Field of the invention

5 The present invention relates to the use of dual NK ₁ ZNK ₂ receptor antagonists for the treatment of functional gastrointestinal disorders, such as functional dyspepsia.

Background of the invention

I Q

Functional gastrointestinal disorders, such as functional dyspepsia, can be defined in accordance with Thompson WG, Longstreth GF, Drossman DA, Heaton KW, Irvine EJ, Mueller-Lissner SA. C. Functional Bowel Disorders and Functional Abdominal Pain. In: Drossman DA, Talley NJ, Thompson WG, Whitehead WE, Coraziarri E, eds. Rome II: 15 Functional Gastrointestinal Disorders: Diagnosis, Pathophysiology and Treatment. 2 ed. McLean, VA: Degnon Associates, Inc.; 2000:351-432 and Drossman DA, Corazziari E, Talley NJ, Thompson WG and Whitehead WE. Rome II: A multinational consensus document on Functional Gastrointestinal Disorders. Gut 45(Suppl.2), II1-II81.9-1-1999.

2Q The neurokinins, also known as the tachykinins, comprise a class of peptide neurotransmitters which are found in the peripheral and central nervous systems. The three principal tachykinins are Substance P (SP), Neurokinin A (NKA) and Neurokinin B (NKB). At least three receptor types are known for the three principal tachykinins. Based upon their relative selectivities favouring the agonists SP, NKA and NKB, the receptors 5 are classified as neurokinin 1 (NK ₁ ), neurokinin 2 (NK ₂ ) and neurokinin 3 (NK ₃ ) receptors, respectively.

Functional gastrointestinal (GI) disorders such as irritable bowel syndrome (IBS) and functional dyspepsia are characterized by increased visceral sensations manifested as 30 abdominal discomfort or even pain. The tachykinins, which bind to neurokinin (NK) receptors, are present in neurons that transmit signals from the GI tract to the brain.

The object of the present invention was to find a new way for the treatment of functional gastrointestinal disorders, such as functional dyspepsia.

Outline of the invention

The present invention is directed to the use of dual NKi/NK ₂ receptor antagonists for the manufacture of a medicament for the treatment of functional gastrointestinal disorders.

One aspect of the present invention is the use of dual NKi/NK ₂ receptor antagonists for the manufacture of a medicament for the treatment of functional dyspepsia.

Still a further aspect of the present invention is directed to the use of a compound of formula I

for the manufacture of a medicament for treatment of functional gastrointestinal disorders, wherein

Het is an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring having at least one nitrogen atom;

Rl is hydrogen, hydroxy, Ci-C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl or C ₂ -C ₄ alkynyl;

R2 and R3 is each and independently selected from hydrogen, Ci-C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, Ci-C ₄ alkoxy, halogen and cyano, provided that R2 and R3 may not both be hydrogen;

R4 is Ci -C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl or C ₂ -C ₄ alkynyl;

Ar is an optionally substituted aromatic ring system selected from pyridinyl; 1- naphthyl; 5,6,7,8-tetrahydro-l-naphthyl; quinolinyl; 2,3-dihydro-l,4-benzodioxinyl; 1,3-benzodioxolyl; 5,6,7,8-tetrahydroquinolinyl; 5,6,7,8-tetrahydroisoquinolinyl;

5 ,6,7, 8-tetrahydroquinazolin-4-yl; 1 -benzo [b] thiophen-7-yl; 1 -benzo [b]thiophen-4-yl; l-benzo[b]thiophen-3-yl; isoquinolinyl; quinazolinyl; and indan-4-yl; or Ar is substituted phenyl;

or an enantiomer thereof or any salt thereof.

In a still further aspect of the invention, the functional gastrointestinal disorder is functional dyspepsia.

In one embodiment of the present invention, the heterocyclic ring Het is connected to the rest of the molecule at one of the nitrogen atoms of the ring. Examples of such heterocyclic rings are optionally substituted l,4-dioxa-8-azaspiro[4.5]decano; optionally substituted piperidino; optionally substituted azepano; optionally substituted pyrrolidino; optionally substituted morpholino; optionally substituted oxazepano; optionally substituted thiomorpholino; optionally substituted thiazepano; and optionally substituted piperazino.

