ZA200605308B - Stereoisomeric compounds and methods for the treatment of gastrointestinal and central nervous system disorders - Google Patents

Description

DESCRIPTION

STEREOISOMAERIC COMPOUNDS AND METHODS FOR THE TREATMENT OF

GASTROINTESTINAL AND CENTRAL NERVOUS SYSTEM DISORDERS

This appl&cation claims priority from U.S. Provisiornal Patent Application number 60/534,892, file January 7, 2004, and US. Provisionaml Patent Application number 60/560,938, filed April 9, 2004.

Background of Invention

Cisapride is one of a class of compounds known as benzamide derivatives, the par-ent compound of wwhich is metoclopramide. U.S. Patent Mos. 4,962,115 and 5,057,525 (collectively "Vaan Daele" and incorporated by reference in their entireties) disclose N—(3- hydroxy-4-piperiidenyl) benzamides of cisapride. Van Daele discloses that these compourads, the pharmaceutically acceptable acid addition salts thereof ard the stereochemically isomeeric forms thereof, st-imulate the motility of the gastrointestinal system.

As a class, these benzamide derivatives have seve=ral prominent pharmacologzical actions. The prominent pharmacological activities of the beenzamide derivatives are du«e to their effects on the neuronal systems which are modulated bey the neurotransmitter serotonin.

The role of sereotonin, and thus the pharmacology of the Wbenzamide derivatives, has been broadly implicated in a variety of conditions for many yearss. Thus, research has focused on locating the production and storage sites of serotonin as “well as the location of seroteonin receptors in thes human body in order to determine the comnection between these sites and various disease states or conditions.

In this regard, it was discovered that a major siste of production and storagse of serotonin is thes enterochromaffin cell of the gastrointestina_1 mucosa. It was also discow ered that serotonin mas a powerful stimulating action on intestina 1 motility by stimulating intestinal smooth muscles, speeding intestinal transit, and decreasingz absorption time, as in diarrhea.

This stimulatin_g action is also associated with nausea and veomiting.

Because= of their modulation of the serotonin neuronal system in the gastrointestinal tract, many of the benzamide derivatives are effective anti —emetic agents and are commnonly used to control vomiting during cancer chemotherapy or racdiotherapy, especially when highly emetogenic compounds such as cisplatin are used. This action is almost certainly the result of the ability cf the compounds to block the actions of se=rotonin (SHT) at specific sites of action, called the SHT3-receptor, which was classically designated in the scientific literature as the serotonin M-receptor. Chemotherapy amd radiation therapy may induce nausea and vomiting by the release of serotonin from damaged enterochromaffin cells in the gastrointestinal tract. Release of the neurotransrmitter serotonin stimulates both afferent vagal nerve fibers (thus initiating the vomiting reflex) and serotonin receptors in the chemoreceptor trigger zone of the area postrema region of the brain. The anatomical site for this action of the benzamide derivatives, and whether such action is central (CNS), peripheral, or a combination thereof, remains unresolved (Barrmes et al., J. Pharm. Pharmacol. 40: 586-588, 1988). Cisapride, like the other benzamide derivatives would appear to be an effective anti- emetic agent based on its ability to modulate the activity of serotonin at the SHT3 receptor.

A second prominent action of the benzamide derivatives is ‘in augmenting: gastrointestinal smooth muscle activity from the esophagus through the proximal small bowel, thus accelerating esophageal and small intestinal transit as well as facilitating gastric= emptying and increasing lower esophageal sph_incter tone (Decktor et al., Eur. J. Pharmacol — 1S 147: 313-316, 1988). Although the benzammide derivatives are not cholinergic receptor= agonists per se, the aforementioned smooth nuscle effects may be blocked by muscarinic receptor blocking agents such as atropine or neuronal transmission inhibitors of the tetrodotoxin type which affect sodium channel s. Similar blocking activity has been reported for the contractile effects of serotonin in the smmall intestine. It is currently believed that thea primary smooth muscle effects of the benzammide derivatives are the result of an agonist action upon a new class of serotonin recept-ors referred to as SHT,4 receptors which ares located on interneurons in the myenteric plexuss of the gut wall. Activation of these receptor=s subsequently enhances the release of acetylczholine from parasympathetic nerve terminal=s located near surrounding smooth muscle fibe=rs, and it is the combination of acetylcholinee with its receptors on smooth muscle membaranes which is the actual trigger for musclee contraction.

A discussion of various SHT receptors , including the SHT, receptor can be found im, for example, U.S. Patent Nos. 6, 331,401 and «6,632,827, which are incorporated by reference herein in their entirety.

Cisapride has been used primarily to treat gastroesophageal reflux disease (GERDD).

This disease is characterized as the backwevard flow of the stomach contents into thme esophagus. One of the most important factors in the pathogenesis of gastroesophageal reflumx

MBHB Ref. No. «04-972-C ARYX-020-PC disease is a reduiaction in the pressure barrier due to the faXlure of the lower esophageal sphincter. Failures of the lower esophageal sphincter can arise= due to a low basal pressure, sphincter relaxation, or to a non-compensated increase in intragastric pressure. Other factors in the pathogeneesis of the disease are delayed gastric emptying, insufficient esophageal clearing due to immpaired peristalsis and the corrosive nature of the reflux material which can damage esophagzeal mucosa, Cisapride is thought to strengthen the anti-reflux barrier and improve esophaggeal clearance by increasing the lower esophageal sphincter pressure and enhancing peristzaltic contractions.

Because of its activity as a prokinetic agent, cisapride would also appear to be useful to treat dyspepssia, gastroparesis, constipation, post-operativee ileus, and intestinal pseudo- obstruction. Dzyspepsia is a condition characterized by arm impairment of the power or function of digesstion that can arise as a symptom of a primary gastrointestinal dysfunction or as a complication due to other disorders such as appendicitis, gallbladder disturbances, or malnutrition. eGastroparesis is a paralysis of the stomach brought about by a motor abnormality in —the stomach or as a complication of disease=s such as diabetes, progressive systemic sclerosis, anorexia nervosa or myotonic dystrophmy. Constipation is a condition characterized by infrequent or difficult evacuation of feces ressulting from conditions such as lack of intestinaml muscle tone or intestinal spasticity. Post-opeerative ileus is an obstruction in the intestine dwmue to a disruption in muscle tone followirmg surgery. Intestinal pseudo- obstruction is =a condition characterized by constipation, c-olicky pain, and vomiting, but without evidence of physical obstruction.

Drug to=xicity is an important consideration in the treatment of humans and animals.

Toxic side effects (adverse effects) resulting from the administration of drugs include a variety of conditions which range from low grade fever to edeath. Drug therapy is justified only when the Wbenefits of the treatment protocol outweigh thee potential risks associated with the treatment. The factors balanced by the practitioneer include the qualitative and quantitative im pact of the drug to be used as well as the reswmilting outcome if the drug is not provided to the= individual. Other factors considered incluade the physica! condition of the patient, the dissease stage and its history of progression, =and any known adverse effects associated withm a drug.

Drug eRimination is typically the result of metabolic activity upon the drug and the subsequent excoretion of the drug from the body. Metabolic activity can take place within the

MBHB Ref. No. 04-972-C ARYX-020-PC vascular supply and/or within cellular compartments or organs. The liver is a principal site o»f drug metabolism. The metabolic process cam be categorized into synthetic and nonsyntheti- © reactions. In nonsynthetic reactions, the drag is chemically altered by oxidation, reductiorn, hydrolysis, or any combination of the aforementioned processes. These processes ar © collectively referred to as Phase I reactions.

In Phase II reactions, also known ass synthetic reactions or conjugations, the parert drug, or intermediate metabolites thereof, are combined with endogenous substrates to yield an addition or conjugation product. Metabolites formed in synthetic reactions are, typicall=y, more polar and biologically inactive. Asa mresult, these metabolites are more easily excrete=d via the kidneys (in urine) or the liver (in bi le). Synthetic reactions include glucuronidatiomn, amino acid conjugation, acetylation, sulfocomnjugation, and methylation. .

More than 90% of a dose of cisapriede is metabolized by oxidative N-dealkylation =at the piperidine nitrogen or by aromatic hydroxylation occurring on either the 4-fluorophenoxmy or benzamide rings.

The administration of cisapride to a_ human has been found to cause serious adversse effects including CNS disorders, increased systolic pressure, interactions with other druges, diarrhea, and abdominal cramping. Further, it has been reported that intravenomus administration of cisapride demonstrates tthe occurrence of additional adverse effects n ot experienced after oral administration of cissapride (Stacher et al. [1987] Digestive Diseas es and Sciences 32(11):1223-1230). It is beli=eved that these adverse effects are caused by thhe metabolites that result from the oxidatives dealkylation or aromatic hydroxylation of tlhe compound which occurs in the cytochrom=e P450 detoxification system. Cisapride is al:s0 subject to a number of undesirable drug/drumg interactions that are also a result of metabolissm by the cytochrome P450 system.

Between July 1993 and Decennber 1999, cisapride (PROPULSID, Janss=en

Pharmaceutica Products, L.P.) was reporte=dly associated with at least 341 serious cardi ac arrhythmias. These arrhythmias include ventricular tachycardia, ventricular fibrillation, torsades de pointes, and QT prolongation. Eighty (80) deaths have been reported. As a resmlt of these adverse effects, the product was voluntarily withdrawn from the open market in the

United States; however, the drug is available through an investigational limited acce=ss program.

MBHB Ref. No. 04-972-C ARYX-020-PC

The safety of SHT reeceptor agonists with gastrointestinal (GI) prokinetic activity has been limited due to cardiac effects (prolongation of QTc intervals, tamchycardia, torsades de pointes) and adverse drug irateractions due to hepatic cytochrome P-450 metabolism. A GI prokinetic agent of this class that lacks these liabilities would be ve=ry valuable in several therapeutic areas including GERD and gastric emptying disorder=s. Certain cisapride derivatives have been described in U.S. Pat. No. 6,552,046 and WO 0M /093849 (incorporated by reference herein in thesir entireties), however further compourands with even more advantageous properties woumld be desirable.

It has now been discovered that certain stereoisomers of one stach esterified structural and/or functional analog of cisapride have distinct and particularly advantageous properties.

Brief Summary

The subject inventiomn provides compounds and compositions of formula (X), which stereoisomeric esterified cissapride analogs, for the safe and effectiv e treatment of various gastrointestinal disorders including, but not limited to, gastroparesis, gastroesophageal reflux and related conditions. The compounds of the subject invention are amlso useful in treating a variety of conditions involvimg the central nervous system.

The compounds of th_e invention comprise compounds of formuzmla X:

Nal

Ses fo [d i, ha

R: RR,

Xx) and pharmaceutically acceptable salts thereof, wherein the bonds at positions 3 and 4 are cis relative to each other;

L is C1-Cs alkyl)- (3n one aspect, {C;-Cs alkyl)-), —(C,-C¢ alBcyl)-C(O)}-, or -C(O)- (C,-Cs alkyl)-, wherein each of the alkyl groups is optionally substitute=d with 1 or 2 groups that are independently halogen, C,-C, alkoxy, or OH and wherein one carbon in the alkyl portion of L may be replaced by ~N(Ro)-;

R, is halogen;

R, is amino, NH(C)-C, alkyl) or N(C;-Cq alkyl)(Ci-Cs alkyl);

Rj; is OH or C,-C alkkoxy;

MBHB Ref. No. 04-972-C ARYX-020-PC

R, is H or methyl; and

Rs is =0-C3-C; cycloalkyl, -O-heterocycloamlkyl, heterocycloalkyl, aryl, -Q-aryl, -

N(Rs)-(Co-Cs alkyl)-C(O)-aryl, or -N(Rs)-Co-Cs al kyl-aryl, -O-heteroaryl, -N(Rg)-C-C¢(0)- heteroaryl, or -N(Rg)-Co-Cs alkyl-heteroaryl, wherein each of the cyclic groups is unsubstituted or substituted at one or more substitiatable positions with C;-Ce alkyl, C1-Cs alkoxy, halogen, C;-Cs haloalkyl, C;-Ce haloalkox_y, hydroxyl, hydroxy-C;-Ca-alkyl, amino,

NH(C;-Cs alkyl), -N(C,-Cs alkyD)(C1-Ce alkyl), -CCo-Ce alkyl)-C(O)R1, or ~0-(Co-Cs alkyl)-

C(O)Ry;, methylsulfone, Co-Ce-sulfonamide, or NaO; wherein

Ro at cach occurrence is independently H or C3-C4 alkyl;

Ry is C;-Cs alkyl, OH, or

Ry; is C1-Cs alkoxy, optionally substituted with 1 or 2 groups that are independently

C,-C, alkoxy, amino, -NH(C,-Cs alkyl), 2N(C-Ces alkyl)(C:-Cs alkyl), (Co-Cs alkyl)-

C(O)N(Ro)-heterocycloalkyl, -O-heterocycloalkylk, -C1-Cs(O)N(Rg)-heteroaryl, or heteroaryl, wherein the heterocycloalkyl groups are op-tionally substituted with 1, 2, or 3 groups that are independently halogen, Ci-Ce alkwyl, C1-Cs alkoxy, hydroxy, hydroxy C,-Cs alkyl, Ci-Ce alkoxycarbonyl, -CO,H, CF, or OCF; the heteroaryl group is optionally substituted with 1, 2, or 3 groups that are independently halogen, C-Ce alkyl, C,-C.¢ alkoxy, hydroxy, hydroxy C;-Cs alkyl, Ci-

Cs alkoxycarbonyl, -COzH, CF, or OCF3 ; or

Ry; is —O-heterocycloalkyl wherein the hesterocycloalkyl is optionally substituted with 1, 2, or 3 groups that are independently halogen, C,-Cs alkyl, C;-Cs alkoxy, hydroxy, hydroxy C,-Cs alkyl, Ci-Cs alkoxycarbonyl, -CO®;H, CF3, or OCF;; and

Rag is C1-Cs alkoxy (preferably Cy-C4 alkoxy, more preferably methoxy), or OH.

The invention also encompasses compositions comprising at least one compound of formula (X) and at least one pharmaceutically acceptable excipient, adjuvant, carrier, or solvent.

The compounds of formula (X) are useful in the treatment or prevention of gastroesophageal reflux disease and substantial®y reduce adverse effects associated with the administration of cisapride. These adverse effexcts include, but are not limited to, diarrhea, abdominal cramping and elevations of blood presssure and heart rate.

Wea 2005/068461 PCT/US2005/0020510

MHABHB Ref. No. 04-972-C ARYX-020-PCC

Additionally, the compounds and commpositions of the invention are useful in tre=ating emesis and other conditions, including but not limited to dyspepsia, gastropa resis, comnstipation, post-operative ileus and intestinal pseudo-obstruction. As an added be=nefit, adlverse effects associated with the administration of cisapride are also reduced in these meethods of treatment.

Advantageously, the compounds of the subject invention are ligands for the 5HT, receptor and, accordingly, can be used to tweat conditions mediated through this receptor.

Thhese receptors are located in several areas ofthe central nervous system and the modulation off these receptors can be used to effect desired modulations of the CNS.

Advantageously, the subject invention provides stereoisomeric compounds ~which contain an ester moiety that does not detract from the ability of these compounds to pro-vide a therapeutic benefit, but which makes them rmore susceptible to degradation by serum aand/or cytosolic esterases, thereby avoiding the cytochrome PA450 drug detoxification system associated with adverse effects caused by cisapride and reducing the incidence off such aedverse events.

The subject invention further provides methods of treatment comprisimag the a-dministration of the compounds of formu la (X) and therapeutically effective amouants to isndividuals in need of treatment for gastroesophageal reflux disease, dyspepsia, gastroparesis, constipation, post-operative ileus, and intestinal pseudo-obstruction; and related conditions.

Advantageously, the therapeutic compounds of the subject invention are stzable in s=torage and provide for safer metabolism of the drugs as compared to other drugs; the=refore, tBhe compounds of the subject invention can be used with a lower incidence of side effe-cts and teoXicity.

In a further aspect, the subject invention pertains to the breakdown pmroducts (preferably metabolic breakdown products) which are formed when the ther=apeutic <ompounds of the subject invention are acted upon by esterases. These breakdown products an be used as described herein to monitor the clearance of the therapeutic compounds froma patient.

In yet a further aspect, the subject invention provides methods for synthesizzing the therapeutic stereoisomeric compounds of the subject invention, as well as internmaediates wuseful in preparing the compounds of interest.

MBHB Ref. No. 04-972-C ARYX-020-PC

Brief Description of the Dravevings

Figure 1 is a graph representing the Concentration-Resgponse Curves for 5-HT4 Receptor

Agonism of ATI-7505, serotonin, Cisapride, and ATI-7500.

