for the particular example of forming a Boc-protected chiral intermediate (VI1) using a chiral oxirane. The reaction is equally useful for the preparation of non-chiral 10 compounds of formula (VI). Scheme C OH (VI') 20 As shown in Scheme C, abenzylamine 2 is contacted with at least one equivalent of a cliiral oxirane 1 in a non-polar diluent such as hexane or toluene to form the 2-hydroxypropylamine 3. The reaction is typically conducted at room temperature for 15 between about 12 and about 24 hours, or until the reaction is substantially complete. The intermediate 3 is typically reacted with a slight excess of di-ter?-butyl dicarbonate (commonly (Boc)2O), for example, about 1.1 equivalents, under a hydrogen atmosphere in the presence of a transition metal catalyst to provide the Boc protected intermediate (VI'). The reaction is typically conducted at ambient temperature for between about 8 to about 24 hours. A process for preparing intermediates of formula (TV) is shown in Scheme D. H2N Scheme D (VIII) (IV) WO 2005/100350 PCT/US2005/011393 ¥ ihe protected aminoazabicyclooctane, or commonly, aminotropane 5 is first reacted with the substituted quinolinone carboxylic acid (VET). Typically, this reaction is conducted by first converting (VET) to an acid chloride by contacting (VTH) with at least one equivalent, preferably between about 1 and about 2 equivalents of an activating agent, 5 such as thionyl chloride or oxalyl chloride in an aromatic diluent, such as toluene, benzene, xylene, or the like. The reaction is typically conducted at a temperature ranging from about 80 °C to about 120 °C for about 15 minutes to about 4 hours, or until the reaction is substantially complete. The acid chloride solution is typically added to a biphasic mixture of about 1 10 equivalent of the aminotropane 5 to form a protected intermediate, which is extracted by standard procedures. The biphasic mixture of 5 is generally prepared by dissolving 5 in an aromatic diluent, such as used above, and adding an aqueous solution containing an excess of base, such as sodium hydroxide or potassium hydroxide, preferably about 2 to 5 equivalents of base. 15 Alternatively, the amide coupling of intermediate 5 with the carboxylic acid (VET) can be performed in the presence of a coupling agent such as 1,3 dicyclohexylcarbodiimide (DCC), 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), or benzotriazol-1-yloxytripyrrolidino-phosphonium hexafluorophosphate (PyBop), optionally combined with l-hydroxy-7-azabenzotriazole (HOAt), as described 20 above for the amide coupling of intermediate (in) with a carboxylic acid. In yet another alternative, the amide coupling of intermediate 5 with the carboxylic acid (VET) can be performed by converting (VTO) to an activated ester, also described above. The protecting group P1 is removed by standard procedures to provide an intermediate of formula (IV). For example when the protecting group is Boc, typically 25 removal is by treatment with an acid, such as trifluoroacetic acid, providing the acid salt of the intermediate. The acid salt of intermediate (TV) can be converted to the free base, if desired, by conventional treatment with base. The protecting group Cbz, for another example, is conveniently removed by hydrogenolysis over a suitable metal catalyst such as palladium on carbon. 30 The protected aminotropane 5 employed in the reactions described in this application is prepared from readily available starting materials. For example, when the protecting group P1 is Boc, the protected aminotropane 5' is prepared by the procedure illustrated in Scheme E. 17 WO 2005/100350 PCT/US2005/011393 Scheme E NH2 5- As described in detail in Example la below, to prepare the protected intermediate 5', first, 2,5-dimethoxy tetrahydrofuran 6 is contacted with between about 1 and 2 equivalents, preferably about 1.5 equivalents of benzyl arnine and a slight excess, for 5 example about 1.1 equivalents, of 1,3-acetonedicarboxylic acid 7 in an acidic aqueous solution in the presence of a buffering agent such as sodium hydrogen phosphate. The reaction mixture is heated to between about 60 and about 100 °C to ensure decarboxylation of any carboxylated intermediates in the product, 8-benzyl-8-azabicyclo[3.2.1]octan-3-one 8, commonly ]V-benzyltropanone. 1 0 The intermediate 8 is typically reacted with a slight excess of di-te?Y-butyl dicarbonate (commonly (Boc^O), for example, about 1.1 equivalents, under a hydrogen atmosphere in the presence of a transition metal catalyst to provide the Boc protected intermediate 9, 3-oxo-8-azabicyclo[3,2.1]octane-8-carboxylic acid te?t-butyl ester. The reaction is typically conducted at ambient temperature for about 12 to about 72 hours. 15 Finally, intermediate 9 is contacted with a large excess, for example at least about 25 equivalents, of ammonium formate in an inert diluent, such as methanol, in the presence of a transition metal catalyst to provide the product 5'in the endo configuration with Mgh stereospecificity, for example endo to exo ratio of >99:1 . The reaction is typically conducted at ambient temperature for about 12 to about 72 hours or until the reaction is 20 substantially complete. It is advantageous to add the ammonium formate reagent in portions. For example, intermediate 9 is contacted with an initial portion of ammonium formate of about 15 to about 25 equivalents. After an interval of about 12 to about 36 hours, an additional portion of about 5 to about 10 equivalents of ammonium formate is added. The subsequent addition can be repeated after a similar interval. The product 5' 25 can be purified by conventional procedures, such as alkaline extraction. 18 WO 2005/100350 PCT/TJS2005/011393 In an alternative method of synthesis, compounds of formula (T) are prepared by coupling the substituted quinolinone carboxylic acid (VIS) with an intermediate of formula (IX) as illustrated in Scheme F. Scheme F H2N (IX) (vin) The reaction of Scheme F is typically conducted under the amide coupling conditions described above for the reaction of the carboxylic acid (VHI) with intermediate 5. Intermediates of formula (TX) can be prepared by deprotecting an intermediate of 10 formula (X): P2-NH (X) where P2 represents an amino-protecting group. Intermediates of formula (X) can be prepared from readily available starting 15 materials using procedures analogous to the alkylation and other reactions described above and/or using alternative reactions well known to those skilled in the art. For example, intermediate (X) can be prepared using an intermediate 10 f) P2-NH 10 which may be formed by protecting the amino nitrogen of the aminoazobicyclooctane 5 20 with amino-protecting group P2 and then removing P1 from the nitrogen of the azabicyclooctane group. Protecting groups P1 and P2 are chosen such that they are removed under different conditions. For example when P1 is chosen as Boc, then Cbz can be used as P2. Substituting the protected aminotropane 10 for intermediate (TV) in the reactions described above for the preparation of intermediate (TO) provides intermediates 25 of formula (X). 19 WO 2005/100350 PCT/US2005/011393 In yet another method of synthesis, compounds of formula (I) in which R3 is hydrogen, represented below as formula (T), can be prepared as illustrated in Scheme G. Scheme G N-R4 -N Intermediate (XT) may contain a chiral center, as shown explicitly for the protected 5 oxirane intermediate (VII). Typically, intermediate (TV) is contacted with between about 1 and about 2 equivalents of the oxirane intermediate (XT) in a polar diluent, such as ethanol to form the product (I'). Intermediate (IV) can be supplied in salt form hi which case a slight molar excess of alkaline base is included in the reaction mixture prior to the addition of the 10 oxirane. The reaction is typically conducted at a temperature of about 60 °C to about 100 °C for between about 1 and about 3 hours, or until the reaction is substantially complete. The product can be isolated by crystallization from an inert diluent as the free base or as an acid salt. Intermediates of formula (XI) can be prepared by reaction of the oxirane 15 intermediate 1, illustrated in Scheme C, with the secondary amine HNR4R5. Typically, an aqueous solution of the amine HNR4R5 containing about 1 equivalent of abase, such as sodium hydroxide, lithium hydroxide, cesium hydroxide, or potassium hydroxide, is contacted with between about 1.5 and about 2.5 equivalents of the oxirane intermediate 1. The reaction is typically conducted at a temperature of between about 0 °C and about 20 10 °C for between about 12 and about 30 hours, or until the reaction is substantially complete. The quinolinone carboxylic acid (VET) is readily prepared by procedures similar to those reported in the literature in Suzuki et al, Heterocycles, 2000,53, 2471-2485 and described in the examples below. 25 The reagents L'—R2, L'—R3, L—R4, H2NR5, and HNR4R5 are available commercially or are readily prepared by standard procedures from common starting materials. 20 WO 2005/100350 PCT/US2005/011393 '"*r Further details regarding specific reaction conditions and other procedures for preparing representative compounds of the invention or intermediates thereto are described in the examples below. Accordingly, in a method aspect, the invention provides a process for preparing a 5 compound of formula (T), or a salt or stereoisomer thereof, the process comprising: (a) reacting a compound of formula (HT): with compound of the formula L—R4 wherein L is a leaving group, or L—R4 represents HO-C(O)R7; or 10 (b) reacting a compound of formula (VET): (Vffl) O with a compound of formula (IX): (DC) to provide a compound of formula (T), or a salt or stereoisomer thereof. 15 The invention further provides a compound of formula (TO), or a salt or stereoisomer or protected derivative thereof, wherein R1, R2, R3, and R5 are defined as in formula (I). In an additional method aspect, the invention provides a process for preparing a compound of formula (I1) wherein R1, R2, R4, and R5 are defined as in formula (I), or a 20 salt or stereoisomer thereof, the process comprising reacting a compound of formula (IV): 21 WO 2005/100350 PCT/US2005/011393 (IV) or a salt thereof with a compound of formula (XT): (XT) to provide a compound of formula (I1) or a salt or stereoisomer thereof. 5 Pharmaceutical Compositions The quinoh'none-carboxamide compounds of the invention are typically administered to a patient in the form of a pharmaceutical composition. Such pharmaceutical compositions maybe administered to the patient by any acceptable route 10 of administration including, but not limited to, oral, rectal, vaginal, nasal, inhaled, topical (including transdermal) and parenteral modes of administration. Accordingly, in one of its compositions aspects, the invention is directed to a pharmaceutical composition comprising a pharmaceutically-acceptable carrier or excipient and a therapeutically effective amount of a compound of formula (!) or a 1 5 pharmaceutically acceptable salt thereof. Optionally, such pharmaceutical compositions may contain other therapeutic and/or formulating agents if desired. The pharmaceutical compositions of the invention typically contain a therapeutically effective amount of a compound of the present invention or a pharmaceutically-acceptable salt thereof. Typically, such pharmaceutical compositions 20 will contain from about 0. 1 to about 95% by weight of the active agent; preferably, from about 5 to about 70% by weight; and more preferably from about 10 to about 60% by weight of the active agent. Any conventional carrier or excipient may be used in the pharmaceutical compositions of the invention. The choice of a particular carrier or excipient, or 25 combinations of carriers or excipients, will depend on the mode of administration being used to treat a particular patient or type of medical condition or disease state, m this 22 WO 2005/100350 PCT/US2005/011393 H* regard, the preparation of a suitable pharmaceutical composition for a particular mode of administration is well within the scope of those skilled in the pharmaceutical arts. Additionally, the ingredients for such compositions are commercially-available from, for example, Sigma, P.O. Box 14508, St. Louis, MO 63178. By way of further illustration, 5 conventional formulation techniques are described in Remington: TJie Science and Practice of Pharmacy, 20th Edition, Lippincott Williams & White, Baltimore, Maryland (2000); and H.C. Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition, Lippincott Williams & White, Baltimore, Maryland (1999). Representative examples of materials which can serve as pharmaceutically 10 acceptable carriers include, but are not limited to, the following: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as com starch and potato starch; (3) cellulose, such as macrocrystalline cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; 15 (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's 20 solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical compositions. The pharmaceutical compositions of the invention are typically prepared by thoroughly and intimately mixing or blending a compound of the invention with a pharmaceutically-acceptable carrier and one or more optional ingredients. If necessary or 25 desired, the resulting uniformly blended mixture can then be shaped or loaded into tablets, capsules, pills and the like using conventional procedures and equipment. The pharmaceutical compositions of the invention are preferably packaged in a unit dosage form. The term "unit dosage form" refers to a physically discrete unit suitable for dosing a patient, i.e., each unit containing a predetermined quantity of active agent 30 calculated to produce the desired therapeutic effect either alone or in combination with one or more additional units. For example, such unit dosage forms may be capsules, tablets, pills, and the like. 23 \VO 2005/100350 PCT/US2005/011393 V In a preferred embodiment, the pharmaceutical compositions of the invention are suitable for oral administration. Suitable pharmaceutical compositions for oral administration maybe in the form of capsules, tablets, pills, lozenges, cachets, dragees, powders, granules; or as a solution or a suspension in an aqueous or non-aqueous liquid; 5 or as an oil-in-water or water-in-oil liquid emulsion; or as an elixir or syrup; and the like; each containing a predetermined amount of a compound of the present invention as an active ingredient. When intended for oral administration in a solid dosage form (i.e., as capsules, tablets, pills'and the like), the pharmaceutical compositions of the invention will typically 10 comprise a compound of the present invention as the active ingredient and one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate. Optionally or alternatively, such solid dosage forms may also comprise: (1) fillers or extenders, such as starches, microcrystalline