In further embodiments of the present invention Het is piperidino optionally substituted with hydroxy, hydroxyalkyl, oxo, methylthio, methylsulfinyl, methylsulfonyl, cyano, 1,3- dioxolan-2-yl, C ₁ -C ₄ alkoxy, amino optionally mono or disubstituted with C ₁ -C ₄ alkyl, C ₃ - C ₄ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, acylamino optionally N-substituted with Ci- C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, (Ci-C ₄ alkylsulfonyl)amino optionally N-substituted by Ci-C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl,

one or two fluoro atoms or disubstituted by Ci-C ₄ alkyl and hydroxy; pyrrolidino optionally substituted at its three position by fluoro, hydroxy or oxo; morpholino or thiomorpholino optionally substituted at its sulfur atom by one or two oxygen; or piperazino optionally substituted at the 4-nitrogen atom by C ₁ -C ₄ alkyl.

Ar may optionally be substituted at one or more of its carbon atoms by one or more groups independently selected from cyano, halogen, C ₁ -C ₄ alkyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, C ₁ -C ₄ alkoxy, nitro, trifluoromethoxy, difluoromethoxy, trifluoromethyl, Ci-C ₄ alkylsulfinyl, C]-C ₄ alkylsulfonyl, Ci-C ₄ alkylthio, trifluoromethylsulfonyloxy, Ci- C ₄ alkyl sulfonyl or Ci-C ₄ acyl.

In one embodiment of the present invention, Ar is phenyl substituted by one or more groups independently selected from cyano, halogen, Ci-C ₄ allcyl, C ₃ -C ₄ cycloalkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, Ci-C ₄ alkoxy, nitro, trifluoromethoxy, difluoromethoxy, Ci-C ₄ alkylsulfinyl, Ci-C ₄ alkylsulfonyl, Ci-C ₄ alkylthio, trifluoromethylsulfonyloxy, Cj-C ₄ alkyl sulfonyl or Ci-C ₄ acyl.

In a further embodiment of the present invention, Ar is phenyl substituted in its 3- and 5- position by groups independently selected from halogen, Ci-C ₄ alkyl, Ci-C ₄ alkoxy, cyano and nitro. Optionally, Ar may additionally also be substituted in its 2- and/or 4-position by a group independently selected from halogen, Ci-C ₄ alkyl and Ci-C ₄ alkoxy.

In one embodiment of the present invention, Rl is hydrogen.

In one embodiment of the present invention, R2 and R3 are both chloro or one is fluoro and the other is hydrogen. In a further embodiment of the invention, R2 and R3 are both chloro and attached in the three and four position of the phenyl ring or R2 is fluoro attached in the four position and R3 is hydrogen.

In one embodiment of the present invention, R4 is methyl.

In one embodiment of the present invention, the compound of formula I is the S- enantiomer or the racemate. In a further embodiment of the invention, the compound of formula I is the S-enantiomer.

In one embodiment of the present invention, the compound of formula I is the R- enantiomer or the racemate. In a further embodiment of the invention, the compound of formula I is the R-enantiomer.

A further aspect of the invention relates to compounds of formula I, wherein Het is tbiomorpholino, morpholino or oxidothiomorpholino; Rl is H;

R2 is fluoro and R3 is hydrogen, fluoro being preferably in para position; Ar is 3-cyano-5,6,7,8-tetrahydro-l-naphthyl; and R4 is as defined above.

