CN1918157B

CN1918157B - Stereoisomeric compounds and methods of treating gastrointestinal and central nervous system disorders

Info

Publication number: CN1918157B
Application number: CN2005800046038A
Authority: CN
Inventors: I·欧文; M·帕姆; C·贝克尔; C·鲁宾斯; P·德鲁加拉
Original assignee: ARYx Therapeutics Inc
Current assignee: Na Ang Reed Appleton LLC; Rene New LLC
Priority date: 2004-01-07
Filing date: 2005-01-07
Publication date: 2010-06-23
Anticipated expiration: 2025-01-07
Also published as: ZA200605308B; CN1918157A

Abstract

The present invention relates to stereoisomeric compounds of formula (X): wherein the variables are as defined herein, and safe, effective treatment of a variety of conditions including, but not limited to, gastroparesis, gastroesophageal reflux disease, and related conditionsComposition for treating gastrointestinal diseases. The compositions of the present invention are also useful for treating a variety of conditions associated with the central nervous system.

Description

Stereoisomeric compounds and methods of treating gastrointestinal and central nervous system disorders

This application claims priority from U.S. provisional patent application No. 60/534,892 filed on 7/1/2004 and U.S. provisional patent application No. 60/560,938 filed on 9/4/2004.

Background

Cisapride (cisapride) belongs to benzamide derivatives, and the parent compound thereof is metoclopramide. U.S. patent nos. 4,962,115 and 5,057,525 (inventor "Van Daele", incorporated herein by reference in its entirety) disclose N- (3-hydroxy-4-piperidylidene) benzamide of cisapride. These compounds, their pharmaceutically acceptable acid addition salts, and their stereochemically isomeric forms, all of which stimulate the motility of the gastrointestinal system, are disclosed by Van Daele.

As a class of drugs, benzamide derivatives have a number of significant pharmacological effects. The significant pharmacological activity of benzamide derivatives is due to their effect on the nervous system regulated by the neurotransmission mediator serotonin. The effects of serotonin, and thus the pharmacological properties of benzamide derivatives, have been indirectly demonstrated over the years in a wide range of applications under a variety of conditions. Therefore, the present study is directed to locating the sites of serotonin production and storage, as well as the location of serotonin receptors in the human body, in order to confirm the relationship between these locations and various disease states or conditions.

In this regard, it has been found that the primary site of production and storage of serotonin is the enterochromaffin cells of the gastrointestinal mucosa. Serotonin has also been found to strongly stimulate intestinal motility by stimulating intestinal smooth muscle, accelerating intestinal transit, decreasing absorption time, similar to what occurs in diarrheal conditions. The irritation is also associated with nausea and vomiting.

Because of their ability to modulate the serotonin nervous system in the gastrointestinal tract, many benzamide derivatives are potent antiemetics and are commonly used to control emesis during cancer chemotherapy or radiotherapy, particularly when using a homoemetogenic compound such as cisplatin. This effect is almost certainly termed 5HT₃The result of the blocking of the serotonin (5HT) action at a specific site of action by substances of the receptor (the canonical name of which in the scientific literature is the serotonin M-receptor). Because of the release of serotonin from injured enterochromaffin cells in the gastrointestinal tract, nausea and vomiting may result from chemotherapy and radiation therapy. The release of neurotransmitter serotonin stimulates serotonin receptors not only into the vagal nerve fibers (i.e., initiates the vomiting reflex), but also in chemoreceptor-inducing regions of the rearmost region of the brain. The anatomical location of the effects of benzamide derivatives, whether they act on the Central Nervous System (CNS) or the peripheral nervous system, or both, is unknown. (Barnes et al, J.Pharm.Pharmacol.40: 586-. Like other benzamide derivatives, cisapride is due to its 5HT₃The ability to modulate serotonin activity at the receptor and thus may be useful as an antiemetic agent.

A second significant effect of benzamide derivatives is to increase the motility of smooth muscles of the gastrointestinal tract through the anterior segment of the small intestine via the esophagus, thereby promoting esophageal and small intestine transit, and to promote gastric emptying and increase esophageal sphincter tone (Decktor et al, Eur.J. Pharmacol.147: 313-316, 1988). Although benzamide derivatives are not cholinergic receptor agonists per se, the above effects on smooth muscle may be mediated by muscarinic receptor blockers (e.g. atropine) or tetrodotoxin-like neurotransmitters that affect sodium channelsBlocking by transfusion inhibitor. Similar blocking behavior of serotonin contraction in the small intestine has been reported. Recently, it is believed that the major smooth muscle actions of benzamide derivatives are directed to a novel class of serotonin receptors (termed 5HT)₄Receptor, on the interneuron of the gut wall myenteric plexus). Agonism of these receptors results from the binding of acetylcholine to its receptors on the smooth muscle membrane, which in turn enhances the release of acetylcholine from the parasympathetic nerve terminals of the surrounding smooth muscle fibers. Smooth muscle membranes are the real motive force for muscle contraction.

For the inclusion of 5HT₄A discussion of various 5HT receptors, including receptors, can be found in U.S. Pat. Nos. 6,331,401 and 6,632,827, which are incorporated herein by reference in their entirety.

Initially, cisapride was used to treat gastroesophageal reflux disease (GERD). The disease is characterized by the regurgitation of food in the stomach into the esophagus. One of the most important factors in the pathogenesis of gastroesophageal reflux disease is the reduction of the pressure barrier caused by the failure of the lower esophageal sphincter. Lower esophageal sphincter failure may be caused by basal depression, sphincter relaxation, or an unbalanced increase in gastric pressure. Other factors of the pathogenesis of the disease include: delayed gastric emptying, inadequate esophageal emptying due to impaired peristalsis, and corrosive reflux materials that may damage the esophageal mucosa. Cisapride is believed to strengthen the anti-reflux barrier and improve esophageal emptying by increasing lower esophageal sphincter pressure and increasing peristaltic contractions.

Due to its actuating force, cisapride can also be used to treat dyspepsia, gastroparesis, constipation, postoperative ileus, intestinal pseudo-obstruction, and the like. A condition characterized by dyspepsia is an impaired digestive function or capacity, which may be a symptom of major gastrointestinal dysfunction, or a complication of other diseases such as appendicitis, gallbladder disturbances, or malnutrition. Gastroparesis is gastric paralysis caused by gastric motility disorder or diabetes, progressive systemic sclerosis, anorexia nervosa or myotonic dystrophy complications. Constipation is a condition characterized by little or difficult excretion resulting from conditions such as a lack of intestinal muscle contractile function or intestinal spasms. Postoperative ileus is an intestinal blockage resulting from muscle damage after surgery. Intestinal pseudo-obstruction is characterized by constipation, colicky pain and vomiting, but there is no evidence of mechanical obstruction.

Drug toxicity is an important consideration in the treatment of humans and animals. Toxic side effects (side effects) resulting from the administration of drugs include various degrees of morbidity, ranging from low fever to death. Drug treatment is only valid if the positive effects of the treatment regimen exceed the potential risks of the associated treatment. Factors that a physician balances include the qualitative effect of the drug used, the quantitative effect, and the corresponding outcome that would result if the drug were not provided to the individual. Other considerations include the physical state of the patient, the stage of disease progression, the medical history, and any known drug-related side effects.

Drug elimination is generally the result of metabolic activity of the drug and subsequent excretion of the drug from the body. Metabolic activity may occur within blood vessels and/or in cellular metabolic regions or organs. The liver is the major site of drug metabolism. Metabolic processes can be divided into synthetic and non-synthetic reactions. In non-synthetic reactions, the drug undergoes a chemical change by oxidation, reduction, hydrolysis, or any combination of the above. These processes are collectively referred to as the first stage reaction.

In a second phase reaction (also referred to as a synthesis reaction or binding reaction), the parent drug or an intermediate metabolite thereof binds to the endogenous substrate to form an addition or binding product. The metabolites formed in the synthetic reaction are generally more polar and biologically inactive. Thus, these metabolites are more easily excreted via the kidney (in urine) or liver (in bile). Synthetic reactions include glucuronidation (glucuronidation), amino acid binding, acetylation, sulfo-binding (sulfonation), and methylation.

More than 90% of cisapride doses are metabolized by oxidative N-dealkylation at the piperidine nitrogen or by aromatic hydroxylation occurring at the 4-fluorophenoxy or benzamide ring.

It has been found that administration of cisapride to humans results in serious side effects including CNS disorders, elevated systolic blood pressure, interaction with other drugs, diarrhea, and abdominal cramps. In addition, intravenous injection of cisapride has been reported to have other side effects that do not occur with its oral route (Stacher et al [1987] diagnostic Diseases and Sciences 32 (11): 1223-1230). It is believed that these side effects are caused by metabolites resulting from oxidative dealkylation or aromatic hydroxylation of a compound produced in the cytochrome P450 detoxification system. Cisapride is also listed among many deleterious drug/drug interactions that are the result of metabolism by the cytochrome P450 system.

Cisapride (PROPULSID, janssen pharmaceutical Products, l.p.) was reported to be associated with at least 341 severe arrhythmias during months 7-1999 to 12-1999 in 1993. These arrhythmias include ventricular tachycardia, ventricular fibrillation, torsades de pointes, and long QT syndrome. A total of 80 cases of death were reported. Because of these side effects, the product was automatically withdrawn from the U.S. public market and the drug was now available only through research with limited access procedures.

5HT with Gastrointestinal (GI) motivational effects due to side effects on the heart (prolongation of long QT interval, tachycardia, torsades de pointes) and deleterious drug interactions due to cytochrome P-450 metabolism in the liver₄Receptor agonists are limited in safety. This class of GI motility agents without these susceptibility would be of great value in several therapeutic areas, including GERD and gastric emptying disorders. Although certain cisapride derivatives have been described in U.S. patent No. 6,552,046 and WO 01/093849 (the entire contents of which are incorporated herein by reference), it would be desirable to have more advantageous compounds.

It has now been found that certain stereoisomers of an esterified structural and/or functional analogue of cisapride possess unique, and particularly advantageous properties.

Disclosure of Invention

The present invention provides compounds and compositions of structural formula (X), which are stereoisomerically esterified cisapride analogs, useful for the safe and effective treatment of various gastrointestinal disorders, including, but not limited to, gastroparesis, gastroesophageal reflux, and related conditions. The compounds of the present invention are also useful in the treatment of a variety of conditions associated with the central nervous system.