cellulose, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, 15 polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and/or sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and/or glycerol monostearate; (8) absorbents, such as 20 kaolin and/or bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and/or mixtures thereof; (10) coloring agents; and (11) buffering agents. Release agents, wetting agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the 25 pharmaceutical compositions of the invention. Examples of pharmaceutically-acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfate sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; 30 and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like. Coating agents for tablets, capsules, pills and like, include those used for enteric coatings, such as cellulose acetate phthalate (CAP), polyvinyl acetate phthalate (PVAP), hydroxypropyl methylcellulose 24 WO 2005/100350 PCT/US2005/011393 pfithalate, methacrylic acid-methacrylic acid ester copolymers, cellulose acetate trimellitate (CAT), carboxymethyl ethyl cellulose (CMEC), hydroxypropyl methyl cellulose acetate succinate (HPMCAS), and the like. If desired, the pharmaceutical compositions of the present invention may also be 5 formulated to provide slow or controlled release of the active ingredient using, by way of example, hydroxypropyl methyl cellulose in varying proportions; or other polymer matrices, liposomes and/or microspheres. In addition, the pharmaceutical compositions of the present invention may optionally contain opacifying agents and may be formulated so that they release the active 10 ingredient only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients. Suitable liquid dosage forms for oral administration include, byway of 15 illustration, pharmaceutically-acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. Such liquid dosage forms typically comprise the active ingredient and an inert diluent, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (esp., 20 cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Suspensions, in addition to the active ingredient, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, 25 and mixtures thereof. Alternatively, the pharmaceutical compositions of the invention are formulated for administration by inhalation. Suitable pharmaceutical compositions for administration by inhalation will typically be in the form of an aerosol or a powder. Such compositions are generally administered using well-known delivery devices, such as a metered-dose 30 inhaler, a dry powder inhaler, a nebulizer or a similar delivery device. When administered by inhalation using a pressurized container, the pharmaceutical compositions of the invention will typically comprise the active ingredient 25 WO 2005/100350 PCT/US2005/011393 ^r and a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. Additionally, the pharmaceutical composition may be in the form of a capsule or cartridge (made, for example, from gelatin) comprising a compound of the invention and 5 a powder suitable for use in a powder inhaler. Suitable powder bases include, by way of example, lactose or starch. The compounds of the invention can also be administered transdermally using known transdermal delivery systems and excipients. For example, a compound of the invention can be admixed with permeation enhancers, such as propylene glycol, 10 polyethylene glycol monolaurate, azacycloalkan-2-ones and the like, and incorporated into a patch or similar delivery system. Additional excipients including gelling agents, emulsifiers and buffers, may be used in such transdermal compositions if desired. The following formulations illustrate representative pharmaceutical compositions of the present invention: 15 Formulation Example A Hard gelatin capsules for oral administration are prepared as follows: Ingredients Amount 20 . —^ Compound of the invention 50 mg Lactose (spray-dried) 200 mg Magnesium stearate 10 mg 25 ' Representative Procedure: The ingredients are thoroughly blended and then loaded into a hard gelatin capsule (260 mg of composition per capsule). WO 2005/100350 PCT/US2005/011393 Formulation Example B Hard gelatin capsules for oral administration are prepared as follows: Ingredients Amount Compound of the invention 20 mg Starch 89 mg Microcrystalline cellulose 89 mg , ^ Magnesium stearate 2 mg Representative Procedure: The ingredients are thoroughly blended and then passed through a No. 45 mesh U.S. sieve and loaded into a hard gelatin capsule (200 mg of composition per capsule). 15 Formulation Example C Capsules for oral administration are prepared as follows: Ingredients Amount 20 . _ Compound of the invention 10 mg Polyoxyethylene sorbitan monooleate 50 mg Starch powder 250 mg 25 ' Representative Procedure: The ingredients are thoroughly blended and then loaded into a gelatin capsule (310 mg of composition per capsule). 27 WO 2005/100350 PCT/US2005/011393 ^* Formulation Example D Tablets for oral administration are prepared as follows: Ingredients Amount 5 Compound of the invention 5 rng Starch 50 mg Macrocrystalline cellulose 35 mg Polyvinylpyrrolidone (10 wt. % in water) 4 mg Sodium carboxymethyl starch 4.5 mg 10 Magnesium stearate 0.5 mg Talc 1 mg Representative Procedure: The active ingredient, starch and cellulose are passed 15 through aNo. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resulting powders, and this mixture is then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50-60°C and passed throughaNo. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc (previously passed through a 20 No. 60 mesh U.S. sieve) are men added to the granules. After mixing, the mixture is compressed on a tablet machine to afford a tablet weighing 100 mg. Formulation Example E Tablets for oral administration are prepared as follows: 25 Ingredients Amount Compound of the invention 25 mg Macrocrystalline cellulose 400 mg 30 Silicon dioxide fumed lOmg Stearic acid 5 mg Representative Procedure: The ingredients are thoroughly blended and then 35 compressed to form tablets (440 mg of composition per tablet). 28 WO 2005/100350 PCT/US2005/011393 %V Formulation Example F Single-scored tablets for oral administration are prepared as follows: Ingredients Amount Compound of the invention 15 mg Cornstarch 50 mg Croscarmellose sodium 25 mg Lactose 120 mg 10 Magnesium stearate 5mg Representative Procedure: The ingredients are thoroughly blended and compressed 15 to form a single-scored tablet (215 mg of compositions per tablet). Formulation Example G A suspension for oral administration is prepared as follows: 20 Ingredients Amount Compound of the invention 0.1 g Fumaric acid 0.5 g Sodium chloride 2.0 g 25 Methyl paraben 0.15 g Propyl paraben 0.05 g Granulated sugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum k (Vanderbilt Co.) 1.0 g 30 Flavoring 0.035 mL Colorings 0.5 mg Distilled water q.s. to 100 mL 35 Representative Procedure: The ingredients are mixed to form a suspension containing 10 mg of active ingredient per 10 mL of suspension. 29 WO 2005/100350 PCT/US2005/011393 V Formulation Example H A dry powder for administration by inhalation is prepared as follows: Ingredients Amount Compound of the invention 1.0 mg Lactose 25 mg 10 Representative Procedure: The active ingredient is micronized and then blended with lactose. This blended mixture is then loaded into a gelatin inhalation cartridge. The contents of the cartridge are administered using a powder inhaler. 15 Formulation Example I A dry powder for administration by inhalation in a metered dose inhaler is prepared as follows: Representative Procedure: A suspension containing 5 wt. % of a compound of the invention and 0.1 wt. % lecithin is prepared by dispersing 10 g of active 20 compound as micronized particles with mean size less than 10 [am in a solution formed from 0.2 g of lecithin dissolved in 200 mL of demineralized water. The suspension is spray dried and the resulting material is micronized to particles having a mean diameter less than 1.5 urn. The particles are loaded into cartridges with pressurized l,l,l>2-tetrafluoroethane. 25 Formulation Example J An injectable formulation is prepared as follows: Ingredients Amount 30 . Compound of the invention 0.2 g Sodium acetate buffer solution (0.4 M) 40 mL HC1 (0.5 N) or NaOH (0.5 N) q.s. to pH 4 35 Water (distilled, sterile) q.s. to 20 mL Representative Procedure: The above ingredients are blended and the pH is adjusted to 4 ± 0.5 using 0.5 N HC1 or 0.5 N NaOH. 40 30 WO 2005/100350 PCT/TJS2005/011393 ^H Formulation Example K Capsules for oral administration are prepared as follows: Ingredients Amount 5 Compound of the Invention 4.05 mg Microcrystalline cellulose (AvicelPH 103) 259.2 mg Magnesium stearate 0.75 mg 10 Representative Procedure: The ingredients are thoroughly blended and then loaded into a gelatin capsule (Size #1, White, Opaque) (264 mg of composition per capsule). Formulation Example L 15 Capsules for oral administration are prepared as follows: Ingredients Amount Compound of the Invention 8.2 mg Microcrystalline cellulose (Avicel PH 103) 139.05 mg 20 Magnesium stearate 0.75 mg Representative Procedure: The ingredients are thoroughly blended and then loaded into a gelatin capsule (Size #1, White, Opaque) (148 mg of composition per 25 capsule). It will be understood that any form of the compounds of the invention, (i.e. free base, pharmaceutical salt, or solvate) that is suitable for the particular mode of administration, can be used in the pharmaceutical compositions discussed above. 30 Utility The quinolinone-carboxamide compounds of the invention are 5-HT4 receptor agonists and therefore are expected to be useful for treating medical conditions mediated by 5-HT4 receptors or associated with 5-HT4 receptor activity, i.e. medical conditions 35 which are ameliorated by treatment with a 5-HT4 receptor agonist. Such medical conditions include, but are not limited to, irritable bowel syndrome (IBS), chronic constipation, functional dyspepsia, delayed gastric emptying, gastroesophageal reflux disease (GEKD), gastroparesis, post-operative ileus, intestinal pseudo-obstruction, and 31 WO 2005/100350 PCT/US2005/011393 ^(rug-induced delayed transit. In addition, it has been suggested that some 5-HT4 receptor agonist compounds may be used in the treatment of central nervous system disorders including cognitive disorders, behavioral disorders, mood disorders, and disorders of control of autonomic function. 5 hi particular, the compounds of the invention increase motility of the gastrointestinal (GI) tract and thus are expected to be useful for treating disorders of the GI tract caused by reduced motility in mammals, including humans. Such GI motility disorders include, by way of illustration, chronic constipation, constipation-predominant irritable bowel syndrome (C-IBS), diabetic and idiopathic gastroparesis, and functional 10 dyspepsia. hi one aspect, therefore, the invention provides a method of increasing motility of the gastrointestinal tract in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a pharmaceutically-acceptable carrier and a compound of the invention. 15 When used to treat disorders of reduced motility of the GI tract or other conditions mediated by 5-HT4 receptors, the compounds of the invention will typically be administered orally in a single daily dose or in multiple doses per day, although other forms of administration may be used. The amount of active agent administered per dose or the total amount administered per day will typically be determined by a physician, in 20 the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like. Suitable doses for treating disorders of reduced motility of the GI tract or other 25 disorders mediated by 5-HT4 receptors will range from about 0.0007 to about 20 mg/kg/day of active agent, including from about 0.0007 to about 1 mg/kg/day. For an average 70 kg human, this would amount to from about 0.05 to about 70 mg per day of active agent. hi one aspect of the invention, the compounds of the invention are used to treat 30 chronic constipation. When used to treat chronic constipation, the compounds of the invention will typically be administered orally in a single daily dose or in multiple doses per day. Preferably, the dose for treating chronic constipation will range from about 0.05 to about 70 mg per day. 32 WO 2005/100350 PCT/US2005/011393 ^j w hi another aspect of the invention, the compounds of the invention are used to treat irritable bowel syndrome. When used to treat constipation-predominant irritable bowel syndrome, the compounds of the invention will typically be administered orally in a single daily dose or in multiple doses per day. Preferably, the dose for treating constipation-5 predominant irritable bowel syndrome will range from about 0.05 to about 70 mg per day. hi another aspect of the invention, the compounds of the invention are used to treat diabetic gastroparesis. When used to treat diabetic gastroparesis, the compounds of the invention will typically be administered orally in a single daily dose or in multiple doses per day. Preferably, the dose for treating diabetic gastroparesis will range from about 0.05 10 to about 70 mg per day. hi yet another aspect of the invention, the compounds of the invention are used to treat functional dyspepsia. When used to treat functional dyspepsia, the compounds of the invention will typically be administered orally in a single daily dose or in multiple doses per day. Preferably, the dose for treating functional dyspepsia will range from about 0.05 15 to about 70 mg per day. The invention also provides a method of treating a mammal having a disease or condition associated with 5-HT4 receptor activity, the method comprising administering to the mammal a therapeutically effective amount of a compound of the invention or of a pharmaceutical composition comprising a compound of the invention. 