A further aspect of the invention is the use of any of the following compounds for the treatment of functional gastrointestinal disorders, such as functional dyspepsia:

3,5-Dichloro-N-[(2 ₁ S)-2-(3,4-dichlorophenyl)-4-(3-thiomorpholin-4-ylazeti din-l-yl)butyl]- N-methylbenzamide;

3,5-Dibromo-N-[(25)-2-(4-fluorophenyl)-4-(3-morpholin-4-y lazetidin-l-yl)butyl]-N- methylbenzamide ;

N-[(2S)-2-(3,4-Dichlorophenyl)-4-(3-thiomorpholin-4-ylaze tidin-l-yl)butyl]-3 ₅ 5-difluoro- N-methylbenzamide;

N-[(25)-2-(3,4-Dichlorophenyl)-4-(3-thiomorpholin-4-ylaze tidrn-l-yl)butyl]-N-methyl-3,5- bis(trifluoromethyl)benzamide;

5-Cyano-N-[(25)-2-(3,4-dichlorophenyl)-4-(3-thiomoφholin-4- ylazetidin-l-yl)butyl]-N- methyl- 1 -benzothiophene-7-carboxamide;

3 -Cyano-N-[(2S)-2-(3 ,4-dichlorophenyl)-4-(3 -thiomorpholin-4-ylazetidin- 1 -yl)butyl]-N- methylbenzamide;

3 -Cyano-N- [(25)-2-(3 ,4-dichloropheny l)-4-(3 -thiomorpholin-4-ylazetidin- 1 -yl)butyl] -N- methyl-5,6,7,8-tetrahydronaphthalene- 1 -carboxamide;

2-Cyano-N-[(25)-2-(3,4-dichlorophenyl)-4-(3-thiomoφholin -4-ylazetidin-l-yl)butyl]-N- methylquinoline-4-carboxamide;

3-Cyano-N-[2-(4-fluorophenyl)-4-(3-thiomorpholin-4-ylazet idin-l-yl)butyl]-N-methyl- 5,6,7,8-tetrahydronaphthalene-l-carboxamide;

N-[2-(4-Fluorophenyl)-4-(3-thiomoφholin-4-ylazetidin-l-y l)butyl]-N-methyl-3,5- bis(trifluoromethyl)benzamide;

7-Chloro-N-[(25)-2-(3,4-dichlorophenyl)-4-(3-thiomoφholi n-4-ylazetidin-l-yl)butyl]-N- methyl-2,3-dihydro-l,4-benzodioxine-5-carboxamide;

N- {(25)-2-(3 ,4-Dichlorophenyl)-4-[3 -( 1 -oxidothiomoφholin-4-yl)azetidin- 1 -yl]butyl} -2- methoxy-N-methylquinoline-4-carboxamide;

3-Fluoro-N-[(2.S)-2-(4-fluorophenyl)-4-(3-moφholin-4-yla zetidin-l-yl)butyl]-N-methyl-5- (trifluoromethyl)benzamide;

3 -Cyano-N- {2-(4-fluorophenyl)-4-[3 -(4-hydroxypiperidin- 1 -yl)azetidin- 1 -yljbutyl} -N- methyl-5,6,7,8-tetrahydronaphthalene- 1 -carboxamide;

N-[4-[3-(l,4-Dioxa-8-azasρiro[4.5]dec-8-yl)azetidin-l-yl]-2 -(4-fluorophenyl)butyl]-iV- methyl-3,5-bis(trifluoromethyl)benzamide;

N-{(26)-2-(3,4-Dichlorophenyl)-4-[3-(4-fluoropiperidin-l- yl)azetidin-l-yl]butyl}-N- methyl-3,5-bis(trifluoromethyl)benzamide;

iV-{(25)-2-(3,4-Dichlorophenyl)-4-[3-(4-hydroxypiperidin- l-yl)azetidin-l-yl]butyl}-N- methyl-3,5-bis(trifluoromethyl)benzamide;

3 -Cyano-N- {(2S)-2-(3,4-dichlorophenyl)-4-[3-(3-hydroxypyrrolidin- 1 -yl)azetidin- 1 - yljbutyl} -iV-methyl- 1 -naphthamide;

N-{(25 ^f )-2-(4-Fluorophenyl)-4-[3-(4-fluoropiperidin-l-yl)azet idin-l-yl]butyl}-N-methyl- 3 ,5-bis(trifluoromethyl)benzamide;