Figure 2 is a graph representing gastric emptying in fed dogs. The data shown are normalized to the averaged vehicle control times of MMC return values. Values represent mean + SEM of 5 dogs. *p <0.05 versus vehicle controls

Figure 3 is a graph representing the metabolism of ATI-75505 and ATI-7500, with and without the CYP450 dependent Cofactor, NADPH. The pelots show mean and SD pM concentrations of ATI-7505 and ATI-7500. ATI-7505 (2 | aM) was incubated with human microsomal protein (1 mg) in the presence or absence of MJADPH regenerating system (cofactor).

Detailed Disclosure

In a further aspect, the invention provides compounds of Formula (X), wherein

Rs is ~O-C3-Cg cycloalkyl, -O-heterocycloalkyl, h eterocycloalkyl, wherein the heterocycloalkyl group is selected from piperidinyl, piper-azinyl, pyrrolidinyl, aza-bicyclo- octyl, in certain embodiments aza-bicyclo[2.2.2]octyl, azaa-bicyclo[3.2.1]octyl, aza-bicyclo- nonyl, aza-bicyclo-decyl, indolinyl, morpholinyl, thiomomrpholinyl, S,S- dioxothiomorpholinyl, and imidazolidinyl, -O-aryl, -N(Re)-C(0)-aryl, or ~N(Rg)-Co-Cs alkyl- aryl, wherein each of the cyclic groups is unsubstituted omr substituted at one or more substitutable positions with C;-Cs alkyl, C,-Cs alkoxy, halogen, C,-Cs haloalkyl, Ci-Cs haloalkoxy, hydroxyl, hydroxy-C;-C,-alkyl, amino, -NHEC,-Cs alkyl), -N(C,-Cs alkyl)(Ci-Cs alkyl), -C(O)R yy, or NO; wherein

Rs at each occurrence is independently H or Ci-C 4 alkyl; and

Ru is C1-Ce alkyl, OH, or

Ry; is C;-Ce alkoxy, optionally substituted with 1 or 2 groups that are independently

C,-C4 alkoxy, amino, -NH(C1-Cq alkyl), -N(C-Cs alkyl)-(C,-Cs alkyl), -C(O)N(Ry)- heterocycloalkyl, heterocycloalkyl or heteroaryl, whereir the heterocycloalkyl group is selected fromm pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, aza-bicyclo-octyl, in certain embodiments aza-

Wa 2005/068461 PCT/US2005/000510

MEHB Ref. No. 04-972-C ARYX-020-PC bicyclo[2.2.2]octyl, aza-bicyclo[3.2.1]octyl, a=za-bicyclo-nonyl and aza-bicyclo-decyl, wherein the heterocycloalkyl groups are optiomally substituted with 1, 2, or 3 groups that are independently halogen, C,-Cs alkyl, CC alkoxy, hydroxy, hydroxy Ci-Cs alkyl, C,-Cs alkoxycarbonyl, -CO:H, CF3, or CQCF,, the heteroaryl group is selected from pyridyl, pyrimidyl, quinolinyl, isoquinolinyl, and indolyl, wherein the heteroaryl groups are optionally substituted with 1, 2, or 3 groups that are independently halogen, C;-Cs alkyl, C1-Cs alkoxy, hydroxy, hydroxy C)-Cs alkyl, Ci-Cs alkoxycarbonyl, -COzH, CF3, or OCF;; or

Ry; is —O-heterocycloalkyl wherein the heter ocycloalky! is selected from piperidinyl, py-rrolidinyl, imidazolidinyl, morpholinyl, aza-bicy clo-octyl, in certain embodiments aza- bicyclo[2.2.2]octyl, aza-bicyclo[3 2.1]octyl, aza-bicyclo-nonyl, aza-bicyclo-decyl, and tegrehydrofuranyl, and wherein each heterocycloalkyl group is optionally substituted with 1, 2, or 3 groups that are independently halogen, C;-C alkyl, C;-Cq alkoxy, hydroxy, hydroxy

C 3-Cs alkyl, C1-Cs alkoxycarbonyl, -CO:H, CFs3, or OCF.

In another aspect, the invention provides compounds of Formula (X), wherein R; is chloro.

In yet another aspect, the invention provides compounds of Formula (X), wherein R2 is amino.

In still another aspect, the invention provides compounds of Formula (X), wherein Rs; is methoxy.

In another aspect, the invention provides compounds of Formula (X), wherein Ra is H om methyl.

In still yet another aspect, the invention prov ides compounds of Formula (X), wherein

R., is chloro; R; is amino; Rs is methoxy; and Ry is F or methyl.

In yet another aspect, the invention provides compounds of Formula (X), wherein R; iss chloro; R; is amino; Rs is methoxy; Re is H, and LL is —(C4-Cs alkyl)-C(O)-.

In another aspect, the invention provides cormpounds of formula (X), wherein two or ne1ore previously described aspects are combined.

In another aspect, the invention provides cornpounds of Formula (XI), which are compounds of formula (X) wherein L is —{CH>)s-CCO)-:

MBHB Ref. No. 04-972-C ARYX-020-PC

NOs Rg

Ry o {i 0

NE ld Re OCH;

R2 Ry xD.

In yet still another aspect, the invention provides compounds of formula (XI), wherein

RR; is chloro; R; is amino; Rj is methoxy; and R; is H or methyl.

In still another aspect, the invention provides compounds of formula (XT), wherein Rs i s-O-heterocycloalkyl, wherein the heterocycloalkyl group is selectezd from aza-bicyclo-octyl, & n certain embodiments 1-aza-bicyclof2.2.2Joct-3-yl or 8-aza-bicycBo[3.2.1]oct-3-yl, aza- bicyclo-nonyl, aza-bicyclo-decyl, where the aza nitrogen, is optionamlly substituted with methyl or ethyl; and Ry is H or methyl.

In still yet another aspect, the invention provides compound s of formula (XI), wherein

WR; is -O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from piperidinyl, -piperazinyl, or pyrrolidinyl, each of which is unsubstituted or substituted at one or two positions with groups that are independently C;-Cs alkyl, C1-Cs alk oxy, halogen, C-C4 “haloalkyl (in one aspect, CF3), Ci-Cq haloalkoxy (in one aspect OC Fs), hydroxyl, hydroxy

C.-C, alkyl, amino, -NH(C;-Cs alkyl), -N(C;-C4 alkyl)(C1-Ca alkyl), -(Ci-Cs alkyD)-C(O)Ry, or NO; and Ry is H or methyl.

In yet another aspect, the invention provides compounds of formula (XI), wherein Rs is -O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from indolinyl, morpholinyl, thiomorpholinyl, S,S-dioxothiomorpholinyl, and imicazolidiny!, each of which is unsubstituted or substituted at one or two positions with groups that are independently C;-

C, alkyl, C1-C, alkoxy, halogen, C;-Cs haloalkyl (in one aspect, CE=3), C;-C4 haloalkoxy (in one aspect OCF3), hydroxyl, hydroxy Ci-Cs alkyl, amino, -NH(C,—C, alkyl), -N(C1-Cs4 alkyl)(C1-C4 alkyl), (Co-Cs alkyl)-C(O)Ri;, or NO; and Ry is H omr methyl.

In yet another aspect, the invention provides compounds of= formula (XI), wherein Rs is -O-phenyl, N(Rg)-(Co-Cs alkyl)-C(O)-phenyl, or -N(Rg)-Co-Ca4 alkyl-phenyl, wherein the phenyl group is substituted with one or two groups that are indepemdently C,-Cs alkyl, Ci-Cs alkoxy, halogen, C;-Cj haloalkyl (in one aspect, CFs), C1-Ca haloaslkoxy (in one aspect

OCF), hydroxyl, hydroxy Ci-Cs alkyl, amino, -NH(C,-Cs alkyl), ~N(C,-C, alkyl)(C;-Cs alkyl), -(Co-Cs alkyl)-C(O)R11, or NO;; and R, and Ry are indepen~dently H or methyl.

MBHB Reef. No. 04-972-C ARY>=-020-PC

In another aspect, the invention provides compounds of formula (XI), wh_erein Ry is

H.

In yet another aspect, the invention provides compounds of formula (XI),- wherein Ryj is C;-Cs alkoxy, optionally substituted with 1 or 2 groups that are independently C;-C4 alkoxy, armino, -NH(Cy-Cs alkyl), -N(C,-Cs alkyl(C,-Cs alkyl), -(Co-Cs alkyl)-C=(O)N(R9)- heterocyc loalkyl, or heterocycloalkyl wherein the heterocycloalky! group is selected from pyrrolidimyl, piperidinyl, piperazinyl, and morpholinyl, wherein the heterocycloamlkyl groups are optiommally substituted with 1, 2, or 3 groups that are independently halogen, CC;-C; alkyl,

C,-Ce alkoxy, hydroxy, hydroxy C-Cs alkyl, C1-Cs alkoxycarbonyl, -COzH, CF, or OCF.

In another aspect, the invention provides compounds of formula (XI), wimerein two or more prewiously described aspects are combined.

In another aspect, the invention provides compounds of Formula (X10), i.e, compounds of formula (X), of the formula:

Py >< Ris 0 a (Seve alkyl COR

Ry 43 XX

NE OCH, Rus

SX Ra

R, Rs 1s xm), wherein RRs is H, C)-Cs alkyl, C;-Cs alkoxy, halogen, Ci-Cs haloalkyl (in one asspect CFs),

C,-Cg hal oalkoxy ( in one aspect OCF3), hydroxyl, hydroxy C,;-Cq alkyl, amino, -NH(C;-Ce alkyl), -NI(C,-Cs alkyl)(Cy-Cs alkyl), methylsulfone, Co-Cs-sulfonamide or NO, and Ry¢ is H or —0-(Co-Cs alkyl)-C(O)R11. In another aspect, Ris is H.

Inn yet another aspect, the invention provides compounds of formula (XID), wherein Rs and Rs are independently H or methyl and Ry, is OH.

Irm still yet another aspect, the invention provides compounds of formula (XII), wherein ER, and Ry are independently H or methyl and Ry, is C-C¢ alkoxy, optionally substitutesd with 1 or 2 groups that are independently C,-C4 alkoxy, amino, -NH&C,-C; alkyl), -N(C,-Ce. alkyl)(C1-Cs alkyl), -(Co-Cs alkyl)-C(O)N(Ry)-heterocycloalkyl, or heterocycsloalkyl wherein the heterocycloalkyl group is selected from aza-bicycl o-octyl, in certain ermbodiments 1-aza-bicyclo[2.2.2]Joct-3-yl or 8-aza-bicyclo[3.2.1]oct-3-3w], aza- bicyclo-n onyl, aza-bicyclo-decyl, where the aza nitrogen is optionally substitute=d with methyl

MBHB Ref. No. 04-972-C ARYX-020-PC or ethyl, pyrrolidinyl, piperidinyl, piperazinyl, and rnorpholinyl, wherein the heterocycloalliyl groups are optionally substituted with 1, 2, or 3 growups that are independently halogen, Cy- Cs alkyl, C,-Cs alkoxy, hydroxy, hydroxy C:-Ce alkyl, C;-Cs alkoxycarbonyl, -COzH, CFs, or~

OCF;, and R4 and Ry are independently H or methy 1. In another aspect, Rs, Rg, and Ry; ares as previously defined and Ris is H, Ry is chloro; Rs is amino; and Rj is methoxy.

In yet still another aspect, the invention prowides compounds of formula (XII), wherein R4 and Ry are independently H or methyl and Ru is C,-Cs alkoxy, optionally substituted with 1 or 2 groups that are independently C,-Cq alkoxy, amino, -NH(C)-Cs alkyl),

N(C)-Cs alkyl)(C;-Cs alkyl), or heteroaryl, wherein the heteroaryl group is selected from pyridyl, pyrimidyl, quinolinyl, isoguinoliny}, and irdolyl, wherein the heteroaryl groups amre optionally substituted with 1, 2, or 3 groups that are= independently halogen, C,-Cs alkyl, Cs-

Cs alkoxy, hydroxy, hydroxy Ci-Cs alkyl, C,-Cs alkoxycarbonyl, -CO;H, CF3, or OCF3; amnd

R4 and Ry are independently H or methyl. In anothmer aspect, Rs, Ro, and Ry are as previorusly defined and Rs is H, R; is chloro; R; is amino; andl Rj is methoxy.

In still another aspect, the invention providees compounds of formula (XII), wherei.nat least one of Rs and Ry is H.

In another aspect, the invention provides compounds of formula (XII), wherein two or more previously described aspects are combined.

In another aspect, the invention provides compounds of Formula (XIII), i.e., compounds of formula (XII), of the formula:

Re Rig o CYC

Rt jo» 7 (Cos alyFCOR

NER OCHy R-s 2X Ra

R, Rs xm , wherein R;s is H, C;-Cs alkyl, C,-C¢ alkoxy, halo_gen, C1-Cs haloalkyl (in one aspect CF =3),

C,-Ce haloalkoxy ( in one aspect OCF), hydroxy, hydroxy C1-Cs alkyl, amino, -NH(C1—Cs alkyl), -N(Ci-Cs alkyl)(C,-Cs alkyl), or methylsulifone, Co-Ce-sulfonamide, NO,, and Ries is H or =0Co-Cs alkyl)-C(O)R11. In another aspect, R;s is H.

In yet another aspect, the invention provides compounds of formula (XID, whereein

MBHB Ref. No. 04-972—C ARYX-020-PC

R, and Rs are independemntly H or methyl, and Ry: is OH, C,-C. alkoxy @Cin another aspect, Ci-

C; alkoxy), or C;-C; alkcoxy-Cy-Cs alkoxy-. In another aspect, Rs, Rg, a-nd Ry, are as previously defined and RR, is chloro; R, is amino; and Rj is methoxy.

In still yet anothe=r aspect, the invention provides compounds of formula (XIN), wherein Rs and Ro are inmdependently H or methyl, and Ry, is C,-C, alkoxy substituted with amino, -NH(C;-Cs alkylD), -N(Ci-Ce alkyl)(C1-Cs alkyl), aza-bicyclo-ocstyl, in certain embodiments 1-aza-bicy=clo[2.2.2]oct-3-yl or 8-aza-bicyclo[3.2.1Joct-3—Yl, aza-bicyclo-nonyl, aza-bicyclo-decyl, wheree the aza nitrogen is optionally substituted with. methyl or ethyl; and

Ry is H or methyl, pyrro lidinyl, piperidinyl, morpholinyl, pyridyl, or {(*Cy-Cs alkyl)-C(O)NH- pyrid-4-yl. In another a=spect, Ra, Rs, and Ry; are as previously defined and R, is chloro; Ra is amino; and R; is methomy.

In still another a spect, the invention provides compounds of for—mula (XTID), wherein

R, and Ry are independe=ntly H or methyl, and Ry; is C;-Ca alkoxy subsstituted with amino, -NH(C;-Cs alkyl), or -NI(C1-Ce alkyl)(C:1-Cs alkyl). In another aspect, “Ry, Rg, and Ry are as previously defined and “R,; is chloro; R; is amino; and Rj is methoxy.

In yet another asspect, the invention provides compounds of formmula (XIII), wherein

R, and Rg are independ: ently H or methyl, and Ry; is C;-Cq alkoxy subsstituted with pyrrolidiny}, piperidiny~1, morpholinyl, pyridyl, or —~Co-Cs alkyl)-C(ONH-pyrid-4-yl. In another aspect, Rs, Rg, :and Ry; are as previously defined and R; is chloro; R; is amino; and

Rj is methoxy. ’

In still another aspect, the invention provides compounds of foermula (XIII), wherein at least one of Ry and RRgis H.

In another aspect, the invention provides compounds of formu 1a (XIII), wherein two or more previously described aspects are combined .

In another aspe -ct, the invention provides compounds of formu_la (XIV), i.e. compounds of formula. (X), of the formula:

MBHB Ref. No. 04-972-C ARYX-020-PC " _ Le

A Ser atkyl)-C(O)Ry4

R, 0 fo) Rs =

SFY OCH;

Ry Ry } (XIV) wherein R;s iss H, C,-Cs alkyl, C1-Cs alkoxy, halogen, C 1-Cs haloalkyl (in one aspect CF3),

C,-Cs haloalkeoxy ( in one aspect OCF), hydroxyl, hydr-oxy Ci-Cs alkyl, amino, -NH(C,-Cs alkyl), -N(C,-=C; alkyl)(C,-Cs alkyl), methylsulfone, Co—Cq-sulfonamide, or NO, and Ry is FH or -O-(Co-Cs =alkyl)-C(O)Ry;. In another aspect, Ris is HH.