20 As described above, compounds of the invention are 5-HT4 receptor agonists. The invention further provides, therefore, a method of agonizing a 5-HT4 receptor in a mammal, the method comprising administering a compound of the invention to the mammal, hi addition, the compounds of the invention are also useful as research tools for investigating or studying biological systems or samples having 5-HT4 receptors, or for 25 discovering new 5-HT4 receptor agonists. Moreover, since compounds of the invention exhibit binding selectivity for 5-HT4 receptors as compared with binding to receptors of other 5-HT subtypes, particularly 5-HT3 receptors, such compounds are particularly useful for studying the effects of selective agonism of 5-HT4 receptors in a biological system or sample. Any suitable biological system or sample having 5-HT4 receptors may be 30 employed in such studies which may be conducted either in vitro or in vivo. Representative biological systems or samples suitable for such studies include, but are not limited to, cells, cellular extracts, plasma membranes, tissue samples, mammals (such as mice, rats, guinea pigs, rabbits, dogs, pigs, etc.) and the like. 33 WO 2005/100350 PCT/US2005/011393 ^ta* ^ In this aspect of the invention, a biological system or sample comprising a 5-HT4 receptor is contacted with a 5-HT4 receptor-agonizing amount of a compound of trie invention. The effects of agonizing the 5-HT4 receptor are then determined using conventional procedures and equipment, such as radioligand binding assays and 5 functional assays. Such functional assays include ligand-mediated changes in intracellular cyclic adenosine monophosphate (cAMP), ligand-mediated changes in activity of the enzyme adenylyl cyclase (which synthesizes cAMP), ligand-mediated changes in incorporation of analogs of guanosine triphosphate (GTP), such as [35S]GTPyS (guanosine 5'-O-(y-thio)triphosphate) or GTP-Eu, into isolated membranes via receptor 10 catalyzed exchange of GTP analogs for GDP analogs, ligand-mediated changes in free intracellular calcium ions (measured, for example, with a fluorescence-linked imaging plate reader or FLEPR® from Molecular Devices, Inc.), and measurement of mitogen activated protein kinase (MAPK) activation. A compound of the invention may agonize or increase the activation of 5-HT4 receptors in any of the functional assays listed above, 15 or assays of a similar nature. A 5-HT4 receptor-agonizing amount of a compound of the invention will typically range from about 1 nanomolar to about 500 nanomolar. Additionally, the compounds of the invention can be used as research tools for discovering new 5-HT4 receptor agonists. In this embodiment, 5-HT4 receptor binding or functional data for a test compound or a group of test compounds is compared to the 20 5-HT4 receptor binding or functional data for a compound of the invention to identify test compounds that have superior binding or functional activity, if any. This aspect of the invention includes, as separate embodiments, both the generation of comparison data (using the appropriate assays) and the analysis of the test data to identify test compounds of interest. 25 Among other properties, compoimds of the invention have been found to be potent agonists of the 5-HT4 receptor and to exhibit substantial selectivity for the 5-HT4 receptor subtype over the S-HTs receptor subtype in radioligand binding assays. Further, compounds of the invention have demonstrated superior pharmacokinetic properties in a rat model. Compounds of the invention are thus expected to be highly bioavailable upon 30 oral administration. In addition, these compounds have been shown not to exhibit an unacceptable level of inhibition of the potassium ion current in an in vitro voltage-clamp model using isolated whole cells expressing the hERG cardiac potassium channel. The voltage-clamp assay is an accepted pre-clinical method of assessing the potential for 34 WO 2005/100350 PCT/US2005/011393 pharmaceutical agents to change the pattern of cardiac repolarization, specifically to cause, so-called QT prolongation, which has been associated with cardiac arrhythmia. , (Cavero et al., Opinion on Pharmacotherapy, 2000,1, 947-73, Fermini et a/., Nature Reviews Drug Discovery, 2003, 2, 439-447) Accordingly, pharmaceutical compositions 5 comprising compounds of the invention are expected to have an acceptable cardiac profile. There properties, as well as the utility of the compounds of the invention, can be demonstrated using various in vitro and in vivo assays well-known to those skilled in the art. Representative assays are described in further detail in the following examples. 10 EXAMPLES The following synthetic and biological examples are offered to illustrate the invention, and are not to be construed in any way as limiting the scope of the invention. 15 In the examples below, the following abbreviations have the following meanings unless otherwise indicated. Abbreviations not defined below have their generally accepted meanings. Boc = fer£-butoxycarbonyl (BocfeO = di-to-^-butyl dicarbonate 20 DCM = dichloromethane DMF = Af,W-dimethylformamide DMSO = dimethyl sulfoxide EtOAc = ethyl acetate mCPBA = m-chlorobenzoic acid 25 MeCN = acetonitrile MTBE = tert-butyl methyl ether PyBop = benzotriazol-1-yloxytripyrrolidino- phosphonium hexafmorophosphate Rf = retention factor 30 RT = room temperature TFA = trifluoroacetic acid THF = tetrahydrofuran Reagents (including secondary amines) and solvents were purchased from 35 commercial suppliers (Aldrich, Fluka, Sigma, etc.), and used without further purification. Reactions were run under nitrogen atmosphere, unless noted otherwise. Progress of reaction mixtures was monitored by thin layer chromatography (TLC), analytical high performance liquid chromatography (anal. HPLC), and mass spectrometry, the details of 35 WO 2005/100350 PCT/US2005/011393 are given below and separately in specific examples of reactions. Reaction mixtures were worked up as described specifically in each reaction; commonly they were purified by extraction and other purification methods such as temperature-, and solvent-dependent crystallization, and precipitation. In addition, reaction mixtures were routinely 5 purified by preparative HPLC: a general protocol is described below. Characterization of reaction products was routinely carried out by mass and ^-NMR spectrometry. For NMR measurement, samples were dissolved in deuterated solvent (CD3OD, CDC13, or DMSO-^eX and 'H-NMR spectra were acquired with a Varian Gemini 2000 instrument (300 MHz) under standard observation conditions. Mass spectrometric identification of 1 0 compounds was performed by an electrospray ionization method (ESMS) with an Applied Biosystems (Foster City, CA) model API 150 EX instrument or an Agilent (Palo Alto, CA) model 1 100 LC/MSD instrument. Water content is determined by Karl Fischer titration using a Brinkmann (Westbury, NY) Metrohm Karl Fischer Model 813 coulometer. 15 Example 1: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(l^JJ^jR^S-p-hydroxy-S^methanesulfonyl-methyl-amino)propyI]-8-azabicyclo [3.2. 1 ] oct-3-yl} amide a. Preparation of 8-benzyl-8-azabicyclor3.2.1]octan-3-one 20 Concentrated hydrochloric acid (30 mL) was added to a heterogeneous solution of 2,5-dimethoxy tetrahydrofuran (82.2 g, 0.622 mol) in water (170 mL) while stirring. In a separate flask cooled to 0°C (ice bath), concentrated hydrochloric acid (92 mL) was added slowly to a solution of benzyl amine (100 g, 0.933 mol) in water (350 mL). The 2,5-dimethoxytetrahydrofuran solution was stirred for approximately 20 min, diluted with 25 water (250 mL), and then the benzyl amine solution was added, followed by the addition of a solution of 1,3-acetonedicarboxylic acid (100 g, 0.684 mol) in water (400 mL) and then the addition of sodium hydrogen phosphate (44 g, 0.3 1 mol) in water (200 mL). The pH was adjusted from pH 1 to pH ~ 4.5 using 40% NaOH. The resulting cloudy and pale yellow solution was stirred overnight. The solution was then acidified to pH 3 from pH 30 7.5 using 50% hydrochloric acid, heated to 85 °C and stirred for 2 hours. The solution was cooled to room temperature, basified to pH 12 using 40% NaOH, and extracted with DCM (3 x 500 mL). The combined organic layers were washed with brine, dried 36 WO 2005/100350 PCT/US2005/011393 (MgSCU), filtered and concentrated under reduced pressure to produce the crude title intermediate as a viscous brown oil (52 g). To a solution of the crude intermediate in methanol (1000 mL) was added di-tert-butyl dicarbonate (74.6 g, 0.342 mol) at 0 °C. The solution was allowed to warm to room 5 temperature and stirred overnight. The methanol was removed under reduced pressure and the resulting oil was dissolved in dichloromethane (1000 mL). The intermediate was extracted into 1 M H3PO4 (1000 mL) and washed with dichloromethane (3 x 250 mL) The aqueous layer was basified to pH 12 using aqueous NaOH, and extracted with dichloromethane (3 x 500 mL). The combined organic layers were dried (MgSO^, 10 filtered and concentrated under reduced pressure to produce the title intermediate as a viscous, light brown oil. 'H-NMR (CDC13) 6 (ppm) 7.5-7.2 (m, 5H, CeHs), 3.7 (s, 2H, CH2Ph), 3.45 (broad s, 2H, CH-NBn), 2.7-2.6 (dd, 2H, CH2CO), 2.2-2.1 (dd, 2H, CH2CO), 2.1-2.0 (m, 2H, CH2CH2), 1.6 (m, 2H, CH2CH2). (m/z): [M+H]+ calcd for Ci4HnNO 216.14; found, 216.0. 15 b. Preparation of 3-oxo-8-azabicyclor3.2.noctane-8-carboxylic acid terr-butyl ester To a solution of 8-benzyI-8-azabicyclo[3.2.1]octan-3-one (75 g, 0.348 mol) in EtOAc (300 mL) was added a solution of di-tert-butyl dicarbonate (83.6 g, 0.383 mol, 1.1 eq) in EtOAc (300 mL). The resulting solution and rinse (100 mL EtOAc) was added to a 1 L Parr hydrogenation vessel containing 23 g of palladium hydroxide (20 wt.% Pd, 20 dry basis, on carbon, -50% wet with water; e.g. Pearlman's catalyst) under a stream of nitrogen. The reaction vessel was degassed (alternating vacuum and N2 five times) and pressurized to 60 psi of H2 gas. The reaction solution was agitated for two days and recharged with H2 as needed to keep the H2 pressure at 60 psi until the reaction was complete as monitored by silica thin layer chromatography. The black solution was then 25 filtered through a pad of Celite® and concentrated under reduced pressure to yield the title intermediate quantitatively as a viscous, yellow to orange oil. It was used in the next step without further treatment. !H NMR (CDC13) 6(ppm) 4.5 (broad, 2H, CH-NBoc), 2.7 (broad, 2H, CH2CO), 2.4-2.3 (dd, 2H, CH2CH2), 2.1 (broad m, 2H, CH2CO), 1.7-1.6 (dd, 2H, CH2CH2), 1.5 (s, 9H, (CH3)3COCON)). 37 WO 2005/100350 PCT/US2005/011393 l«r c. Preparation of (lS3R,5R}-3-arawo-8-azabicvclo\3.2Aloctane-8-carboxvlic acid tert- butyl ester To a solution of the product of the previous step (75.4 g, 0.335 mol) in methanol (1 L) was added ammonium formate (422.5 g, 6.7 mol), water (115 mL) and 65 g of 5 palladium on activated carbon (10% on dry basis, -50% wet with water; Degussa type E101NE/W) under a stream of Na while stirring via mechanical stirrer. After 24 and 48 hours, additional portions of ammonium formate (132g, 2.1 mol) were added each time. Once reaction progression ceased, as monitored by anal. HPLC, Celite® (>500g) was added and the resulting thick suspension was filtered and then the collected solid was 10 rinsed with methanol (~500 mL). The filtrates were combined and concentrated under reduced pressure until all methanol had been removed. The resulting cloudy, biphasic solution was then diluted with 1M phosphoric acid to a final volume of ~1.5 to 2.0 L at pH 2 and washed with dichloromethane (3 x 700 mL). The aqueous layer was basified to pH 12 using 40% aq. NaOH, and extracted with dichloromethane (3 x 700 mL). The 15 combined organic layers were dried over MgSC>4, filtered, and concentrated by rotary evaporation, then high-vacuum leaving 52 g (70%) of the title intermediate, commonly 7V-Boc-e« 20 1H, CHNH2), 2.1-2.05 (m, 4H), 1.9 (m, 2H), 1.4 (s, 9H, (CH3)3OCON), 1.2-1.1 (broad, 2H). (m/z): [M+H]+ calcd for CnH^NaOa) 227.18; found, 227.2. Analytical HPLC (isocratic method; 2:98 (A:B) to 90:10 (A:B) over 5 min): retention time = 3.68 min. d. Preparation of l-isopropvl-2-oxo-1.2-dihvdroquinoline-3-carboxvlic acid First, acetone (228.2 mL, 3.11 mol) was added to a stirred suspension of 2-25 aminophenylmethanol (255.2 g, 2.07 mol) and acetic acid (3.56 mL, 62 mmol) in water (2 L) at room temperature. After 4 h, the suspension was cooled to 0 °C and stirred for an additional 2.5 h and then filtered. The solid was collected and washed with water and the wet solid cooled and dried by lyophilisation to yield 2,2,-dimethyl-l ,4-dihydro-2fiT-benzo[l,3]oxazine (332.2 g, 98 %) as an off-white solid. }H NMR (CDC13; 300MHz): 30 1.48 (s, 6H, C(CH3)2), 4.00 (bs, 1H, KB), 4.86 (s, 2H, CEfe), 6.66 (d, 1H, ArH), 6.81 (t, 1H, ArH), 6.96 (d, 1H, ArH), 7.10 (t, 1H, ArH). A solution of 2,2,-dirnethyl-l,4-dihydro-2F-benzo[l,3]oxazine (125 g, 0.77 mol) in THF (1 L) was filtered through a scintillation funnel and then added dropwise via an 38 WO 2005/100350 PCT/US2005/011393 V addition funnel, over a period of 2.5 h, to a stirred solution of 1.0 M LiAffiU in THF (800 mL) at 0 °C. The reaction was quenched by slow portionwise addition of Na2SO4-10H2O (110 g), over a period of 1.5 h, at 0 °C. The reaction mixture was stirred overnight, filtered and the solid salts were washed thoroughly with THF. The filtrate was 5 concentrated under reduced pressure to yield 2-isopropylaminophenyhnethanol (120 g, 95 %) as a yellow oil. 'HNMR (CDC13; 300MHz): 1.24 (d, 6H, CH(CH3)2), 3.15 (bs, IH, OH), 3.61 (sept, IH, CH(CH3)2), 4.57 (s, 2H, CH,), 6.59 (t, IH, ArH), 6.65 (d, IH, ArH), 6.99 (d, IH, ArH), 7.15 (t, IH, ArH). Manganese dioxide (85 % 182.6 g, 1.79 mol) was added to a stirred solution of 10 2-isopropylaminophenyhnethanol (118 g, 0.71 mol) in toluene (800 mL) and the reaction mixture was heated to 117 °C for 4 h. The reaction mixture was allowed to cool to room temperature overnight and then filtered through a pad of Celite which was eluted with toluene. The filtrate was concentrated under reduced pressure to yield 2-isopropylaminobenzaldehyde (105 g, 90 %) as an orange oil. !H NMR (CDC13; 15 300MHz): 1.28 (d, 6H, CH(CH3)2), 3.76 (sept, IH, CH(CH3)2), 6.65 (t, IH, ArH), 6.69 (d, IH, ArH), 7.37 (d, IH, ArH), 7.44 (t, IH, ArH), 9.79 (s, IH, CHO). 2,2-Dimethyl-[l,3]dioxane-4,6-dione, commonly Meldrum's acid, (166.9 g, 1.16 mol) was added to a stirred solution of 2-isopropylaminobenzaldehyde (105 g, 0.64 mol), acetic acid (73.6 mL, 1.29 mol) and ethylenediamine (43.0 mL, 0.64 mol) in 20 methanol (1 L) at 0 °C. The reaction mixture was stirred for 1 h at 0 °C and then at room i' temperature overnight. The resulting suspension was filtered and the solid washed with methanol and collected to yield the title intermediate, l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid (146 g, 98 %) as an off-white solid. *H NMR (CDC13; 300MHz): 1.72 (d, 6H, CH(CHj)2), 5.50 (bs, IH, CH(CH3)2), 7.44 (t, IH, ArH), 7.75-7.77 25 (m, 2H, ArH), 7.82 (d, IH, ArH), 8.89 (s, IH, CH). e. Preparation of (IS,3R,5J?)-3-ri-isopropvl-2-oxo-l,2-dihydroquinoHne-3-carbonvl)amino]-8-azabicvclor3.2.11octane-8-carboxvlic acid tert-butvl ester Thionyl chloride (36.6 mL, 0.52 mol) was added to a stirred suspension of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid (80 g, 0.35 mol) in toluene 30 (600 mL) at 85 °C and the reaction mixture then heated to 95 °C for 2 h. The reaction mixture was cooled to room temperature and then added over 25 min to a vigorously stirred biphasic solution of (l^S^jS^-S-amino-S-azabicyclop^.lJoctane-acid tert-bntyl ester (78.2 g, 0.35 mol) and sodium hydroxide (69.2 g, 1.73 mol) in 39 WO 2005/100350 PCT/US2005/011393 toluene/water (1:1) (1L) at °C. After 1 h, the layers were allowed to separate and the organic phase concentrated under reduced pressure. The aqueous phase was washed with EtOAc (1 L) and then (500 mL) and the combined organic extracts used to dissolve the concentrated organic residue. This solution was washed with 1M H3PO4 (500 mL), sat. 5 aq. NaHCO3 (500 mL) and brine (500 mL), dried over MgSO4, filtered and concentrated under reduced pressure to yield the title intermediate (127.9 g, approx. 