3-Cyano-N-[(25)-2-(4-fluorophenyl)-4-(3-morpholin-4-ylaze tidin-l-yl)butyl]-N-methyl- 5,6,7,8-tetrahydronaphthalene-l-carboxamide;

N- {(25)-2-(4-fluorophenyl)-4-[3-(4-hydroxypiperidin- 1 -yl)azetidin- 1 -yljbutyl} -N-methyl- 3,5-bis(trifluoromethyl)benzamide;

3,5-Dichloro-N-[(25)-2-(4-fluorophenyl)-4-(3-morpholin-4- ylazetidin-l-yl)butyl]-N- methylbenzamide;

3-Cyano-N-[(2,S)-2-(3,4-dichlorophenyl)-4-(3-morpholin-4- ylazetidin-l-yl)butyl]-iV- methyl- 1 -naphthamide;.

3-Cyano-N-[(25)-2-(3,4-dichlorophenyl)-4-(3-thiomorpholin -4-ylazetidin-l-yl)butyl]-N- methyl- 1 -naphthamide;

3-Cyano-N-{(2S)-2-(3,4-dichlorophenyl)-4-[3-(l,4-dioxa-8-aza spiro[4.5]dec-8-yl)azetidin- 1 -yljbutyl} -N-methyl- 1 -naphthamide;

3 -Cyano-N- {(2-S)-2-(3,4-dichlorophenyl)-4-[3 -(4-hydroxypiperidin- 1 -yl)azetidin- 1 - 5 yljbutyl} -N-methyl- 1 -naphthamide;

3 -Cyano-N- [2-(4-fluorophenyl)-4-(3 -thiomorpholin-4-ylazetidin- 1 -yl)butyl] -N-methyl- 1 - naphthamide;

i o 3-Cyano-N-[2-(4-cyanophenyl)-4-(3 -thiomorpholin-4-ylazetidin- 1 -yl)butyl] -N-methyl- 1 - naphthamide;

3-Cyano-N-{(2 ₁ S)-2-(3,4-dichlorophenyl)-4-[3-(l,l-dioxidothiomorphol in-4-yl)azetidin-l- yl]butyl} -N-methyl- 1 -naphthamide;

15

3-Cyano-N- {(2-S)-2-(4-fluorophenyl)-4-[3-(4-hydroxypiperidm- 1 -yl)azetidin- 1 -yljbutyl} - N-methyl-5,6,7,8-tetrahydronaphthalene-l-carboxamide;

3-Cyano-N-ethyl-N-[(25)-2-(4-fluorophenyl)-4-(3-morpholm- 4-ylazetidin-l-yl)butyl]- 20 5,6,7,8-tetrahydronaphthalene-l-carboxamide;

3-Cyano-N-[(25)-4-[3-(l,4-dioxa-8-azaspiro[4.5]dec-8-yl)a zetidin-l-yl]-2-(4- fluorophenyl)butyl]-N-methyl-5,6,7,8-tetrahydronaphthalene-l -carboxamide;

25 3-cyano-N-[(2.S)-2-(4-fluorophenyl)-4-(3-morpholin-4-ylazeti din-l-yl)butyl]-N-methyl-l- naphthamide;

3-Cyano-N-{(25)-2-(4-fluorophenyl)-4-[3-(l,4-oxazepan-4-y l)azetidin-l-yl]butyl}-N- methyl-5,6,7,8-tetrahydronaphthalene-l-carboxamide;

30

3-Fluoro-N-[(2,S)-2-(4-fluorophenyl)-4-(3-morpholin-4-ylazet id.in-l-yl)butyl]-N-methyl- 5,6,7,8-tetraliydronaphthalene-l-carboxamide;

3,5-Dibromo-N-{(25)-2-(4-fluorophenyl)-4-[3-(4-hydroxypip eridin-l-yl)azetidin-l- yl]butyl}-N-methylbenzatnide;