In still another aspect, the invention provides commpounds of formula (XIV), wherein

Rs and Ro are independently H or methyl, and Ru is OHM, C,-C, alkoxy (in another aspect, C1 —

C; alkoxy) or C,-C; alkoxy-C;-C; alkoxy-. In another amspect, Rs, Ro, and R, are as previously defined and R; is chioro; R2 is amino; and R= is methoxy. In still another aspect, at least one of” R; and Ry is H.

In yet still another aspect, the invention providess compounds of formula (XIV), wherein Rs amd Ry are independently H or methyl, and ER; is C1-Ca alkoxy substituted with amino, -NH(C-Ce alkyl), -N(C1-Cs alkyl)(C,-Cs alkyl),. aza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]Joct-3-y! or 8-aza-bicywrclo[3.2.1]oct-3-yl, aza-bicyclo-nonyR, aza-bicyclo-d-ecyl, where the aza nitrogen is optionally ssubstituted with methyl or ethyl; and

Ry, is H or methyl, pyrrolidinyl, piperidinyl, morpholiny~l, pyridyl, or <(Co-Ce alkyl)-C(O)NH_- pyrid-4-yl. Im another aspect, Rs, Rs, and Ry, are as pre=viously defined and R, is chloro; Rz us amino; and R= is methoxy.

In still another aspect, the invention provides co. mpounds of formula (XIV), wherein

Rs and Ry are independently H or methyl, and Ri is C,—C, alkoxy substituted with amino, -NH(C-Cs al kyl), or -N(C,-Cs alkyl)(C1-Cs alkyl). In &another aspect, Rs, Ry, and Ry; are as previously de fined and R is chloro; R; is amino; and R_3 is methoxy.

In yet another aspect, the invention provides cormpounds of formula (XIV), wherein

R, and Ry are: independently H or methyl, and Ri is Cy--Cs atkoxy substituted with pyrrolidinyl, gpiperidinyl, morpholinyl, pyridyl, or (Co—Cs alkyl)-C(O)NH-pyrid-4-yl. In

MBHB Ref. No. 04-972-C ARYX ~020-PC another aspect, Ra, Ro, and Ry; are as previously defined and R, is chloro; R; is armino; and

Rj is methoxy.

In still another aspect, the inventzion provides compounds of formula (XIV), wherein at least one of Rs and Ro is H. "In another aspect, the invention gprovides compounds of formula (XIV), wherein two or more previously described aspects aree combined .

In another aspect, the invention gprovides compounds of formula (XV), i.e, compounds of formula (X) of the formula: ] 3 AE alkyl)-C(O)R44

R, 43 o (7 bm,

R: Rr s

XV) wherein n is 1 or 2. ‘

In still another aspect, the invention provides compounds of formula (XV, wherein

Ry is H or methyl, and Ry, is OH, C;-Cq- alkoxy (in another aspect, C,-C3 alkoxy) orC,-C; alkoxy-C;-C; alkoxy-. In another aspec=t, Rs and Ry, are as previously defined an«d R, is chloro; Rj is amino; and Rj is methoxy. In still another aspect, at least one of R4 and Ry is H.

In yet still another aspect, the inwvention provides compounds of formula (XV), wherein R4 and Rg are independently H or methyl, and Ry; is C;-Cs alkoxy substituted with amino, -NH(C,-Cs alkyl), -N(C,-Cs alkmy)(C1-Cs alkyl), aza-bicyclo-octyl, in cert=ain embodiments 1-aza-bicyclo[2.2.2]oct-3—yl or 8-aza-bicyclo[3.2.1]oct-3-l, aza-biccyclo-nonyl, aza-bicyclo-decyl, where the aza nitrogen is optionally substituted with methyl or ethyl; and

Ry is H or methyl, pyrrolidinyl, piperidimyl, morpholinyl, pyridyl, or -C(O)NH-pwrid-4-yl. In another aspect, Rs, Ro, and Ry; are as previously defined and R; is chloro; R; is arnino; and

R; is methoxy.

In still another aspect, the invention provides compounds of formula (XV», wherein

R, and Ry are independently H or methyl, and Ry; is C1-C4 alkoxy substituted with amino, -NH(C;-Cs alkyl), or -N(Cy-Cs alkyl)(C 1-Cs alkyl). In another aspect, Ry, Ry, andl Ry, are as previously defined and R is chloro; Ry is amino; and Rj is methoxy.

MBHB Ref. No. 04-972-CC ARYX-020-PC

In yet another aspe=ct, the invention provides compounds of formula (X_V), wherein Rs is H or methyl, and Ry; is C;-C, alkoxy substituted with aza-bicyclo-octyl, in certain embodiments 1-aza-bicyc=lo[2.2.2]oct-3-yl or 8-aza-bicyclo[3.2.1]oct-3-Yl, azam-bicyclo-nonyl, aza-bicyclo-decyl, where the aza nitrogen is optionally substituted with methyl or ethyl; and

Ra is H or methyl, pyrroli_dinyl, piperidinyl, morpholinyl, pyridyl, or <(Co-Cé alkyl)-C(O)NH- pyrid-4-yl. In another asgpect, Rs, Ro, and Ry; are as previously defined and Rx is chloro; Ra is amino; and Rj is methoxy.

In another aspect, the invention provides compounds of formula (XV), wherein two or more previously describe=d aspects are combined .

In another aspect, the invention provides compounds according to any one of formulas (X), (XI), XIN (XID, (XIV) or (XV), wherein R;, Ry, and Rj are o-riented on the phenyl ring as follows:

O

“OLY

R: Rs

In another aspect, the invention provides compounds according to any~ one of formulas (X), (XT), (XIDw, (XII), (XIV) or (XV), wherein bond 3 has the “8” configuration and bond 4 has the “R” configuration.

In still another as=pect, the invention provides compounds according to any one of formulas (X), (XT), (XIDw, (XIID, (XIV) or (XV), wherein R,, Raz, and R3 are Oriented on the phenyl ring as follows:

Oo “OL v

R: Ry , and bond 3 has the “S” c=onfiguration and bond 4 has the “R” configuration.

In another aspect, the invention provides compounds according to any one of formulas (X), (XT), (XID, (XID), (XIV) or (XV), wherein bond 3 has the “R’ configuration and bond 4 has the “S” configuration.

In another aspect, the invention provides compounds according to any one of formulas (X), (XT), (XID, (XII), (XIV) or (XV), wherein Ry, Ra, and R; are oriented on the phenyl ring as follows: 16

MBHB Ref. No. 04-972-C ARYX-020-PC . le

CL

Rs Rs _ and bond 3 has the “R” configuration and bond 4 has thme “S” configuration.

Imn still another aspect, the invention provides cosmpounds of formula (X), wherein

R, is ch] oro; R; is amino; Rs is methoxy; Ry is H, and Ry, Ry, and R; have the following orientati-on on the phenyl ring: 0}

CL

Rs Ra ,and

L_ is (C3-Cs alkyl)- wherein one carbon may be= replaced by -N(Ro)-, or ~(C2-Cs alkyt)-C (0)-. In yet another aspect, the R1, R2, and RS are as defined and oriented on the phenyl ring as previously described, Ry is as previously” defined and Rs is-O-heterocycloal kyl, wherein the heterocycloalkyl group is selected from azaa-bicyclo-octyl, in certain embodirments 1-aza-bicyclof2.2.2Joct-3-yl or 8-aza-bicyclo[3.2.1Joct-3-y1, aza-bicyclo-nomnyl, aza-bicy-clo-decyl, where the aza nitrogen is optionally= substituted with methyl or ethyl, piperidiryl, piperazinyl, and pyrrolidinyl, wherein the piperidinyl, piperazinyl, and pyrrolidinyl groups are unsubstituted or substituted at cene or two positions with groups that are independently C-Cs alkyl, C,-C4 alkoxy, halogen, &C;-Cy haloalkyl, C,-C4 haloalkoxy, hydroxy 1, hydroxy C,-Cs alkyl, amino, -NH(C,-C4 alkyl), -N(C;-C, alky)(C,-C, alkyl), (oC;-

Ce alkyl D-C(O)Ry}, or NO,, wherein

FR; is C;-Cg alkoxy, optionally substituted with 1 or 2 groups that are independently

C,;-Cs alkoxy, amino, -NH(C;-Cs alkyl), -N(C;-Cs alky 1)(Ci-Ce alkyl), (Co-Cs alkyl)-

C(O)N(Ry)-hcterocycloalkyl, or heterocycloalkyl wherein the heterocycloalkyl group is selected from aza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3-yl or 8— aza-bicy-clo[3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza-bicyaclo-decyl, where the aza nitrogen iss optional Jy substituted with methyl or ethy!; and Rs is FC or methyl, pyrrolidinyl, piperidiny~-1, piperazimnyl, and morpholinyl, wherein the heterocycloamlkyl groups are optionally substitut=ed with 1, 22, or 3 groups that are independently halogen, C)-Cs alkyl, C,-Cs alkoxy, hydroxy, hydroxy C,-Cs alkyl, C;-Cs alkoxycarbonyl, -CO,H, CIF; or OCF.

In still yet another aspect, the invention provide-s compounds of formula (X), whereein 17

W/ (0 2005/068461 PCT/US2005/000510

MDBHB Ref. No. 04-972-C ARYX-020-PC

R; is chloro; R; is amino; Rs is methoxy; Ry is H, and R;, “R;, and R; have the following orientation on the phenyl ring: 0] “OLY

RS Rj , and

L is «(C3-Cs alkyl)- wherein one carbon may be regplaced by —-N(Ry)-, or |(C2-Cs allecyl)-C(O)-. In yet another aspect, the R), Ry, and R; are as defined and oriented on the ph enyl ring as previously described, Ry is as previously de=fined and Rs is heterocycloalkyl, which is selected from aza-bicyclo-octyl, in certain embodiments 1-aza-bicyclo[2.2.2]oct-3- yl or 8-aza-bicyclo{3.2.1]oct-3-yl, aza-bicyclo-nonyl, aza- bicyclo-decyl, where the aza nitrogen, is optionally substituted with methyl or ethyl.

In still yet another aspect, the invention provides ceompounds of formula (X), wherein

Ry is chloro; Rj is amino; Rs is methoxy; Ry is H, and R;, "Ry, and R; have the following ormentation on the phenyl ring: 0 “OLY

Rs Ra , and

L is =(C3+Cs alkyl)- wherein one carbon may be re-placed by -N(Ry)-, or «(C2-Cs alk<yl)-C(O)-. In yet another aspect, the R), Rz, and R3 are as defined and oriented on the ph enyl ring as previously described, Ry is as previously defined and Rs is-N(Ry)-Co-C4 alkyl- ars] or -N(R)-(Co-Cs alkyl)-C(O)-aryl, wherein the aryl group is unsubstituted or substituted at «one or more substitutable positions with C,-Cs alkyl, C; -Cs alkoxy, halogen, Ci-Cs haJoalkyl, C,-Cs haloalkoxy, hydroxyl, hydroxyalkyl, amimno, -NH(C,-Cs alkyl), -N(C,-C¢ alkzyl)(C)-Cs alkyl), -(Co-Cs alkyl)-C(O)Ry;, or NO. In st-ill another aspect, the aryl group is a phenyl substituted with ~(Co-Cs alkyl)-C(O)Ry; and optionally substituted with 1 or 2 groups independently selected from C;-Cs alkyl, Ci-Cs alkzoxy, halogen, CFs, OCFs, hydroxyl, hydroxyalkyl, amino, -NH(C;-Cj alkyl), -N(C;-«C, alkyl}(C,-Cs alkyl), or NO;, wherein

Ry; is C;-Cs alkoxy, optionally substituted with 1 or 2 groups that are independently

CC, alkoxy, amino, -NH(C;-Cs alkyl), -N{(C:-Cs alkyl)(C1-Cs alkyl), -(Co-Cs alkyl)-

C(=0O)N(Ry)-heterocycloalkyl, or heterocycloalkyl wherein the heterocycloalkyl group is seE ected from pyrrolidinyl, piperidinyl, piperazinyl, and m orpholinyl, wherein the

MBHB Ref. No . 04-972-C ARYX-02 O-PC heterocycloalky™1 groups are optionally substituted with 1, 2, or 3 groups that are independently h alogen, C,-Cs alkyl, Ci-C¢ alkoxy, hydmroxy, hydroxy C,-Cs alkyl, Cy ~Cs alkoxycarbonyl, -CO;H, CF3, or OCF;. In a preferred aspect the -(Co-Cs alkyl}-C(O)e Ri group is attached to position 4 of the phenyl ring.

In still arnother aspect, the orientation of bonds =3 and 4 is as follows: :

NT ve

OCH;

In a preferred aspect, the orientation of bonds 3 and 4 is as follows: or 4 ap 2

OCH;

The invention further provides methods for trealing emesis, dyspepsia, gastrogparesis, constipation, inteestinal pseudo-obstruction, gastroesoph_ageal reflux, or post-operative ileus, the method comprising administering a therapeutically seffective amount of a compou-nd or salt according ofS formula (X) to a patient in need of such treatment.

The subject invention provides compounds that are more susceptible to degraciation by serum and/or cytosolic esterases than cisapride, thus avoiding the adverse effects associated with rmnetabolism by cytochrome P450.

Advantageously, the therapeutic compounds of the subject invention are stablee in storage but have a relatively short half-life in the physicological environment; thercforee, the compounds of th_e subject invention can be used with a Hower incidence of side effectss and toxicity.

In a preferred aspect of the subject invention, the=rapeutic stereoisomeric compounds are provided that= are useful in the treatment of gastroesophageal reflux disease and thzat contain an ester group, which is susceptible to degradatiZ on by esterases, thereby breaking down the compomund and facilitating its efficient removaml from the treated individual. Ina preferred aspect, the therapeutic stereoisomeric compounds are metabolized by the Phmase 1 drug detoxificatieon system.

A further aspect of the subject invention pertainss to the breakdown products (preferably metabolic breakdown products, i.c., metabolites, generally acids of parent esters) 19

MBHB Ref. No. 04-972-C ARYX-020-PC that are produced when the therapeutic compouncs of the subject invention are acted upon by an esterase. The presence of these breakdown preoducts in the urine or serum can be used to monitor the rate of clearance of the therapeutic compound from a patient.

Degradation of the compounds of the subject invention by esterases is particularly advantageous for drug metabolism because these enzymes are ubiquitously distributed and their activity is not dependent on age, gender, or adisease state to the same extent as oxidative hepatic drug metabolism.

The subject invention further provides me=thods of treating disorders, such as gastroesophageal reflux disease comprising the asdministration of a therapeutically effective amount of at least one stereoisomeric structural a-nd/or functional analog of cisapride to an individual in need of treatment. In a specific aspe=ct, the subject invention provides stereoisomeric structural and/or functional analogs of cisapride and pharmaceutical compositions of these esterified compounds.

The subject invention further provides mamterials and methods for the treatment of emesis and such other conditions, including but mot limited to dyspepsia, gastroparesis, constipation, and intestinal pseudo-obstruction, vevhile substantially reducing adverse effects associated with the administration of cisapride.

In a preferred aspect of the subject invention, therapeutic stereoisomeric compounds are provided which are useful in the treatment of gastroesophageal reflux, dyspepsia, gastroparesis, constipation, post-operative ileus, zand intestinal pseudo-obstruction and which contain an ester group which is acted upon by est-erases thereby breaking down the compound and facilitating its efficient removal from the treamted individual.

The subject invention further provides me=thods of synthesizing the unique and advantageous compounds of the subject inventiom. Particularly, methods of producing and purifying such stereoisomeric compounds are tau- ght. Methods of adding such ester moieties and of producing and purifying stereoisomers, aree well known to the skilled artisan and can be readily carried out utilizing the guidance provided herein.

Preferred Compounds

In a preferred aspect, the present inwwention provides isolated stereoisomers of

Compound I, which contains three chiral centers.

MBHB Ref. No. 04-972-C AR YX-020-PC < © HN

H SH

: 3 a~~h © © 0 N o 6-[4-(4-Amino-5-chlaro-2-met-hoxy-benzoytamino)-3-methoxy-piperidin-1-yll-hexanoi ¢ acid 1-aza- bicycio[2.2.2Joct-3-yl ester

Compound

Two of the chiral centers exist in cisapride and norcisapride ancl are in the cis configuration in the active «drugs: na wa pe ’ ° 7 ~~ A o BQ! / (*)-Cisapride ()-Norcisaprides

Thus, for example, pharmaceutically active norcisapride is a racemiic mixture of the two cis enantiomers: cl !