The compounds of the present invention include compounds of structural formula (X):

wherein,

the chemical bonds at the 3-position and the 4-position are in cis form;

l is- (C)₁-C₆Alkyl) - (in one aspect, - (C)₃-C₅Alkyl) -, - (C)₁-C₆Alkyl) -C (O) -, or-C (O) - (C)₁-C₆Alkyl) -, wherein each alkyl group may be optionally substituted with 1 or 2 groups, independently halogen, C₁-C₄Alkoxy or-OH, wherein one carbon atom of the alkyl moiety of L may be substituted by N (R)₉) Replacement;

R₁is halogen;

R₂is amino, NH (C)₁-C₄Alkyl) or N (C)₁-C₄Alkyl) (C₁-C₄Alkyl groups);

R₃is OH or C₁-C₄An alkoxy group;

R₄is H or methyl; and

R₅is-O-C₃-C₈Cycloalkyl, -O-heterocycloalkyl, aryl, -O-aryl, -N (R)₉)-(C₀-C₆Alkyl) -C) O) -aryl or-N (R)₉)-C₀-C₆Alkyl-aryl, -O-heteroaryl, -N (R)₉)-C₁-C₆(O) -heteroaryl or-N (R)₉)-C₀-C₆Alkyl-heteroaryl, wherein each cyclic group may be unsubstituted or may be substituted in one or more substitutable positions, wherein the substituents are selected from C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, C₁-C₆Haloalkyl, C₁-C₆Haloalkoxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), - (C)₀-C₆Alkyl) -C (O) R₁₁or-O-C₀-C₆Alkyl) -C (O) R₁₁Methanesulfonyl, C₀-C₆Sulfonamide or NO₂(ii) a Wherein

R₉Each occurrence is independently hydrogen or C₁-C₄An alkyl group;

R₁₁is C₁-C₆Alkyl, OH, or

R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), - (C)₀-C₆Alkyl) -C (O) N (R)₉) -heterocycloalkyl, -O-heterocycloalkyl, -C₁-C₆(O)N(R₉) -heteroaryl, or heteroaryl, wherein

The heterocycloalkyl group may be optionally substituted with 1, 2 or 3 groups, where the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃；

The heteroaryl group may be optionally substituted with 1, 2 or 3 groups, where the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃；

R₁₁is-O-heterocycloalkyl, wherein heterocycloalkyl may be optionally substituted with 1, 2 or 3 groups, independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃(ii) a And

R₂₀is C₁-C₆Alkoxy (preferably C)₁-C₄Alkoxy, more preferably methoxy), or hydroxy.

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

The compounds of structural formula (X) are useful in the treatment or prevention of gastroesophageal reflux disease and substantially reduce the side effects of cisapride administration. These side effects include, but are not limited to, diarrhea, abdominal cramps, elevated blood pressure, and increased heart rate.

In addition, the compounds and compositions of the present invention may be used to treat emesis and other conditions, including but not limited to dyspepsia, gastroparesis, constipation, post-operative ileus, intestinal pseudo-obstruction, and the like. It has the additional benefit of also reducing the side effects associated with the administration of cisapride.

Advantageously, the compound of the invention is 5HT₄A ligand for a receptor, and therefore,can be used to treat conditions mediated by the receptor. These receptors are located in various regions of the central nervous system and modulation of these receptors can be used to achieve modulation of the CNS.

Advantageously, the present invention provides for the provision of stereoisomeric compounds containing an ester moiety which does not impair the therapeutic efficacy of these compounds, but which makes them more susceptible to degradation by plasma and/or cytosolic esterases, thereby avoiding the cytochrome P450 drug detoxification system which causes the side effects of cisapride, and reducing the incidence of such side effects.

The present invention further provides methods of treating gastroesophageal reflux disease, dyspepsia, gastroparesis, constipation, post-operative and intestinal pseudo-obstruction, and related conditions, comprising administering to an individual in need of such treatment a therapeutically effective amount of a compound of structural formula (X).

Advantageously, the therapeutic compounds of the present invention are stable in storage and safer in their metabolic mechanisms compared to other drugs; thus, the compounds of the present invention have a low incidence of side effects and toxicity when used.

In a further aspect, the invention relates to breakdown products (preferably metabolic breakdown products) formed when the therapeutic compounds of the invention are exposed to an esterase. These breakdown products can be used to monitor clearance of the therapeutic compound from the patient, as described herein.

In a further aspect, the invention provides methods of synthesis of the therapeutic stereoisomeric compounds of the invention, as well as intermediates useful in the preparation of the compounds of interest.

Drawings

FIG. 1 is 5-HT₄Concentration response profiles for the receptor agonists ATI-7505, serotonin, cisapride, and ATI-7500.

Fig. 2 is a graph of gastric emptying in a full dog. The data shown are normalized against MMC regression values averaged transit time. Values represent mean + standard error for 5 dogs. P < 0.05 relative to transport control

FIG. 3 is a metabolic map of ATI-7505 and ATI-7500 in the presence and absence of the CYP450 dependent cofactor NADPH. The graph shows the mean and standard deviation in μ M of the concentration of ATI-7505 and ATI-7500. ATI-7505 concentration (2. mu.M) was incubated with human microsomal protein (1mg) in the presence or absence of NADPH regenerating system (cofactor).

Detailed Description

In a further aspect, the present invention relates to compounds of formula (X), wherein

R₅is-O-C₃-C₈Cycloalkyl, -O-heterocycloalkyl, wherein said heterocycloalkyl group is selected from piperidinyl, piperazinyl, pyrrolidinyl, azabicyclo-octyl, and in certain embodiments is azabicyclo [2.2.2]Octyl, azabicyclo [3.2.1]Octyl, azabicyclo-nonyl, azabicyclo-decyl, indolinyl, morpholinyl, thiomorpholinyl, S-dioxothiomorpholinyl, and imidazolidinyl, -O-aryl, -N (R)₉) -C (O) -aryl, or-N (R)₉)-C₀-C₆Alkyl-aryl, wherein each cyclic group may be unsubstituted or may be substituted at one or more substitutable positions, wherein the substituents are selected from C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, C₁-C₆Haloalkyl, C₁-C₆Haloalkoxy, hydroxy-C₁-C₄-alkyl, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), -C (O) R₁₁Or NO₂(ii) a Wherein

R₉Each occurrence is independently hydrogen or C₁-C₄An alkyl group; and

R₁₁is C₁-C₆Alkyl, hydroxy, or

R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), -C (O) N (R)₉) -heterocycloalkyl, or heteroaryl, wherein

The heterocycloalkyl group is selected from pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azabicyclo-octyl, and in certain embodiments is azabicyclo [2.2.2]Octyl, azabicyclo [3.2.1]Octyl, azabicyclo-nonyl, azabicyclo-decyl, wherein the heterocycloalkyl group can optionally be substituted with 1, 2 or 3 groups, independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃，

The heteroaryl group is selected from the group consisting of pyridyl, pyrimidinyl, quinolinyl, isoquinolinyl, and indolyl, wherein the heteroaryl group is optionally substituted with 1, 2, or 3 groups, wherein the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃(ii) a Or

R₁₁is-O-heterocycloalkyl, wherein heterocycloalkyl is selected from piperidinyl, pyrrolidinyl, imidazolidinyl, morpholinyl, azabicyclo-octyl, and in certain embodiments is azabicyclo [2.2.2]Octyl, azabicyclo [3.2.1]Octyl, azabicyclo-nonyl, azabicyclo-decyl, and tetrahydrofuranyl, wherein each heterocycloalkyl group may optionally be substituted with 1, 2 or 3 groups, the substituents independently being halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃。

In another aspect, the invention provides a compound of formula (X), wherein R₁Is chlorine.

In yet another aspect, the present invention provides compounds of formula (X), wherein R₂Is an amino group.

In yet another aspect, the present invention provides compounds of formula (X), wherein R₃Is methoxy.

In another aspect, the invention provides a compound of formula (X), wherein R₄Is H or methyl.

In yet another aspect, the present invention provides compounds of formula (X), wherein R₁Is chlorine; r₂Is amino; r₃Is methoxy; r₄Is H or methyl.

In yet another aspect, the present invention provides compounds of formula (X), wherein R₁Is chlorine; r₂Is amino; r₃Is methoxy; r₄Is H, and L is- (C)₄-C₆Alkyl) -C (O) -.

In another aspect, the invention provides compounds of formula (X) wherein 2 or more of the foregoing aspects are combined.

In another aspect, the invention relates to compounds of formula (XI) wherein L is- (CH)₂)₅-C (O) -a compound of formula (X):

in yet another aspect, the invention provides a compound of formula (XI), wherein R₁Is chlorine; r₂Is ammoniaA group; r₃Is methoxy; r₄Is H or methyl.

In yet another aspect, the invention provides a compound of formula (XI), wherein R₅is-O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from azabicyclo-octyl,

in yet another aspect, the invention provides a compound of formula (XI), wherein R₅is-O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from azabicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2 [ ]]Oct-3-yl or 8-azabicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl, azabicyclo-decyl, wherein the nitrogen of the aza group may optionally be substituted by methyl or ethyl;

R₄is H or methyl.

In yet another aspect, the invention provides a compound of formula (XI), wherein R₅is-O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from piperidinyl, piperazinyl, or pyrrolidinyl, each of which may be unsubstituted or substituted at 1 or 2 positions, where the substituents are independently C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halogen, C₁-C₄Haloalkyl (in one aspect, it is CF)₃)、C₁-C₄Haloalkoxy (in one aspect, it is OCF)₃) Hydroxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₄Alkyl), -N (C)₁-C₄Alkyl) (C₁-C₄Alkyl), - (C)₁-C₆Alkyl) -C (O) R₁₁Or NO₂(ii) a And R is₄Is H or methyl.

In yet another aspect, the invention provides a compound of formula (XI), wherein R₅is-O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from indolinyl, morpholinyl, thiomorpholinyl, S-dioxothiomorpholinyl and imidazolidinyl, each of which may be unsubstituted or substituted at 1 or 2 positions, wherein the substituents are independently C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halogen, C₁-C₄Haloalkyl (in one aspect, it is CF)₃)、C₁-C₄Haloalkoxy (in one aspect, it is OCF)₃) Hydroxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₄Alkyl), -N (C)₁-C₄Alkyl) (C₁-C₄Alkyl), - (C)₀-C₆Alkyl) -C (O) R₁₁Or NO₂(ii) a And R is₄Is H or methyl.

In yet another aspect, the invention provides a compound of formula (XI), wherein R₅is-O-phenyl, N (R)₉)-(C₀-C₆Alkyl) -C (O) -phenyl, or-N (R)₉)-C₀-C₄Alkyl-phenyl, wherein the phenyl group is substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halogen, C₁-C₄Haloalkyl (in one aspect, it is CF)₃)、C₁-C₄Haloalkoxy (in one aspect, it is OCF)₃) Hydroxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₄Alkyl), -N (C)₁-C₄Alkyl) (C₁-C₄Alkyl), - (C)₀-C₆Alkyl) -C (O) R₁₁Or NO₂；R₄And R₉Independently H or methyl.

In another aspect, the invention provides a compound of formula (XI), wherein R₄H, hydrogen.

In a further aspect, the invention provides a compound of formula (XI), wherein R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), - (C)₀-C₆Alkyl) -C (O) N (R)₉) -, heterocyclic ringAlkyl, or heterocycloalkyl, wherein the heterocycloalkyl group is selected from pyrrolidinyl, piperidinyl, piperazinyl, and morpholinyl, wherein the heterocycloalkyl group can be optionally substituted with 1, 2, or 3 groups, wherein the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃。

In another aspect, the invention provides a compound of formula (XI) wherein 2 or more of the foregoing aspects are combined.

In another aspect, the present invention provides compounds of structural formula (XII), i.e., compounds of structural formula (X) having the structural formula:

wherein R is₁₅Is hydrogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, C₁-C₆Haloalkyl (in one aspect, it is CF)₃)、C₁-C₆Haloalkoxy (in one aspect, it is OCF)₃) Hydroxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), methanesulfonyl, C₀-C₆-sulfamoyl, or NO₂，R₁₆Is H or-O- (C)₀-C₆Alkyl) -C (O) R₁₁. In another aspect, R₁₅Is H.

In yet another aspect, the present invention provides compounds of formula (XII), wherein R₄And R₉Independently is H or methyl, R₁₁Is a hydroxyl group.