3-Bromo-N-[(2 ₁ S)-2-(4-fluorophenyl)-4-(3-morpholin-4-ylazetidin-l-yl )butyl]-5-iodo-N- methylbenzamide;

3-Cyano-N-[2-(4-fluoro-2-methylphenyl)-4-(3-morpliolin-4- ylazetidin-l-yl)butyl]-N- methyl-5,6,7,8-tetrahydronaphthalene-l-carboxamide;

6-Cyano-N-[(2 ₁ S)-2-(4-fluorophenyl)-4-(3-moφholin-4-ylazetidin-l-yl )butyl]-N- methylindane-4-carboxamide;

3-Cyano-N-{(2S)-2-(3,4-dichlorophenyl)-4-[3-(l-oxidothiom orpholin-4-yl)azetidin-l- yljbutyl} -N-methyl- 1 -naphthamide;

3 -Cyano-N- {2-(4-cyanophenyl)-4-[3-( 1 -oxidothiomorpholin-4-yl)azetidin- 1 -yljbutyl} -N- methyl- 1 -naphthamide;

3,5-Dichloro-N-{(25)-2-(3,4-dichlorophenyl)-4-[3-(l-oxido thiomoφholin-4-yl)azetidin-l- yl]butyl} -N-methylbenzamide;

N- [(2S)-2-(3 ,4-Dichlorophenyl)-4-(3 -oxidothiomorpholin-4-ylazetidin- 1 -yl)butyl] -N- methyl-3,5-bis(trifluoromethyl)benzamide;

3-Cyano-N- {(25)-2-(3 ,4-dichlorophenyl)-4-[3-( 1 -oxidothiomoφholin-4-yl)azetidin- 1 - yl]butyl} -N-methyl-5 ,6,7,8-tetrahydronaphthalene- 1 -carboxamide;

3-Cyano-N-{2-(4-fluorophenyl)-4-[3-(l-oxidothiomorpholin-4-y l)azetidin-l-yl]butyl}-N- methyl- 1 -naphthamide;

3-cyano-N-{2-(4-fluorophenyl)-4-[3-(l-oxidothiomorpholin- 4-yl)azetidin-l-yl]butyl}-N- methyl-5,6,7,8-tetrahydronaphthalene- 1 -carboxamide;

N-{2-(4-Fluorophenyl)-4-[3-(l-oxidothiomorpholin-4-yl)aze tidin-l-yl]butyl}-N-methyl- 3 , 5 -bis(trifluoromethyl)benzamide ;

3-Cyano-Ν-{(2S)-2-(3,4-dichlorophenyl)-4-[3-(4-oxopiperi din-l-yl)azetidin-l-yl]butyl}- N-methy 1- 1 -naphthamide ;

N- {2-(4-fluorophenyl)-4-[3-(4-oxopiperidin- 1 -yl)azetidin- l-yl]butyl} -N-methyl-3 ,5- bis(trifluoromethyl)benzamide;

3-Cyano-Ν-{(2S)-2-(3,4-dichlorophenyl)-4-[3-(4-fluoropip eridin-l-yl)azetidin-l- yl]butyl} -N-methyl- 1 -naphthamide;

3 -Cyano-N- {2-(4-fluorophenyl)-4- [3 -(4-fluoropiperidin- 1 -yl)azetidin- 1 -yljbutyl} -N- methyl-5,6,7,8-tetrahydronaphthalene-l-carboxamide;

3-Cyano-N-{(2>S)-2-(3,4-dichlorophenyl)-4-[3-(4-methyl piperazin-l-yl)azetidin-l- yl]butyl} -N-methyl- 1 -naphthamide;

N-[(2iS)-4-[3-(4-Acetylpiperazin-l-yl)azetidin-l-yl]-2-(3 ,4-dichlorophenyl)butyl]-3-cyano- N-methyl- 1 -naphthamide;

3-Cyano-N-[(25)-4-[3-(4-cyanopiperidin-l-yl)azetidin-l-yl ]-2-(3,4-dichlorophenyl)butyl]- N-methyl- 1 -naphthamide;

or an enantiomer thereof or any salt thereof.