HoN HN

NH “O) _0 © To AO Og \NH / / (->-Norcisapride (+)-Norcisapride

In one aspect, the current invention is particularly concerned with the configuration at the third chiral center, in thee quinuclidinol moiety. This group is eliminated din the conversion to the acid metabolite refer-red to herein as + Compound II: ci

Hp N ~0 0 p Ne~~Aoon

Compound I1

MB=HB Ref. No. 04-972-C ARYX-020-PC

The preferred Compound I stereoisomers of the presemt invention are made by conjugating R or S quinuclidinol to (+)- or (-)-norcisapride, givings Compounds HI, IV, V and

VL

Cl

HN § (R,

Toot

GN ~~A

LO Og N o / (3R .4S,3'R)-6-[4-(4-Amino-5-chioro-2-methoxy-benzoylamine)-3-methoxy-pE peridin-1-yi]-hexanoic acid 1-aza- bicyclo{2.2.2]oct-3-yl ester compound IIL: (-)(R)-compound I 1

HN

H (R) N

No, &

S, (25,4R,3'R)-6-{4-(4-Amino-S-chloro-2-methoxy-benzoylamino)-3-methoxy-pipe=ridin-1-yil-hexanolc acid 1- aza-bicyclof2.2.2]Joct-3-yl ester compound IV: (+)}(R)-compound I

Cl y=

NH

T) sd i

O (o] N pd 7 (R 0 (3R ,4S,3'S)-6-[4-(4-Amino-5-chloro-2-methoxy-benzoylamino)-3-methoxy -piperidin-1-ylj-hexancic acid 1- aza-bicyclo[2.2.2]oct-3-yl ester compound V: (-)(S)-compound

MBHB Ref. No. 04-972-C | ARYX-020-PC

HN PY

OY 7

A200 7 LoL (38.4R 35)-6-{4-{4-Amino-5-chioro-2-methoxy-bo szoplarino) & methoxy piperidin-1 -yl}hexanoic acid t- compound VI: (+)(S)-compound 1

In a preferred aspect, the subject inventiosn pertains to stereoisomerically isolated compounds, and compositions comprising the cosmpounds. The isolated stereoisomeric forms of the compounds of the invention are substantia lly free from one another (i.e., in stereoisomeric excess). In other words, the “R” forms of the compounds are substantially free from the “S” forms of the compounds and amre, thus, in stereoisomeric excess of the “S” forms. Conversely, “S” forms of the compounds are substantially free of “R” forms of the compounds and are, thus, in stereoisomeric excesss of the “R” forms. In one aspect of the invention, the isolated stereoisomeric compounds are in at least about 80% stereoisomeric excess. In a preferred aspect, the compounds ares in at least about 90% stereoisomeric excess.

In a more preferred aspect, the compounds are in. at least about 95% stereoisomeric excess.

In an even more preferred aspect, the compounds are in at least about 97.5% stereoisomeric excess. In a most preferred aspect, the compounds are in at least about 99% stereoisomeric excess. Similarly, the “(+)” and “(-)” forms of thme compounds are also provided in stereoisomeric excess.

As described herein, the various stereoiscmmers have particular unexpected properties that, advantageously, can be used to customize treatment for a particular set of circumstances.

Thus, for example, compounds containing the (3 R)-isomer in the quinuclidinyl ester moiety, i.e., compounds III and IV, are rapidly metabolized by esterases in human plasma, whereas

MBHB Ref. No. 04-972-C ARYX-020-PC compounds containing the (3’S)-i somer of quinuclidinol, i.e., compounds V znd VI, undergo a much slower metabolism.

Thus, the (3’R)-isomers of compound I can be used when a short-durzation of action is preferred, for example stimulation: of gastric motility in an acute episode, suc=h as pulsatile administration to patients with acute gastroparesis, or in acute gastroesophageal reflux.

Another advantage of rapid metabolism by esterases to an substantially less amctive metabolites, i.e., compound II, is the very low probability of drug-drug interamctions and toxicity. Therefore these short-acting (R)-isomers can be advantageously use=d as an intravenous formulation for treating gastroesophageal reflux in premature ne~wborn who notoriously are not able to metabolize drugs as well as adults because their CC YP450 system is not fully developed. In these newborn, a drug having rapid metabolism by a system other than CYP450, ¢.g., esterases, is a great advantage. On the other hand, the (37S)-isomers of compound I are best used in chromic situations of the same ailments, for exarnple gastroparesis in diabetic patients ©r cancer patients under opiates, or in chromic gastroesophageal reflux in patients who need 24-hour coverage.

In addition to their differences in metabolic fate, these separate isomers also have different binding affinities for thes 5-HT4 receptor, thus suggesting different sactivities as well, and therefore different therapeutic uses. Thus, in a decreasing order of affinity for the 5-HT4 receptor, the isomers can be ranked as follows (in parentheses are the bindin_g constant Ki values); compound IV (1.4nM), compound VI (3.4nM), compound II (28nN), and compound V (72nM). These binding experiments were performed using the= radiolabel displacement method described im standard textbooks and easily reproducible by persons skilled in the art of molecular biology.

As a conclusion to these considerations: when the 3 and 4 positions zare cis relative to each other, compound I is a mixture of 4 isomers, consisting of 2 pairs of en_antiomers. The first pair of enantiomers is (+)(R)-compound I and (-)(S)-compound I (compeounds IV and V, respectively), the second pair of enantiomers is (-)(R)-compound I and (+)(S8)-compound I (compounds III and VI, respectively). Within each enantiomeric pair, each sseparate enantiomer has different properti es regarding both their rate of hydrolysis bys esterases and regarding their affinity at the 5-FIT4 receptor. These different properties give them separately advantageous therapeutic uses which are not interchangeable, i.e., which arez specific to each isomer, and which are not applicable to the racemic mixture. These differermces of affinity at

- MBHB Ref. No. 04--972-C ARYX-020-PC the receptor and these differences in metabolic rates are not predictable and neithesris it possible to dissect these properties when testing the racemic mixture.

Definitions

As used herein, the term “alkyl” includes those alkyl groups of a designed number of carbon atoms. Alk=y! groups may be straight, or branched. Examples of “alkyl” imnclude methyl, ethyl, propzy], isopropyl, butyl, iso-, sec- and tert-butyl, pentyl, hexyl, hepmtyl, 3- ethylbutyl, and the like. If the number of carbon atoms is not specified, the subjeact “alkyl” moiety has from 1 to 6 carbons.

The term “alkoxy represents an alkyl group of indicated number of carbown atoms attached to the pare=nt molecular moiety through an oxygen bridge. Examples of alkoxy groups include, for example, methoxy, ethoxy, propoxy and isopropoxy.

By “aryl” is meant an aromatic carbocyclic group having a single ring (e.£2., phenyl) that is optionally fimsed or otherwise attached to other aromatic hydrocarbon ringss or non- aromatic hydrocarbon rings. “Aryl” includes multiple condensed rings in which zat least one is aromatic, (e.g., 1 ,2,3,4-tetrahydronaphthyl, naphthyl), wherein each ring is optionally mono-, di-, or trisulbstituted with the groups identified below, as well as multiple rings that are not fused, such as, for example, biphenyl or binaphthyl. Preferred aryl groupss of the present invention ame phenyl, 1-naphthyl, 2-naphthyl, indanyl, indenyl, dihydronamphthyl, fluoreny), tetralinyk or 6,7,8,9-tetrahydro-5H-benzofa]cycloheptenyl. More preferred are phenyl, biphenyl, and naphthyl. Most preferred is phenyl. The aryl groups hereimn are unsubstituted or, as specified, substituted in one or more substitutable positions with various groups. For exampele, such aryl groups may be optionally substituted with, for example, C;-

Cs alkyl, C,-Cs alkoxy, halogen, hydroxy, cyano, nitro, amino, mono(C;-Ce)alkyRamino, di(C,-Cs)alkylamim o, C5-Csalkenyl, C,-Cealkynyl, C,-Cs haloalkyl, C;-Cs haloalleoxy, amino(C,-Ce)alkyl, mono(C;-Cs)alkylamino(Ci-Cs)alkyl or di(C;-Ce)alkylamino®(C;-

Ce)alkyl.

The term “haloalkoxy” refers to an alkoxy group substituted with at least mone halogen atom and optionally further substituted with at least one additional halogen atom, where each halogen is indepenclently F, Cl, Br or I. Preferred halogens are F or Cl. Preferred haloalkoxy groups contain 1-6 carbons, more preferably 1-4 carbons, and still more preferably 1-2 carbons. “Haloalkoxy” includes perhaloalkoxy groups, such as OCF; or OCF,CF=.

MB-HB Ref. No. 04-972-C ARYX-020-PC

The term "heteroaryl" refers to an aromm atic ring system containing at least ore hete=roatom selected from nitrogen, oxygen, aned sulfur. The heteroaryl ring may be used or othearwise attached to one or more heteroaryl ri ngs, aromatic or non-aromatic hydroc=arbon ringzs or heterocycloalkyl rings. Examples of heteroaryl groups include, for example=, pyridyl, pyri midinyl, quinolinyl, benzothienyl, indolyl, indolinyl, pyridazinyl, pyrazinyl, isoiradolyl, isoqguinolyl, quinazolinyl, quinoxalinyl, phthalazzinyl, imidazolyl, isoxazolyl, pyrazoly®, oxazzolyl, thiazolyl, indolizinyl, indazolyl, benzOthiazolyl, benzimidazolyl, benzofuramxyl, fura_nyl, thienyl, pyrroly), oxadiazolyl, thiadiazo=1yl, benzo[1,4]Joxazinyl, triazolyl, tetreazolyl, isothiazolyl, naphthyridinyl, isochromanyl, chrasmanyl, tetrahydroisoquinolinyl, isoincdolinyl, isobwenzotetrahydrofuranyl, isobenzotetrahydrothhienyl, isobenzothienyl, benzoxazolyl, pyri dopyridinyl, benzotetrahydrofuranyl, benzostetrahydrothienyl, purinyl, benzodioxo-lyl, tria=ziny], pteridinyl, benzothiazolyl, imidazopy~-ridinyl, imidazothiazolyl, dihyvdrobenzisoxazinyl, benzisoxazinyl, benzox=azinyl, dihydrobenzisothiazinyl, ben=zopyranyl, benzothiopyranyl, chromonyl, chromanonyl, pyridinyl-N-oxide, tetraahydroquinolinyl, dihydroquinolinyl, dihyd-roquinolinonyl, dihydroisoquinolinon_yl, dihy~drocoumarinyl, dihydroisocoumarinyl, isoSndolinonyl, benzodioxanyl, benzoxaz=olinonyl, pyrrolyl N-oxide,, pyrimidinyl N-oxide, pyrida_zinyl N-oxide, pyrazinyl N-oxide, quiinolinyl

N-oxide, indolyl N-oxide, indofinyl N-oxide, issoquinolyl N-oxide, quinazolinyl N-oxide, quiraoxalinyl N-oxide, phthalazinyl N-oxide, immidazolyl N-oxide, isoxazolyl N-oxide. oxazolyl

N-oxide, thiazolyl N-oxide, indolizinyl N-oxide, indazolyl N-oxide, benzothiazolyl NF -oxide, benzzimidazolyl N-oxide, pyrrolyl N-oxide, oxa=diazolyl N-oxide, thiadiazolyl N-oxide, triazwolyl N-oxide, tetrazolyl N-oxide, benzothicpyranyl S-oxide, benzothiopyranyl S,,S- dioxxide. Preferred heteroaryl groups include pyridyl, pyrimidyl, quinolinyl, indolyl, pyrrolyl, fura ny], thienyl, and imidazolyl. More preferresd heteroaryl groups include pyridyl, pyrrolyl, and indolyl. The heteroaryl groups herein are Lansubstituted or, as specified, substituted in one or more substitutable positions with various groups. For example, such heteroaryl groups may~ be optionally substituted with, for examplee, C,-Cs alkyl, C,-Cs alkoxy, halogens hydaroxy, cyano, nitro, amino, mono(C,-Ce)alk mylamino, di(C-Cs)alkylamino, C,-Csamlkenyl,

C,-C alkynyl, C;-Cs haloalkyl, C,-Cs haloalko=xy, amino(C;-Cs)alkyl, mono(C;-

Ce)alkylamino(C;-Ce)alkyl or di(C;-Cs)alkylanziino(Cy-Ce)alkyl.

The term "heterocycloalkyl” refers to a ring or ring system containing at least one hete-roatom that is preferably selected from nitrogen, oxygen, and sulfur, whe=rein said

MBHB Ref. No. 04-972-C ARYX-020-PC heteroatom is in a non-aromatic ring. The heterocycloalky® ring is optionally fused to or otherwise attached to other heterocycloalkyl rings and/or neon-aromatic hydrocarbon rings and/or phenyl rings. Preferred heterocycloalkyl groups haves from 3 to 7 members. More preferred heterocycloalkyl groups have 5 or 6 members. Examples of heterocycloalky! groups include, for example, aza-bicyclo[2.2.2]octyl, aza-bi cyclo[3.2.1]octyl, morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholimyl S,S-dioxide, piperazinyl, homopiperazinyl, pyrrolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, tetrahydrofuranyl, tetrahydrothienyl, homopiperidinyl, homomorpholimnyt, homothiomorpholinyl, homothiomorpholinyl S,S-dioxide, oxazolidinonyl, dihyciropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydrofuryl, dihydropyranyl, tetrahydrothienyl S-oxide, tetrahydrothienyl S,S-dioxide and homothiomorpholinyl S-oxide.

Preferred heterocycloalkyl groups include aza-bicyclo[2.2.2]octyl, aza-bicyclo[3.2.1]octyl, piperidinyl, piperazinyl, pyrrolidinyl, thiomorpholiny~l, S,S-dioxothiomorpholinyl, morpholinyl, and imidazolidinyl. =~ More preferred are aza-bicyclo[2.2.2]octyl, aza- bicyclo[3.2.1]octyl, piperidinyl, piperazinyl, pyrrolidinyl, inmidazolidinyl, and morpholinyl.

The heterocycle groups herein are unsubstituted or, as specified, substituted in one or more substitutable positions with various groups. For example, smch heterocycle groups may be optionally substituted with, for example, C;-Cs alkyl, Ci-C¢ allkoxy, halogen, hydroxy, cyano, nitro, amino, mono(C;-Cs)alkylamino, di(Ci-Ce)alkylamino, Cz-Cs alkenyl, C>-Cs alkynyl,

C,-Cs haloalkyl, C;-C¢ haloalkoxy, amino(C;-Cg)alkyl, mono=(C;-Cg)alkylamino(C;-Cs)alkyl, di(C,-Ce¢)alkylamino(C,-Cg)alkyl or =O. .

The term "pharmaceutically acceptable salts” or "a p=harmaceutically acceptable salt thereof" refer to salts prepared from pharmaceutically acceptable non-toxic acids or bases including inorganic acids and bases and organic acids and ba ses. Since the compound of the present invention is basic, salts may be prepared from pharmaceutically acceptable non-toxic acids. Suitable pharmaceutically acceptable acid addition salts for the compound of the present invention include acetic, benzenesulfonic (besylate), beenzoic, camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic, glutamic, hydrobromic, Bhydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pa_moic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, and the like. Preferred acid addition salts are the chloride and sulfate salts. In the most preferred aspect, structumral and/or functional analogs of cisapride are administered as the free base or as the mono or d ihydrochloride salt. -

MBHB Ref. No. 04-—-972-C ARYX-020-—-PC

As used he=rein, the terms “treatment” and “retreating” encompass propinylactic administration of thee compound or a pharmaceutical com . position comprising the compound (“prophylaxis”) as =well as remedial therapy to reduce eor eliminate a disease or dilisorder mentioned herein. Prophylactic administration is intendeed for prevention of disordeers and may be used to treat: a subject that is at risk of having or suaffering from one or more disorders mentioned herein. Thus, as used herein, the term "tr-eatment”, or a derivative whereof, contemplates partia_l or complete inhibition of the stasted disease state, when ana active ingredient of the irmvention is administered prophylactic ally or following the onset= of the disease state for wh_ich such active ingredient of the is administered. "Prophylaxis" reefers to administration of thes active ingredient(s) to a mammal to gorotect the mammal from an; y of the disorders set forth heerein, as well as others.

The term “thmerapeutically effective amount" referss to an amount necessary to achieve a derived therapeuti c effect such as: 1) an amount sufficie=nt to alleviate reflux diseas -e, 2) an amount sufficient to alleviate nausea and vomiting, or 3) an amount sufficient to allmeviate a condition caused by= gastrointestinal motility dysfunction. Therapeutically effective ammounts of structural and/or functional analogs of cisapride are ermcompassed by the above-described dosage amounts andB dose frequency schedule.