84 %) as a yellow solid. JHNMR (CDC13): 1.47 (s, 9H), 1.67 (d, 6H), 1.78-1.84 (m, 2H), 2.04-2.18 (m, 6H), 4.20-4.39 (m, 3H), 5.65 (bs, 1H), 7.26 (dd. 1H), 7.63 (m, 2H), 7.75 (dd, 1H), 8.83 (s, 1H), 10.63 (d, 1H). 10 f. Preparation of l-isopropvl-2-oxo-l,2-dihvdroquinoline-3-carboxylic acid t(lS,3R,5R)-8-azabicyclof3.2.1]oct-3-vl) amide TFA (300 mL) was added to a stirred solution of the product of the previous step (127.9 g) in CBbClz (600 mL) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h and then concentrated under reduced pressure. The oily 15 brown residue was then poured into a vigorously stirred solution of ether (3 L) and a solid precipitate formed immediately. The suspension was stirred overnight and then the solid collected by filtration and washed with ether to yield the title intermediate as its trifluoroacetic acid salt (131.7 g, 86% over two steps) as a light yellow solid. 1H NMR (CDC13): 1.68 (d, 6H), 2.10 (d, 2H), 2.33-2.39 (m, 4H), 2.44-2.61 (m, 2H), 4.08 (bs, 2H), 20 4.41 (m, 1H), 5.57 (bs, 1H), 7.31 (m. 1H), 7.66 (m, 2H), 7.77 (d, 1H), 8.83 (s, 1H), 9.38 (bd, 2H), 10.78 (d, 1H). g. Preparation of 3-hydroxv-3'-{[l-isopropvl-2-oxo-l,2-dihydroquinolin-3-yl)carbonvl]amino>spirofazetidine-l,8'-(liS'.3J?.5J?V8-azabicyclor3.2.11octane (Intermediate (V) with R1 = H. R2 = isopropvl) 25 2-Bromomethyloxirane (10.72 mL, 129.5 mmol) was added to a stirred solution of 1 -isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-&-aza-bicyclo[3.2.1]oct-3-yl}amide trifluoroacetic acid salt (14.65 g, 43.2 mmol) in ethanol (150 mL) at room temperature. The reaction mixture was stirred for 36 h, at which time a solid precipitate formed. The solid was collected by filtration and washed with ethanol 30 (70 mL) to yield the title intermediate as the bromide salt (8.4 g). (m/z): [M]+ calcd for C23H30N3O3 396.23; found, 396.5. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.13min. 40 WO 2005/100350 PCT/US2005/011393 f h. Preparation of l-isopropvl-2-oxo-l,2-dihvdroquinolinone-3-carboxylic acid B^. 1 loct-3-vll amide 3-Hydroxy-3'-{[l-isopropyl-2-oxo-l)2-dihydroquinolin-3- yl)carbonyl]amino}spiro[azetidine-l,8'-(l'Sr,3^,5JR)-8-aza-bicyclo[3.2.1]octane bromide 5 (678 mg, 1.4 mmol) was dissolved in ethanol (10 mL), and then methylamine (41% solution in water) (5 1 0 \iL, 8.0 mmol) was added. The mixture was heated at 80 °C for 16 h, and then concentrated under reduced pressure to give the title intermediate as a crude oil, which was used directly in the following step. i. Synthesis of l-isopropvl-2-oxo-l,2-dihvdroquinolinone-3-carboxylic acid {(\S3R5R)-10 8- f 2-hvdroxy-3 -(methanesulfonvl-methvl-amino)propyll -8-azabicvclo [3.2. 1 ]oct-3 -yl) amide The product of the previous step was dissolved in dichloromethane (10 mL), and then i38-diazabicyclo[5.4.0]undec~7-ene (763 \\L, S.lmmol) was added, and the mixture was stirred under nitrogen and cooled to 0 °C. Methanesulfonylchloride (132 pL, 15 1 .7 mmol) was added and the mixture was stirred at 0 °C for 30 min. The reaction was quenched by the addition of water, and concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1 : 1) (1 0 mL) and purified by HPLC chromatography. The purified fractions were lyophilized yielding the title compound as the trifluoroacetic acid salt (340 mg). (m/z): [M+H]+ calcd for CajHseWsS, 505.25; 20 found, 505.4. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.17 min. Example 2: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoImone-3-carboxylic acid {(l,S;3#,5/0-8-[2-methoxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yI}amide 25 a. Preparation of 3-methoxv-3'- (IT -isopropvl-2-oxo-l ,2-dihvdroquinolin-3- (Intermediate (V) with R1 - H. R2 = isopropvl. R3 =methvl) Potassium-tert-butoxide (1.63 g, 14.5 mmol) was added to a stirred suspension of 3-hydroxy-3'-{[l-isopropyl-2-oxo-l,2-dihydroquinolin-3- 30 yl)carbonyl]amino}spiro[azetidine-l,8'-(lJSl,3^,5J?)-8-azabicyclo[3.2.1]octane bromide (3.45 g, 7.25 mmol) in dichloromethane (100 mL) at room temperature. After 2 min, methyl iodide (0.477 mL, 7.61 mmol) was added to the reaction mixture. After 30 min, water (2 mL) was added to quench the reaction and the reaction mixture concentrated WO 2005/100350 PCT/US2005/011393 trifluoroacetic acid salt (2.1 g). (m/z): [Mf calcd for C24H32N3C>3, 410.24; found 410.5. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.36min. b. Preparation of l-isopropvl-2-QXo-l,2-dihvdroquinolinone-3-cafboxvlic acid |(l)$',3^?.5j?)-8-r2-methoxv-3-methvlaminopropyn-8-azabicvclo[3.2.noct-3-vllaniide 5 3-Methoxy-3'-{[l-isopropyl-2-oxo-l,2-dihydroquinolin-3- yl)carbonyl]amino}spiro[azetidine-l,8l-(l»S'33-R,5^)-8-aza-bicyclo[3.2.1]octane trifluoroacetic acid salt (410 mg, 0.84 mmol) was dissolved in ethanol (10 mL), and then methylamine (41% solution in water, 320 uL, 5 mmol) was added. The mixture was heated at 80 °C for 16 h, and then concentrated under reduced pressure to give the product 10 as a crude oil which was used directly in the following step. c. Synthesis of l-isopropvl-2-oxo-l,2-dihvdroquinolinone-3-carboxvlicacid |(1S,3R.5R)- 8-r2-methoxv-3-(methanesulfonvl-m6thvl-amino)propyl1-8-azabicyclo[3.2.11oct-3- yllamide The product of the previous step (53.6 nig, 0.12 mmol) was dissolved in 15 dichloromethane (1.0 mL), and then l,8-diazabicyclo[5.4.0]undec-7-ene (89,7 \\L, 0.6 mmol) was added, and the mixture was stirred under nitrogen and cooled to 0°C. Methanesulfonylchloride (18.6 JJ.L, 0.24 mmol) was added and the mixture was stirred at 0°C for 30 min. The mixture was quenched by the addition of water, and concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) 20 (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (45.8 mg). (m/z): [M+H]+ calcd for C26H38N4O5S, 519.27; found 519.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.72 min. Example 3: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxyIic 25 methoxypropyl]-8-azabicycIo[3.2.1]oct-3-yI}amide a. Preparation of l-isopropvl-2-oxo-1.2-dihydroqurnolme-3-carboxvlic acid 8-(2-methoxv-3-[(pvridin-3-vlmethvI)arDino]propvl>-8-azabicvclo[3.2.1]oct-3-vl> amide 3 -Methoxy-3 '- { [ 1 -isopropyl-2-oxo- 1 ,2-dihydro quinolin-3- 30 yl)carbonyl]amino}spiro[azetidine-l581-(15',3^,5JR)-8-aza-bicyclo[3.2.1]octane (410 mg, 0.84 mmol) was dissolved in ethanol (10 mL), and then 3-aminomethylpyridine (153 pL, 1.5 mmol) was added. The mixture was heated at 60 °C for 16 h, and then concentrated under reduced pressure to give the title intermediate as a crude oil which was used directly in the next step. 42 WO 2005/100350 PCT/US2005/011393 W - b. Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxvlic acid ((l^S^S^VS- r3-(methanesulfonvl-pyridin-3-vlmethvl-amiiioV2-methoxvpropvl]-8-azabicyclo[3 .2. 1 loct-3-yll amide The product of the previous step (102.6 mg, 0.2 mmol) was dissolved in 5 dichloromethane (1 .0 mL ) and then l,8-diazabicyclo[5.4.0]undec-7-ene (1 19.6 JJ.L, 0.8 mmol) was added, and the mixture was stirred under nitrogen and cooled to 0°C. Methanesulfonylchloride (30. 1 uL, 0.4 mmol) was added and the mixture was stirred at 0°C for 30 min. The reaction was quenched by the addition of water, and the mixture was concentrated to dryness under reduced pressure. The product was taken up in acetic *^ 1 0 acid/water (1 : 1 ) (1 .5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (63.5 mg). (m/z): [M+H]+ calcd for CaiKUiNsOsS, 596.29; found 596.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.35 min. Example 4: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3- 15 carboxylic acid {(l1S',3/?,5^)-8-[2-hydroxy-3-methanesulfonylamino)propyl]-8- azabicyclo[3.2.1]oct-3-yl}amide a. Preparation of l-isopropyl-2-oxo-l,2-dihvdroquinoline-3 -carboxylic acid 8-[3-(l,3-dioxo-L3-dihvdroisoindol-2-vl)-2-hydroxypropy]-8-azabicvclor3.2.noct-3- yl} amide 20 l-Isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxyh'c acid {(lS,3R,5K)-8-aza- bicyclo[3.2.1]oct-3-yl} amide (3.39 g, lOmmol) was dissolved in ethanol (40 mL), and then2-oxiranyhnethylisoindole-l,3-dione, commonly, epoxypropylphthalimide, (3.05 g, 15 mmol) was added. The mixture was heated at 80 °C for 36 h, cooled, and then concentrated under reduced pressure. The resulting oil was flash chromatographed (SiC>2, 25 eluting with a 9:1 solution of dichloromethane/methanol) to give the title intermediate (4.95 g) as a white solid which was used directly in the next step. b. Preparation of l-isopropyl-2-oxo-l,2-dihvdroquinolinone-3-carboxvlic acid The product of the previous step (4.95 g, 9.13 mmol) was dissolved in ethanol 30 (40 mL), and then hydrazine (860 |xL, 27.4 mmol) was added. The mixture was refluxed for 16 h, and then cooled to room temperature. The mixture was filtered, and the filtrate concentrated to give the title intermediate as a crude oil, which was used directly without further purification. \VO 2005/100350 PCT/US2005/011393 V c. Synthesis of l-isopropvl-2-oxo-l,2-dihydroquiiiolinone-3-carboxylic acid ((1S,3R,5R)-8-r2-hvdroxy-3-methanesulfonylamino)propvl]-8-azabicvclor3.2.1]oct-3-vl> amide The product of the previous step (82 mg, 0.2 romol) was dissolved in dichloromethane (1.0 mL), then l,8-diazabicyclo[5.4.0]undec-7-ene (60 \iL, 0.4 mmol) 5 was added, and the mixture was stirred under nitrogen and cooled to -78°C. Methanesulfonylchloride (15.5 joL, 0.2 mmol) was added and the mixture was stirred and allowed to warm to room temperature over 30 min. The reaction was quenched by the addition of water, and the mixture was concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC 10 chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (70.7 mg). (m/z): [M+H]+ calcd for CwHw^OsS, 491.23; found 491 .2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.43 min. Example 5: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic 1 5 hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl} amide a. Preparation of l-isopropvl-2-oxo-l,2-dihvdroquinoline-3-carboxylic acid 8-{2-hydroxy-3-[(pyridm-3-ylmethyl')arnino]propyl}-8-azabicvclo[3.2.1]oct-3-vn amide 3-Hydroxy-3 '- { [ 1 -isopropyl-2-oxo- 1 ,2-dihydroquinolin-3 - yl)carbonyl]amino}spiro[azetidine-l,8'-(liS',3JR,5J?)-8-aza-bicyclo[3.2.1]octane bromide 20 (505 mg, 1.27 mmol) was dissolved in ethanol (10 mL), and then 3-(aminomethyl)-pyridine (193 [iL, 1.9 mmol) was added. The mixture was heated at 80 °C for 16 h, and then concentrated under reduced pressure to give the title intermediate as a crude oil which was used directly in the following step. b. Synthesis of l-isopropyl-2-oxo-lT2-dihydroquinoline-3-carboxylic acid |(liS',3J?,5j?)-8- 25 [3-(memaiiesulfonyl-pyridin-3-ytoiemyl-arninoV2-hydroxypropyl]-8- azabicyclo[3 .2. l]oct-3-yl} amide The product of the previous step (42.5 mg, 0.08 mmol) was dissolved in dichloromethane (1.0 mL), l,8-diazabicyclo[5.4.0]undec-7-ene (74.8 \\L, 0.5 mmol) was added, and the mixture was stirred under nitrogen and cooled to 0 °C. 30 Methanesulfonylchloride (6.1 uL, 0.08 mmol) was added and the mixture was stirred at 0 °C for 30 min. The reaction was quenched by the addition of water, and concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1 :1) (1 .5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (22.8 mg). (m/z): [M+H]+ calcd 44 WO 2005/100350 PCT/US2005/011393 V for CsoHagNsOsS, 582.28; found 582.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.78 min. Example 6: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(l^SJZjS^-S-J^H-hydroxy-S-Cmethanesulfonyl-methyl- 5 amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide a. Preparation of dS'j-l-fbenzyl-niethvl-aminol-B-chloropropan^-ol N-Benzylmethylamine (13.95 mL, 108.lmm.ol) and (,S)-2-chloromethyloxirane5 commonly (iS)-epichlorohydrin (8.48 mL, 108.1 mmol) were dissolved in hexane (40 mL), and stirred for 16 h. The solution was then flash chromatographed (SiCh, eluting with 10 10 % methanol/90 % dichloromethane). Fractions containing product were concentrated to give the title intermediate as an oil (19.7 g). ^-NMR (DMSO d6, 299.96 MHz): 5 (ppm) 2.01 (s, 3H), 2.2-2.4 (m, 2H), 3.21-3.5 (m, 3H), 3.53-3.6 (m, 1H), 3.65-3.75 (m, 1H), 4.95 (d, 1H), 7.0-7.25 (m, 5H). (m/z): [M+H]+ calcd for CnHi6ClNO, 214.10; found 214.1. 15 b. Preparation of ((iS^-3-chloro-2-hydroxvpropvDmethylcarbamic acid tert-lontyl ester (5^-l-(benzyl-methyl-amino)-3-chloropropan-2-oi (8.4 g, 39.3 mmol) was dissolved in ethyl acetate (75 mL), and then di-tert-butyl dicarbonate (9.3 g, 43.23 mmol) and palladium hydroxide (2.5 g) were added. The mixture was shaken for 12 h under hydrogen (60 atm). The mixture was filtered through a bed of Celite®, and concentrated 20 to dryness under reduced pressure. The resulting oil was filtered through silica, eluting with hexane, followed by dichloromethane, followed by diethyl ether. The ether layer was concentrated to give the title intermediate as an oil (7.1 g). JH-NMR (DMSO dt, 299.96 MHz): 5 (ppm) 1.35-1.46 (s, 9H), 2.81-2.85 (s, 3H), 2.95-3.1 (m, 1H), 3.3-3.6 (m, 3H), 3.67-3.85 (m, 1H), 5.25-5.4 (m, 1H). (m/z): [M+H-Bocf calcd for C9Hi8ClNO3, 25 123.10; found 123.1. c. Preparation of ((J?V2-hydroxv-3-|3-f(l-isopropvl-2-oxo-1.2-dihydroquinoline-3-carbonyl)amino]-('liS',3J?,5jR)-8-azabicvclo[3.2.11oct-8-vl)propvl)methvlcarbamicacid tert-butyl ester (((S)-3-chloro-2-hydroxypropyl)methylcarbamic acid toi-butyl ester (335 mg, 30 1.5 mmol) and l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-&-aza-bicyclo[3.2.1]oct-3-yl}amide (339 mg, 1.0 mmol), were dissolved in methanol (5 mL) and then JVjJV-diisopropylethylamine (523 pL, 3.0 mmol) was added. The mixture was heated at 80 °C for 16h, and then concentrated under reduced pressure. The resulting oil 45 WO 2005/100350 PCT/US2005/011393 V was flash chromatographed (SiOa, eluting with 10 % methanol/90 % dichloromethane). Fractions containing product were concentrated to give the title intermediate as a white solid (0.5 g). (m/z): [M+Hf calcd for C^BUaN^s, 527.33; found 527.6. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.75 min. d. Preparation of l-isopropyl-2-oxo-l,2-dihvdroquinoline-3-carboxvh'c acid 8-f(^-2-hydroxy-3-methylaminopropyl]-8-azabicyclo[3.2.11oct-3-vl>amide ((J?)-2-hydroxy-3- {3-[(l -isopropyl-2-oxo-l ,2-dihydroquinoline-3- tert-butyl ester (575 mg, 1.09 mmol) was dissolved in dichloromethane (5 mL) and then 10 trifluoroacetic acid (5 mL) was added slowly. The mixture was stirred for 30 min, and then concentrated under reduced pressure. The resulting oil was triturated with diethyl ether, and then filtered. The precipitate was dried under vacuum to give the title intermediate as the trifluoroacetic acid salt (0.68 g). (m/z): [M+H]+ calcd for €248.34^403, 427.27; found 427.2. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 3.40 15 min. e. Synthesis of l-isopropvl-2-oxo-1.2-dibvdroquinoline-3-carboxvlic acid [f^V2-hydroxy-3-(methanesulfonyl-methvl-amino')propvl1-8-aza-bicyclo[3.2.1]oct-3-yll amide l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylicacid {(lS,3R,5R)-S-[(S)-2-20 hydroxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (1.03 g, 1.57 mmol) was dissolved hi dichloromethane (6.0 mL), l,8-diazabicyclo[5.4.0]undec-7-ene (747 (J.L, 5.0 mmol) was added, and the mixture was stirred under nitrogen and cooled to 0 °C. Methanesuifonylchloride (124.4 uL, 1 .6 mmol) was added and the mixture was stirred at 0 °C for 30 min. The reaction was quenched by the addition of water, and the mixture 25 was concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (5 mL) and purified by HPLC chromatography. The purified fractions were lyophihzed to give the title compound as the trifluoroacetic acid salt (0.38 g). (m/z): [M+H]+ calcd for CksBfo^UOsS, 505.25; found 505.4. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.17 min. Free base: 'H-NMR (DMSO d6, 30 299.96 MHz): 5 (ppm) 1.40-1.68 (d, 6H), 1.81-2.02 (br s, 4H), 2.02-2.18 (br m, 2H), 2.22-2.36 (d, 2H), 2.78-2.90 (2 s, 6H), 2.91-3.04 (m, 1H), 3.10-3.30 (m, 4H), 3.61-3.78 (br s, 1H), 4.02-4.17 (m, 1H), 4.71-4.79 (br s, 1H), 5.2-5.8 (br s, 1H), 7.3-7.4 (t, 1H), 7.67-7.78 (t, 1H), 7-82-7.94 (d, 1H), 7.98-8.02 (d, 1H), 8.80 (s, 1H), 10.37-10.40 (d, NH). 46 WO 2005/100350 PCT/US2005/011393 Example 7: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxy!ic acid{(liS',3JR,55)-8-[(5)-2-hydroxy-3-(methanesulfonyl-methyl-ainino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl} amide 5 Following the procedure of Example 6, with the substitution of (R)-2- chloromethyloxirane for (5)-2-chloromethyloxirane in step a, the following intermediates and the title compound were prepared. (J?>l-(benzyl-methyl-amino)-3-chloropropan-2-ol: ^-NMR (DMSO d6, 299.96 MHz): 5 (ppm) 2.01 (s, 3H), 2.2-2.4 (m, 2H), 3.21-3.5 (m, 3H), 3.53-3.6 (m, 1H), 3.65-10 3.75 (m, 1H), 4.95 (d, 1H), 7.0-7.25 (m, 5H). (m/z): [M+H]+ calcd for CnHi6ClNO5 214.10; found 214.1. ((jR)-3-chloro-2-hydroxy-propyl)methylcarbamic acid tert-butyl ester: ^-NMR (DMSO d6, 299.96 MHz): & (ppm) 1.35-1.46 (s, 9H), 2.81-2.85 (s, 3H), 2.95-3.1 (m, 1H), 3.3-3.6 (m, 3H), 3.67-3.85 (m, 1H), 5.25-5.4 (m, 1H). (m/z): [M+H-Bocf calcd for 15 C9Hi8ClNO3, 123.10; found 123.1. ((S)-2-hydroxy-3 - {3- [( 1 -isopropyl-2-oxo- 1 ,2-dihydroquinoline-3-carbonyl)amino]-(liS',37?,5JR)-8-azabicyclo[3.2.1]oct-8-yl}propyl)methylcarbamicacid tert-butyl ester: (m/z): [M+H]+ calcd for C29H42N4O5, 527.33; found 527.6. Retention time (anal. HPLC: 2-50% MeCN/H20 over 5 min) = 4.75 min. 20 1 -isopropyl-2-oxo- l,2-dihydroquinoline-3-carboxylic acid {(15,3jR:,5J?)-8-[(R)-2- hydroxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide: (m/z}: [M+H]+ calcd for C24H34N4O3, 427.27; found 427.2. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 3 .40 min. l-isopropyl-2-oxo-l,2-dihydroqurnoline-3-carboxylicacid {(liS',3^!,5JR)-8-[()y)-2-25 hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl} amide: (m/z): [M+H]+ calcd for €25^6^058, 505.25; found 505.4. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.17 min. Example 8: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3- carboxylic acid {(l^,3i?,5^)-8-[2-hydroxy-3-[methyl-(pyridine-4- 30 carbonyl)amino]propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide 1 -isopropyl-2-oxo- l,2-dihydroquinolinone-3-carboxylic acid hydroxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (71 mg, 0.17 mmol) was dissolved in DMF (0.5 mL), and then JV,7V-diisopropylethylamine (88,9 uL, 47 WO 2005/100350 PCT/US2005/011393 V 0.51 mmol) was added. Next, a solution of isonicotinic acid (41.8 mg, 0.34 mmol) and PyBOP (177 mg, 0.34 mmol) in DMF was added. The resulting mixture was shaken at room temperature for 30 min, and then concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC 5 chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (30.8 mg). (m/z): [M+H]+ calcd for C3oH37N5O43 532.29; found 532.7. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.11 min. Example 9: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3- carboxylic acid {(l^S^^^-S-Jl-hydroxy-S-Ifpyridine^- 10 carbonyl)amino]propyl]-8-azabicyclo [3.2.1] oct-3-yl} amide l-Isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylicacid {(lS,3R,5R)-8-[2-hydroxy-3-aminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (82.4 mg, 0.2 mmol) was dissolved in DMF (0.5 mL), and then W, AMiisopropylethylamine (69.7 \\L, 0.4 mmol) was added. Next, a solution of isonicotinic acid (49.2 mg, 0.4 mmol) and PyBOP 15 (208.2 mg, 0.4 mmol) in DMF was added. The resulting mixture was shaken at room temperature for 30 min, and then concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (82.1 mg). (m/z): [M+Hf calcd for C29H35N5O4, 518.28; 20 found 518.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.20 min. Example 10: Synthesis of l-isopropyI-2-oxo-l,2-dihydroquinoIine-3-carboxylic acid {(lS,3R,5K)-8- [3-(acetyl-methyl-amino)-2-methoxypropyl]-8-azabicyclo[3.2.1]oct-3-yI}amide 1 -Isopropyl-2-oxo-l,2-dilrydroquinolinone-3-carboxylic acid {(l«S,3.R,5.R)-8-[2-25 methoxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (53.6 mg, 0.12 mmol) was dissolved hi DMF (1.0 mL), and then N.N-diisopropylethylamine (104. uL, 0.6 mmol) was added. The mixture was stirred under nitrogen and cooled to 0°C. Acetyl chloride (17.1 uL, 0.24 mmol) was added and the mixture was stirred at 0°C for 30 min, and then concentrated to dryness under reduced pressure. The product was taken up in 30 acetic acid/water (1:1) (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (40.4 mg). (m/z): [M+H]+ calcd for C27H3sN4O4, 483.30; found 483.2. Retention tune (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.54 min. 48 WO 2005/100350 PCT/US2005/011393 V Example 11. Synthesis of l-isopropyl~2-oxo-l,2-dihydroquinoIinone-3-carboxylic acid {(15,31?,510-8-[2Hmethoxy-3-[methyI-(pyriduie-4-carbonyl)ammo]propyl]-8-azabicycIo[3.2.1]oct-3-yl}amide l-Isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylicacid {(lS,3R,5K)-8-[2-5 methoxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (53.6 mg, 0.12mmol) was dissolved in DMF (1.0 mL), and then TV; N-ffisopropylemylamine (104.5 ^iL, 0.6 mmol) was added. The mixture was stirred under nitrogen and cooled to 0°C. Isonicotinyl chloride (42.7 mg, 0.24 mmol) was added and the mixture was stirred at 0 °C for 30 min, and then concentrated to dryness under reduced pressure. The product was 10 taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (54.4 mg). (m/z): [M+H]+ calcd for C31H39N5O4, 546.31; found 546.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.29 min. Example 12. Synthesis of l-isopropyI-2-oxo-l,2-dihydro-quinoline-3- 15 carboxylic acid {(l£,3^,5^)-8-[3-(acetyl-pyridm-3-ylmethyl-amino)-2- methoxypropyI]-8-azabicyclo[3.2.1]oct-3-yl}amide l-Isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid (8-{2-methoxy-3-[(pyridin-3-yhnethyl)amino]propyl} -8-azabicyclo[3.2.1 ]oct-3 -yl)amide (102.6 mg, 0.2 mmol) was dissolved in DMF (1.0 mL), and then 7V,AT-diisopropylethylamine (139.4 20 jiL, 0.8 mmol) was added. The mixture was stirred under nitrogen and cooled to 0°C. Acetyl chloride (28.5 fiL, 0.4 mmol) was added and the mixture was stirred at 0°C for 30 min, and then concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (33.2 25 mg). (m/z): [M+Hf calcd for CaaHuNsO^ 560.33; found 560.4. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.27 min, Example 13. Synthesis of l-isopropyl-2-oxo-l,2-dihydro-quinoline-3-carboxylic acid {(l£,3U,5^)-8-[3-acetylamino-2-hydroxypropyI]-8-azabicy clo [3.2.1] oct-3-yl} amide 30 l-Isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(lS,3R,5R)-8-[2- hydroxy-3-aminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (82.4 mg, 0.2mmol) was dissolved in DMF (0.5 mL), and then JV*,7V-diisopropylethylamine (69.7 jiL, 0.4 mmol) was added. Next, a solution of acetic acid (22.7 uL, 0.4 mmol) and PyBOP (208.2 mg, 0.4 mmol) in DMF was added. The resulting mixture was shaken at room temperature for 49 WO 2005/100350 PCT/US2005/011393 30 min, and then concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1 .5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title compound as the trifluoroacetic acid salt (79.2 mg). (m/z): [M+H]+ calcd for C^Hs^lSLA, 455.27; found 455.2. Retention 5 time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.33 min. Example 14. Synthesis of l-isopropyl-2-oxo-l,2-dihydro-quinoline-3-carboxylic acid {(lS£R,5R)-8- [3-(acetyl-methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yI}amide l-Isopropyl-2-oxo-l,2-dmydroqumolinone-3-carboxylicacid {(lS,3R,5R)-B-[2-10 hydroxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (43 mg, 0.1 mmol) was dissolved in DMF (1.0 mL), and then J\T,2V-diisopropylethylamine (87.1 pL, 0.5 mmol) was added. The mixture was stirred under nitrogen and cooled to 0 °C. Acetyl chloride (17.8 uL, 0.25 mmol) was added and the mixture was stirred at 0 °C for 30 min. The mixture was concentrated to dryness under reduced pressure. The product was taken 15 up in acetic acid/water (1 : 1) (1 .5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title intermediate as the trifluoroacetic acid salt (17.1 mg). (m/z): [M+H]+ calcd for C26H36N4O4, 469.28; found 469.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.49 min. Example 15. Synthesis of l-isopropyl-2-oxo-l,2-dihydro-quinoline-3- 20 carboxylic acid {(15,3JR,5J?)-8-[3-(formyl-methyl-amino)-2-hydroxypropyl]-8- azabicyclo[3.2.1]oct-3-yI}amide l-Isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylicacid hydroxy-3-methylammopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (43 mg, 0.1 mmol) was dissolved in ethyl formate (1.0 mL). The mixture was heated at 65 °C for 16 h, and 25 men concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1 .5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title intermediate as the trifluoroacetic acid salt (22.7 mg). (m/z): [M+Hf calcd for Cis^N^A, 455.27; found 455.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.37 min. 50 WO 2005/100350 PCT/US2005/011393 V Example 16. Synthesis of l-isopropyl-2-oxo-l,2-dihydro-quinoline-3-carboxylic acid {(Itf^S^-S-p-formylammo-l-hydroxypropyri-S-azabicyclo[3.2.1]oct-3-yl}amide 1 -Isopropyl-2-oxo-l32-dihydroquinolinone-3-carboxylic acid {(lS,3R,5R~)-8-[2-5 hydroxy-3-aminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (41.2 mg, 0.1 mmol) was dissolved in ethyl formate (1.0 mL). The mixture was heated at 65°C for 16 h, and then concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title intermediate as the trifluoroacetic acid salt (37.5 mg). 10 (m/z): [M+Hf calcd for Ca^N^, 441.25; found 441.2. Retention time (anal. HPLC: 5-40% MeCN/H2O over 4 min) = 2.89 min. Example 17. Synthesis of l-isopropyl-2-oxo-l,2-dihydro-quinoIine-3-carboxylic acid {(l^S^jS^-S-f^-S-Cacetyl-methyl-amino)^-hydroxypropyl]-8-azabicycIo [3.2.1] oct-3-yl} amide 15 l-Isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-8-[(S)-2- hydroxy-3-methylaminopropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide (200 mg, 0.3 mmol) was dissolved in DMF (1.0 mL), and then W, Af-diisopropylethylamine (160.3 JJ.L, 0.92 mmol) was added. Next, a solution of acetic acid (17.3 pL, 0.3 mmol) and PyBOP (159 mg, 0.3 mmol) in DMF was added. The resulting mixture was shaken at room 20 temperature for 30 min, and then concentrated to dryness under reduced pressure. The product was taken up in acetic acid/water (1:1) (1.5 mL) and purified by HPLC chromatography. The purified fractions were lyophilized to give the title intermediate as the trifluoroacetic acid salt (130 mg). (m/z): [M+H]+ calcd for CzeHseN^, 469.28; found 469.5. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 3.94 min. 25 Example 18. Synthesis of l-isopropyl-2-oxo-l,2-dihydro-quinoline-3- carboxylic acid {(l£3^5tf)-84(^)-3 fr» WO 2005/100350 PCT/US2005/011393 (97.5 mg). (m/z): [M+H]+ calcd for C^Hs^C^, 455.27; found 455.2. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 3.87 min. Example 19: Synthesis of 5-bromo-l-isopropyl-2-oxo-l,2-dihydroquinoline-3- carboxylicacid{(15',3JR,5JR)-8-[(J?)-2-hydroxy-3-(methanesulfonyl-metliyl- 5 amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl} amide To a solution of l-isopropyl-2-oxo-l ,2-dihydroquinoline-3-carboxylic acid {(1^3^,5^)-8-[(^)-2-hydroxy-3-(methanesulfonyl-methyI-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl} amide (500 mg, 0.99 mmol) dissolved in a mixture of acetonitrile (5 mL) and acetic acid (10 mL) was added bromine (0.30 mL, 5.9 mmol). 1 0 The mixture was stirred at ambient temperature for 12 h, and concentrated under reduced pressure, yielding a pale yellow oily residue. The residue was dissolved in 20% acetonitrile in water (0.5% TFA) (5 mL), and purified by HPLC. The title compound was obtained as a major product and isolated as a tifluoroacetic acid salt (200 mg) ^-NMR (CD3OD): 5 (ppm) 8.64 (s, 1H), 7.97 (s, 1H), 7.72-7.70 (m, 2H), 4.18 (br m, 2H), 4.0 (br 15 s, 1H), 3.2-3.0 (m), 2.88 (s, 3H), 2.79 (s, 3H), 2.5-2.2 (m, 2H), 1.56 (d, 6H). (m/z): [M+H]+ calcd for C25H35BrN4O5S, 583.16; found 583.4. Retention time (anal. HPLC: 10-50% MeCN/H20 over 6 min) - 3.90 min. Example 20: Alternative synthesis o l,2-dihydroquinoline-3-carbonyl)amino]-(liS'^i?,5JR)-8-azabicyclo[3.2.1]oct-8- 20 yl}propyl)methylcarbamic acid tert-butyl ester a. Preparation of methyl-ffl-l-oxiranyhnethvlcarbamic acid te?-t-butyl ester ((jS)-3-chloro-2-hydroxypropyl)-methylcarbamic acid te;-t-butyl ester (2.23 g, 10 mmol) was dissolved in THF (30 mL), and then an aqueous sodium hydroxide solution (0.48 g in 10 mL water) was added. The mixture was stirred for 2 h. The mixture was 25 then concentrated under reduced pressure to remove most of the THF, and the remaining aqueous solution was extracted into ethyl acetate, washing with water. The product was dried over sodium sulfate, filtered and concentrated under reduced pressure to give the title intermediate as an oil (1.6 g). (m/z): [M+Na]+ calcd for C9Hi7NO3, 210.10; found 210.1. JH-NMR (DMSO d6, 299.96 MHz): 5 (ppm) 1.32-1.42 (s, 9H), 2.69-2.73 (m, 1H), 30 2.75-2.85 (s, 3H), 2.95-3.05 (br s, 1H), 3.10-3.15 (dm, 1H), 3.16-3.21 (d, 1H), 3.37-3.51 (m,2H). WO 2005/100350 PCT/US2005/011393 V b. Synthesis of ((J?V2-hvdroxv-3-(3-rd-isopropvl-2-oxo-1.2-dihvdroqiiiiioline-3-carbonYnamino]-aS.3J?.5J?V8-azabicvclor3.2.11oct-8-vl>proT>vnmethvlcarbamicacid tert-butyl ester Methyl-(5)-l-oxiranylmethylcarbamic acid tert-butyl ester (9.53 g, 51.1 mmol), 5 and l-isopropyl-2-oxo-l,2-dihydroqumoline-3-carboxylic acid {(lS,3#,5jR)-8-aza-bicyclo[3.2.1]oct-3-yl}amide (8.7 g, 25.6 mmol), were dissolved in methanol (100 mL). The mixture was heated at 80 °C for 2h, and then concentrated under reduced pressure. The resulting oil was taken up in ethyl acetate and washed with saturated aqueous sodium bicarbonate, followed by saturated aqueous sodium chloride. The organics were dried 10 over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting oil was flash chromatographed (SiOa, eluting with 10% methanol/90% dichloromethane). Fractions containing product were concentrated to give the title intermediate as a white solid (13.5 g). (m/z): [M+H]+ calcd for C29H42N4O5, 527.33; found 527.6. Retention time (anal. HPLC: 2-50% MeCN/H2O over 5 min) = 4.75 min. 15 Example 21: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3- carboxylic acid {(15,3/?,5JR)-8-[2-hydroxy-3-(methanesulfonyl-ethylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide Following Hie procedure of Example 1 replacing methylamine with ethylamine in step h, the title compound was prepared, (m/z): [M+H]+ calcd for C26H38N4OsS, 519.28; 20 found 519.2. Retention time (anal. HPLC: 10-70% MeCN/H2O over 6 min) = 2.91 min. Example 22: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3- carboxylicacid{(15,3jR,5i?)