The compounds useful in accordance with the present invention are capable of forming salts with various inorganic and organic acids and such salts are also within the scope of this invention. Examples of such acid addition salts include acetate, adipate, ascorbate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, citrate, cyclohexyl sulfamate, ethanesulfonate, fumarate, glutamate, glycolate, hemisulfate, 2- hydroxyethylsulfonate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, hydroxymaleate, lactate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nitrate, oxalate, palmoate, persulfate, phenylacetate, phosphate, picrate, pivalate, propionate, quinate, salicylate, stearate, succinate, sulfamate, sulfanilate, sulfate, tartrate, tosylate (p-toluenesulfonate), and undecanoate. Non-toxic physiologically acceptable salts are preferred, although other salts are also useful, such as in isolating or purifying the product.

Pharmaceutically acceptable salts may be prepared from the corresponding acid in conventional manner. Non-pharmaceutically-acceptable salts may be useful as intermediates and as such are another aspect of the present invention.

Acid addition salts may also be in the form of polymeric salts such as polymeric sulfonates.

The salts may be formed by conventional means, such as by reacting the free base form of the product with one or more equivalents of the appropriate acid in a solvent or medium in which the salt is insoluble, or in a solvent such as water, which is removed in vacuo or by freeze drying or by exchanging the anions of an existing salt for another anion on a suitable ion-exchange resin.

Compounds of formula I have one or more chiral centres, and it is to be understood that the invention encompasses all optical isomers, enantiomers and diastereomers. The compounds according to formula (I) can be in the form of the single stereoisomers, i.e. the

single enantiomer (the R-enantiomer or the S-enantiomer) and/or diastereomer. The compounds according to formula (I) can also be in the form of a racemic mixture, i.e. an equimolar mixture of enantiomers.

It is to be understood that the present invention also relates to the use of any and all tautomeric forms of the compounds of formula I.

Some compounds can exist as a mixture of conformational isomers. The compounds useful in accordance with this invention comprise both mixtures of, and individual, conformational isomers.

Consequently, the present invention is directed to the use of a dual NKi/NK ₂ receptor antagonist for the manufacture of a medicament for the treatment of functional gastrointestinal disorders, such as functional dyspepsia.

Functional dyspepsia refers to pain or discomfort centered in the upper abdomen. Discomfort may be characterized by or combined with upper abdominal fullness, early satiety, bloating or nausea. Etiologically, patients with functional dyspepsia can be divided into two groups: 1- Those with an identifiable pathophysiological or microbiologic abnormality of uncertain clinical relevance (e.g. Helicobacter pylori gastritis, histological duodenitis, gallstones, visceral hypersensitivity, gastroduodenal dysmotility) 2- Patients with no identifiable explanation for the symptoms.

Functional dyspepsia can be diagnosed according to the following:

At least 12 weeks, which need not be consecutive within the preceding 12 months of

1- Persistent or recurrent dyspepsia (pain or discomfort centered in the upper abdomen) and

2- No evidence of organic disease (including at upper endoscopy) that is likely to explain the symptoms and

3- No evidence that dyspepsia is exclusively relieved by defecation or associated with the onset of a change in stool frequency or form.

Functional dyspepsia can be divided into subsets based on distinctive symptom patterns, such as ulcer-like dyspepsia, dysmotility-like dyspepsia and unspecified (non-specific) dyspepsia.

Currently existing therapy of functional dyspepsia is largely empirical and directed towards relief of prominent symptoms. The most commonly used therapies still include antidepressants.

Unless stated otherwise, the term "alkyl" includes straight as well as branched chain C\ .4 alkyl groups, for example methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl or t- butyl. One or more of the hydrogen atoms of the alkyl group may be substituted for a fluorine atom, such as in difluoromethyl or trifluoromethyl.

As used herein, C ₃ -C ₄ cycloalkyl is a cyclic alkyl such as cyclopropyl or cyclobutyl. The cycloalkyl may also be unsaturated. One or more of the hydrogen atoms of the cycloalkyl group may be substituted for a fluorine atom.