A "mammal " may be, for example, a mouse, rat_, pig, horse, rabbit, goat, ccow, cat, dog, or human. In aa preferred aspect, the mammal is a human.

The term “imndividual(s)” is defined as a single nmammal to which is administered a compound of the pmresent invention. The mammal may be, for example, a mouse, mrat, pig, horse, rabbit, goat, ccow, cat, dog, or human. In a preferrec aspect, the individual is a Fauman.

The term "essterified cisapride” means therapeutic «compounds of the subject irm vention that are structural ard/or functional analogs of cisapride, vovhich contain a hydrolysable= group, generally an ester, “that does not detract from the abilityw of these compounds to provide a therapeutic benefit, but which makes these compounds mmore susceptible to degradeation by hydrolases, particularly serum and/or cytosolic esterases, zand which reduces the intcra_ction of the cytochrome P-4-50 drug detoxification system with tlhe cisapride compounds. EZ sterase- mediated metabolis of esterified cisapride compounds re=duces the role of the cytochzxome P- 450 drug detoxificamtion system in cisapride metabolism and reduces or eliminates adverse effects caused by cissapride.

MBHB Ref. No. 0#4.972-C ARYX-020-PC

The term “structural analog” as used herein means that a descriibed compound shares structural characteristics with a parent compound. For example, az structural analog of cisapride may share one or more structural characteristics with the parent cisapride compound, such &as a substituted aryl ring connected to a piperdine ring through an amide linker, but differ sstructurally in other ways, such as the inclusion or de=letion of one or more other chemical moieties.

The term “functional analog” as used herein means that a describecd compound shares a functional characteristic with a parent compound. For example, a functional analog of cisapride may sha re few, if any, structural characteristics with cisapridlle, but affect a similar function, for example, 5-HT,4 agonism.

The term ™'adverse effects” includes, but is not limited to, gastrointestinal disorders such as diarrhea, abdominal cramping, and abdominal grumbling; tiredness; headache; increased systolic pressure; death; ventricular tachycardia; ventricular fam brillation; torsades de pointes; QT prokongation; increased heart rate; neurological and CNS disorders; and interaction of cis=apride with other drugs given concurrently such zas but not limited to digoxin, diazeparm, ethanol, acenocoumarol, cimetidine, ranitidinee, paracetamol, and * propranolol.

The term * "gastroesophageal reflux disease” as used herein me=ans the incidence of, and the symptoms of, those conditions causing the backward flow of the stomach contents into the esophagus=.

The terms “eliciting an anti-emetic effect” and "anti-emetic the=rapy” as used herein mean providing rezlief from or preventing the symptoms of nausea aand vomiting induced spontaneously or associated with emetogenic cancer chemotherapy or irmradiation therapy.

The term "®reating a condition caused by gastrointestinal motilitye dysfunction” as used herein means treaating the symptoms and conditions associated witha this disorder which include, but are mot limited to, gastroesophageal reflux disease, dys; pepsia, gastroparesis, constipation, post-soperative ileus, and intestinal pseudo-obstruction.

The term '""prokinetic" as used herein means the enhancement of peristalsis in, and thus the movement through the gastrointestinal tract.

The term “dyspepsia® as used herein means a condition characterized by an impairment of the power or function of digestion that can arise as a s=ymptom of a primary

MBHB Ref. No. 04-972-C ARYX-0920-PC gastrointestinal dysfunction or as a c~omplication due to other disorders such as apspendicitis, gallbladder disturbances, or malnutrition.

The term "gastroparesis” as wused herein means a paralysis of the stomach brought about by a motor abnormality in thhe stomach or as a complication of disease=s such as diabetes, progressive systemic sclerosais, anorexia nervosa, or myotonic dystrophy.

The term "constipation" as useed herein means a condition characterized by infrequent or difficult evacuation of feces resulting from conditions such as lack of intestirnal muscle tone or intestinal spasticity.

The term "post-operative ileus" as used herein means an obstruction in th_e intestine due to a disruption in muscle tone folBowing surgery.

The term ‘intestinal pseudlo-obstruction" as used herein means a condition characterized by constipation, colicky pain, and vomiting, but without evidence of physical obstruction.

Preparation of Compounds

The chemical synthesis of v-arious analogs of cisapride can be performmed by the methods described in European Patesnt Application No. 0,076,530 A2 published Apr. 13, 1983, U.S. Pat. Nos. 4,962,115 and 5,057,525 and in Van Daele et al,, Drug Development

Res. 8: 225-232 (1986), the disclosuraes of which are incorporated herein by referermce in their entireties, and modified by the incorporation of an ester group at a point conven ient in the synthesis of the disclosed ‘compourds. Exemplary, non-limiting synthesis sczhemes for certain esterified cisapride analogs of the subject invention are provided in WO 01/€093849.

The invention is illustrated further by the following examples, which are not to be construed as limiting the invention in scope or spirit to the specific procedures descmribed in them. Those having skill in the art will recognize that the starting materials may be- varied and additional steps employed to prodiiuce compounds encompassed by the inventiom, as demonstrated by the following exampwles. Those skilled in the art will also recogniz=e that it may be necessary to utilize different s-olvents or reagents to achieve some of the above transformations. In some cases, protection of reactive functionalities may be necessary to achieve the above transformations. Ins general, such need for protecting groups, as well as the conditions necessary to attach and remove such groups, will be apparent to those sk-illed in the art of organic synthesis. When a protecting group is employed, deprotection step> may be required. Suitable protecting groups aand methodology for protection and deprotecti_on such-

MBHB Ref. No. 04—972-C ARYX-020-PC as those described ir Protecting Groups in Organic Synthesis by T—. Greene are well known and appreciated in tthe art.

Unless othemwise specified, all reagents and solvents are of standard commercial grade and are usedll without further purification. The appropri ate atmosphere to run the reaction under, for @example, air, nitrogen, hydrogen, argon and thhe like, will be apparent to those skilled in the zxrt.

Example 1

Preparation eof 6-[4R-(4-amino-5-chloro-2-methoxy-benzomylamino)-3S-methoxy- piperidin-1-yl}-hexamnoic acid 1-aza-bicyclo[2.2.2]oct-3'R-yl ester, dihydrochloride salt (ATI- 7505 Dihydrochlori«de Salt)

MBHB Ref. No. 04-972-C ARYX-020-PC

L-DBT ° NH (Dibenzoyl-L- cl 0 Tartaric Acid) o of Sy oO OBz

YY, 3&0 A I on

HN OMe HN OMe OMe OBz O

Rac-Norcisapride Crude (+)-Norcisapride DBT Salt 1. Recrystallization a i C™ 2. Salt breakup N' Y ——— OMe

HN OMe (+)-Norcisapride Base

Br NN OH 0 oh s

HN OMe OMe

HOR O/,R © R&T

N BOR} S [o} N

Lag H -—

HoN OMe OMe o OR

NTT ,

H -

HN ome OTe 2HCI

Step1: Resolution of Racemic Norcisapridie (-)-Dibenzoyl-L-tartaric acid ((-)-DBsT, about 1 part by weight) was dissolved in ethanol and filtered to remove residual particu lates. Separately, racemic norcisapride {about 0.8 part by weight) was dissolved in a mixtures of ethanol and water and then filtered. The filtrate was heated to about 75°C before addin g the (-)-DBT solution. After stirring at this temperature for about 30 minutes, the mixture was slowly cooled for several hours to about 5 °C and the product salt was collected under vacuum filtration and washed with EtOH/H;O mixture. The wetcake was recrystallized fromm EtOH/H,O by heating to about 79 °C and slow

MBHB Ref. No . 04-972-C ARYX-020-PCC cooling to about § °C as before. The product was collected on a vacuum filter and washead with EtOH/H,O= to give a wetcake.

The weetcake was suspended in water and the pH was adjusted to about 12 using 7% (W/W) aq. NaOH. The resulting suspension was stimred for about 3 hours at room temperature before filtering under vacuum and washing the solid material with water and drying under vacuum. The product was then retreatezd with (-)-DBT to form the salt by thee same general preocedure described above. The isolat-ed salt was then neutralized with aq.

NaOH as described above. The product was isolatec] on a filter and dried as before to pro vide (+)-norcisapride= base (about 0.25 parts by weight). “The e.e. by chiral HPLC analysis was=s about 100% (+)—norcisapride. The optical rotation wevas about +5° (methanol; 25°C and 589 nm), confirmming the positive isomer of norcisap Tide.

Step 2: Coupl_iing with Ethyl 6-bromohexanoate (+)-Nosrcisapride (about 1 part by weight), potassium carbonate (about 0.48 part by weight) and poteassium iodide (about 0.063 part by weight) were suspended in anhydrous "USP ethanol. Ethyl 6—bromohexanoate (about 0.76 part bss weight) was added slowly to the suspension at ro-om temperature. The mixture was heated to reflux until completion of thes reaction. Subsesquent cooling to room temperature tlhe reaction mixture was filtered to remove, e.g., inorganic solids, and the filtrate was concentrated under reduced pressure to- about one-half tlhe volume. The product was precipi tated by slowly adding the crude mateerial to cold water (about 13 parts by weight) with rapid stirring. The precipitate was filtered under vacuum amnd washed with water and then repre=cipitated twice more by dissolution imn anhydrous ethanmol and slow addition into cold water as before. The resulting wetcake was washed with n-lmeptane and resuspended in ethyl acetate and n-heptane (1:9; v/v) and stirred for about 1 hour and before filtering and drying undesr vacuum to yield 0.73 parts by weig'ht of the coupled product as a white solid.

Step 3: Coupling with (R)-3-Quinuclidinol and Dihydrochloride Salt Formation

The estxer (1 part by weight) and (R)-3-Quiruclidinol (about 1.12 part by weight® were suspended in toluene before slowly adding titarium (IV) ethoxide (about 0.5 part by weight) to the s&timred suspension. The mixture was heated to about 91 °C under a stream of nitrogen, and pamrtial vacuum was applied to the flaskz through a distillation apparatus in or-der to azeotropically~ remove the ethanol. Additional tolwmene was added as needed to maintair a

MBHB Ref_. No. 04-972-C ARYX-020-PC minimum solvent volume in the flask. The reaction was considered complete after about 33 hours.

Tae mixture was cooled to about room temperature and extracted five times with water. The organic layer was concentrated under reduced pressure and the resulting residue was redisso Tved in EtOH//PrOH (about 1:1 v/v) and then filtered tiarough a 0.45 micron membrane filter to remove any particulates. Concentrated hydroch.. loric acid was added slowly to thme stirred filtrate to precipitate out the desired product ass the dihydrochloride salt.

The resultimg suspension was stirred for several hours at room temperature and collected under vacuwam filtration and rinsed with EtOH/iPrOH (1:1; v/v) tom provide 0.53 part by weight of tthe crude product salt.

Cwmrude dihydrochloride salt was resuspended in ethanol armd heated to reflux before cooling to room temperature over about 1 hour. The product was collected under vacuum filtration ard rinsed with ethanol and then air-dried. The solids we=re resuspended in ethanol and warmead to about 55 °C to give a clear solution before adding vovarm isopropanol and the product wa_s allowed to precipitate by slow cooling to room tempemrature. The resulting suspension was stirred for several hours before vacuum filtering ard rinsing with, e.g., isopropano 1. The product was vacuum dried, initially at room temperature for several hours and then at about 55 °C until a constant weight was achieved.

Example 2=

Pmreparation of (R)-quinuclidin-3-y] 6-((3 S,4R)-4-(4-amirmo-2-chloro-6- methoxybe=nzamido)-3-methoxypiperidin-1-yl)hexanoate

Step 1: Symnthesis of ethyl 4-(dibenzylamino)-3-methoxypiperidine=-1-carboxylate (1): 0] 0

Not BnBr, K,CO,, Kl Not ne DMF, A ne

OCH; OCH; : racemic 1

To a solution of racemic ethyl 4-amino-3-methoxypiperidizme-1-carboxylate (1 part by mole) in D MF were added benzyl bromide (about 2.2 part by mole), potassium carbonate (about 2.4 ~part by mole) and potassium iodide (about 0.2 part by anole) respectively. The reaction waas heated to about 80 °C. After about 6 hours, the react: ion was slowly diluted with

MBB Ref. No. 04-972-C ARYX-020-PC water (about 12 parts by volume) and extracted with, for example, ethyl acetate. The Organic laye x was washed with brine and then dried over anhy®;. Na;SO,. Subsequent filtration and concentration of the solvent provided the 1 as the yellow-orange oil (1 part by mole).

Step» 2. Synthesis of N,N-dibenzyl-3-methoxypiperidizn-4-amine (2): 0} oot NaOH, PrOHA NH ne A ne

OCH, OCHs 1 2

To a solution of 1 was added NaOH (about 1 © part by mole) in isopropanol znd the mixture was stirred and heated to reflux. After abovLat 3 to about 5 hours, the reactison was cooled to room temperature and the alcoholic solvermt was removed via rotary evapOration.

The mixture was diluted with water and extracted wish ethyl acetate. The organic laywer was brined washed before drying over anhyh. Na;SO,. =Subsequent filtration and concertration of thie solvent provided a crude oil which was purifiecd over SiO; (CHCl; : MeOH: NWH,0H; (about) 15:1:0.01) to furnish 2.

Step 3. Synthesis of (3S,4R)-N,N-dibenzyl-3-methoxzypiperidin-4-amine (3):

Lr 1. (DBT, EtOH/HAL0 $i

BnzN 2. aq. NaOH BN" :

OCH, OCH, 2 3 (-)-Dibenzoyl-L-tartaric acid (about 1.2 part bZy weight) is dissolved in ethanol before slowly adding to a solution of 2 (about 1 part by weighht). The solution is gently warmed and then allowed to cool to room temperature to crystalli ze the sait product. The salt is filtered and washed with EtOH/H,O before suspending in water and basifying by adding ag. NaOH (7% , wt/wt) until the pH reaches about 12. The suspe=nsion is stirred vigorously at rt =and the solid is filtered away, washed with water and vacuum dried to furnish the cis-isomer 3

Step 4. Synthesis of ethyl 6-((3S,4R)4-(dibenzylami no)-3-methoxypiperidin-1-yl)hem<anoate (4):

MBHB “Ref. No. 04-972-C ARYX-020-PC

Cy Ethyl bromohexanoate OY OEt

BrgN" K,CO,, Ki, DMF, A Bn,N"" , o

OCH; OCH 3 3 4 “Mo a solution of 3 (1 part by mole) in DMF are added ethy=1 bromohexanoate (about 1.2 part “by mole), potassium carbonate (about 1.4 part by mole) and potassium iodide (about 0.2 part “by mole) respectively. The reaction is then heated to 80 °C. After about 8 h, the reaction is slowly diluted with water (about 12 part by volume) ard extracted with ethyl acetate. The organic layer is washed with brine and then dried ower anhyd. Na;S0a.

Subsequent filtration and concentration of the solvent furnishes thae crude material.

Purification over SiO; and gives the alkylated material 4.

Step S. Synthesis of (R)-quinuclidin-3-yl 6-((38,4R)-4-(dibenzyl=amino)-3-methoxypiperidin-

I-yDhex_anoate (5):

Shes TOE), (RFQuinuciidinol OYE

Bn NY v 0 toluene, A 8nN* < o N

OCH, «OCH; 4 3

Titanium tetraethoxide is added to a mixture of 4 (1 paar by mole) and (R)-(-)-3- quinucliclinol (1 part by mole) in toluene. The reaction mixture iss equipped with a dean-stark apparatu=s before heating to about 90 °C and partial vacuum is ther applied (additional toluene is added as needed to main the requisite solvent level). The mixture is then cooled to rt and the reaction is diluted with ethyl acetate and then water is added to the resulting mixture. The organic 1. ayer is separated, brine washed, dried over anhyd. Na; SOB, filtered and concentrated.

Purification over SiO, gives the enantiomerically enriched §.

Step 6. Synthesis of (R)-quinuclidin-3-yl 6-((3S,4R)-4-armino-3-methoxypiperidin-1- yDhexaneoate (6):

Chats 4D Hj, Pd/C, EtOH LOTTE

BN" N OCH,

OCH; 5 6

MBHB Ref. No. 04-972-C ARYX-020-PaC

A solution of § (1 part by mole) in ETOH is added to a reaction flask contamining patladium on carbon (about 0.2 part by mole). The mixture is then evacuated of air bmefore subjecting to hydrogenolysis condition by usimg atmospheric H,. Upon completion oof the reaction, the palladium is filtered off under a prad of celite followed by EtOH washes. The filtrated is concentrated via rotary evaporation to furnish 6.