In yet another aspect, the present invention provides compounds of formula (XII), wherein R₄And R₉Independently is H or methyl, R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), - (C)₀-C₆Alkyl) -C (O) N (R)₉) -heterocycloalkyl, or heterocycloalkyl group, wherein the heterocycloalkyl group is selected from azabicyclooctyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl, pyrrolidinyl, piperidinyl, piperazinyl, and morpholinyl, wherein the heterocycloalkyl groups may optionally be substituted by 1, 2, or 3 groups, wherein the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃，R₄And R₉Independently H or methyl. In another aspect, R₄、R₉And R₁₁As defined above, R₁₅Is hydrogen, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In another aspect, the invention provides compounds of formula (XII), wherein R₄And R₉Independently is H or methyl, R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), or heterocycloalkyl, wherein the heterocycloalkyl group is selected from pyridyl, pyrimidinyl, quinolinyl, isoquinolinyl, and indolyl, wherein the heterocycloalkyl group can be optionally substituted with 1, 2, or 3 groups, wherein the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxyRadical, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃，R₄And R₉Independently H or methyl. In another aspect, R₄、R₉And R₁₁As defined above, R₁₅Is hydrogen, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the present invention provides compounds of formula (XII), wherein R₄And R₉Is H.

In another aspect, the invention provides compounds of structural formula (XII), wherein 2 or more of the foregoing aspects are combined.

In another aspect, the invention provides a compound of formula (XIII), i.e., a compound of formula (XII) having the formula:

wherein R is₁₅Is hydrogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, C₁-C₆Haloalkyl (in one aspect, it is CF)₃)、C₁-C₆Haloalkoxy (in one aspect, it is OCF)₃) Hydroxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), or methanesulfonyl, C₀-C₆-sulfamoyl, NO₂，R₁₆Is H or-O- (C)₀-C₆Alkyl) -C (O) R₁₁. In another aspect, R₁₅Is H.

In yet another aspect, the invention provides compounds of formula (XIII), wherein R₄And R₉Independently is H or methyl, R₁₁Is OH, C₁-C₄Alkoxy (on the other hand, it is C₁-C₃Alkoxy), or C₁-C₂alkoxy-C₁-C₃Alkoxy-. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIII), wherein R₄And R₉Independently is H or methyl, R₁₁Is substituted C₁-C₄Alkoxy, where the substituent is amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), aza-bicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl; and R is₄Is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, or- (C)₀-C₆Alkyl) -C (O) NH-pyridin-4 yl; in another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIII), wherein R₄And R₉Independently is H or methyl, R₁₁Is by amino, -NH (C)₁-C₆Alkyl), or-N (C)₁-C₆Alkyl) (C₁-C₆Alkyl) substituted C₁-C₄An alkoxy group. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIII), wherein R₄And R₉Independently is H or methyl, R₁₁Is pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, or- (C)₀-C₆Alkyl) -C (O) NH-pyridin-4-yl substituted C₁-C₄An alkoxy group. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIII), wherein R₄And R₉Is H.

In another aspect, the invention provides compounds of formula (XIII) wherein 2 or more of the foregoing aspects are combined.

In another aspect, the invention provides a compound of formula (XIV), i.e., a compound of formula (X) of formula:

In a further aspect, the present invention provides a compound of formula (XIV), wherein R₄And R₉Independently is H or methyl, R₁₁Is OH, C₁-C₄Alkoxy (on the other hand, it is C₁-C₃Alkoxy), or C₁-C₂alkoxy-C₁-C₃Alkoxy-. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy. In yet another aspect, R₄And R₉Is H.

In yet another aspect, the invention provides compounds of formula (XIV), wherein R₄And R₉Independently is H or methyl, R₁₁Is substituted C₁-C₄Alkoxy, where the substituent is amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), aza-bicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl; and R is₄Is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, or- (C)₀-C₆Alkyl) -C (O) NH-pyridin-4 yl. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIV), wherein R₄And R₉Independently is H or methyl, R₁₁Is by amino, -NH (C)₁-C₆Alkyl), or-N (C)₁-C₆Alkyl) (C₁-C₆Alkyl) substituted C₁-C₄An alkoxy group. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIV), wherein R₄And R₉Independently is H or methyl, R₁₁Is pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, or- (C)₀-C₆Alkyl) -C (O) NH-pyridin-4-yl substituted C₁-C₄An alkoxy group. On the other hand, in the case of a liquid,R₄、R₉and R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the invention provides compounds of formula (XIV), wherein R₄And R₉Is H.

In another aspect, the invention provides compounds of formula (XIV) wherein 2 or more of the foregoing aspects are combined.

In another aspect, the present invention provides a compound of formula (XV), i.e., a compound of formula (X) having the formula:

wherein n is 1 or 2.

In yet another aspect, the present invention provides compounds of formula (XV), wherein R₄Is H or methyl, R₁₁Is OH, C₁-C₄Alkoxy (on the other hand, it is C₁-C₃Alkoxy), or C₁-C₂alkoxy-C₁-C₃Alkoxy-. In another aspect, R₄And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy. In yet another aspect, R₄And R₉Is H.

In yet another aspect, the present invention provides compounds of formula (XV) wherein R₄And R₉Independently is H or methyl, R₁₁Is substituted C₁-C₄Alkoxy, where the substituent is amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), aza-bicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl,Azabicyclo-decyl, wherein the heteroatom may be optionally substituted by methyl or ethyl; and R is₄Is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, or-C (O) NH-pyridin-4 yl. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the present invention provides compounds of formula (XV) wherein R₄And R₉Independently is H or methyl, R₁₁Is by amino, -NH (C)₁-C₆Alkyl), or-N (C)₁-C₆Alkyl) (C₁-C₆Alkyl) substituted C₁-C₄An alkoxy group. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the present invention provides compounds of formula (XV) wherein R₄Independently is H or methyl, R₁₁Is substituted C₁-C₄Alkoxy where the substituent is aza-bicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl; and R is₄Is H or methyl, pyrrolidinyl, piperidinyl, morpholinyl, pyridinyl, or- (C)₀-C₆Alkyl) -C (O) NH-pyridin-4 yl. In another aspect, R₄、R₉And R₁₁As defined above, R₁Is chlorine; r₂Is amino; r₃Is methoxy.

In yet another aspect, the present invention provides compounds of formula (XV) wherein 2 or more of the foregoing aspects are combined.

In another aspect, the invention provides a compound according to any one of structural formulae (X), (XI), (XII), (XIII), (XIV) or (XV), wherein R₁、R₂And R₃The positions on the benzene ring are as follows:

in another aspect, the invention provides a compound according to any one of structural formulae (X), (XI), (XII), (XIII), (XIV), or (XV), wherein the bond at position 3 is in the "S" configuration and the bond at position 4 is in the "R" configuration.

In yet another aspect, the invention provides a compound according to any one of structural formulae (X), (XI), (XII), (XIII), (XIV) or (XV), wherein R₁、R₂And R₃The positions on the benzene ring are as follows:

and the bond at position 3 is in the "S" configuration and the bond at position 4 is in the "R" configuration.

In another aspect, the invention provides a compound according to any one of structural formulae (X), (XI), (XII), (XIII), (XIV), or (XV), wherein the bond at position 3 is in the "R" configuration and the bond at position 4 is in the "S" configuration.

and the bond at position 3 is in the "R" configuration and the bond at position 4 is in the "S" configuration.

In yet another aspect, the present invention providesA compound of formula (X) wherein R₁Is chlorine; r₂Is amino; r₃Is methoxy; r₄Is H, and R₁、R₂And R₃The positions on the benzene ring are as follows:

and is

L is- (C)₃-C₅Alkyl) -, one of the carbon atoms of which may be replaced by-N (R)₉) -or- (C)₂-C₆Alkyl) -C (O) -substitution. In yet another aspect, R₁、R₂And R₃As defined above and the position thereof on the phenyl ring is as defined above, R₄As defined above, R₅is-O-heterocycloalkyl, wherein the heterocycloalkyl group is selected from aza-bicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, aza-bicyclononyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl, piperidinyl, piperazinyl, and pyrrolidinyl, wherein piperidinyl, piperazinyl, and pyrrolidinyl may be unsubstituted or may be substituted in 1 or 2 positions, wherein the substituents are independently C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halogen, C₁-C₄Haloalkyl, C₁-C₄Haloalkoxy, hydroxy-C₁-C₄Alkyl, amino, -NH (C)₁-C₄Alkyl), -N (C)₁-C₄Alkyl) (C₁-C₄Alkyl), - (C)₀-C₆Alkyl) -C (O) R₁₁Or NO₂Wherein

R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups, wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), -C (O) N (R)₉) -heterocycloalkyl, or heterocycloalkyl group, wherein said heterocycloalkyl group is selected from azabicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl, 8-azabicyclo [3.2.1]Oct-3-yl, azabicyclo-nonyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl; and R is₄Is H or methyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl; wherein the heterocycloalkyl group is optionally substituted with 1, 2 or 3 groups, wherein the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃。

In yet another aspect, the present invention provides compounds of formula (X), wherein R₁Is chlorine; r₂Is amino; r₃Is methoxy; r₄Is H, and R₁、R₂And R₃The positions on the benzene ring are as follows:

and is

L is- (C)₃-C₅Alkyl) -, one of the carbon atoms of which may be replaced by-N (R)₉) -or- (C)₂-C₆Alkyl) -C (O) -substitution. In yet another aspect, R₁、R₂And R₃As defined above and the position thereof on the phenyl ring is as defined above, R₄As defined above, R₅Is a heterocycloalkyl group selected from aza-bicyclo-octyl, which in certain embodiments is 1-azabicyclo [2.2.2]Oct-3-yl or 8-aza-bicyclo [3.2.1]Oct-3-yl, aza-bicyclononyl, azabicyclo-decyl, wherein the heteroatoms may optionally be substituted by methyl or ethyl.

and is

L is- (C)₃-C₅Alkyl) -, one of the carbon atoms of which may be replaced by-N (R)₉) -or- (C)₂-C₆Alkyl) -C (O) -substitution. In yet another aspect, R₁、R₂And R₃As defined above and the position thereof on the phenyl ring is as defined above, R₄As defined above, R₅is-N (R)₉)-C₀-C₄Alkyl-aryl or-N (R)₉)-(C₀-C₆Alkyl) -C (O) -aryl, wherein the aryl group may be unsubstituted or may be substituted in 1 or more substitutable positions, wherein the substituent is C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, C₁-C₆Haloalkyl, C₁-C₆Haloalkoxy, hydroxy, hydroxyalkyl, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), - (C)₀-C₆Alkyl) -C (O) R₁₁Or NO₂. In yet another aspect, the aryl group is- (C)₀-C₆Alkyl) -C (O) R₁₁Substituted phenyl, which may also be optionally substituted with 1 or 2 groups, wherein the substituents are independently selected from C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, CF₃、OCF₃Hydroxy, hydroxyalkyl, amino, -NH (C)₁-C₄Alkyl), -N (C)₁-C₄Alkyl) (C₁-C₄Alkyl), or NO₂Wherein

R₁₁Is C₁-C₆Alkoxy, which may be optionally substituted with 1 or 2 groups,wherein the substituents are independently C₁-C₄Alkoxy, amino, -NH (C)₁-C₆Alkyl), -N (C)₁-C₆Alkyl) (C₁-C₆Alkyl), - (C)₀-C₆Alkyl) -C (O) N (R)₉) -heterocycloalkyl, or heterocycloalkyl wherein said heterocycloalkyl group is selected from pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl, wherein said heterocycloalkyl group may be optionally substituted with 1, 2 or 3 groups, wherein the substituents are independently halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, hydroxy-C₁-C₆Alkyl radical, C₁-C₆Alkoxycarbonyl, -CO₂H、CF₃Or OCF₃. In a preferred aspect, - (C)₀-C₆Alkyl) -C (O) R₁₁The group is attached at the 4-position of the benzene ring.