As used herein, C ₂ -C ₄ alkenyl is a straight or branched alkenyl group, for example vinyl. One or more of the hydrogen atoms of the alkenyl group may be substituted for a fluorine atom.

As used herein, C ₂ -C ₄ alkynyl is a straight or branched alkynyl group, for example ethynyl. One or more of the hydrogen atoms of the alkynyl group may be substituted for a fluorine atom.

As used herein, C ₁ -C ₄ hydroxyalkyl is a hydroxyalkyl group comprising 1-4 carbon atoms and a hydroxyl group. One or more of the hydrogen atoms of the hydroxyalkyl group may be substituted for a fluorine atom.

5 The term "alkoxy" as used herein, unless stated otherwise includes C ₁ -C ₄ alkoxy groups, for example methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, s-butoxy or t- butoxy. One or more of the hydrogen atoms of the alkoxy group may be substituted for a fluorine atom.

o The term "alkylthio" as used herein, unless stated otherwise includes C ₁ -C ₄ alkylthio groups, for example methylthio, ethylthio, n-propylthio, i-propylthio, n-butylthio, i- butylthio, s-butylthio or t-butylthio. One or more of the hydrogen atoms of the alkylthio group may be substituted for a fluorine atom.

s In this specification, unless stated otherwise, the term "halogen" includes chloro, bromo, fiuoro and iodo.

In this specification, unless stated otherwise, the term "alkyl sulfonyl" includes C ₁ -C ₄ alkyl sulfonyl groups, for example methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, i- o propylsulfonyl, n-butylsulfonyl, i-butylsulfonyl, s-butylsulfonyl or t-butylsulfonyl.

In this specification, unless stated otherwise, the term "alkylsulfinyl" includes Ci-C ₄ alkyl sulfinyl groups, for example methylsulfmyl, ethylsulfmyl, n-propylsulfmyl, i- propylsulfinyl, n-butylsulfinyl, i-butylsulfinyl, s-butylsulfinyl or t-butylsulfinyl. 5

In this specification, unless stated otherwise, the term "acyl" includes C ₁ -C ₄ acyl groups, for example formyl, acetyl, propionyl, butyryl and isobutyryl.

As used herein, the term "free base" means the compound in its neutral form, i.e. when the o compound is not present as a salt.

For the purpose of this invention, the term "antagonist" should be understood as including full antagonists, inverse agonists, non-competitive antagonists or competitive antagonists, as well as partial antagonists, whereby a "partial antagonist" should be understood as a compound capable of partially, but not fully, in-activating the NK-I and NK-2 receptors.

For the purpose of this invention, the term "dual NK-I /NK-2 antagonist" should be understood as including a compound with antagonistic activity at the NKj receptor within the interval 7-9 for the pKβ and within the interval of 7-9 for the pKjg at the NK 2 receptor.

The present invention is directed to the use of any dual NKi/NK ₂ receptor antagonist that has a therapeutic effect in functional gastrointestinal disorders, such as functional dyspepsia.

The term "therapy" and/or "treatment" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.

The term "therapeutic effect" is defined herein as an effect favourable in the context of the therapy and/or treatment of functional gastrointestinal disorders, such as functional dyspepsia.

The compounds useful in accordance with the present invention can be synthesized as described in WO2004/110344 A2.

Pharmaceutical formulations

For clinical use, the dual NK ₁ ZNK ₂ receptor antagonists are in accordance with the present invention suitably formulated into pharmaceutical formulations for oral administration.

Also rectal, parenteral or any other route of administration may be contemplated to the skilled man in the art of formulations. Thus, the dual NKiMEC ₂ receptor antagonists are formulated with at least one pharmaceutically and pharmacologically acceptable carrier or adjuvant. The carrier may be in the form of a solid, semi-solid or liquid diluent.