Step 7. Synthesis of (R)-quinuclidin-3-yl 6-((3S,4R)-4-(4-amino-2-chloro-6- methoxybenzamido)-3-methoxypiperidin-1-yl)h exanoate (7):

CHO ©

CARS Q HN cl CHO © Chl =)

EN o J eococ, vr oe Ne,

OCH, HN Cl 6 ?

To a solution of, for example, ethyl chloroformate (1 part by mole) in THF at atoout 0 oC is added the benzoic acid (1 part by mole) in portions. The mixture is warmed to rt for about 1 h before cooling to about 0 °C and adding dropwise a solution of 6 (1 part by role).

The reaction is then warmed to rt. Upon completion of the reaction, reaction is quench.ed by addition of a sat’d solution of NaHCO3 and extracting over EA. The organic layer is brine washed, dried over anhyd. Na,;SO,, filtered and concentrated to furnish the desired product 7.

Example3

Alternate synthesis of ATI-7505:

Oy, R 0 TY ©

HN ome OMe 2HCI

Under acidic conditions, 1-benzylpipericlin-4-one (1) and hydrobromic acid are reacted in the presence of acetic acid to generate N-benzyl-3-bromopiperidin-4-one (2).

Treatment of 2 with a sodium methoxide and methanol solution provides 1-benzyl-4.4- dimethoxypiperidin-3-ol (3). [The presence of the beta-amino group negates the possibi: lity of a Favorskii-type reaction.] Methylation of th.e hydroxy! group is done using a hydridee base followed by treatment with iodomethane in the presence of DMF as the solvent to fumissh compound 4.

MBHB Ref. No. 04-972-C : ARWX-020-PC

Lhe AcOH, HBr (aq.) NBn NaOMe/MeOH, tt NBn 0 fo) MeO

Br MeO OH 1 2 3

NaH, Met NBn 1% HSOr 4, A NBn NaNH3CN, NH, OAC NBn

DMF MeO " TT weon.a HN

OMe OMe Omi 4 5 6 (-}-0BT Cer BOC,0, “THF Coe Pd/C, MeOH H, Cy

BIOHH0 nN BocHN" BocHN™"

OMe OMe OMe 7 8 ]

HO.,, 6-Bromohexanenitrile - Oy S Ch eS

K2CO3, DMF, A BocHN" : H* BocHN' [o] N

OMe OMe 10 1"

OMe O

Iso oO... (o]

IA, Shil 4 PNT BL anamr ses

HON o N~ EtOCOCI, TEA, THF

OMe A 12

Subsequent acetal hydrolysis using 1% sulfuric acid in the presence of heat yields a piperidine S, which can then undergo a reductive amination using, for example, sodium cyanoborohydride and ammoniurm acetate in methanol to yield 1-benzyl-3-mesthoxypiperidin- 4-amine (6). At this stage, 6 can undergo a chiral resolution technique=. This can be accomplished, for example, using (-)-DBT or other variant of tartaric acid in_ the presence of the suitable solvent to afford exclusively asymmetrically pure compound 7. Boc group protection of the primary amine in 7 can be accomplished using Boc amhydride in the presence of THF solvent to obtairs 8. A debenzylation reaction by hydrogeno lysis using Pd/C in methanol in the presence of atmospheric hydrogen gas set the stage for the= alkylation step.

Treatment of 6-bromohexanenitxrile in the presence of mild base and DMF generates compound 10. A nitrile to ester conversion using (R)-quinuclidinol in the pmresence of dilute acid generates 11. Subsequent rermoval of the Boc group using TFA furnishe=s the free amine,

MBHB Ref. No. 04-972-C ARYX-020-PC which can unclergo a coupling reaction with requisite benzoic sacid in the presence of a coupling reagemt such as ethyl chloroformate to afford ATI-7505 a_s an enantiomerically pure material. 1. Ethyl 6—~Bromohexanoate HO. Q

KoCCOD,, DMF, OEt 1. o — 1CCHOMRE Sh — NN 13:AT7S05 2.TFA HN" ° THOE=H),, Tol., 4

OMe 2. Am 13

OM eO SEAS NS Carisburg esterasse 13: ATI-7505

HN a OMe 1“

Alternatively, compound 9 can be alkylated using ethyl 6-bromohe=xanoate in the presence of mild base. Sumbsequent removal of the Boc group yields compound 13. Titanium mediated transesterificattion of 13 using (R)-quinuclidinol and titanium tetra_ethoxide in toluene solvent generates ATE-7505. Carlsburg esterase hydrolyzes esters that aare of the S-configuration, therefore leaving intact esters that are of the R configuration . Therefore treatment of diasteriomeric mixtures of 14 with the Carlsburg esterase may alsos yield ATI-7505.

Example 4 (+) ane (-)-norcisapride can be made from its racemic mixture by resolution of the enantiomers umsing conventional means such as optically resolvi_ng acids, according to the method described in US Patent 6,147,093, or in “Enantiomers, Raacemates and Resolutions”, by J. Jacques, A. Collet, and S.H. Wilen (Wiley-Interscience, Ndew York, NY), or in S.H.

Wilen et al., Tetrahedron (1977) 33:2725.

The 4 isomers were obtained in low-mg amounts by using preparative column chromatography followed by evaporation of the solvent. This me=thod is useful for preparing small amount_s for analytical and characterization purposes. Thais is a standard separation method used routinely in analytical labs in order to isolate and cha_racterize metabolites.

Possib le synthetic routes to Compound IV, Compound V1 and (+)-Compound II are described below using (+)-norcisapride as a starting material. THhe routes to Compound III,

MBHB Ref. No. 04-972-C ARY”X-020-PC

Compound V and (-)-Compound II are identical except that they use (-)-no-rcisapride as a starting material.

Example 5

Production of (+)-Compound II, ethyl esaer

A equimolar mixture of (+)-norcisapride and ethyl 6-bromohexanoate= (1 equivalent each), a catalytic amount of KI, and K2€CO; (2 equivalents) in DMF is heatec at about 60 Cc for several hours or until TLC analysis indicates that the reaction is over. A= fter cooling to room temperature, water is added and tlhe mixture is extracted with EtOAc. The combined organic extracts are washed successiveRy with’ water, 10% LiCliagy solution amnd brine, then dried over Na;SO4. Concentration gives (¥)<ompound II, ethyl ester.

Production of (+)-Compound II

A mixture of crude (+)-compoundd II, ethyl! ester, from above (1eq.), KOH (2M, § eq.) in MeOH and THF (enough to dissolve) is stirred at room temperature for apperoximately 1 to 2 hours. The MeOH and THF are removed under vacuum, and the residue is diluted with water. Wash with an organic solvent such as EtOAc. The aqueous layer is acidified to pH ~5 : using HCl. The precipitate is filtered of if and dried to give (+)-Compound II.

Production of Compound IV and Compo-und VI

A mixture of (+)-Compound II (1 eq.), (R)-(-)-3-quinuclidinol HCI salt (1 eq.), EDAC (1 eq.) and DMAP (1 eq.) in DMF iss heated at around 50C overnight. After cooling and diluting with water, the mixture is purified by chromatography or by crystallization to provide Compound IV. Similarly, using: (8)-(+)-quinuclidinol, Compound VI is obtained.

The following compounds ares prepared essentially according tos methods and procedures described above. The compeound names were generated using either ChemDraw

Ultra version 8.03, which is available from Cambridgesoft Corporation or ACD Namepro software, version 6.0. (35)-1-azabicyclo[2.2.2]oct-3-yl 6-{(35,4R)-4-{(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypipeeridin-1-yl}hexanoate; (35)-1-azabicyclo[2.2.2]oct-3-yl 6-{(3R,45)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3 -methoxypipweridin-1-yl}hexanoate; (3R)-1-azabicyclo[2.2.2]oct-3-y1 6-{(3R,48)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino}-3-methoxypipweridin-1-yl }hexanoate; 8-methyl-8-azabicyclo[3 2.1]oct—3-yl 6-{(3S,4R)-4-[(4-amino-S-chloroe-2- methoxybenzoyl)amino]-3-methoxypipweridin-1-yl} hexanoate;

MBHB Ref.

No. 04-972-C ARYX-020-PC= 4-[({(35,4R)-4-[(4-amino-5-ch loro-2-methoxybenzoyl)amino}-3- methoxypiperidin-1-yl}acetyl)amino_ benzoic acid; methyl 4-[({(3S,4R)-4-[(4-ami=no-5-chloro-2-methoxybenzoyl)amino}-3- methoxypiperidin-1-yl}acetyl)amino_Jbenzoate; methyl 4-[({(35,4R)-4-[(4-amimo-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acetyl)amino_Jbenzoate; methyl 4-{({(3S,4R)-4-[(4-amimo-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acetyl)amino_]benzoate; ethyl 4-[({(35,4R)-4-[(4-aminO>-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acetyl)amino Jbenzoate; isopropyl 4-[({(38,4R)-4-[(4-a=mino-5-chloro-2-methoxybenzoyl)amino]- 3-methoxypiperidin-1-yl}acetyl)amimno]benzoate; 2-methoxyethyl 4-[({(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino}-3-methoxypoiperidin-1-yl }acetyl)amino] benzoate; 2-pyrrolidin-1-ylethyl 4-[({(3S5,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypoiperidin-1-yl} acetyl)aminojbenzoate; 1-methylpiperidin-4-yl 4-[({(3 S4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-pyridin-2-ylethy} 4-[({(3S,44R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypoiperidin-1-yl} acetyl)amino]benzoate; 2-(dimethylamino)ethyl 4-[({(35,4R)-4-[(4-amino-5-chloro-2- raethoxybenzoyl)amino}-3-methoxypoiperidin-1-yl}acetyl)amino] benzoate; 1-methylpiperidin-3-yl 4-[({(3 S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypoiperidin-1-yl }acetyl)amino]benzoate; 2-morpholin-4-ylethy! 4-[({(3=S4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 1,4-dimethylpiperidin-4-yl 4-[«({(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypiperidin-1-yl}acetyl)amino]benzoate; 4-[({(3S,4R)-4-[(4-amino-5-chmloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acetyl)amino=]Jbenzoic acid; 2-oxo-2-(piperidin-4-ylamino)methyl 4-[({(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypiperidin-1-yt}acetyl)amino]benzoate; 1-({(35,4R)-4-[(4-amino-5-chE oro-2-methoxybenzoyl)amino)-3- methoxypiperidin-1-yl}acetyl)piperiadine-4-carboxylic acid; methyl 1-({(3S,4R)-4-[(4-amir0-5-chloro-2-methoxybenzoyl)amino}-3- methoxypiperidin-1-yl}acetyl)piperiadine-4-carboxylate; methyl 1-({(35,4R)-4-[(4-amirn0-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl} acetyl)piperidinc-4-carboxylate; methyl 1-({(3S,4R)-4-[(4-amir0-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl} acetyl)piperiedine-4-carboxylate; ethyl 1-({(35,4R)-4-{(4-amino—5-chloro-2-methoxybenzoyl)amino}-3- methoxypiperidin-1-yl}acetyl)piperiadine-4-carboxylate; 2-methoxyethyl 1-({(3S,4R)-4—[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3-methoxypoiperidin-1-yl} acety)piperidine-4- carboxylate;

MBHB Ref. No. 04-972-C ARYX-02 O-PC 4-{[(2-{(3S,4R)-4-[(4-amino-5-chBoro-2-methoxybenzoyl)amino}-3- methoxypiperidin-1-yl}ethyl)(methyl)aminolmethyl} benzoic acid; methyl 4-{[(2-{(3S,4R)-4-[(4-amimo-5-chloro-2-methoxybenzoyl)amino]- 3-methoxypiperidin-1-yl}ethyl)(methyl)amino]methyl} benzoate; methyl 4-{[(2-{(3S,4R)~4-[(4-amimo-5-chloro-2-methoxybenzoylamino]- 3-methoxypiperidin-1-yl}ethyl)amino)m ethyl} benzoate; isopropyl 4-{[(2-{(35.4R)-4-[(4-arnino-5-chloro-2- methoxybenzoyl)amino}-3-methoxypipe ridin-1- yl}ethyl)amino]methyl} benzoate; ethyl 4-{[(2-{ (35,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl} ethyl)amino]methyl} benzoate Dihydrochloride; (3R)-1-azabicyclo[2.2.2]oct-3-yl &-{[(2-{(35,4R)-4-[(4-amino-5-chloro- 2-methoxybenzoyl)amino}-3-methoxypiperidin-1- yl}ethyl)amino]carbonyl} benzoate; (R)-quinuclidin-3-yl 6-((3S,4R)-4—(4-amino-5-chloro-2- methoxybenzamido)-3-methoxypiperidin-1-yl)hexanoate; or 6-((3 S,4R)-4-(4-amino-5-chloro-2 -methoxybenzamido)-3- methoxypiperidin-1-yl)hexanoic acid

Formulation, Administration, and Uses

Dosage rates and routes of administration of the disclosed compounds are similar to those already used in the art and known to the skilled artisan (see, for example, P=hysicians' : Desk Reference, 54th Ed., Medical Economics Company, Montvale, NJ, 2000).

The magnitude of a prophylactic or therapeutic dose of structural and/or —functional analog of cisapride in the acute or chronic management of diseases and/or disorders described herein will vary with the severity of the condition to be treated, and the route of administration. The dose, and perhaps thes dose frequency, will also vary according ®&o the age, body weight, and response of the individiual patient. In general, the total daily dose= range for structural and/or functional analogs of ciisapride, for the conditions described herei_n, is from about 1 mg to about 200 mg, in single or divided doses. Preferably, a daily dose rarmge should be between about 5 mg to about 100 mg, in single or divided doses, while most pre=ferably, a : daily dose range should be between about 5 mg to about 75 mg, in single or divideed doses. It is preferred that the doses are administered from 1 to 4 times a day. In managing tiie patient, the therapy should be initiated at a lower dose, perhaps about 5 mg to about 10 mg, and increased up to about 50 mg or higher depending on the patient's global response. Ya is further recommended that children, and patients over 65 years, and those with impairead renal or hepatic function, initially receive low doses, and that they be titrated based on individual response(s) and blood level(s). It may be necessary to use dosages outside these ranges in

TWO 2005/068461 PCT/US2005/0005100

MABHB Ref, No. 04-972-C : ARYX-020-PC scome cases as will be apparent to those skilled in the art. Further, it is noted that the clinician or treating physician will know how and when to interrupt, adjust, or terminate therapy in conjunction with individual patient response.

The compounds of the subject invention camn be formulated according to known m _ethods for preparing pharmaceutically useful compositions. Formulations are described in detail in a number of sources which are well known =and readily available to those skilled in th.e art. For example, Remington's Pharmaceutical Science by E.W. Martin describes formulations which can be used in connection with the subject invention. In general, the compositions of the subject invention are formulates such that an effective amount of the bioactive compound(s) is combined with a suitable carrier in order to facilitate effective acAministration of the composition.

The compositions of the subject invention inc-lude compositions such as suspensions, sco lutions and elixirs; aerosols; or carriers such as star—ches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like, in the case off oral solid preparations (such as powders, capsules, and tablets) with the oral solid preparations being preferred over the oral liquid preparations. A preferred oral solid preparation is capsules. The most preferred oral ssolid preparation is tablets. Preferred armounts of active ingredient (i.e., an structural andor functional analog of cisapride) in a sco lid dosage form are about § mg, 10 mg, and 25 mg.

Further, acceptable carriers can be either so lid or liquid. Solid form preparations inaclude powders, tablets, pills, capsules, cachets, suppositories and dispersible granules. A solid carrier can be one or more substances which may act as diluents, flavoring agents, soo lubilizers, lubricants, suspending agents, binders, pmreservatives, tablet disintegrating agents or— encapsulating materials.

The disclosed pharmaceutical compositions may be subdivided into unit doses coontaining appropriate quantities of the active comp onent. The unit dosage form can be a peackaged preparation, such as packeted tablets, cap sules, and powders in paper or plastic containers or in vials or ampules. Also, the unit dos=age can be a liquid based preparation or fowrmulated to be incorporated into solid food productss, chewing gum, or lozenge.

In addition to the common dosage forms set owut above, the compounds of the present ins vention may also be administered by controlled release means and/or delivery devices such

MBHB Ref. No. 04-972-C ARYX-0820-PC as those described in U.S. Pat. Mos.: 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719, the disclosures of which are hereby incorporated by reference in their entirety.

Any suitable route of administration may be employed for providing the patient with an effective dosage of a structural and/or functional analog of cisapride. For exammple, oral, rectal, parenteral (subcutaneous, in-tramuscular, intravenous), transdermal, and lik e forms of administration may be employed. Dosage forms include tablets, troches, d ispersions, suspensions, solutions, capsules, pagches, and the like.