In yet another aspect, the orientation of the bonds in positions 3 and 4 is as follows:

in a preferred aspect, the orientation of the bonds in positions 3 and 4 is as follows:

the present invention further provides a method of treating emesis, dyspepsia, gastroparesis, constipation, intestinal pseudo-obstruction, gastroesophageal reflux disease, or post-operative ileus, which method comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (X) or a salt thereof.

The present invention provides compounds that are more readily degraded by plasma and/or cytosolic esterases than cisapride, thereby avoiding the side effects associated with cytochrome P450 metabolism.

Advantageously, the therapeutic compounds of the present invention are stable in nature upon storage, but have a relatively short half-life in a physiological environment; thus, the compounds of the present invention have a low incidence of side effects and toxicity when used.

In a preferred aspect of the invention there is provided a therapeutic stereoisomeric compound useful in the treatment of gastroesophageal reflux disease, said compound containing an ester group which is readily degraded by an esterase enzyme thereby decomposing the compound and facilitating its effective elimination from the individual being treated. In a preferred aspect, the therapeutic stereoisomeric compound is metabolized by a first stage drug detoxification system.

In a further aspect, the invention relates to breakdown products (preferably metabolic breakdown products, i.e. metabolites, typically the acid of the parent ester) which are formed when the therapeutic compounds of the invention are exposed to an esterase. The presence of these breakdown products in urine or plasma can be used to monitor the clearance of the therapeutic compound from the patient.

The degradation of the compounds of the invention by esterases is particularly advantageous for drug metabolism, since esterases are distributed everywhere and their activity is less dependent on age, sex or disease state than oxidizing liver drug metabolism.

The present invention further provides a method of treating a disorder, such as gastroesophageal reflux disease, which comprises administering to a subject in need of such treatment a therapeutically effective amount of at least one stereoisomeric structure and/or functional analog of cisapride. In particular aspects, the present invention provides stereoisomeric structural and/or functional analogs of cisapride and pharmaceutical compositions of these esterified compounds.

The present invention further provides materials and methods for treating emesis and other disorders, including but not limited to dyspepsia, gastroparesis, constipation, and intestinal pseudo-obstruction, with substantially reduced side effects associated with the administration of cisapride.

In a preferred aspect of the invention there is provided a stereoisomeric compound useful in the treatment of gastroesophageal reflux disease, dyspepsia, gastroparesis, constipation, post-operative ileus and intestinal pseudo-obstruction, said compound containing an ester group which is acted upon by an esterase enzyme, thereby causing decomposition of the compound and promoting its effective removal from the individual being treated.

The present invention further provides a method for the synthesis of the unique and advantageous compounds of the present invention. In particular, methods of making and purifying such stereoisomeric compounds are taught. Methods for introducing ester moieties and making and purifying stereoisomers are well known to those skilled in the art and can be readily performed by those skilled in the art using the guidance provided herein.

Preferred compounds

In a preferred aspect, the present invention provides isolated stereoisomers of compound I having 3 chiral centers.

6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid-1-aminoheterobicyclo [2.2.2] oct-3-yl ester

Compound I

Cisapride and norcisapride (norcisapride) have 2 chiral centers present in their cis configuration in the active drug.

(±) -cisapride (±) -norcisapride

Thus, for example, pharmaceutically active norcisapride is a racemic mixture of two cis enantiomers.

(-) -norcisapride (+) -norcisapride

In one aspect, the invention is particularly directed to the configuration at the 3 rd chiral center of the quininol moiety. This group elimination is converted to an acid metabolite (referred to herein as compound II).

Compound II

The preferred stereoisomers of compound I of the present invention are prepared by combining quinuclidinol in either the R or S configuration with (+) -or (-) -norcisapride to give compounds of structural formulae III, IV, V and VI.

(3R, 4S, 3' R) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid-1-azabicyclo [2.2.2] oct-3-yl ester

Compound III: (-) - (R) -Compound I

(3S, 4R, 3' R) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid-1-azabicyclo [2.2.2] oct-3-yl ester

Compound IV: (+) (R) -Compounds I

(3R, 4S, 3' S) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid-1-azabicyclo [2.2.2] oct-3-yl ester

Compound V: (-) - (S) -Compound I

(3S, 4R, 3' S) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid-1-azabicyclo [2.2.2] oct-3-yl ester

Compound VI: (+) (S) -Compounds I

In a preferred aspect, the present invention relates to stereoisomerically isolated compounds, and compositions comprising the compounds. The isolated stereoisomeric form of the compound of the present invention is substantially free of the other (i.e., stereoisomeric excess). In other words, the compound of the "R" configuration is almost free of the compound of the "S" configuration, and therefore it has a stereoisomeric excess for the "S" configuration. In contrast, a compound of the "S" configuration contains little compound of the "R" configuration, and therefore, it has a stereoisomeric excess for the "R" configuration. In one aspect of the invention, the stereoisomeric excess of the isolated stereoisomeric compound is at least about 80%. In a preferred aspect, the stereomeric excess of the compound is at least about 90%. In a more preferred aspect, the stereomeric excess of the compound is at least about 95%. In a further preferred aspect, the stereomeric excess of the compound is at least about 97.5%. In a most preferred aspect, the stereomeric excess of the compound is at least about 99%. Similarly, "(+)" and "(-) -forms of the compound also have stereoisomeric excesses.

As described herein, the different stereoisomers have particular, unexpected properties that can be advantageously used in treatments specifically targeted to specific conditions. Thus, for example, compounds containing the (3' R) -isomer in the quinuclidinyl ester moiety (i.e., compounds III and IV) can be rapidly metabolized by esterases in human plasma; however, the metabolism of the quininol (3' S) -isomer-containing compounds (i.e., compounds V and VI) is much slower.

Thus, the (3' R) -isomer of compound I can be used preferably with a rapid onset of action, for example in pulse therapy for patients suffering from acute gastroparesis or acute gastroesophageal reflux disease. Another advantage of rapid metabolism of esterases to produce metabolites with greatly reduced activity (i.e. compound II) is that: it has a very low probability of drug-drug interactions and toxicity. Therefore, these short-acting (R) -isomers can be advantageously used as intravenous formulations for the treatment of gastroesophageal reflux disease in premature newborns who, because their CYP450 system has not yet fully developed, are unable to metabolize the drug as in adults. In neonates, drugs that are rapidly metabolized by systems other than CYP450 (e.g., esterase systems) are of great advantage. On the other hand, the (3' S) -isomer of compound I is most suitable for the treatment of chronic symptoms of the same disease, e.g. gastroparesis in diabetic or cancer patients in the stationary phase, or chronic gastroesophageal reflux disease in patients requiring 24 hours of drug action.

Apart from their differences in metabolic rate, these single isomers have different 5-HT₄Receptors bind and thus also exhibit different activities and therefore have different therapeutic uses. Thus, in 5-HT₄In order of decreasing receptor binding, isomers can be classified as follows (binding constants Ki values in parentheses): compound IV (1.4nM), compound VI (3.4nM), compound III (28nM), and compound V (72 nM). These binding experimentsThis is done using a radiolabelling method as described in standard texts, which can be easily repeated by a person skilled in the art of molecular biology.

Based on these considerations conclusions: when the 3-and 4-positions are in cis with respect to each other, compound I is a mixture of 4 isomers, which consists of two pairs of enantiomers. The first pair of enantiomers was (+) (R) -Compound I and (-) (S) -Compound I (Compounds IV and V, respectively), and the second pair of enantiomers was (-) (R) -Compound I and (+) (S) -Compound I (Compounds III and VI, respectively). In each pair of enantiomers, each single enantiomer has an esterase hydrolysis rate and a 5-HT₄The nature of the receptor varies in its binding capacity. These different properties give them advantageous therapeutic uses that are not interchangeable for each, i.e. they are specific for each isomer and they are not applicable to racemic mixtures. When testing racemic mixtures, differences in the binding capacity of these receptors and differences in these metabolic rates are unpredictable and it is not possible to profile these properties.

Definition of

As used herein, the term "alkyl" includes those alkyl groups having the specified number of carbon atoms. The alkyl group may be straight chain, or branched. Examples of "alkyl" include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyl and the like. If the number of carbon atoms is not specified, the "alkyl" moiety has from 1 to 6 carbon atoms.

The term "alkoxy" represents an alkyl group having the indicated number of carbon atoms attached to the parent molecular moiety through an oxygen bridge. Examples of alkoxy groups include, for example, methoxy, ethoxy, propoxy, isopropoxy.

"aryl" refers to an aromatic carbocyclic group having a single ring (e.g., a benzene ring), which may optionally be fused or attached to other aromatic or non-aromatic hydrocarbon rings. "aryl" includes multiple fused rings (at least one of which is aromatic, e.g. 1, 2)3, 4-tetrahydronaphthyl, naphthyl, and wherein each ring may be optionally mono-, di-, or tri-substituted with groups described below) and an unfused polycyclic group, such as a biphenyl or binaphthyl group. Preferred aryl groups of the invention are phenyl, 1-naphthyl, 2-naphthyl, indanyl, indenyl, dihydronaphthyl, fluorenyl, 1, 2, 3, 4-tetrahydronaphthyl, or 6, 7, 8, 9-tetrahydro-5H-benzo [ a ]]Cycloheptenyl. More preferred are phenyl, biphenyl and naphthyl. Phenyl is most preferred. The aryl groups herein may be unsubstituted or substituted, as specifically indicated, with various groups at one or more substitutable positions. For example, these aryl groups may be optionally substituted with, for example, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, hydroxy, nitrile, nitro, amino, mono (C)₁-C₆) Alkylamino radical, di (C)₁-C₆) Alkylamino radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₆Haloalkyl, C₁-C₆Haloalkoxy, amino- (C)₁-C₆) Alkyl, mono (C)₁-C₆) Alkylamino radical- (C)₁-C₆) Alkyl or di (C)₁-C₆) Alkylamino radical- (C)₁-C₆) An alkyl group.