-8-[(/i:)-2-liydroxy-3-(methanesulfonylamino)- propyl]-8-azabicyclo[3.2.1]oct-3-yI}amide al. Preparation of (>SV2-oxiranvlmethylisoindole-l,3-dione 25 To a cold solution of (S)-l-oxiranylmethanol (5 g, 67.5 mmol) and phthalimide (9.9 g, 67.3 mmol) in tetrahydrofuran (200 mL) in ice bath was added triphenylphosphine (17.9 g, 68.2 mmol) and diethyl azodicarboxylate (12.3 g, 70.6 mmol). The mixture was stirred at 0°C for 2 h and at ambient temperature for 48 h. The mixture was concentrated under vacuum, and the oily residue was purified by flash silica column chromatography, 30 yielding the desired product (10.1 g) as pale yellow solid: Rf = 0.51 in 1:1 EtOAc/hexane. ^-NMR (CDC13, 300MHz): 6 (ppm) 7.76-7.64 (m, 2H), 7.63-7.60 (m, 2H), 3.9-3.8 (dd, 1H), 3.70-3.65 (dd, 1H), 3.15 (m, 1H), 2.70-2.67 (dd, 1H), 2.58-2.55 (dd, 1H). 53 WO 2005/100350 PCT/US2005/011393 bl. Synthesis of l-isopropyl-2-oxo-l,2-dmydroqumolmone-3-carboxvlic acid azabicyclo[3 .2. lloct-3-yU amide Following the procedure of Example 4, replacing racemic 2-oxiranylmethyl-5 isoindole-l,3-dione with the chiral intermediate of the previous step, the trifluoroacetate salt of the title compound was prepared, (mlz): [M+H]+ calcd for 02^34^058, 491 .24; found 491.4. Retention time (anal. HPLC: 10-50% MeCN/H2O over 6 min) = 3.69 min. !H-NMR (CD3OD, 300MHz): 8 (ppm) 8.67 (s, 1H), 7.74-7.73 (m, 3H), 7.7-7.6 (dt, 1H), 7.3-7.2 (t, 1H), 4.2 (br s, 2H), 4.0 (br m, 2H), 3.2-2.9 (m, 4H), 2.8 (s, 3H), 2.6-2.3 (br m, 10 6H), 2.2-2.1 (br m, 2H), 1.57-1.55 (d, 6H). The title compound was also prepared by the following procedure. a2. Preparation of JV-((^)-oxiranyhnethvl)methanesulfonamide To a cold solution of methanesulfonamide (10 g, 0.105 mol) in water (100 mL) in an ice bath was added sodium hydroxide as pellets (8.4 g, 0.21 mol) and then 15 (iS)-2-chloromethyloxirane (12.4 g, 0.158 mol). The mixture was stirred at the same temperature for 2 h, and at room temperature for 12 h and then concentrated hydrochloric acid (18 mL) was added. The product was isolated by extracting the aqueous layer with dichloromethane (2 x 300 mL). The organic layer was dried over MgSO4 and then evaporated to dryness, yielding a colorless liquid (2.5 g), which was used directly in the 20 next step. b2. Synthesis of l-isopropyl-2-oxo-L2-dihvdroquinolinone-3-carboxylic acid azabicyclo[3.2.1]oct-3-yl) amide To a solution of l-isopropyl-2-oxo-l,2-dihydroquinoHne-3-carboxylic acid 25 {(lS3#,5tf)-8-aza-bicyclo[3.2.1]oct-3-yl}amide (TFA salt; 4 g, 8.8 mmol) in methanol (150 mL) was added A^JV-diisopropylethylamuie (1.7 mL, 9.5 mmol), and the product of the previous step (2.5 g, 18.2 mmol). The mixture was stirred at 80 °C for 2 days. After being concentrated under vacuum, the residue was purified by preparative HPLC, yielding the trifluoroacetate salt of the title compound (1.3 g). (mlz): [M+H]+ calcd for 30 C24H34N4O5S, 491 .24; found 491 .4. Retention time (anal. HPLC: 10-50% MeCN/H2O over 6 min) = 3.69 min. 54 WO 2005/100350 PCT/US2005/01 1393 Example 23: Synthesis of l-isopropyI-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(lS,3jR,5JZ)-8-[2-hydrosy-3-(l,l-dioxo-2-isothiazolidinyl)propyI]-8-azabicyclo[3.2.1]oct-3-yl} amide a. Preparation of l-isopropyl-2-oxo-l,2-dihvdroquinoliBone-3-carboxylic acid 5 {(l»S',3J?,5J?V8-r2-hvdroxv-3-(3-chloropropanesulfonvlamino^propyl]-8- azabicyclo[3 .2. 11oct-3-yl> amide To a cold solution of l-isopropyl-2-oxo-l32-dihydroquinolinone-3-cafboxylic acid {(15',3JR,5JR)-8-[2-liydroxy-3-aminopropyl]-8-azabicyclo[3 .2. 1 Joct-3-yl} amide TFA salt (0.125 g, 0.195 mmol) in dichloromethane (2 mL) in an ice bath was added N,N- 10 diisopropylethylamine (0.119 mL, 0.683 mmol) and 3-chloropropanesulfonyl chloride (0.025 mL, 0.205 mmol). After stirring at 0 °C for 2 h, the mixture was stirred at room temperature overnight. It was diluted with dichloromethane (50 mL), and washed with brine and saturated NaHCOa solution. After drying over MgSCU, the organic solution was evaporated to dryness, yielding an oily residue. The crude product was used directly in 15 next step. b. l-isopropyl-2-oxo-L2-dihydroquinolinone-3-carboxyric acid hydroxy-3-(l.l-dioxo-2-isothiazolidinyl)propvl1-8-azabicyclo[3.2.1]oct-3-yl|amide To a solution of l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(16',3JR,5JR)-8-[2-hydroxy-3-(3-chloropropanesulfonylamino)propyl]-8- 20 azabicyclo[3.2.1]oct-3-yl}amide (100 mg, 0.18 mmol) in anhydrous DMF (3 mL) was added potassium carbonate (75 mg, 0.56 mmol). The reaction mixture was shaken at 85 C for 12 h, and concentrated under vacuum. The residue was dissolved in dichloromethane (50 mL), and washed with saturated NaHCOs. After drying over MgSC>4, the filtrate was evaporated to dryness, and the residue was purified by preparative 25 HPLC to yield the title compound, (mfz): [M+H]+ calcd for C26H36N4O5S, 5 1 7.26; found 517.3. Example 24: Synthesis of l-isopropyl-2-oxo-lj2-dihydroquinoline-3- carboxylicacid{(15l,3JR,5jR)-8-[CR)-2-hydroxy-3-(methanesuIfonyl-methyI- 30 amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide a. Prepfflationof(l>S'.3J?,5J?)-3-[l-isopropyl-2-oxo-l,2-dihvdroquinoline-3-carbonvl)amino]-8-azabicvclo[3.2.1]octane-8-carboxylic acid te/t-butyl ester In a 3 L flask, l-isopropyl-2-oxo-l,2-dihydroquinolme-3-carboxylic acid (112.4 g, 0.486 mol, 1.1 eq) was suspended in toluene (1 L). The mixture was heated to 85 °C and 55 \VO 2005/100350 PCT/US2005/011393 %' thionyl chloride (86.74 g, 0.729 mol) was added dropwise over 70 min. The mixture was heated at 95 °C for 1.5 h with stirring and then allowed to cool to room temperature. In a separate 12 L flask, (lS,3J?,5£)-3-arruno-8-azabicycIo[3.2.1]octane-8-carboxylic acid fert-butyl ester (100.0 g, 0.442 mol, 1 eq) was suspended in toluene (1 L) 5 and 3 M NaOH (4 eq) was added. The mixture was stirred at room temperature for 10 min and then cooled to about 5 °C. The acid chloride solution was added slowly with stirring over 40 min keeping the internal temperature below 10 °C. The mixture was stirred at 3-5 °C for 30 min and the layers were allowed to separate overnight. The toluene layer (~2.5 L) was collected, concentrated to about half (-1.2 L) by rotary 10 evaporation, and used directly in the next step. b. Preparation of l4sopropyl-2-oxo-l,2-dmydroquinoline-3-carboxylic acid J(IS,3R,5R)~ 8-azabicyclo[3.2. l]oct-3-yU amide To the toluene solution prepared in the previous step (~1.2 L) was added trifiuoroacetic acid (200 mL) over 20 min at 20 °C with stirring. The mixture was stirred 15 at 20 °C for 2 h. Water (1.55 L) was added and the mixture was stirred for 30 min at 20 °C. After 30 min, the mixture separated into three layers. The bottom layer (~350 mL), a viscous brown oil, contained the crude intermediate. To a 12 L flask charged with MTBE (2.8 L), the crude brown oil was added over 1 h at 1-2 °C with stirring. The suspension was stirred at the same temperature for 1 h and 20 then filtered. The filtrate was washed with MTBE (2 x 300 mL) and dried under vacuum at room temperature for 4 days to provide the trifiuoroacetate salt of the title intermediate (163.3 g) as a pale yellow powder. c. Preparation of 7/-methyl-Ar-[f)S)-2-oxiran-2-ylrnethyl]rne1:hanesulfonamide A 12 L flask was charged with water (1 L) followed by the addition NaOH (50% 25 in water, 146.81 g, 1.835 mol). The beaker containing NaOH was washed with water (2 x 500 mL) and the washings were added to the flask. The mixture was stirred at room temperature for 10 min and cooled to ~8 °C. (A^methyl)methanesulfonamide (200.2 g, 1.835 mol) in water (500 mL) was added over 5 min. The mixture was stirred for 1 h at ~4 °C and (^-l-chloromethyloxirane (339.6 g, 3.67 mol) was added. The mixture was 30 stirred for 20 h at 3-4 °C. Dichloromethane (2 L) was added and the mixture was stirred for 30 min at 5-10 °C. The two layers were allowed to separate over 10 min and collected. The organic layer (~2.5 I) was added back to the 12 L flask and washed with 1 M H3P04 (800 mL) and brine (800 mL). Dichloromethane was removed by rotary 56 WO 2005/100350 PCT/US2005/011393 %e evaporation. To the crude product, toluene (400 mL) was added and removed by rotary evaporation. After three additional cycles of the toluene process, the title intermediate was obtained (228.2 g) which was used without further purification in the next step. d. Synthesis of l-isopropyl-2-oxo-1.2-dihydroquinoline-3-carboxvlic acid {(lS,3R,5R}-8-5 [ffl^^-hydroxy-S^methanesulfonvl-methyl-amino^propylj-S-aza-bicvclofS^.lloct-S-yl) amide hi a 3 L flask, l-isopropyl-2-oxo-l,2-dihydroquinohne-3-carboxylic acid {(l,S;3£,5JO-8-azabicyclo[3.2.1]oct-3--yl}amide trifluoroacetate (105.0 g, 0.232 mol) was suspended in absolute ethanol (400 mL). To this suspension, NaOH (50 % in water, 10 0.243 mol. 1.05 eq) dissolved in absolute ethanol (100 mL) was added at room temperature. The beaker containing the NaOH was washed with ethanol (2 x 50 mL) and the washings were added to the reaction mixture. After 30 min of stirring, a solution of ^-memyl-JV'-[(5)-2-oxiran-2-yhiiethyl]methanesulfonamide (62.0 g, 1.5 eq) in absolute ethanol (100 mL) was added. The mixture was refluxed for 2 h, cooled to room 15 temperature and seed crystals of the title compound were added. After about 5 min of stirring a white solid formed. The mixture was cooled to 3-5 °C and stirred for 2 h. The white solid was filtered and the wet cake was washed with cold absolute ethanol (3 x 50 mL). The solid was dried under vacuum at 30 °C for 60 h to provide the title compound (93.8 g, water content by Karl Fischer method 2.03 %). *H NMR (CDC13) 20 8 ppm 10.52 (d, 1H), 8.83 (s, 1H), 7.75 (d, 2H), 7.64-7.60 (m, 2H), 7.28-7.26 m, 1H), 4.33-4.26 (m, 2H), 3.78-3.75 (m, 1H), 3.27-3.20 (m, 4H), 3.01 (s, 3H), 2.88 (s, 3H), 2.58-2.53 (m, 1H), 2.30-1.81(m, 11H), 1.68 (d, 6H). The seed crystals were obtained from a previous preparation of the title compound by the method of this example at smaller scale, in which crystallization occurred 25 spontaneously. Example 25: Synthesis of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylicacid{(15,3/?551?)-8-[CR)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-a/a-bicyclo[3.2.1]oct-3-yl} amide hydrochloride In a 1 L flask, l-isopropyl-2-oxo-l,2-dihydroquinonne-3-carboxylic acid 30 {(16r,3J?,5^)-8-[(^)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide (34.7 g, 0.069 mol) was suspended in absolute ethanol (210 mL). Concentrated HC1 (1.1 eq) was added at room temperature with stirring. The mixture was stirred at reflux for 30 min and cooled to room temperature and stirred for 57 WO 2005/100350 PCTAJS2005/011393 2 h. The solid was filtered and the wet cake was washed with cold absolute ethanol (3 x 50 mL). The solid was dried under vacuum at 30 °C for 48 h to provide the title compound (34.5 g, 93.7 % yield, water content by Karl Fischer method 0.13%). Example 26: Synthesis of citric acid salt of l-isopropyl-2-oxo-l,2- 5 dihydroquinoline-3-carboxylic acid {(15,3jR,5jR)-8-[(l?)-2-hydroxy-3- (methanesulfonyl-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl} amide l-Isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5K)-8-[(R)-2-hydroxy-3-(methanesulfonyl-methyl-arnino)propyl]-8-aza-bicyclo[3.2.1 ]oct-3-yl} amide (0.1 g, 0.2 mmol) was suspended in ethanol (1 mL). To this suspension was added a 1M 10 solution of citric acid in ethanol (0.072 mL, 0.072 mmol, 0.33 eq). The mixture was briefly sonicated until clarity, capped, and then allowed to sit overnight. The cap was then removed and the mixture was allowed to evaporate under ambient conditions until solids were observed. The mixture was then recapped and allowed to sit for 72 h. The resulting solid was filtered and washed with cold ethanol to give the title compound as a 15 solid (74.3 mg). Example 27: Synthesis of acid salts of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(l^SJZjS^-S-^jR^-hydroxy-a-(methanesulfonyI-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl} amide Following the procedure of Example 26, the acid salts of l-isopropyl-2-oxo-l,2-20 dihydroquinoline-3-carboxylic acid {(l^S^S^-S-KJi^-hydroxy-S-tmemanesulfonyl-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide listed below in Table DI were prepared in solid form using the indicated equivalents of acid. Table El: Acid Salts Acid No. of equivalents of acid Product weight (mg) adipic 0.5 48.5 phosphoric 0.5 86.6 sulfuric 0.5 27.0 tartaric 0.5 66.3 malic 0.5 25.3 hydrobromic 1 62.9 58 WO 2005/100350 PCT/US2005/011393 W Example 28: Synthesis of methanesulfonic acid salt of l-isopropyl-2-oxo-l,2-dihydroquittoline-3-carboxylicacid{(15,3Jl?,5JR)-8-[(12)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyll-8-aza-bicyclol3.2.1]oct-3-yl} amide To a solution of l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid 5 {(15,3^,5^)-8-[(^)-2-hydroxy-3-(methanesulfonyl-methyl-aniino)propyl]-8-aza- bicyclo[3.2.1]oct-3-yl}amide (0.1 g, 0.2 nunol) in 50 % acetonitrile/water (1 mL) was added a 1M solution of methanesulfonic acid in ethanol (0.2 mL, 0.2 mmol, 1 eq). The mixture was then frozen and lyophilized to dryness ovenu'ght. The resulting solid was dissolved in isopropanol (1 mL) with gentle wanning and allowed to cool. The resulting 10 solid was collected by filtration and washed with cold isopropanol to give the title compound as a solid (90 mg). Example 29: Synthesis of acid salts of l-isopropyl-2-oxo-l,2-dihydroquinoline-S-carboxylicacidi^^S^jS^-S-^^-hydroxy-S-(methanesulfonyl-methyl-amino)propyl]-8-aza-bicycIo[3.2.1]oct-3-yl} amide 15 Following the procedure of Example 28, the acid salts of l-isopropyl-2-oxo-l,2- dihydroquinoline-3-carboxylic acid {(15',3jR,57?)-8-[(^?)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide listed below in Table IV were prepared in solid form using the indicated equivalents of acid. Table IV: Acid Salts Acid No. of equivalents of acid Product weight (mg) fumaric 1 107.2 benzoic 1 105.0 (7?)