One aspect of the inventiomn provides a method of treating gastroesophageal reflux disease in a mammal, while substamtially reducing the concomitant adverse effects associated with the administration of cisapridee, which comprises administering to a human in need of such treatment, a therapeutically effective amount of a structural and/or functional analog of cisapride, or a pharmaceutically acc=eptable salt thereof. A preferred aspect is the treatment of gastroesophageal reflux disease in mumans.

Another aspect of the inven tion provides a composition for the treatment of a human suffering from gastroesophageal re flux disease, which comprises a therapeutically effective amount of a structural and/or functi -onal analog of cisapride, or a pharmaceutically acceptable salt thereof.

Yet another aspect of the present invention provides a method of elicitimg an anti- emetic effect in a mammal, while substantially reducing the adverse effects associated with the administration of cisapride, whiich comprises administering to a mammal in ne=ed of such anti-emetic therapy, a therapeutical Ry effective amount of structural and/or functioral analogs of cisapride, or a pharmaceuticallzy acceptable salt thereof. Preferably, the ma_mmal is a human.

In an additional aspect, the poresent invention encompasses an anti-emetic ccomposition for the treatment of a mammal in need of anti-emetic therapy, which cosmprises a therapeutically effective amount of a structural and/or functional analog of cisagpride, or a pharmaceutically acceptable salt thesreof.

A further aspect of the pressent invention includes a method of treating am condition caused by gastrointestinal motility «dysfunction in a mammal which comprises administering to a mammal in need of treatment —for gastrointestinal motility dysfunction, a therapeutically effective amount of a structural ancl/or functional analog of cisapride, or a pharmaaceutically acceptable salt thereof. Conditions caused by gastrointestinal motility dysfunction include,

MBHB Ref. Neo. 04-972-C ARYX-020-PC but are not limited to, dyspepsia, gastroparesis, constipation, p ost-operative ileus, and intestinal pseucdo-obstruction. Preferably, the mammal is a human.

The ob servation that cisapride enters the central nervous sy”stem and binds to SHT, receptors indiczates that cisapride may have centrally-mediated effects. Cisapride is a potent ligand at SHT 4 receptors, and these receptors are located in sevesral areas of the central nervous systerm. Modulation of serotonergic systems has a variegty of behavioral effects.

Accordingly, ®he compounds of the subject invention can be used in the treatment of: 1) cognitive disomrders, including but not limited to Alzheimer's diseases; 2) behavioral disorders, including but not limited to schizophrenia, mania, obsessive-compulsive disorder, and psychoactive substance use disorders; 3) mood disorders, inclu-ding but not limited to depression ancd anxiety; and 4) disorders of control of autonomic fianction, including but not limited to esse=ntial hypertension and sleep disorders.

Accorclingly, the present invention also provides methocds of treating cognitive, behavioral, mmood, or autonomic function control disorders in a mammal comprising the administratiom of a therapeutically effective amount of structural and/or functional analog of cisapride, or a_ pharmaceutically acceptable salt thereof. Preferably, the mammal is a human.

ATI-7505 Birds with High Affinity to S-HT¢ Receptors

The 5—HT4 receptor is known to be the major receptor subtype involved in the prokinetic act-ivity of cisapride in the gut. ATI-7505 has a high binding affinity for 5-HTy4 receptor, witha alow nanomolar ICsg. As shown in Table 1, the affi nity of ATI-7505 for the 5.HT,4 receptor was 18-fold greater than cisapride and at least 360—fold greater than the

ATI-7505 mamjor metabolite, ATI-7500.

Table= 1. 5-HT", Receptor Binding 5-HT, Receptor

Guinea Pig Striatum

Comgpound 1Cgo (NM) K; (nM) Ny

ATI-7505 8.3 1.4 0.7

ATI-7500 >3,000 >500 —_

Cisaporide 150 24.9 0.8 ny, Hi I coefficient. .

S-HT o receptor prototypic reference antagonist HIGR 113808 (0_70 nM)

MBHB IRef. No. 04-972-C ARYX-020-PC

ATI-75C35 is a Highly Potent Partial Agonist at Human 5-_HT4 Receptor

AARYx performed in vitro assays based on adenylyl cyclase stimulation in cells engineer—ed to stably express human 5-HT4 receptor. ATI-7" 505 proved to be a highly potent 5-HT4 receptor agonist, whereas its major metabolite, ATI- 7500 was relatively weak (F igure 1 and Table 2). The estimated ECso of ATI-7505 (4 nM) w-as approximately 10-fold lower than that of cisapride (49 nM), and approximately 100-fold lower than that of ATI-7500 (395 nM). Baased on its estimated Eng value, ATI-7505 had 85%% of the efficacy of S-HT (serotonmin) (Table 2) , demonstrating that ATI-7505 is a partial agonist of HT, receptors.

Table 2.

Potenecy and Efficacy (Intrinsic Activity) at Huma 5-HT, Receptor —— Potency___________ Efficacy

Compeound ECg PECs % of SHT (serotonin) 5-HT (-serotonin) 45 7 NA

ATI-7505 4 8.45 85

ATI-7500 395 6.40 81

Cisaprride 49 7 77

ECso, CO Ncentration causing 50% maximal increase in adenylyl cyclase activity

PECs. Negative logarithm of the ECs

ATI-75 05 Accelerates Gastric Emptying in Fed DogsTo Characterize the effects of ATI- 7505 omn gastric emptying, experiments were performed En a post-prandial model involving consciomus dogs instrumented with sets of strain gauge trarsducers placed on the stomach and small toowel. The objective of the experiments was “to measure the time required for migratizng motor contractions (MMCs) to return to base line following ingestion of a solid meal. 2A drug-induced shortening of MMC return time inclicated an early end of the digestive period edue to accelerated gastric emptying. Immediately after completion of an MMC in the mid-smmall intestine, various doses of test drugs (vehicle, ATI-7505, or cisapride) were infused. intravenously (TV) over 20 minutes. At the end «of the drug infusion, the dogs were fed a mmeal. Gut contractions were recorded for a minimumm of 60 minutes prior to the start of the drug infusion to establish the fasting state and to Identify the onset of MMC in the duodermum, and at least 30 minutes after the return of MEMC in the duodenum. Quantitative companmrisons of the treatments were based on the time of” MMC return as an index of gastric emptying following ingestion of a solid meal. As swemmarized in Figure 2, ATI-7505 significantly shortened the time of MMC return, indBicating an acceleration of gastric emptyi ng in normal fed dogs. Cisapride showed a similar pattern of action.

MBHB Ref. No. 04-972-C : AaRYX-020-PC

ATI-7505 Increases Gastric and Smazdl Intestinal Motor Activity with Negligible Effect on

Colonic Activity

Experiments were performed im fasted, conscious dogs to evaluate tie gastric, small intestinal and colonic motor activity of ATI-7505 compared to cisapride. A specific goal was to determine the dose sizes of ATI-7505 (IV and PO) that most closely mimm ic the pattern and magnitude of contractile activity caused by cisapride at typical therapeutic loses in dogs (0.5 mg/kg IV; 1 mg/kg PO).

When given IV and PO, both ATI-7505 and cisapride caused prokin _etic effects in the dog gut. The onset of action typically occurred within 1 to 2 minutes and 258 to 30 minutes following IV and PO administration, respectively. The effect of ATI-7505 on gastric and small intestinal motor activity mimicked cisapride. Like cisapride, ATI-750 5 appeared to cause dose-dependent stimulation of antral and small bowel contractility with relatively little effect on colonic motor activity. The perokinetic effects caused by ATI-7505< in the upper GI tract occurred along with a small but s ignificant (p < 0.05) increase in the frequency of giant migrating contractions (GMC).

ATI-7505 was not associated with the development of retrograde gizant migrating contractions (RGC). Like cisapride, ATI-7505 had a minimal effect on mig=rating motor complex (MMC) characteristics in the antrum as well as proximal, mid and distal small intestine. With regard to MMC frequesncy and phase 111 duration, only one =significant difference was noted: PO ATI-7505 imcreased MMC frequency in the proxi=imal small intestine relative to the controls. The dogs tolerated the IV and PO doses of ATI-7505 well and exhibited no side effects such as d iarrhea, anorexia, or weight loss.

Overall, the results showed that on a mg/kg-basis, ATI-7505 was apgproximately twice as potent as cisapride. In addition, the actions of ATI-7505, like those of cissapride, were consistent with a mechanism involving the facilitation of acetylcholine relezase from enteric neurons rather than a direct smooth muscle action. In conclusion, ATI-7505 increases gastric and small intestinal motor activity in a cisapride-like manner with minimal-tte-no effect on colonic activity.

The Metabolism of ATI-7505 is CYF?450-Independent

Based on data from pooled hurman microsomes, ATI-7505 undergoe=s biotransformation to a single metabolite, ATI-7500, which does not appear ®o be subject to further metabolism. The conversion of ATI-7505 to ATI-7500 was not depe=ndent on the

MBHB Ref. No. 04-9-72-C ARYX-020-PC presence of NADPH. Thus the major biotransformation pathway for AT1—7505 occurs independently of CYF®450 enzymes.

ATI-7505 Does Not Mnhibit CYP450 Enzymes

To test the potential for ATI-7505 and/or its main metabolite, ATI1-72500 to act as

CYP450 inhibitors, thmese two molecules were screened using Gentest Sup>ersomes™.

Consistent with publisshed reports, cisapride had significant inhibitory act- ivity against

CYP450 enzyme isoforms, CYP3A4, 2D6 and to a lesser extent 2C9. Ne=ither ATI-7505 nor its primary metabolites, ATI-7500 displayed significant inhibitory activity— against these three

CYP450 isoforms, no~r against a panel of other isoforms known to play a - role in drug metabolism.

ATI-7505 Has Negliggible Affinity for the Cardiac Channel, Ix,

The rapidly activating delayed rectifier potassium (K+) current in . humans (human Ir) is a K+ channel that i=s encoded by the human-ether-a-go-go-related gene (hERG). Cisapride is known to produce CQT interval prolongations via a blockade of Ix, and it was therefore of interest to determine ®fATI-7505 and ATI-7500 have important inhibitor—y effects on human

Ix. The test system was mammalian HEK-293 cells expressing the hERCS K+ channels, in which the potassium ecurrent was measured by whole cell patch-clamp technique. The ranking of the ICs values was: cisapride (9.5 nM) > ATI-7505 (24,521 nM) > ATT1-7500 (204,080 nM) (Table 3). Overa_li, the findings indicate that ATI-7505 has a significcantly lower pro- arrhythmic potential ®han cisapride and suggest that both ATI-7505 and AATI-7500 have negligible affinity for— human Ix, channels. :

Table 3.

Inhibition of I, Activity

Activity of I, inHEK Cells __

Compound % lx, control ICs (10,000 nM)

ATI-7505 78.0 24. 521

ATI-7500 88.9 204 080

Cisapride 0 9.5

Data are normalized te % control tail I, (current elicited without drug or vehicle present)

MBHB Ref. NO. 04-972-C ARYX-020-PC

ATI-7505 Doe=s not Induce Important ElectrophysioBogical Changes in Guinea Pig

Hearts

The cardiac electrophysiological effects of ATI—7505 were examined in isolated, perfused guinea pig hearts. The study examined ATI-75505, ATI-7500 and cisapride, all of which were each tested at concentrations up to 10,000 mM. The no observed effect level (NOEL) was d=efined as the highest concentration of tesst compound not showing a response that was signifSicantly different from baseline (p < 0.05). The following 6 cardiac parameters were tested: (17) QT interval; (2) MAPDg; (3) SA interwal; (4) QRS interval; (5) AH interval; and (6) HV. While ATI-7505 was a very weak modulator of cardiac electrophysiologic parameters, its. metabolite, ATI-7500 entirely lacked eleectrophysiological activity (Table 4).

The NOEL for= ATI-7500 was > 10,000 nM for the enti_re set of 6 cardiovascular parameters.

Since cisapridee had a NOEL of 10 nM for the combine~d set of 6 cardiac parameters tested, while ATI-7505 had a combined NOEL of 1,000 nM, #ATI-7505 appears to lack the potency of cisapride in modulating cardiac electrophysiologic parameters. Overall, the findings demonstrate that ATI-7505 is significantly safer than c-isapride with regard to the potential to induce important cardiac electrophysiologic fluctuatiors.

Table 4.

Cardiac Electrophysiologic Parameters in Isolated Perfused ys

Electropthysiological No Obse=rved Effect Level (NOEL)

Parameteer Cisapride ATI-7505 ATI-7500

QT Interwal 10 1,000 > 10,000

MAPDg, 10 1,000 > 10,000

SA Interv~al 100 > 10,000 > 10,000

QRS Inte=rval 1,000 > 10,000 > 10,000

AH Interwal 1,000 > 10,000 > 10,000

HVinterwal ~~ 1000 = 1000 >10.000

Combine d Parameters 10 1,000 > 10,000

All molecules were tested at baseline, 10, 100, 1,0€00, and 10,000 nM.

Other thar for values reported as > 10,000 nM, a s=ignificant difference (p < 0.05) from baseline vavas observed when the molecule was tessted ata 10-fold higher

Metabolism imn human microsomal preparations

The metabolism of these compounds was studied in pooled human microsomes inthe presence and absence of the Cytochrome P-450 cofac~tor NADPH and both the disappearance of parent ancl the appearance of the corresponding =acid metabolite, i.c., the corresponding compound-II isomer, monitored with time.

EMBHB Ref. No. 04-972-C ARYX-020-PC

As shown in Table 5, Compounds III and IV were rapidly hydrolyzed by esterase to ®heir respective metabolites (+) and (-)-Compound II. Thee metabolism was not dependent on &CYP450 since the rate of hydrolysis was independent on NADPH presence, which is a mecessary cofactor for CYP450 to function. In contrast, (£)-S Compounds V and VI sappeared to be quite stable with time under the same conditions. In this experiment, the =amount of substrate (compounds III, IV, V, and VI) rema-ining in the reaction after 5, 60, and 90 minutes were evaluated by a tandem HPLC-MS method. This remaining amount was correlated with the appearance of the metabolite comppound II. The sum of remaining substrate and compound I was constant over time amad equal to the amount of starting 0 material at time zero, therefore indicating that hydrolyssis was the only metabolic reaction taking place. of NADPH cofactor. The remaining amount of test com _pound and the appearance of the metabolite compound II were monitored over 90 minutes.

Bp cl WO = PO ompound (ng/mL) [(ng/mL Compowund (ng/mL) {ng/mL 5 Bis Pp p33 P2o 5 Bad © bos [as ps2 pos Ns Bid

G0 li69 __ lio4 Bes pro 15 [334

Metabolism in fresh human blood.

Test compounds were dissolved in DMSO to make 12.5 mM stock solution and diluted with water to a final concentration of 2.5mM (DIMMSO/H20 =20/80). Fresh blood was collected into heparinized tubes from 3 human donors and blood was stored on ice until incubation. Separate aliquots of blood from each donor were pipetted into 1.5 mL centrifuge tubes and the tubes were pre-incubated in a shaking waater bath at 37°C for 5 minutes. The reaction was initiated by the addition of 10 pL of the apgpropriate test compound stock to each tube (final concentration = 100 pM). Incubations weree quenched after 0, 5, 15, 30 and 60 minutes, by the addition of acetonitrile (750 mL), cen®trifuged at 12,000 rpm for 2 minutes and the supernatant analyzed on an Agilent 1100 HPLC system. Separations were accomplished on a Keystone Intersil ODS2, 250X4.6mom, 5 m column. The aqueous mobile phase consisted of 20 mM ammonium acetate buffer (pH 5.7) and the organic phase acetonitrile. A gradient was used: initial condition consisted of 20% acetonitrile for 1

MBHB Ref. No. 04-9722.C ARYX-020-PC minute. The acetonitrile concentration was increased linearly to 90% over the next 8 minutes and held there for 1 minute. The system was then recycle=d to initial conditions over the= course of 1 minute andl held there for 4 minutes before the neext injection. The peak area for the parent peak was d_etermined by monitoring absorbance at 240, 254 and 290 nM. The= results were expressed as amount of initial compound remain ing and data subjected to kinetic analysis using WinNorLin. The half-lives for the individual compounds are given below frm

Table 6.