The term "haloalkoxy" refers to an alkoxy group substituted with at least one halogen atom, and optionally further substituted with at least one other halogen atom, wherein each halogen atom is independently F, Cl, Br, or I. Preferred halogens are F or Cl. Preferred haloalkoxy groups contain 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms, and most preferably 1 to 2 carbon atoms. "Haloalkoxy" includes perhaloalkoxy groups, e.g. OCF₃Or OCF₂CF₃。

The term "heteroaryl" refers to an aromatic ring system containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. The heteroaromatic ring may be fused or linked to one or more heteroaromatic rings, aromatic or non-aromatic hydrocarbon rings, or heterocycloalkyl rings. Examples of heteroaryl groups include, for example, pyridyl, pyrimidinyl, quinazo, and the likeLinyl, benzothienyl, indolyl, indolinyl, pyridazinyl, pyrazinyl, isoindolyl, isoquinolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, imidazolyl, isoxazolyl, pyrazolyl, oxazolyl, thiazolyl, indolizinyl, indazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, furyl, thienyl, pyrrolyl, oxadiazolyl, thiadiazolyl, benzo [1, 4 ] diazanyl, thiadiazolyl, and mixtures thereof]Oxazinyl, triazolyl, tetrazolyl, isothiazolyl, 1, 5-diazanaphthyl, isochromanyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazinyl, pyridopyridyl, benzotetrahydrofuranyl, benzothiophenyl, purinyl, benzodioxolyl, triazinyl, pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl, dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl, dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, chromonyl, chroman-4-one, pyridyl N-oxide, tetrahydroquinolinyl, dihydroquinolinyl, dihydroquinolinonyl, dihydroisoquinolinonyl (dihydroisoquinolinonyl), Dihydrocoumarinyl, dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl, benzoxazolinonyl (benzoxazolinonyl), pyrrolyl N-oxide, pyrimidinyl-N-oxide, pyridazinyl N-oxide, pyrazinyl N-oxide, quinolinyl-N-oxide, indolyl-N-oxide, indolinyl-N-oxide, isoquinolinyl-N-oxide, quinazolinyl N-oxide, quinoxalinyl N-oxide, naphthyridinyl N-oxide, imidazolyl N-oxide, isoxazolyl-N-oxide, oxazolyl N-oxide, thiazolyl N-oxide, indolizinyl N-oxide, indazolyl-N-oxide, benzoxazolinyl, Benzothiazolyl N-oxide, benzimidazolyl N-oxide, pyrrolyl N-oxide, oxadiazolyl N-oxide, thiadiazolyl N-oxide, triazolyl N-oxide, tetrazolyl N-oxide, benzothiopyranyl S, S-dioxide. Preferred heteroaryl groups include pyridyl, pyrimidinyl, quinolinyl, indolePhenyl, pyrrolyl, furyl, thienyl, and imidazolyl. More preferred heteroaryl groups include pyridyl, pyrrolyl and indolyl. Heteroaryl groups herein may be unsubstituted or substituted, as specifically indicated, with various groups at one or more substitutable positions. For example, these heteroaryl groups may be optionally substituted with, for example, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, hydroxy, nitrile, nitro, amino, mono (C)₁-C₆) Alkylamino radical, di (C)₁-C₆) Alkylamino radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₆Haloalkyl, C₁-C₆Haloalkoxy, amino- (C)₁-C₆) Alkyl, mono (C)₁-C₆) Alkylamino radical- (C)₁-C₆) Alkyl or di (C)₁-C₆) Alkylamino radical- (C)₁-C₆) An alkyl group.

The term "heterocycloalkyl" refers to a ring or ring system containing one heteroatom preferably selected from nitrogen, oxygen and sulfur, wherein said heteroatom is located in a non-aromatic ring. The heterocycloalkyl ring may be optionally fused or linked to other heterocycloalkyl rings and/or non-aromatic hydrocarbon rings and/or benzene rings. Preferred heterocycloalkyl groups are 3 to 7 membered rings. More preferred heterocycloalkyl groups are 5 to 6 membered rings. Examples of heterocycloalkyl radicals include, for example, aza-bicyclo [2.2.2]Octyl, aza-bicyclo [3.2.1]Octyl, morpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S, S-dioxide, piperazinyl, homopiperazinyl, pyrrolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, tetrahydrofuranyl, tetrahydrothienyl, homopiperidinyl, homomorpholinyl, homothiomorpholinyl S, S-dioxide, oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidyl, dihydrofuranyl, 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, thiomorpholinyl, S-dioxothiomorpholinyl, morpholinyl, and imidazolidinyl. More preferred heterocycloalkyl groups include aza-bicyclo [2.2.2]Octyl, aza-bicyclo [3.2.1]Octyl, piperidinyl, piperazinyl, pyrrolidinyl, imidazolidinyl, and morpholinyl. The heterocycloalkyl group herein may be unsubstituted or substituted at one or more substitutable positions with various groups as specifically noted. For example, these heterocyclic groups may be optionally substituted with, for example, C₁-C₆Alkyl radical, C₁-C₆Alkoxy, halogen, hydroxy, nitrile, nitro, amino, mono (C)₁-C₆) Alkylamino radical, di (C)₁-C₆) Alkylamino radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₆Haloalkyl, C₁-C₆Haloalkoxy, amino- (C)₁-C₆) Alkyl, mono (C)₁-C₆) Alkylamino radical- (C)₁-C₆) Alkyl or di (C)₁-C₆) Alkylamino radical- (C)₁-C₆) Alkyl, or ═ O.

The term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" refers to a salt prepared from a pharmaceutically acceptable, non-toxic acid or base, including inorganic and organic acids and bases. Since the compounds of the present invention are basic, salts can be prepared from pharmaceutically acceptable, non-toxic acids. Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention include acetic acid, benzenesulfonic (benzenesulfonate), benzoic, camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic and the like. Preferred acid addition salts are chloride and sulfate salts. In a most preferred aspect, the structural and/or functional analog of cisapride is administered as the free base or as the mono-or dihydrochloride salt.

As used herein, the terms "treatment" and "treating" include prophylactic administration ("prophylaxis") of a compound or a pharmaceutical composition comprising the compound, as well as remedial treatment to reduce or eliminate a disease or condition mentioned herein. Prophylactic administration is intended to prevent a disease, which can be used to treat a subject at risk of suffering from or suffering from one or more of the diseases mentioned herein. Thus, as used herein, the term "treatment" or derivatives thereof refers to the partial or total inhibition of the disease state to which the active ingredients of the present invention are administered prophylactically, or when administered after the onset of the disease state to which these active ingredients are administered. "preventing" refers to administering an active ingredient to a mammal to protect the mammal from any of the diseases described herein as well as other diseases.

The term "therapeutically effective dose" refers to an amount necessary to achieve the desired therapeutic effect, for example: 1) an amount sufficient to alleviate reflux disease; 2) an amount sufficient to reduce nausea and vomiting; 3) an amount sufficient to alleviate the disease caused by gastric motility dysfunction. The above dose and dosing frequency table includes therapeutically effective amounts of structural and/or functional analogs of cisapride.

A "mammal" can be, for example, a mouse, rat, pig, horse, rabbit, sheep, cow, cat, dog, or human. In a preferred aspect, the mammal is a human.

The term "subject" refers to a single mammal administered with a compound of the present invention. A "mammal" can be, for example, a mouse, rat, pig, horse, rabbit, sheep, cow, cat, dog, or human. In a preferred aspect, the mammal is a human.

The term "esterified cisapride" refers to therapeutic compounds of the present invention that are structural and/or functional analogs of cisapride, which contain hydrolyzable groups (usually esters) that do not reduce the ability of these compounds to provide therapeutic benefit, but that render these compounds more susceptible to degradation by esterases, particularly plasma and/or cytosolic esterases, thereby reducing the interaction of the cisapride compound with the cytochrome P450 drug detoxification system. Esterase-mediated metabolism of esterified cisapride compounds reduces the effect of the cytochrome P450 drug detoxification system in cisapride metabolism and reduces or eliminates the side effects caused by cisapride.

The term "structural analog" as used herein means that the compound has the same structural characteristics as the parent compound. For example, structural analogs of cisapride may share one or more structural features with the parent compound of cisapride (e.g., a substituted aromatic ring that is both attached to the piperidine ring through an amide linker), but may also differ structurally, e.g., by the inclusion or deletion of one or more chemical moieties.

The term "functional analog" as used herein means that the compound has the same functional properties as the parent compound. For example, a functional analog of cisapride may have few, if any, of the same structural features as cisapride, but performs a similar function, e.g., 5-HT₄An agonist.

The term "side effects" includes, but is not limited to, gastrointestinal disorders such as diarrhea, abdominal cramps, and borborygmus; fatigue; headache; the systolic pressure rises; death; ventricular tachycardia; ventricular fibrillation; torsade de pointes ventricular velocity and long QT syndrome; an increase in heart rate; neurological and CNS disorders; and cisapride interactions with other drugs such as, but not limited to, digoxin, diazepam, alcohol, acetonitrocoumaryl alcohol, cimetidine, ranitidine, acetaminophen, and propranolol.

The term "gastroesophageal reflux disease" as used herein refers to the onset and symptoms of those conditions that cause the reflux of food from the stomach into the esophagus.

The terms "induce an antiemetic effect" and "antiemetic treatment" as used herein refer to the reduction or prevention of symptoms of nausea and vomiting caused by or associated with emetic cancer chemotherapy or radiotherapy.

The term "treating a disease caused by gastric motility dysfunction" as used herein refers to treating symptoms and conditions associated with gastric motility dysfunction, including, but not limited to, gastroesophageal reflux disease, dyspepsia, gastroparesis, constipation, post-operative ileus, and intestinal pseudo-obstruction.

The term "prokinetic" as used herein refers to enhancing the motility, and thus the movement, of the gastrointestinal tract.

The term "dyspepsia" is used to refer to an impairment of digestive ability or function that may cause major gastrointestinal dysfunction or a complication that is or is caused by other disorders such as appendicitis, gallbladder disease or malnutrition.

The term "gastroparesis" is used to refer to gastric paralysis that is caused by an abnormal gastric motility or as a complication of diseases such as diabetes, progressive systemic sclerosis, anorexia or myotonic dystrophy.

The term "constipation" is used to refer to conditions such as a lack of intestinal muscle contractile function or intestinal spasms resulting in little or difficult excretion.

The term "post-operative ileus" as used herein refers to blockage of the intestine due to decreased muscle tone following surgery.

The term "intestinal pseudo-obstruction" as used herein refers to a condition characterized by constipation, colic and vomiting, but without evidence of mechanical obstruction.

Preparation of the Compounds

The chemical synthesis of various cisapride analogs can be performed according to the methods described in the following references and modified by the introduction of an ester group at a convenient position in the synthesis of the disclosed compounds: european patent application No. 0,076,530 a2, U.S. patent nos. 4,962,115 and 5,057,525, published on 13/4/1983, and Daele et al, drug development res.8: 225-232(1986), the entire contents of which are incorporated herein by reference. An exemplary, non-limiting synthetic scheme for certain esterified cisapride analogs of the present invention is described in WO 01/093849.

The invention is further illustrated by the following examples, which are not intended to limit the scope or spirit of the invention to the particular methods described in these examples. One skilled in the art will recognize that the starting materials may be varied and additional steps may be employed to prepare the compounds encompassed by the present invention, as described in the following examples. One skilled in the art will also recognize that different solvents or reagents may be necessary to effect the above transformations. In some cases, it may be necessary to protect the reactive functional groups in order to achieve the above transformations. In general, such a need for protecting groups, as well as the conditions necessary to attach and remove such protecting groups, will be apparent to those skilled in the art of organic synthesis. When protecting groups are used, a deprotection step may be required. Suitable Protecting Groups and methods of protection and deprotection, such as those described by protective Groups in Organic Synthesis, T.Greene, are well known in the art and are appreciated in the art.

All reagents and solvents were of standard commercial grade and used without further purification unless otherwise indicated. Suitable atmospheres for carrying out the reaction, such as air, nitrogen, hydrogen, argon, and the like, will be apparent to those skilled in the art.