-mandelic 1 96.1 20 Assay 1: Radioligand Binding Assay on 5-HT4(C) Human Receptors a. Membrane Preparation 5-HT4(C) HEK-293 (human embryonic kidney) cells stably-transfected with human 5-HT4(c) receptor cDNA (Bmax = ~ 6.0 pmol/mg protein, as determined using [3H]-GR113 808 25 membrane radioligand binding assay) were grown in T-225 flasks in Dulbecco's Modified Eagles Medium (DMEM) containing 4,500 mg/L D-glucose and pyridoxine hydrochloride (GIBCO-Invitrogen Corp., Carlsbad CA: Cat #11965) supplemented with 10% fetal bovine serum (FBS) (GBCO-hivitrogen Corp.: Cat #10437), 2 mM L-glutamine and (100 units) penicilHn-(100 p,g) streptomycin/ml (GIBCO-Invitrogen Corp.: Cat #15140) in 59 WO 2005/100350 PCT/US2005/011393 w a 5% CC>2, humidified incubator at 37 °C. Cells were grown under continuous selection pressure by the addition of 800 jag/mL geneticin (GIBCO-Invitrogen Corp.: Cat #10131) to the medium. Cells were grown to roughly 60-80% confluency ( For the membrane preparation, cell pellets were resuspended in ice-cold 50 mM 10 4-(2-hydroxyethyl)-1 -piperazineethanesulphonic acid (HEPES), pH 7.4 (membrane preparation buffer) (40 mL/total cell yield from 30-40 T225 flasks) and homogenized using a polytron disrupter (setting 19, 2 x 10 s) on ice. The resultant homogenates were centrifuged at 1200 g for 5 min at 4°C. The pellet was discarded and the supernatant centrifuged at 40,000 g (20 min). The pellet was washed once by resuspension with 15 membrane preparation buffer and centrifugation at 40,000 g (20 min). The final pellet was resuspended in 50 mM HEPES, pH 7,4 (assay buffer) (equivalent 1 T225 flask/1 mL). Protein concentration of the membrane suspension was determined by the method of Bradford (Bradford, 1976). Membranes were stored frozen in aliquots at -80 °C. 20 b. Radioligand Binding Assays Radioligand binding assays were performed in 1.1 mL 96- deep well polypropylene assay plates (Axygen) in a total assay volume of 400 |J.L containing 2 ug membrane p'rotein in 50 mM HEPES pH 7.4, containing 0.025% bovine serum albumin (BSA). Saturation binding studies for determination of K^ values of the radioligand were 25 performed using [3H]-GR113 808 (Amersham Inc., Bucks, UK: Cat #TRK944; specific activity -82 Ci/mmol) at 8-12 different concentrations ranging from 0.001 nM - 5.0 nM. Displacement assays for determination of pKj values of compounds were performed with [3H]-GR113808 at 0.15 nM and eleven different concentrations of compound ranging fromlOpM-lOOuM. 30 Test compounds were received as 10 mM stock solutions in DMSO and diluted to 400 uM into 50 mM HEPES pH 7.4 at 25°C, containing 0.1% BSA, and serial dilutions (1:5) then made in the same buffer. Non-specific binding was determined in the presence 60 WO 2005/100350 PCT/US2005/011393 % of 1 uM unlabeled GR113808. Assays were incubated for 60 min at room temperature, and then the binding reactions were terminated by rapid filtration over 96-well GF/B glass fiber filter plates (Packard BioScience Co., Meriden, CT) presoaked in 0.3% polyethyleneimine. Filter plates were washed three times with filtration buffer (ice-5 cold 50mM HEPES, pH7.4) to remove unbound radioactivity. Plates were dried, 35 uL Microscint-20 liquid scintillation fluid (Packard BioScience Co., Meriden, CT) was added to each well and plates were counted in a Packard Topcount liquid scintillation counter (Packard BioScience Co., Meriden, CT). Binding data were analyzed by nonlinear regression analysis with the GraphPad 10 Prism Software package (GraphPad Software, Inc., San Diego, CA) using the 3-parameter model for one-site competition. The BOTTOM (curve minimum) was fixed to the value for nonspecific binding, as determined in the presence of 1 uM GR113808. Kj values for test compounds were calculated, in Prism, from the best-fit ICso values, and the K Test compounds having a higher pKj value in this assay have a higher binding . affinity for the 5-HT4 receptor. The compounds of the invention which were tested in this 20 assay had a pKj value ranging from about 6.3 to about 9.0, typically ranging from about 6.5 to about 8.5. Assay 2: Radioligand Binding Assay on 5-HTaA Human Receptors: Determination of Receptor Subtype Selectivity a. Membrane Preparation 5-HT^A 25 HEK-293 (human embryonic kidney) cells stably-transfected with human 5-HTaA receptor cDNA were obtained from Dr. Michael Bruess (University of Bonn, GDR) (Bmax = ~ 9.0 pmol/mg protein, as determined using [3H]-GR65630 membrane radioligand binding assay). Cells were grown in T-225 flasks or cell factories in 50% Dulbecco's Modified Eagles Medium (DMEM) (GffiCO-Invitrogen Corp., Carlsbad, CA: 30 Cat #11965) and 50% Ham's F12 (GIBCO-Invitrogen Corp.: Cat #11765) supplemented with 10% heat inactivated fetal bovine serum (FBS) (Hyclone, Logan, UT: Cat 61 WO 2005/100350 PCT/US2005/011393 1^SH30070.03) and (50 units) penicillin-(50 ng) streptomycin/ml (GDBCO-Invitrogen Corp.: Cat #15140) in a 5% CO2, humidified incubator at 37 °C. Cells were grown to roughly 70-80% confluency ( 10 b. Radioligand Binding Assays '. Radioligand binding assays were performed in 96-well polypropylene assay plates in a total assay volume of 200 uL containing 1.5-2 p,g membrane protein in 50 mM HEPES pH 7.4, containing 0.025% BSA assay buffer. Saturation binding studies for determination of K^ values of the radioligand were performed using [3H]-GR65630 15 (PerkinElmer Life Sciences Inc., Boston, MA: Cat #NET1 Oil, specific activity-85 Ci/mmoi) at twelve different concentrations ranging from 0.005 nM to 20 nM. Displacement assays for determination of pK, values of compounds were performed with [3H]-GR65630 at 0.50 nM and eleven different concentrations of compound ranging from 10 pM to 100 uM. Compounds were received as 10 mM stock solutions in DMSO (see 20 section 3.1), diluted to 400 uM into 50 mM HEPES pH 7.4 at 25°C, containing 0.1% BSA, and serial (1:5) dilutions then made in the same buffer. Non-specific binding was determined in the presence of 10 uM unlabeled MDL72222. Assays were incubated for 60 min at room temperature, then the binding reactions were terminated by rapid filtration over 96-well GF/B glass fiber filter plates (Packard BioScience Co., 25 Meriden, CT) presoaked in 0.3% polyethyleneimine. Filter plates were washed three times with filtration buffer (ice-cold 50mM HEPES, pH7.4) to remove unbound radioactivity. Plates were dried, 35 uL Microscint-20 liquid scintillation fluid (Packard BioScience Co., Meriden, CT) was added to each well and plates were counted in a Packard Topcount liquid scintillation counter (Packard BioScience Co., Meriden, CT). 30 Binding data were analyzed using the non-linear regression procedure described above to determine Kj values. The BOTTOM (curve minimum) was fixed to the value for 62 \VO 2005/1 00350 PCT/US2005/011393 nonspecific binding, as determined in the presence of 10 uM MDL72222. The quantity [L] in the Cheng-Prusoff equation was defined as the concentration [3H]-GR65630. Selectivity for the 5-HT4 receptor subtype with respect to the S-HTs receptor subtype was calculated as the ratio Kj(5-HT3A)/Ki(5-HT4(C)). The compounds of the 5 invention which were tested in this assay had a S-HiyS-HTa receptor subtype selectivity ranging from about 50 to about 8000, typically ranging from about 100 to about 4000. Assay 3: Whole-cell cAMP Accumulation Flashplate Assay with HEK-293 cells expressing human S-BTT^c) Receptors hi this assay, the functional potency of a test compound was determined by 10 measuring the amount of cyclic AMP produced when HEK-293 cells expressing 5-HT4 receptors were contacted with different concentrations of test compound. a. Cell Culture HEK-293 (human embryonic kidney) cells stably-transfected with cloned human 5-HT4(C) receptor cDNA were prepared expressing the receptor at two different densities: 1 5 (1) at a density of about 0.5-0.6 pmol/nig protein, as determined using a [3H]-GR1 13808 membrane radioligand binding assay, and (2) at a density of about 6.0 pmol/mg protein. The cells were grown in T-225 flasks in Dulbecco's Modified Eagles Medium (DMEM) containing 4,500 mg/L D-glucose (GIBCO-Invitrogen Corp.: Cat #1 1965) supplemented with 10% fetal bovine serum (FBS) (GIBCO-Invitrogen Corp.: Cat #10437) and (100 20 units) penicillin-(100 u.g) streptomycin/ml (GIBCO-Invitrogen Corp.: Cat #15140) hi a 5% COa, humidified incubator at 37°C. Cells were grown under continuous selection pressure by the addition of geneticin (800 i^g/mL: GIBCO-hivitrogen Corp.: Cat #10131) to the medium. b. Cell Preparation 25 Cells were grown to roughly 60-80% confluency. Twenty to twenty-two hours prior to assay, cells were washed twice, and fed, with serum-free DMEM containing 4,500 mg/L D-glucose (GIBCO-Invitrogen Corp.: Cat #1 1965). To harvest the cells, the media was aspirated and 10 mL Versene (GIBCO-hivitrogen Corp.: Cat #15040) was added to each T-225 flask. Cells were incubated for 5 min at RT and then dislodged from 30 the flask by mechanical agitation. The cell suspension was transferred to a centrifuge tube containing an equal volume of pre-warmed (37°C) dPBS and centrifuged for 5 min at 1000 rpm. The supernatant was discarded and the pellet was re-suspended in pre-warmed 63 \VO 2005/100350 PCT/US2005/011393 %r (37°C) stimulation buffer (lOmL equivalent per 2-3 T-225 flasks). This time was noted and marked as time zero. The cells were counted with a Coulter counter (count above 8 (am, flask yield was 1-2 x 107 cells/flask). Cells were resuspended at a concentration of 5 x 105 cells/ml inpre-warmed (37°C) stimulation buffer (as provided in the flashplate 5 kit) and preincubated at 37°C for 10 min. cAMP assays were performed in a radioimmunoassay format using the Flashplate Adenylyl Cyclase Activation Assay System with 125I-cAMP (SMP004B, PerkinElmer Life Sciences Inc., Boston, MA), according to the manufacturer's instructions. Cells were grown and prepared as described above. Final cell concentrations in 10 the assay were 25 x 103 cells/well and the final assay volume was 100 uL. Test compounds were received as 10 mM stock solutions in DMSO, diluted to 400 uM into 50 mM HEPES pH 7.4 at 25°C, containing 0.1% BSA, and serial (1:5) dilutions then made in the same buffer. Cyclic AMP accumulation assays were performed with 11 different concentrations of compound ranging from 10 pM to 100 uM (final assay 15 concentrations). A 5-HT concentration-response curve (10 pM to 100 uM) was included on every plate. The cells were incubated, with shaking, at 37°C for 15 min and the reaction terminated by addition of 100 ul of ice-cold detection buffer (as provided in the flashplate kit) to each well. The plates were sealed and incubated at 4°C overnight. Bound radioactivity was quantified by scintillation proximity spectroscopy using the 20 Topcount (Packard BioScience Co., Meriden, CT). The amount of cAMP produced per mL of reaction was extrapolated from the cAMP standard curve, according to the instructions provided in the manufacturer's user manual. Data were analyzed by nonlinear regression analysis with the GraphPad Prism Software package using the 3-parameter sigmoidal dose-response model (slope 25 constrained to unity). Potency data are reported as pECso values, the negative decadic logarithm of the ECso value, where ECso is the effective concentration for a 50 % maximal response. Test compounds exhibiting a higher pEC5o value in this assay have a higher potency for agonizing the 5-HT4 receptor. The compounds of the invention which were 30 tested in this assay, for example, in the cell line (1) having a density of about 0.5-0.6 pmoymg protein, had a pECso value ranging from about 7.0 to about 9.0, typically ranging from about 7.5 to about 8.5. 64 \VO 2005/100350 PCT/US2005/011393 Assay 4: In viti-o Voltage Clamp Assay of Inhibition of Potassium Ion Current in Whole Cells Expressing the hERG Cardiac Potassium Channel CHO-K1 cells stably transfected with hERG cDNA were obtained from Gail Robertson at the University of Wisconsin. Cells were held in cryogenic storage until 5 needed. Cells were expanded and passaged hi Dulbecco's Modified Eagles Medium/F12 supplemented with 10 % fetal bovine serum and 200 u.g/mL geneticin. Cells were seeded onto poly-D-lysine (100 p,g/mL) coated glass coverslips, in 35 mm2 dishes (containing 2 mL medium) at a density that enabled isolated cells to be selected for whole cell voltage-clamp studies. The dishes were maintained in a humidified, 5% CC>2 environment at 37°C. 10 Extracellular solution was prepared at least every 7 days and stored at 4°C when not in use. The extracellular solution contained (mM): NaCl (137), KC1 (4), CaCl2 (1.8), MgCIa (1), Glucose (10), 4-(2-hydroxyethyl)-l-piperazineethanesulphonic acid (HEPES) (10), pH 7.4 with NaOH. The extracellular solution, in the absence or presence of test compound, was contained in reservoirs, from which it flowed into the recording chamber 15 at approximately 0.5 mL/min. The intracellular solution was prepared, aliquoted and stored at -20°C until the day of use. The intracellular solution contained (mM): KC1 (130), MgCla (1), ethylene glycol-bis(beta-amuioethyl ether) N,N,N',N'-tetfa acetic acid salt (EGTA) (5), MgATP (5), 4-(2-hydroxyethyl)-l-piperazineethanesulphonic acid (HEPES) (10), pH 7.2 with KOH. All experiments were performed at room temperature (20-22°C). 20 The coverslips on which the cells were seeded were transferred to a recording chamber and perfused continuously. Gigaohm seals were formed between the cell and the patch electrode. Once a stable patch was achieved, recording commenced in the voltage clamp mode, with the initial holding potential at -80 mV. After a stable whole-cell current was achieved, the cells were exposed to test compound. The standard voltage 25 protocol was: step from the holding potential of -80 mV to +20 mV for 4.8 sec, repolarize to -50 mV for 5 sec and then return to the original holding potential (-80 mV). This voltage protocol was run once every 15 sec (0.067 Hz). Peak current amplitudes during the repolarization phase were determined using pClamp software. Test compounds at a concentration of 3 |jM were perfused over the cells for 5 minutes, followed by a 5-minute 30 washout period in the absence of compound. Finally a positive control (cisapride, 20 nM) was added to the perfusate to test the function of the cell. The step from -80 mV to +20 mV activates the hERG channel, resulting in an outward current. The step back to -50 mV 65 WO 2005/J00350 PCT/US2005/011393 results in an outward tail current, as the channel recovers from inactivation and deactivates. Peak current amplitudes during the repolarization phase were determined using pCLAMP software. The control and test article data were exported to Origin® (OriginLab 5 Corp., Northampton MA) where the individual current amplitudes were normalized to the initial current amplitude in the absence of compound. The normalized current means and standard errors for each condition were calculated and plotted versus the time course of the experiment. Comparisons were made between the observed K current inhibitions after the 10 five-minute exposure to either the test article or vehicle control (usually 0.3 % DMSO). Statistical comparisons between experimental groups were performed using a two-population, independent t-test (Microcal Origin v. 6.0). Differences were considered significant at p The smaller the percentage inhibition of the potassium ion current in this assay, 15 the smaller the potential for test compounds to change the pattern of cardiac repolarization when used as therapeutic agents. The compounds of the invention which were tested in this assay at a concentration of 3 jiM exhibited an inhibition of the potassium ion current of less than about 20 %, typically, less than about 15%. Assay 5: /« vitro Model of Oral Bioavailability: Caco-2 Permeation Assay 20 The Caco-2 permeation assay was performed to model the ability of test compounds to pass through the intestine and get into the blood stream after oral administration. The rate at which test compounds in solution permeate a cell monolayer designed to mimic the tight junction of human small intestinal monolayers was determined. 