Table 6 1 Diastereomeric Configuration

Norcis “half” _| Quinuclindol “ha Af’ | Half-life (min

Sea tT] 1203

Subjet2 | | 00000001 1037

Sweets | | ez;

MeanxtsD | | 0 0000| 105%14l wv | + | ® 0 0000

Subject1 | | | 847

Swjez | | | ser

Sujets | |] 8%

Meamzsb | | | sseo0m

Vv 1s

Sweet] | | | >e0mm

Swetz | |__| >emn

Subjects | |] >60min_

M+ Ts 1

Sweet | [| comm

Subject2

Subject |__

It should be understood that the examples and amspects described herein are foor illustrative purposes only and that various modifications ox- changes in light thereof will oe suggested to persons skilled in the art and are to be includecd within the spirit and purview eof this application and the scope of the appended claims. Further, all patents, patemnt applications, provisiconal applications, and publications referred to or cited herein are incorporated by refer—ence in their entirety to the extent tiey are not inconsistent with tEne explicit teachings of this specification.

MBHB Ref. No. 04-972-C ARYX-020-PC

The invention and the manner and process of making and using it, are now described in such full, clear, concise and exact terms as to enab-le any person skilled in the art to which it pertains, to make and use the same. It is to be understood that the foregoing describes preferred aspects of the invention and that modifications may be made therein without

S departing from the spirit or scope of the invention as set forth in the claims. To particularly point out and distinctly claim the subject matter regarded as invention, the following claims conclude this specification.

Claims (1)

1. ~ MBHB Ref. No. 04-972-C ARYX-020-PC What is claimed is:

   1. A compound of the formula: Re R2 Ro x) and pharmaceutically acceptable salts thesreof, wherein the bonds at positions 3 and 4 are «cis relative to each other; Lis —(C)-Cs alky})- (in one aspect=, {C3-Cs alkyl)-), {Ci-Cs alkyl)-C(0)-, or -C(OD~ (C1-Cs alkyl)-, wherein each of the alkyl groups is optionally substituted with l or 2 groupss that are independently halogen, C,-C, alk=oxy, or OH and wherein one carbon in the alkyl portion of L may be replaced by -N(Ro)-= R, is halogen; R; is amino, NH(C,-C4 alkyl) or I=(C,-C, alkyl)(C;-C4 alkyl); Rj is OH or C;-C, alkoxy; R4 is H or methyl; and Rs is “0-C3-C; cycloalkyl, -O-hetterocycloalkyl, heterocycloalkyl, aryl, -O-aryl, - N(Rg)~(Co-Cs alkyl)-C(O)-aryl, or -N(Ry )-Co-Cs alkyl-aryl, -O-heteroaryl, -N(Rg)-C1-Cs(®0)- heteroaryl, or -N(Rs)-Co-Cs alkyl-heterozaryl, wherein each of the cyclic groups is unsubstituted or substituted at one or more substitutable positions with C,-Cs alkyl, Ci-Cs alkoxy, halogen, C,-Cs haloalkyl, C1-Cs Shaloalkoxy, hydroxyl, hydroxy-C;-Cs-alkyl, amirmo, -NH(C,-Cs alkyl), -N(C,-Cs alkyl)(Ci-Ces alkyl), -(Co-Cs alkyl)-C(O)Rn, or -0-(Co-Cs alkxyl)- C(O)R}, methylsulfone, Co-Cs-sulfonamide, or NO; wherein Rs at each occurrence is independently H or C;-C, alkyl; Ry; is C,-Cs alkyl, OH, or Ry; is C,-Cs alkoxy, optionally sLabstituted with 1 or 2 groups that are independently C,-C. alkoxy, amino, -NH(C;-Cs alkyl), -N(C;-Cs alky!)(Ci-Cs alkyl), -(Co-Cs alkyl)- C(O)N(Ro)-heterocycloalkyl, -O-heteroczycloalkyl, -C;-Cs(O)N(Ry)-heteroaryl, or heteroa_ryl, wherein

   MBHB Ref. No. 04-922-C ARYX-020-PC the heterocycloalkyl groups are optionally substituted wi_th 1, 2, or 3 groups that are indepe-ndently halogen, C,-Cs alkyl, C,-Cs alkoxy, hydreoxy, hydroxy C-Cs alkyl, C;-Cs alBkoxycarbonyl, -CO;H, CF;, or OCF3, the heteroaryl group is optionally substituted with 1, 2, oer 3 groups that are independently halogen, C-Cs alkyl, Ci-Cs alkoxy, hydroxy, hyQroxy Ci-Cs alkyl, Ci- Cs alkoxycarbednyl, -CO;H, CFs, or OCF; or Ry is —O-hetemrocycloalky! wherein the heterocycloalkyl is optionally substituted with 1, 2, or 3 groups that zare independently halogen, C,-Cs alkyl, C;-C, alk-oxy, hydroxy, hydroxy Ci-Cs alkyl, &C,-C; alkoxycarbonyl, -CO,H, CF3, or OCF3; anA Rao is C1-Ces alkoxy (preferably Ci-C4 alkoxy, more preferably rmethoxy), or OH.

   2. A compound according to claim 1, wherein : R, is chloro.

   3. A compound according to claim 1, wherein R; is amino.

   4. A compound aaccording to claim 1, wherein Rs is methoxy.

   5. A compound according to claim 1, wherein Rs is H.

   6. A compound according to claim 1, wherein R, is chloro; R; is amino; Rj is methoxy; R, is H, and R, Ry, and R; nave the following orientation on the ph enyl ring: 0] Rs Rs

   7. A Compouncd according to claim 6, wherein

   MBHB Ref. No. 04-972-C ARYX-020-PC L is «(C3-Cs alkyl)- wherein one camrbon may be replaced by -N(Ro)-, or —~(C2-Cs alkyl)-CEO)-

   8. A compound according to claim 7, wherein 3 Rj is-O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from aza— bicyclo-octyl, aza-bicyclo-nonyl, a=a-bicyclo-decyl, where the aza nitrogen is optionally substituted with methyl or ethyl, is optionally substituted with methyl or ethyl, piperidiny 1, piperazinyl, and pyrrolidinyl, whereein the piperidinyl, piperazinyl, and pyrrolidinyl groupes are unsubstituted or substituted at cone or two positions with groups that are independently Ci- 0 Cs alkyl, C;-C, alkoxy, halogen, C2-C4 haloalkyl, Ci-C4 haloalkoxy, hydroxyl, hydroxy CCCs alkyl, amino, -NH(C,-Cy alkyl), -NE(C,-C4 alkyl)(C1-Cs alkyl), (Co-Cs alkyl)-C(O)R1y, or~ NO, wherein R, is C,-Cs alkoxy, option ally substituted with 1 or 2 groups that are independen_tly C,-C4 alkoxy, amino, -NH(C1-Cs alkyl), -N(C;-Cs alkyl)(C,-Cs alkyl), -(Co-Cs alkyl)- C(O)N(Rg)-heterocycloalkyl, or he=terocycloalkyl wherein the heterocycloalkyl group is selected from pyrrolidinyl, piperid iny}, piperazinyl, and morpholinyl, wherein the : heterocycloalkyl groups are optioraally substituted with 1, 2, or 3 groups that are independently halogen, Ci-Cs alkyl, C1-Cs alkoxy, hydroxy, hydroxy C-Ce alkyl, C,-Cs alkoxycarbonyl, -CO-H, CFs, or OCFs.

   0

   9. A compound according to claim 7, wherein Rs is heterocycloalkyl, which is selected from 1-aza-bicyclo[2.2.2Joct-3-yl, and 8-aza- bicyclo[3.2.1]oct-3-yl, where the mitrogen in the 8-aza-bicyclo[3.2.1]oct-3-yl group is optionally substituted with methyl or ethyl. '5

   10. A compound according to claim 7, wherein Rs is -N(R9)-Co-Cs alkyl-zaryl or -N(Rg)-C(O)-aryl, wherein the aryl group is unsubstituted or substituted at one or more substitutable positions with C;-Cs alkyl, C,-aC;s alkoxy, halogen, C,-C haloalkyl. C,-Cs haloalkoxy, hydroxyl, hydroxyalkyl, amino, -NJH(C;- Cs alkyl), -N(C1-Cs alkyl}(C:-Cs alkyl), -(Co-Cs alkyl)-C(O)Ry, or NO».

   11. A compound according to claim 10, wherein

   MBHB Ref. No. 04-972-C ARYX -020-PC the aryl group is a phenyl! substituted with -(Co-Cs alkyl)-C(O)R,; and optionally substituted with 1 or 2 groupes independently selected from C,-Cg alkyl, C1-Cs alkoxy, halogen, CF3, OCF3, hydroxyl, hydroxyalkyl, amino, -NH(C;-C; alkyl), -N(C,-Cas alkyl)(C;- C, alkyl), or NO, and Ry; is C;-Cs alkoxy, optionally substituted with 1 or 2 groups that are inde=pendently C-C4 alkoxy, amino, -NH(C,-Cs alkyl), -N(C,-Cs alky!)(C,-Cs alkyl), (Co-Ce allxyl)- C(O)N(Ry)-heterocycloalkyl , or heterocycloalkyl wherein the heterocycloalkyl group is selected from pyrrolidinyl, peiperidinyl, piperazinyl, and morpholinyl, wherein thes heterocycloalkyl groups are optionally substituted with 1, 2, or 3 groups that are independently halogen, C,-(C alkyl, C-Cs alkoxy, hydroxy, hydroxy C:-Cs alkyl, C-Cs alkoxycarbonyl, -CO;H, CFs, or OCFs.

   12. A compound according to claim 11, wherein the -(Co-Cs alkyl)-C(O)R,) g=roup is attached to position 4 of the phenyl ring.

   13. A compound according to claim 1 that is (35)-1-azabicyclo[2.2.2]oct-3-y1 6-{(3S5,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3 —methoxypiperidin-1-yl} hexanoate; (35)-1-azabicyclo[2.2 2]oct-3-yl 6-{(3R,45)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3 —methoxypiperidin-1-yl}hexanoate; (3R)-1-azabicyclo[2-2.2]oct-3-y] 6-{(3R,4S)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino}-3 —methoxypiperidin-1-yi}hexanoate; 8-methyl-8-azabicyc-10[3.2.1Joct-3-y! 6-{(3S,4R)-4-[(4-amino-5-chloro-2— methoxybenzoyl)amino]-3 —methoxypiperidin-1-yl} hexanoate; 4-[({(3S,4R)-4-[(4-ammino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acestyl)amino]benzoic acid; methyl 4-[({(35,4R)—4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acestyl)amino]benzoate; methyl 4-[({(3S,4R)—4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acestyl)amino]benzoate; methyl 4-[({(35,4R)—4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acestyl)amino]benzoate; ethyl 4-[({(35,4R)~4~—[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin-1-yl}acestyl)amino]benzoate; isopropyl 4-[({(3S,4R)-4-{(4-amino-5-chloro-2-methoxybenzoyl)amino]- 3-methoxypiperidin-1-yl} aacetyl)amino]benzoate; 2-methoxyethyl 4-[( {(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)amino]-3 —methoxypiperidin-1-yl}acetyl)amino]benzoate;

   MBHB Ref.

   No. 04--972-C ARYX-0200-PC 2-pyrrolidin—1-ylethy! 4-[({(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)armino}-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 1-methylpipezridin-4-yl 4-[({(35,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)arnino}-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 2-pyridin-2-wlethyl 4-{({(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)armino)-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 2-(dimethyla.mino)ethyl 4-[({(3S.4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)arnino]-3 -methoxypiperidin-1-yl}acetyl)amino]benzoate; : 1-methylpiperidin-3-yl 4-[({(3S.,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)arnino]-3 -methoxypiperidin-1-yl}acetyl)amino]benzoate; 2-morpholin—4-ylethyl 4-[({(35,4R)-4-[(4-amino-5-chloro-2- methoxybenzoy})armino}-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 1 4-dimethyl piperidin-4-yl 4-[({(3S,4R)-4-[(4-amino-5-chloro-2- . methoxybenzoyl)arnino}-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 4-[({(35,4R)—4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin- Bl -yl }acetyl)amino]benzoic acid; 2-0x0-2-(pip eridin-4-ylamino)ethyl 4-[({(35,4R)-4-[(4-amino-5-chloro-2- methoxybenzoy})arnino]-3-methoxypiperidin-1-yl }acetyl)amino]benzoate; 1-({(3S,4R)~=4-[(4-amino-S5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin- 1 -yl} acetyl)piperidine-4-carboxylic acid; methyl 1-({(35,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin- 1 -yl}acetyl)piperidine-4-carboxylate; methyl 1-({(35,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino}-3- methoxypiperidin-R -yl} acetyl)piperidine-4-carboxylate; methyl 1<({(354R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin- B -yl} acetyl)piperid ine-4-carboxylate; ethyl 1-({(355,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]-3- methoxypiperidin- 1 -yl} acetyl)piperidine-4-carboxylate; 2-methoxyetdhyl 1<({(35,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)arnino)}-3 -methoxypiperidin-1-yi} acetyl)piperidine-4- carboxylate; 4-{[(2- {(3S,44R)4-{(4-amino-S-chloro-2-methoxybenzoyl)amino}-3- methoxypiperidin- -yl}ethyl)(methyl)amino]methyl} benzoic acid; methyl 4-{[(2-{ (35,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)armnino}-3-methoxypiperidin-1- yl}ethyl)(methyl)arnino]methyl}benzoate; methyl 4-{[(2-{(35,4R)~4-[(4-amino-5-chloro-2- methoxybenzoyl)arnino]-3-methoxypiperidin-1- yl}ethyl)amino]me-thyl}benzoate; isopropyl 4- € [(2- {(3S,4R)-4-[(4-amino-5-chloro-2- methoxybenzoyl)armino]-3-methoxypiperidin-1- yl}ethyl)amino]me-thyl} benzoate; ethyl 4-{[(2- {(38,4R)-4-[(4-amino-5-chloro-2-methoxybenzoyl)amino]- 3-methoxypiperidir-1-yl} ethyl)amino]methyl} benzoate Dihydrochloride; (3R)-1-azabiccyclof2.2.2]oct-3-yl 4-{[(2-{(3S,4R)-4-{(4-amino-5-chloro- 2-methoxybenzoyl@amino}-3-methoxypiperidin-1- yl}ethyl)amino]car@bony!} benzoate;

   MBHB Ref. No. 04-972.C ARYX-0220-PC {R)-quinuclidin-3-yl 6-((3S,4R)-4-(4-amino-5-chloro-2- methoxy benzamido)-3-methioxypiperidin-1 -yl)hexanoate; or 6-((3S,4R)-4-(4-amin 0-5 <chloro-2-methoxybenzamido)-3- methoxypiperidin-1-yl)hexanoic acid (ATI-7500).

   14. A compound according to claim 1 that is (R)-guinuclidin-3-yl 6-((3S,4R)~4-4-amino- 5-chloro-2-methoxybenzamido)-3-methoxypiperidin- 1-y )hexanoate.

   15. A salt according to claim | that is [R)-quinuclidin-3-yl 6-((3S,4R)-4-(4-amirmo-5- chloro-2-methoxybenzamido)-3-methoxypiperidin-I-yl)hexanoate dihydrochloride.

   16. A composition comprising a compound or pharmaceutically acceptable salt Of claim 1 and at least one pharmaceutically acceptable carrier, solvent, adjuvant, or excipient.

   17. Use of a compound or sait according to claim 1 in the manufacture of a rmedicament for the treatment of emesis, dys pepsia, gastroparesis, constipation, intestinal pseudcobstruction, gastroesophageal reflux, or post-operative ileus.

   18. Use according to claim 17, wherein the medicament is administered intram venously.

   20. Use of a compound or salt according to claim 14 in the manufacture of a medicament for the treatment of emesis, dyspepsia. gastroparesis, constipation, intestinal pseudosobstruction,

   19. A composition comprising at least one of (R)-quinuclidin-3-y} 6-((3S,4R)-4-( 4- amino-5-chloro-2-methox yberezamido)-3-methoxypiperidin-1-ylhexanoate and (R)- quinuclidin-3-yl 6-((3S,4R)-4—(4-amino-5-chloro-2-methoxybenzamido)-3-methox ypmiperidin- I-yhhexanoate Dihydrochloridle and at least one pharmaceutically acceptable carrier, solvent, adjuvant, or excipient. gastroesophageal reflux, or post-operative ileus.

   21. A compound accordi ng to claim 1, substantially as herein described and ~exemplified and/or described with reference to the accompanying tigures.

   22. A salt according to claim (5, substantially as herein described and exemplified and/or described with reference to the accompanying figures. 58 AMENDED SHEET

   23. A composition according to claim 16 or 19, substantially as herein descri bed and exemplified and/or described with reference to the accompanying figures .

   24. Use according to claim 17 or 20, substamtially as herein described and exemplified and/or described with reference to the acccompanying figures. 59 AMENDED SHEET