Example 1

Preparation of 6- [4R- (4-amino-5-chloro-2-methoxy-benzoylamino) -3S-methoxy-piperidin-1-yl ] -hexanoic acid-1-azabicyclo [2.2.2] oct-3' R-yl ester dihydrochloride (ATI-7505 dihydrochloride)

Step 1: resolution of racemic norcisapride

(-) -DBT, about 1 part by weight, was dissolved in ethanol and filtered to remove insoluble particles. In addition, racemic cisapride (about 0.8 parts by weight) was dissolved in a mixture of ethanol and water, and then filtered. The filtrate was heated to about 75 ℃ and then (-) -DBT solution was added. Stirring is carried out at this temperature for about 30 minutes, the mixture is slowly cooled to 5 ℃ over several hours, the product salt is collected by vacuum filtration and washed with an ethanol/water mixture. The wet cake was recrystallized from ethanol/water by heating to about 79 deg.C, cooling to about 5 deg.C (as above). The product was collected by vacuum filtration and washed with an ethanol/water mixture to give a wet cake product.

The wet cake product was suspended in water and the pH adjusted to about 12 with 7% (W/W) aqueous NaOH. The resulting suspension was stirred at room temperature for about 3 hours, then filtered under vacuum, the solid material was washed with water and dried under vacuum. The product was then treated again with (-) -DBT in the same manner as described above to form a salt. The isolated salt was then neutralized with aqueous NaOH as described above. The product was isolated by filtration and dried to yield (+) -norcisapride base (about 0.25 parts by weight). The e.e. value was about 100% (+) -norcisapride by manual HPLC analysis. The optical rotation was about +5 ° (methanol; 25 ℃, 589nm), and norcisapride was confirmed to be a dextrorotatory isomer.

Step 2: coupling with ethyl 6-bromohexanoate

(+) -norcisapride (about 1 part by weight), potassium carbonate (about 0.48 part by weight) and potassium iodide (about 0.063 part by weight) were suspended in anhydrous USP ethanol. To this suspension was slowly added ethyl 6-bromohexanoate (about 0.76 parts by weight) at room temperature. The mixture was heated to reflux until the reaction was complete. Subsequently, the reaction mixture was cooled to room temperature, filtered to remove inorganic solids and the like, and the filtrate was concentrated under reduced pressure to half the original volume. The crude product was slowly added to cold water (about 13 parts by weight) with rapid stirring, and the product precipitated. The precipitate was vacuum filtered, washed with water, then dissolved in absolute ethanol as described above, slowly added with cold water and reprecipitated twice. The resulting wet cake was washed with n-heptane and resuspended in ethyl acetate/n-heptane (1: 9; v/v), stirred for about 1 hour, filtered and dried in vacuo to give 0.73 parts by weight of the coupling product as a white solid.

And step 3: coupling with (R) -3-hydroxyninginol and formation of the dihydrochloride

The ester (1 part by weight) and (R) -3-hydroxyningitol (about 1.12 parts by weight) were suspended in toluene, and then titanium (IV) ethoxide (about 0.5 part by weight) was slowly added to the suspension under stirring. The mixture was heated to about 91 ℃ under a stream of nitrogen and the flask portion was evacuated by a distillation apparatus to form an azeotrope to remove ethanol. Additional toluene was added to maintain a minimum solvent volume in the flask, if needed. The reaction was considered complete after about 33 hours.

The mixture was cooled to room temperature and extracted 5 times with water. The organic layer was concentrated under reduced pressure and the resulting residue was redissolved in EtOH/iPrOH (ca. 1: 1v/v) and then filtered through a 0.45 micron filter to remove any particles. To the filtrate was slowly added concentrated hydrochloric acid with stirring to precipitate the desired product as the dihydrochloride salt. The resulting suspension was stirred at room temperature for several hours, collected by vacuum filtration and washed with EtOH/iPrOH (1: 1; v/v) to give 0.53 parts by weight of crude product salt.

The crude dihydrochloride salt was again suspended in ethanol, heated to reflux for more than 1 hour, and then cooled to room temperature. The product was collected by vacuum filtration and rinsed with ethanol, then air dried. The solid was resuspended in ethanol and heated to about 55 ℃ to give a clear solution, then warm isopropanol was added and the mixture was slowly cooled to room temperature and the product precipitated out. The resulting suspension is stirred for several hours, then filtered under vacuum and washed with, for example, isopropanol. The product was dried under vacuum, first at room temperature for several hours and then at about 55 ℃ to constant weight.

Example 2

Preparation of (R) -quinuclidin-3-ol-6- ((3S, 4R) -4- (4-amino-2-chloro-6-methoxybenzamide) -3-methoxypiperidin-1-yl) hexanoate

Step 1: synthesis of 4- (dibenzylamino) -3-methoxypiperidine-1-carboxylic acid ethyl ester (1):

to a solution of racemic 4-amino-3-methoxypiperidine-1-carboxylic acid ethyl ester (1 molar part) in DMF were added benzyl bromide (about 2.2 molar parts), potassium carbonate (about 2.4 molar parts), and potassium iodide (about 0.2 molar part), respectively. The reaction was heated to 80 ℃. After 6 hours, the reaction mixture (about 12 parts by volume) is diluted slowly with water and extracted, for example, with ethyl acetate. The organic layer was washed with brine and then with anhydrous Na₂SO₄And (5) drying. Then filtered and the solvent was concentrated to give 1(1 molar part) as an orange-yellow oil.

Step 2: synthesis of N, N-dibenzyl-3-methoxypiperidine-4-amine (2):

to the solution of 1 was added a solution of NaOH (about 10 molar parts) in isopropanol, the mixture was stirred and heated to reflux. After about 3 hours to about 5 hours, the reaction was cooled to room temperature and the alcohol solvent was removed by rotary evaporation. The mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with brine and then with anhydrous Na₂SO₄And (5) drying. Then filtering, concentrating the solvent to obtain crude oil, and using SiO to make it₂(CH₂Cl₂∶MeOH∶NH₄OH; (ca.) 15: 1: 0.01) to give 2.

Step 3, (3S, 4R) -N, N-dibenzyl-3-methoxypiperidine-4-amine (3):

(-) -dibenzoyl-L-tartaric acid (about 1.2 parts by weight) was dissolvedIn ethanol, then a solution of 2 (about 1 part by weight) was slowly added. The solution was gradually heated and then cooled to room temperature to crystallize the salt product. The salt product was filtered and washed with EtOH/H₂O washed, then suspended in water, basified by addition of aqueous NaOH (7%, wt/wt) and pH adjusted to 12. The suspension was stirred vigorously at room temperature, the solid was separated by filtration, washed with water and dried under vacuum to give cis-isomer 3.

Step 4. Synthesis of Ethyl 6- ((3S, 4R) -4- (dibenzylamino) -3-methoxypiperidin-1-yl) hexanoate (4):

to a solution of 3(1 molar parts) in DMF were added ethyl bromohexanoate (about 1.2 molar parts), potassium carbonate (about 1.4 molar parts), and potassium iodide (about 0.2 molar parts), respectively. The reaction was then heated to 80 ℃. After 8 hours, the reaction mixture was diluted slowly with water (ca. 12 parts by volume) and extracted with ethyl acetate. The organic layer was washed with brine and then with anhydrous Na₂SO₄And (5) drying. Then filtered and the solvent concentrated to give the crude product. Using SiO to make it₂Purification to obtain alkylated product 4.

Step 5. Synthesis of (R) -quinuclidin-3-ol-6- ((3S, 4R) -4- (dibenzylamino) -3-methoxypiperidin-1-yl) hexanoate (5):

to a mixture of 4(1 molar part) and (R) - (-) -3-quinuclidinol (1 molar part) in toluene was added titanium tetraethoxide. The reaction mixture was charged to a dean-stark apparatus, then heated to 90 ℃ and partially evacuated (plus toluene if necessary to maintain the necessary solvent level). The mixture was cooled to room temperature, and the reaction solution was diluted with ethyl acetate, followed byTo the resulting mixture was added water. The organic layer was separated, washed with brine and dried over anhydrous Na₂SO₄Drying, filtering and concentrating. SiO 2₂After purification, enantiomerically enriched 5 was obtained.

Step 6. Synthesis of (R) -quinuclidin-3-ol-6- ((3S, 4R) -4-amino-3-methoxypiperidin-1-yl) hexanoate (6):

a 5(1 molar part) ethanol solution was added to a reaction flask containing palladium on carbon (about 0.2 molar part). The mixture is deaerated and freed of air by means of H₂The atmosphere subjects the mixture to hydrogenolysis conditions. After the reaction was completed, palladium was filtered off through a celite pad, and then washed with ethanol. The filtrate was concentrated by rotary evaporation to give 6.

Step 7. Synthesis of (R) -quinuclidin-3-ol-6- ((3S, 4R) -4- (4-amino-2-chloro-6-methoxybenzamide) -3-methoxypiperidin-1-yl) -hexanoate (7):

benzoic acid (1 molar part) is added portionwise to a solution of THF, such as ethyl chloroformate (1 molar part), at 0 deg.C. The mixture was heated to room temperature for 1 hour, then cooled to 0 ℃, and a solution of 6 (1 molar part) was added dropwise. The reaction was then warmed to room temperature. Upon completion of the reaction, saturated NaHCO was added₃The reaction was quenched with aqueous solution and extracted with EA. The organic layer was washed with brine and dried over anhydrous Na₂SO₄Drying, filtering and concentrating to obtain the desired product 7.

Example 3

Alternative ATI-7505 Synthesis methods:

reacting 1-benzylpiperidin-4-one (1) with hydrobromic acid in the presence of acetic acid under acidic conditions to form N-benzyl-3-bromopiperidin-4-one (2). Treatment of 2 with sodium methoxide and methanol solution yielded 1-benzyl-4, 4-dimethoxypiperidin-3-one (3). (the presence of the beta-amino group made the Favorskii reaction impossible.) methylation of the hydroxyl group with hydride base in the presence of DMF solvent by treatment with methyl iodide gave compound 4.

Followed by acetal hydrolysis with 1% sulfuric acid under heating to give piperidine 5, followed by reductive amination of piperidine 5 in methanol using, for example, sodium cyanoborohydride and amine acetate to give 1-benzyl-3-methoxypiperidin-4-amine (6). At this stage, chiral resolution techniques can be used on 6. This can be done, for example, by using (-) -DBT or other variants of tartaric acid in the presence of a suitable solvent to give compound 7 as a pure asymmetric compound. Boc group protection of the primary amine in 7 can be performed using Boc anhydride in the presence of THF solvent to give 8. The debenzylation reaction was achieved by hydrogenolysis using Pd/C in methanol under atmospheric pressure of hydrogen, providing a platform for the alkylation step. Treatment with 6-bromohexanenitrile in the presence of a weak base and DMF affords compound 10. Conversion of the nitrile to the ester using (R) -quinuclidinol under dilute acid conditions yields 11. Subsequent removal of the Boc group with TFA yields the free amine, which can be coupled with the requisite benzoic acid in the presence of a coupling reagent such as ethyl chloroformate to give the enantiomerically pure material ATI-7505.

Alternatively, compound 9 can be alkylated with ethyl 6-bromohexanoate in the presence of a weak base. Subsequent removal of the Boc group provided compound 13. Titanium-catalyzed transesterification of 13 using (R) -quinuclidinol and titanium tetraethoxide in toluene solvent yields ATI-7505. Carlsburg esterase hydrolyzes the S-configured ester leaving an unreacted R-configured ester. Thus, treatment of the diastereomeric mixture of 14 with the Carlsburg esterase also produced ATI-7505.

Example 4

According to U.S. patent No. 6,147,093, or j.jacques, a.collets and s.h.wilen (Wiley internscience, New York, NY) "eneriomers, racemes and solutions", or s.h.wilen et al, Tetrahedron (1977) 33: 2725 by resolution of the enantiomers using conventional means such as optical resolution of the acid, (+) and (-) -norcisapride can be obtained from racemic mixtures of norcisapride.