25 Caco-2 (colon, adenocarcinoma; human) cells were obtained from ATCC (American Type Culture Collection; Rockville, MD). For the permeation study, cells were seeded at a density of 63,000 cells/cm2 on pre-wetted transwells polycarbonate filters (Costai-; Cambridge, MA). A cell monolayer was formed after 21 days in culture. Following cell culture in the transwell plate, the membrane containing the cell monolayer 30 was detached from the transwell plate and inserted into the diffusion chamber (Costar; «mc 4-nepftfiri into the heating block which was WO 2005/100350 l'( 'T/'li.S2il05/01 1.W.! control. The air manifold delivered 95% O?/5% CO2 to each half oi a diffusion chamber and created a laminar flow pattern across the cell monolayer, which was effective in reducing the unstirred boundary layer. The permeation study was performed with test compound concentrations at 5 100 uM and with C-mannitol to monitor the integrity of the monolayer. All experiments were conducted at 37 °C for 60 inin. Samples were taken at 0, 30 and 60 min from both the donor and receiver sides of the chamber. Samples were analy/.ed by HPLC or liquid scintillation counting for test compound and mannitol concentrations. The permeation coefficient (Kp) in cm/sec was calculated. 10 hi this assay, a Kp value greater than about 10x1 0^(> cm/sec is considered indicative of favorable bioavai lability. The compounds of the invention that were tested in this assay exhibited Kp values of between about 10 x 1 0 () cm/sec and about 50 x 1 0"6 cm/sec, typically between about 20 x 10"° cm/sec and about 40 x 10"° crn/sec. Assay 6: Pharmacokinetic Study in the Rat 15 Aqueous solution formulations of test compounds were prepared in 0. 1 % lactic acid at apH of between about 5 and about 6. Male Sprague-Dawley rats (CD strain, Charles River Laboratories, Wilmington, MA) were dosed with test compounds via intravenous administration (IV) at a dose of 2.5 mg/kg or by oral gavage (PO) at a dose of 5 mg/kg. The dosing volume was 1 mL/kg for IV and 2 ml 7kg for PO administration. 20 Serial blood samples were collected from animals pre-dose, and at 2 (IV only), 5, 15, and 30 min, and at 1, 2, 4, 8, and 24 hours post-dose. Concentrations of lest compounds in blood plasma were determined by liquid cbromatography-mass spectrometry analysis (LC-MS/MS) (MDS SCIEX, API 4000, Applied Biosysterns, Foster City, C'A) with a lower limit of quantitation of 1 ng/mL. 25 Standard pharmacokinctic parameters were assessed by non-compartmental analysis (Model 201 for IV and Model 200 for PO) using WinNonlin (Version 4.0. 1 , Pharsight, Mountain View, CA). The maximum in the curve of test compound concentration in blood plasma vs. time is denoted Cinax. The area under the concentration vs. time curve from the time of dosing to the last measurable concentration (AUC(O-t)) 30 was calculated by the linear trape/oidal rule. Oral bioavailability (F(%)), i.e. the dose-normalized ratio of AUC(O-t) forPO administration to AUC(O-t) for IV administration, was calculated as: WO 2005/100350 PCT/liS20 Test compounds which exhibit larger values of the parameters Cmax. AUC(O-t). andF(%) in this assay are expected to have greater bioavailability when administered orally. The compounds of the invention that were tested in this assay had Cmax values 5 typically ranging from about 0.1 to about 0.25 Lig/nrL and AUC(O-t) values typically ranging from about 0.4 to about 0.9 [ig-hr/mL. By way of example, the compound of Example 1 had a Crnax value of 0.17 ug/mL, an AUC(O-t) value of 0.66 fig-hr/mL and oral bioavailability (F(%)) in the rat model of about 35 %. While the present invention has been described with reference to the specific 10 embodiments thereof, it should be understood by those skilled in the art that various changes maybe made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present invention. All such modifications are intended 15 to be within the scope of the claims appended hereto. Additionally, all publications, patents, and patent document^ cited hereirmbove are incorporated by reference herein in full, as though individually incorporated by reference. WE CLAIM: 1. A Quinolinone-carboxamide compounds of formula (I): (Formula Removed) wherein: R1 is hydrogen, halo, hydroxy, C1-4alkyl, or C1-4alkoxy; R2 is C3-4alkyl, or C3-6cycloalkyl; R is hydrogen or C1-3alkyl; R4 is —S(O)2R6 or —C(O)R7; R5 is hydrogen, C1-3alkyl, C2-3alkyl substituted with -OH or C1-3alkoxy, or -CH2-pyridyl; R6 is C1-3alkyl; or, R5 and R6 taken together form C3-4alkylenyl; and R7 is hydrogen, C1-3alkyl, or pyridyl; or a pharmaceutically-acceptable salt or solvate or stereoisomer thereof. 2. The compound as claimed in claim 1 wherein: R5 is hydrogen, C1-3alkyl, C2-3alkyl substituted with -OH or C1-3alkoxy, or -CH2-pyridyl; and R6 is C1-3alkyl. 3 The compound as claimed in claim 2 wherein R1 is hydrogen or halo. 4. The compound as claimed in claim 2 wherein R2 is C3-4alkyl. 5. The compound as claimed in claim 2 wherein R4 is —S(O)2R6. 6. The compound as claimed in claim 5 wherein R4 is —S(O)2CH3. 7. The compound as claimed in claim 2 wherein R4 is —C(O)R7. 8. The compound as claimed in claim 7 wherein R7 is hydrogen or methyl. 9. The compound as claimed in claim 2 wherein R5 is hydrogen or methyl. 10. The compound as claimed in claim 2, wherein: R1 is hydrogen; R is C3-4alkyl or C4-5cycloalkyl; R3 is hydrogen; R4 is —S(O)2R6 or —C(O)R7; R5 is hydrogen or C1-3alkyl; R is C1-3alkyl; and R is hydrogen or C1-3alkyl. 11. The compound as claimed in claim 1 wherein the compound is selected from: 1 -isopropyl-2-oxo-1,2-dihydroquinolinone-3 -carboxylic acid {(1S,3R,5R)-8-[2-hydroxy-3 - (methanesulfonyl-methyl-amino)propyl] -8-azabicyclo [3.2.1] oct-3 -yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[2-methoxy-3- (methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-1,2-dihydroquinoline-3 -carboxylic acid {(1S,3R,5R)-8-[3- (methanesulfonyl-pyridin-3 -ylmethyl-amino)-2-methoxypropyl] -8-azabicyclo [3.2.1] oct-3 -yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[2-hydroxy-3-methanesulfonylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3 -carboxylic acid {(1S,3R,5R)-8-[3- (methanesulfonyl-pyridin-3-ylmethyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1 ]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[(R)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(1 S,3R,5R)-S-[(5)-2-hydroxy-3-(methanesulfonyl-memyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l ,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[2-hydroxy-3-[methyl-(pyridine-4-carbonyl)amino]propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[2-hydroxy-3-[(pyridine-4-carbonyl)amino]propyl]-8-azabicyclo[3.2.1 ]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-methyl- amino)-2-methoxypropyl]-8-azabicyclo[3.2.1 ]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[2-methoxy-3- [methyl-(pyridine-4-carbonyl)amino]propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid {(1 S,3R,5R)-8-[3 -(acetyl-pyridin- 3-ylmethyl-amino)-2-methoxypropyl]-8-azabicyclo[3.2.1 ]oct-3-yl} amide; 1 -isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-acetylamino-2- hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydro-quinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(acetyl-methyl- amino)-2-hydroxypropyl] -8-azabicyclo [3.2.1] oct-3 -yl} amide; 1 -isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid {(1 S,3R,5R)-8-[3-(formyl- methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amid; l-isopropyl-2-oxo-l,2-dihydro-quinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-formylamino-2- hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydro-quinoline-3-carboxylic acid {(1S,3R,5R)-8-[(R)-3-(acetyl- methyl-amino)-2-hydroxypropyl] -8-azabicyclo [3.2.1] oct-3 -yl} amide; 1 -isopropyl-2-oxo-1,2-dihydro-quinoline-3-carboxylic acid {(1S,3R,5R)-8-[(R)-3-(formyl- methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 5-bromo-l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-8-[(R)-2- hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(lS,3R,5R)-8-[2-hydroxy-3- (methanesulfonyl-ethylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-l ,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[(R)-2-hydroxy- 3-(methanesulfonylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; and l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[2-hydroxy-3- (l,l-dioxo-2-isothiazolidinyl)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; and pharmaceutically-acceptable salts and solvates and stereoisomers thereof. 12. The compound as claimed in claim 2, wherein the compound is selected from: 1-isopropy 1-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-8-[2-hydroxy-3- (methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-1,2-dihydroquinoline-3 -carboxylic acid {(1S,3R,5R)-8-[2-hydroxy-3 - (methanesulfonylamino)propyl]-8-aza-bicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[(R)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroqninoline-3-carboxylic acid {(1S,3R,5R)-8-[(S)-2-hydroxy-3-(methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-l ,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-8-[3-(acetyl-methyl-amino)-2-hydroxypropyl] -8-azabicyclo [3.2.1] oct-3 -yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[3-(formyl-methyl-amino)-2-hydroxypropyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1 -isopropyl-2-oxo-1,2-dihydroquinoline-3 -carboxylic acid {(1 S,3R,5R)-8- [(R)-3 -(acetyl-methyl-amino)-2-hydroxypropyl] -8-azabicyclo [3.2.1 ]oct-3 -yl} amide; l-isopropyl-2-oxo-l ,2-dihydroquinoline-3 -carboxylic acid {(lS,3R,5R)-8-[(R)-3-(formyl-methyl-amino)-2-hydroxypropyl] -8-azabicyclo [3.2.1 ]oct-3 -yl} amide; l-isopropyl-2-oxo-l,2-dihydroquinolinone-3-carboxylic acid {(1S,3R,5R)-8-[(R)-2-hydroxy-,3-(methanesulfonylamino)propyl]-8-azabicyclo[3.2.1 ]oct-3-yl}amide; and pharmaceutically-acceptable salts and solvates and stereoisomers thereof. 13. The compound as claimed in claim 12 wherein the compound is selected from: 1 -isopropyl-2-oxo-1,2-dihydroquinoline-3 -carboxylic acid { (1 S,3R,5R)-8- [2-hydroxy-3 - (methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(\S,3R,5R)-8-[(R)-2-hydroxy-3- (methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; l-isopropyl-2-oxo-l,2-dihydroquinoline-3-carboxylic acid {(1S,3R,5R)-8-[(S)-2-hydroxy-3- (methanesulfonyl-methyl-amino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; 1-isopropyl-2-oxo-1,2-dihydroquinoline-3-carboxylic acid {(lS,3R,5R)-8-[(R)-2-hydroxy-3- (methanesulfonylamino)propyl]-8-azabicyclo[3.2.1]oct-3-yl}amide; and pharmaceutically acceptable salts and solvates and stereoisomers thereof. 14. The compound as claimed in any of the preceding claims as and when used in the preparation of a pharmaceutical composition. 15. The compound as claimed in any one of claims 1 to 13 for use in therapy. 16. A process for preparing a compound as claimed in any one of claims 1 to 13, the process comprising: (a) reacting a compound of formula (III): (Formula Removed) with compound of the formula L—R4 wherein L is a halo leaving group in the presence of at least three equivalents of base at a temperature between -100 °C and 30 °C, or wherein L—R4 represents HO-C(0)R in the presence of a coupling agent selected from benzotriazol-1-yloxytripyrrolidino-phosphonium hexafluorophosphate, 1,3 dicyclohexylcarbodiimide, l-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and l-hydroxy-7-azabenzotriazole, and combinations thereof at ambient temperature; or (b) reacting a compound of formula (VIII): (Formula Removed) with a compound of formula (DC): (Formula Removed) in the presence of a coupling agent defined in the previous step; or (c), when R is defined as hydrogen, reacting a compound of formula (IV): (Formula Removed) or a salt thereof with a compound of formula (XI): (Formula Removed) at a temperature between 60 °C and 100 °C to provide a compound of formula (I), or a pharmaceutically-acceptable salt or solvate or stereoisomer thereof. |
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5398-DELNP-2006-Abstract-(29-06-2012).pdf
5398-delnp-2006-Assignment-(22-12-2014).pdf
5398-DELNP-2006-Claims-(29-06-2012).pdf
5398-delnp-2006-Copy-Of-Form-6-(22-12-2014).pdf
5398-delnp-2006-Correspondence Others-(12-04-2013).pdf
5398-delnp-2006-Correspondence Others-(20-03-2014).pdf
5398-delnp-2006-Correspondence Others-(22-12-2014).pdf
5398-DELNP-2006-Correspondence Others-(29-06-2012).pdf
5398-delnp-2006-Correspondence-Others-(15-03-2013).pdf
5398-delnp-2006-correspondence-others.pdf
5398-delnp-2006-description (complete).pdf
5398-delnp-2006-Form-1-(22-12-2014).pdf
5398-delnp-2006-Form-2-(22-12-2014).pdf
5398-delnp-2006-Form-3-(12-04-2013).pdf
5398-DELNP-2006-Form-3-(29-06-2012).pdf
5398-delnp-2006-GPA-(12-04-2013).pdf
5398-delnp-2006-GPA-(22-12-2014).pdf
5398-DELNP-2006-GPA-(29-06-2012).pdf
5398-delnp-2006-pct-request.pdf
5398-delnp-2006-pct-search-report.pdf
5398-DELNP-2006-Petition-137-(29-06-2012).pdf
| Patent Number | 264747 | |||||||||||||||||||||||||||
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| Indian Patent Application Number | 5398/DELNP/2006 | |||||||||||||||||||||||||||
| PG Journal Number | 04/2015 | |||||||||||||||||||||||||||
| Publication Date | 23-Jan-2015 | |||||||||||||||||||||||||||
| Grant Date | 19-Jan-2015 | |||||||||||||||||||||||||||
| Date of Filing | 18-Sep-2006 | |||||||||||||||||||||||||||
| Name of Patentee | THERAVANCE BIOPHARMA R & D IP, LLP | |||||||||||||||||||||||||||
| Applicant Address | 901 GATEWAY BOULEVARD, SOUTH SAN FRANCISCO, CA 94080UNITED STATES OF AMERICA | |||||||||||||||||||||||||||
Inventors:
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| PCT International Classification Number | C07D 451/04 | |||||||||||||||||||||||||||
| PCT International Application Number | PCT/US05/011393 | |||||||||||||||||||||||||||
| PCT International Filing date | 2005-04-06 | |||||||||||||||||||||||||||
PCT Conventions:
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