The 4 isomers can be obtained in milligram quantities by collection by preparative column chromatography followed by evaporation of the solvent. The method can be used to prepare small amounts of samples for analysis and characterization. This is a standard separation method routinely employed by analytical laboratories for the isolation and characterization of metabolites.

The following illustrates possible routes for the synthesis of compound IV, compound VI and (+) -compound II using (+) -norcisapride as starting material. The synthetic routes for compound III, compound V and (-) -compound II are the same except that they use (-) -norcisapride as a starting material.

Example 5

Preparation of ethyl ester of (+) -Compound II

Equimolar amounts of (+) -norcisapride and ethyl 6-bromohexanoate (1 equivalent each), KI of catalyst, and K₂CO₃(2 equivalents) the mixture in DMF was heated to 60 ℃ for several hours or until TLC analysis indicated the reaction was complete. After cooling to room temperature, water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed successively with water, 10% aqueous LiCl and brine, then Na₂SO₄And (5) drying. Concentration gave (+) -Compound II ethyl ester.

Preparation of (+) -Compound II

A mixture of crude ethyl ester of (+) -Compound II (1 eq), KOH (2M, 5 eq) in MeOH and THF (sufficient to dissolve) obtained above was stirred at room temperature for about 1-2 hours. MeOH and THF were removed in vacuo and the residue was diluted with water. Washed with an organic solvent such as EtOAc. The pH of the aqueous layer was adjusted to 5 using HCl. The precipitate was filtered off and dried to give (+) -Compound II.

Preparation of Compound IV and Compound VI

A mixture of (+) -Compound II (1 equivalent), (R) - (-) -3-quinine alkoxide (1 equivalent), EDAC (1 equivalent) and DMAP (1 equivalent) in DMF was heated at about 50 ℃ overnight. After cooling and dilution with water, the mixture is purified by chromatography or crystallization to give compound IV. Similarly, using (S) - (+) -quinine alkoxide, compound VI is obtained.

The following compounds were prepared essentially according to the methods and procedures described above. Nomenclature for compounds generated using ChemDraw Ultraversion 8.03 (available from cambridge soft Corporation) or ACD namepro software, version 6.0.

(3S) -1-azabicyclo [2.2.2] oct-3-ol-6- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } hexanoate;

(3S) -1-azabicyclo [2.2.2] oct-3-ol-6- { (3R, 4S) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } hexanoate;

(3R) -1-azabicyclo [2.2.2] oct-3-ol-6- { (3R, 4S) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } hexanoate;

8-methyl-8-azabicyclo [3.2.1] oct-3-ol-6- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } hexanoate;

4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoic acid;

4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoic acid methyl ester;

ethyl 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-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoic acid isopropyl ester;

2-methoxyethyl 4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

2-pyrrolidin-1-ylethanol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

1-methylpiperidin-4-ol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

2-pyridin-2-ylethanol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

2- (dimethylamino) ethyl 4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

1-methylpiperidin-3-ol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

2-morpholin-4-yl-ethanol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

1, 4-dimethylpiperidin-4-ol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

2-oxo-2- (piperidin-4-ylamino) ethanol-4- [ ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) amino ] benzoate;

1- ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) piperidine-4-carboxylic acid;

1- ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) piperidine-4-carboxylic acid methyl ester;

1- ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) piperidine-4-carboxylic acid ethyl ester;

1- ({ (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } acetyl) piperidine-4-carboxylic acid (2-methoxyethyl) ester;

4- { [ (2- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } ethyl) (methyl) amino ] methyl } benzoic acid;

methyl 4- { [ (2- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } ethyl) (methyl) amino ] methyl } benzoate;

methyl 4- { [ (2- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } ethyl) amino ] methyl } benzoate;

isopropyl 4- { [ (2- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } ethyl) amino ] methyl } benzoate;

ethyl 4- { [ (2- { (3S, 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-ol-4- { [ (2- { (3S, 4R) -4- [ (4-amino-5-chloro-2-methoxybenzoyl) amino ] -3-methoxypiperidin-1-yl } ethyl) amino ] carbonyl } benzoate;

(R) -quinuclidin-3-ol-6- ((3S, 4R) -4- (4-amino-5-chloro-2-methoxybenzamide) -3-methoxypiperidin-1-yl) -hexanoate; or

6- ((3S, 4R) -4- (4-amino-5-chloro-2-methoxybenzamide) -3-methoxypiperidin-1-yl) hexanoic acid.

Formulations, administration and uses

The frequency of administration and route of administration of the disclosed compounds are similar to those already used in the art and known to the skilled artisan (see, e.g., Physicians' Desk Reference, 54th Ed., Medical Economics Company, Montvale, NJ, 2000).

The size of the dose of the structural and/or functional analogs of cisapride in the prevention or treatment of the acute or chronic diseases and/or disorders described herein will vary with the severity of the condition to be treated and the route of administration. The dosage, and possibly the frequency of administration, will also vary according to the age, weight and response of the individual patient. Typically, for the conditions described herein, the total daily dose of cisapride structural and/or functional analogs is from about 1mg to about 200mg in a single or divided dose. Preferably, the daily dose is a single or divided dose of about 5mg to about 100mg, more preferably, the daily dose should be a single or divided dose of about 5mg to about 75 mg. Preferably, administration is 1 to 4 times a day. When treating a patient, it should be started at a lower dose, which may be about 5mg to about 10mg, and gradually increased to about 50mg or higher depending on the overall response of the patient. Children, patients over 65 years of age, and people with impaired renal or hepatic renal function are further advised to use low doses initially and to adjust based on individual response and blood levels. In some cases, it may be necessary to use dosages outside these ranges as will be apparent to those skilled in the art. Furthermore, it should be noted that the clinical or treating physician will know how, when to interrupt, adjust or discontinue treatment based on the individual patient's response.

The compounds of the present invention may be formulated according to known methods for preparing pharmaceutical compositions. Formulations are described in several documents well known and readily available to those skilled in the art. Preparations that can be used in conjunction with the present invention are described, for example, in "Remington's Pharmaceutical Science" of e.w. martin. Generally, the compositions of the present invention are formulated such that an effective amount of the biologically active compound is combined with a suitable carrier to facilitate effective administration of the composition.

The compositions of the present invention include, for example, suspensions, solutions and elixirs; or, in the case where oral solid preparations such as powders, capsules, and tablets are more preferable than oral liquid preparations, carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like. The preferred oral solid preparation is a capsule. The most preferred oral solid formulation is a tablet. Preferred amounts of the active ingredient (i.e., a structural and/or functional analog of cisapride) in solid dosage forms are about 5mg, 10mg, and 25 mg.

In addition, acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances that act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.

The pharmaceutical compositions disclosed herein may be subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form may be a packaged preparation, for example a packaged tablet, capsule, and paper or a powder in a plastic container or vial or ampoule. In addition, the unit dosage form can be a liquid-based formulation, or formulated in combination with a solid food product such as a chewing gum, or a lozenge.

In addition to the common dosage forms described above, the compositions of the present invention may also be used with controlled release means and/or delivery devices, such as those described in U.S. Pat. Nos. 3,845,770, 3,916,899, 3,536,809, 3,598,123, and 4,008,719, the entire disclosures of which are incorporated herein by reference.

Any suitable route of administration may be employed to provide an effective dose of a structural and/or functional analog of cisapride to the patient. For example, oral, rectal, parenteral (subcutaneous, intramuscular, intravenous), transdermal and similar forms of administration may be employed. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, patches and the like.

In one aspect, the present invention provides a method for treating gastroesophageal reflux disease in a mammal with a substantial reduction in the side effects associated with the administration of cisapride, which method 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 acceptable salt thereof. Preferably, the present invention provides a method of treating gastroesophageal reflux disease in a human.

In another aspect, the present invention provides a composition for treating a human suffering from gastroesophageal reflux disease comprising a therapeutically effective amount of a structural and/or functional analog of cisapride, or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present invention provides a method for effecting an antiemetic effect in a mammal while substantially reducing the side effects associated with the administration of cisapride, which comprises administering to a mammal in need of such treatment a therapeutically effective amount of a structural and/or functional analog of cisapride, or a pharmaceutically acceptable salt thereof. Preferably, the mammal is a human.

In another aspect, the present invention is directed to an antiemetic composition for treating a mammal in need of antiemetic therapy comprising a therapeutically effective amount of a structural and/or functional analog of cisapride, or a pharmaceutically acceptable salt thereof.

In a further aspect, the present invention comprises a method for treating a condition resulting from gastric motility dysfunction, which comprises administering to a mammal in need of treatment for gastric motility dysfunction a therapeutically effective amount of a structural and/or functional analog of cisapride, or a pharmaceutically acceptable salt thereof. Diseases caused by gastric motility dysfunction include, but are not limited to, dyspepsia, gastroparesis, constipation, post-operative ileus, and intestinal pseudo-obstruction. Preferably, the mammal is a human.

Cisapride was observed to enter the central nervous system and interact with 5HT₄Receptor binding, suggesting that cisapride may have a centrally mediated effect. Cisapride is 5HT₄Potent ligands for receptors, which are located in several regions of the central nervous system. Modulating the serotonin system has a variety of behavioral effects. Thus, the compounds of the invention are useful in the treatment of: 1) cognitive disorders including, but not limited toAlzheimer's disease; 2) behavioral disorders including, but not limited to, schizophrenia, mania, obsessive-compulsive disorder, and psychoactive substance use disorders; 3) mood disorders including, but not limited to, depression and anxiety; 4) autonomic dysfunction control disorders including, but not limited to, essential hypertension and sleep disorders.

Accordingly, the present invention also provides a method for the treatment of cognitive disorders, behavioral disorders, mood disorders or autonomic dysfunction control disorders in a mammal which comprises administering a therapeutically effective amount of a structural and/or functional analog of cisapride, or a pharmaceutically acceptable salt thereof. Preferably, the mammal is a human.

ATI-7505 and 5-HT₄High affinity binding of receptors

Known 5-HT₄The receptors are the major receptor subtype in the gut associated with cisapride prokinetic activity. ATI-7505 binds to 5-HT₄High binding capacity of the receptor, nanomolar IC₅₀Low. ATI-7505 and 5-HT as shown in Table 1₄The receptor has an 18-fold higher affinity than cisapride and at least 360-fold higher affinity than the major metabolite of ATI-7505, ATI-7500.

Table 1.

5-HT₄Receptor binding

n_HCoefficient of Hill

5-HT₄A standard control antagonist of receptor³H]GR113808(0.70nM)

ATI-7505 is human 5-HT₄High potency partial agonists of receptors

ARYx based cells engineered to stably express human 5-HT₄Receptor) of adenylyl cyclase stimulation. ATI-7505 was shown to be highly effective5-HT₄The receptor agonist, while its major metabolite ATI-7500 is relatively weak (fig. 1 and table 2). Estimated EC of ATI-7505(4nM)₅₀About 10 times lower than cisapride (49nM) and about 100 times lower than ATI-7500(395 nM). Estimate E based thereon_maxThe value ATI-7505 had a potency of 85% of 5-HT (serotonin) (Table 2), which indicates that ATI-7505 is an HT₄Partial agonists of the receptor.

Table 2.

Human 5-HT₄Receptor potency and potency (intrinsic activity)

EC₅₀Concentrations which result in a maximum 50% increase in adenylyl cyclase activity

_pEC50，EC₅₀Negative logarithm of

ATI-7505 promotes gastric emptying in full-bodied dogs

To characterize the effect of ATI-7505 on gastric emptying, a postprandial model experiment was performed on conscious dogs having metered sensors in both the stomach and small intestine. The purpose of this experiment was to measure the time required for transitional motor contractions (MMC) to return to baseline levels after digestion of solid food. The drug-induced reduction in MMC time indicates an early end of digestion time due to accelerated gastric emptying. Once MMC in the small and medium intestine was completed, intravenous infusion (vehicle, ATI-7505 or cisapride) was immediately carried out with different doses of experimental drug over 20 minutes. At the end of the drug infusion, the dogs were allowed to eat. Recording of gut contraction is started at least 60 minutes before starting drug infusion to establish a fasted state and to determine the onset of MMC in the duodenum, recording being performed at least 30 minutes after duodenal MMC recovery. Quantitative comparison of experimental results was performed based on MMC recovery time as index of gastric emptying after digestion of solid food. As summarized in fig. 2, ATI-7505 significantly reduced MMC recovery time, indicating acceleration of gastric emptying in normal satiety dogs. The mode of action of cisapride is similar.

ATI-7505 increased gastric and small intestinal motility with negligible effect on colonic activity

The kinetic activity of ATI-7505 relative to cisapride on the stomach, small intestine and colon was evaluated in fasted, conscious dogs. A particular objective was to determine the dose size of ATI-7505(IV and OP) that most closely mimics the pattern and size of contractile activity caused by typical therapeutic doses (0.5mg/kg IV; 1mg/kg PO) of cisapride in dogs.

Both ATI-7505 and cisapride cause prokinetic activity in the dog gut when administered IV and PO. Following IV and PO administration, action typically begins within 1-2 minutes and 25-30 minutes, respectively. The effect of ATI-7505 on gastric and small intestinal motility was similar to cisapride. Like cisapride, ATI-7505 appears to result in dose-dependent stimulation of antrum and small intestine contractility with relatively low effect on colonic motility. The resulting motile force effects of ATI-7505 in the upper GI tract are accompanied by a slight but significant (p < 0.05) increase in the frequency of massive transitional contractions (GMC).

ATI-7505 was not associated with the occurrence of retrograde massive transitional contractions (RGCs). Similar to cisapride, ATI-7505 had minimal effect on the transitional motor contraction (MMC) characteristics in the antrum and near center, center and terminal small intestine. For MMC frequency and third phase duration, only one significant difference was noted: PO ATI-7505 increased the frequency of MMC in the proximal small intestine compared to control. Dogs tolerated ATI-7505 doses of IV and PO well and showed no side effects such as diarrhea, anorexia or weight loss.

Overall, the results show that ATI-7505 is approximately twice as potent as cisapride on a mg/kg scale. In addition, the action of ATI-7505 is similar to cisapride, and is consistent with mechanisms associated with promoting acetylcholine release from enteric neurons, rather than directly soothing muscle movements. In summary, ATI-7505 increased gastric and small intestinal motility in a manner similar to cisapride with minimal to no effect on colonic activity.

ATI-7505 metabolism is independent of CYP450

Based on the stored (pooled) human microsomal data, ATI-7505 was biotransformed into the single metabolite ATI-7500 which was not metabolized further. The conversion of ATI-7505 to ATI-7500 is not NADPH-dependent. The main bioconversion pathway of ATI-7505 therefore proceeds independently of CYP450 enzymes.

ATI-7505 does not inhibit CYP450 enzyme

To test the potential of ATI-7505 and/or its major metabolite ATI-7500 as CYP450 inhibitors, Gentest Supersomes were used^TMBoth molecules were screened.

Consistent with the published reported results, cisapride has significant inhibitory activity against CYP450 enzyme isomers CYP3A4 and 2D6, and has a low degree of inhibition effect on 2C 9. ATI-7505 and its major metabolite ATI-7500 did not show significant inhibition against all three CYP450 isomers, nor against the other group of isomers (panel) known to play a role in the metabolism mechanism of drugs.

ATI-7505 and cardiac channel I_KrHas negligible affinity

Rapid activation delayed rectifier potassium (K) in humans⁺) Current (Rapid activating delayed rectifier current) (human I)_Kr) Is K encoded by human-ether-a-go-go related gene (hERG)⁺A channel. Cisapride is known to pass through obstruction I_KrCauses prolongation of the QT interval, and it is therefore determined whether ATI-7505 and ATI-7500 are in human I_KrHas important inhibition effect and is very significant. The test system is expression of hERG K⁺Mammalian HEK-293 cells of the channel, in which potassium current was measured by the whole-cell patch clamp technique. IC (integrated circuit)₅₀The order of the values is: cisapride (9.5nM) > ATI-7505(24,521nM) > ATI-7500(204,080nM) (Table 3). In general, the knotThe results showed that ATI-7505 had a significantly lower probability of causing arrhythmia than cisapride, and that ATI-7505 and ATI-750 were comparable to human I_KrThe affinity of the channel is negligible.

Table 3.

I_KrInhibition of Activity

Data against% control tail I_Kr(current caused by absence of drug or carrier) was normalized

ATI-7505 did not cause important electrophysiological changes in the guinea pig heart

The cardiac electrophysiological effects of ATI-7505 were tested in isolated perfused guinea pig hearts. This study examined ATI-7505, ATI-7500 and cisapride, all tested at concentrations up to 10,000 nM. The no-effect level (NOEL) was defined as the highest concentration at which the test compound did not exhibit a response significantly different from baseline (p < 0.05). The following 6 cardiac parameters were tested: (1) the QT interval; (2) MAPD₉₀(ii) a (3) An SA interval; (4) a QRS interval; (5) an AH interval; and (6) HV. Although ATI-7505 is a weak regulator of the electrophysiological parameters of the heart, its metabolite ATI-7500 is completely devoid of electrophysiological activity (Table 4). The NOEL of ATI-7500 was greater than 10,000nM for all 6 cardiovascular parameters. Since the combined NOEL of the 6 measured cardiac parameters for cisapride was 10nM and that of ATI-7505 was 1,000nM, ATI-7505 appeared to have no ability of cisapride to modulate cardiac electrophysiological parameters. Overall, the results show that ATI-7505 is significantly safer than cisapride in terms of potential for causing significant cardiac electrophysiology fluctuations.

Table 4.

Isolation of electrophysiological parameters of perfused guinea pig Heart

All molecules were assayed at 10, 100, 1,000 and 10,000nM baseline.

With the exception of the values reported as > 10,000nM, significant differences from baseline (p < 0.05) were observed when the molecules were measured more than 10-fold.

Metabolism in human microparticulate formulations

The metabolism of these compounds in the mixed human microsomes was studied in the presence and absence of the cytochrome P-450 coenzyme NADPH, and the disappearance of the parent and the appearance of the corresponding acid metabolite (i.e. the corresponding isomer of compound II) were monitored in real time.

As shown in FIG. 5, compounds III and IV are rapidly hydrolyzed by lipase to their respective metabolites (+) and (-) -compound II. Since the rate of hydrolysis is independent of the presence of NADPH, an essential cofactor for CYP450 action, metabolism is not CYP450 dependent. In contrast, (±) -S compounds V and VI appear to be fairly stable over time under the same conditions. In this experiment, the amount of substrate (compounds III, IV, V and VI) remaining from the reaction after 5, 60 and 90 minutes was evaluated by a two-stage HPLC-MS method. This residual amount is related to the appearance of metabolite compound II. The total amount of remaining substrate and compound II was constant over time and equal to the amount of starting material at time zero, thus indicating that hydrolysis is the only metabolic reaction that occurs.

Metabolism in fresh human blood

Test compounds were dissolved in DMSO to make a 12.5mM stock solution, diluted with water to a final concentration of2.5mM(DMSO/H₂O-20/80). Fresh blood was collected from 3 donors into heparinized tubes and the blood was stored in ice until incubation. Aliquots of blood from each donor were pipetted into 1.5mL centrifuge tubes and the tubes were preincubated in 37 ℃ shaking water bath for 5 minutes. To each tube, 10. mu.L of the appropriate test compound stock was added to initiate the reaction (final concentration of 100. mu.M). 0. After 5, 15, 30 and 60 minutes acetonitrile (750mL) was added, the incubation was stopped, centrifuged at 12,000rpm for 2 minutes and the supernatant was taken for analysis on an Agilent 1100 HPLC system. The separation was carried out on a Keystone Intersil ODS2, 250X 4.6mm, 5m column. The aqueous mobile phase consisted of 20mM ammonium acetate buffer (pH 5.7) and the organic phase acetonitrile. The following gradient was used: initially eluted with 20% acetonitrile for 1 min. The acetonitrile concentration increased linearly to 90% over the next 8 minutes, which was held for 1 minute. The system was then returned to the initial condition for 1 minute and held for 4 minutes for the next injection. The absorption at 240, 254 and 290nM was measured and the peak area of the mother peak was determined. Results are expressed as residual amounts of starting compounds and data were subjected to kinetic analysis using WinNonLin. The half-lives of the compounds are shown in table 6.

TABLE 6

It is understood that the embodiments and aspects described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. Further, all patents, patent applications, provisional applications, and publications cited herein are hereby incorporated by reference in their entirety to the extent that they are not inconsistent with the explicit teachings of this specification.

The present invention and the manner of practicing and using it, are described herein in such full, clear, concise and exact terms as to enable any person skilled in the relevant art to which it pertains to practice and use the same. It should be understood that modifications to the preferred aspects of the invention may be made without departing from the spirit and scope of the invention as set forth in the claims. To particularly point out and distinctly claim the subject matter, the following claims conclude this specification.

Claims

1. A compound selected from the group consisting of

(3R, 4S, 3' R) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid 1-aza-bicyclo [2.2.2] oct-3-yl ester.

2. A compound selected from the group consisting of

(3S, 4R, 3' R) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] hexanoic acid 1-aza-bicyclo [2.2.2] oct-3-yl ester.

3. A compound selected from the group consisting of

(3R, 4S, 3' S) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid 1-aza-bicyclo [2.2.2] oct-3-yl ester.

4. A compound selected from the group consisting of

(3S, 4R, 3' S) -6- [4- (4-amino-5-chloro-2-methoxy-benzoylamino) -3-methoxy-piperidin-1-yl ] -hexanoic acid 1-aza-bicyclo [2.2.2] oct-3-yl ester.

5. The compound according to claim 2, wherein the compound is in the form of the dihydrochloride salt, and pharmaceutically acceptable salts thereof.

6. A composition comprising a compound of claim 1.

7. A composition comprising a compound of claim 2.

8. A composition comprising a compound of claim 3.

9. A composition comprising a compound of claim 4.

10. A composition comprising a compound of claim 5.

11. A composition comprising a compound of claim 1 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

12. A composition comprising a compound according to claim 2 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

13. A composition comprising a compound according to claim 3 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

14. A composition comprising a compound according to claim 4 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

15. A composition comprising a compound according to claim 5 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

16. A composition comprising the composition of claim 6 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

17. A composition comprising the composition of claim 7 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

18. A composition comprising the composition of claim 8 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

19. A composition comprising the composition of claim 9 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.

20. A composition comprising the composition of claim 10 and a pharmaceutically acceptable excipient, adjuvant, carrier or solvent.