In the preparation of oral pharmaceutical formulations in accordance with the invention, the dual NKi/NK ₂ receptor antagonist(s) to be formulated is mixed with solid, powdered ingredients such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose derivatives, gelatin, or another suitable ingredient, as well as with disintegrating agents and lubricating agents such as magnesium stearate, calcium stearate, sodium stearyl fumarate and polyethylene glycol waxes. The mixture is then processed into granules or compressed into tablets.

Soft gelatine capsules may be prepared with capsules containing a mixture of the active compound or compounds of the invention, vegetable oil, fat, or other suitable vehicle for soft gelatine capsules. Hard gelatine capsules may contain the active compound in combination with solid powdered ingredients such as lactose, saccharose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives or gelatine.

Dosage units for rectal administration may be prepared (i) in the form of suppositories which contain the active substance(s) mixed with a neutral fat base; (ii) in the form of a gelatine rectal capsule which contains the active substance in a mixture with a vegetable oil, paraffin oil, or other suitable vehicle for gelatine rectal capsules; (iii) in the form of a ready-made micro enema; or (iv) in the form of a dry micro enema formulation to be reconstituted in a suitable solvent just prior to administration.

Liquid preparations for oral administration may be prepared in the form of syrups or suspensions, e.g. solutions or suspensions, containing the active compound and the remainder of the formulation consisting of sugar or sugar alcohols, and a mixture of ethanol, water, glycerol, propylene glycol and polyethylene glycol. If desired, such liquid

preparations may contain colouring agents, flavouring agents, saccharine and carboxymethyl cellulose or other thickening agent. Liquid preparations for oral administration may also be prepared in the form of a dry powder to be reconstituted with a suitable solvent prior to use.

Solutions for parenteral administration may be prepared as a solution of a compound of the invention in a pharmaceutically acceptable solvent. These solutions may also contain stabilizing ingredients and/or buffering ingredients and are dispensed into unit doses in the form of ampoules or vials. Solutions for parenteral administration may also be prepared as a dry preparation to be reconstituted with a suitable solvent extemporaneously before use.

In one aspect of the present invention, the dual NK ₁ ZNK ₂ receptor antagonists may be administered once or twice daily, depending on the severity of the patient's condition.

Pharmacology

Transfection and culturing of cells used in FLIPR and Binding assays

Chinese Hamster Ovary (CHO) Kl cells (obtained from ATCC) are stably transfected with the human NK ₂ receptor (hNK ₂ R cDNA in pRc/CMV, Invitrogen). The cells are transfected with the cationic lipid reagent LIPOFECT AMINE™ (Invitrogen) and selection is performed with Geneticin (G418, Invitrogen) at lmg/ml for the hNK ₂ R transfected cells. Single cell clones are collected by aid of Fluorescence Activated Cell Sorter (FACS), tested for functionality in a FLIPR assay (see below), expanded in culture and cryopreserved for future use. CHO cells stably transfected with human NK ₁ receptors originates from AstraZeneca R&D, Wilmington USA. Human NK ₁ receptor cDNA (obtained from RNA-PCR from lung tissue) is subcloned into pRcCMV (Invitrogen). Transfection is performed by Calcium Phosphate and selection with lmg/ml G418.

The CHO cells stably transfected with hNKiR and hNK ₂ R are cultured in a humidified incubator under 5% CO ₂ , in Nut Mix F12 (HAM) with Glutamax 1, 10% Foetal Bovine

Serum (FBS), 1% Penicillin/Streptomycin (PEST) supplemented with 200μg/ml Geneticin for the hNKiR and hNK ₂ R expressing cells. The cells are grown in Tl 75 flasks and routinely passaged when 70-80% confluent for up to 20-25 passages.

Assessing the Activity of Selected test Compounds to Inhibit Human NK ₁ /NK ₂ Receptor Activation (FLIPR assay)

The activity of a compound useful in accordance with the present invention to inhibit NK]/NK ₂ receptor activation measured as NKi/NK ₂ receptor mediated increase in intracellular Ca ²⁺ is assessed by the following procedure:

CHO cells stably transfected with human NKi or NK ₂ receptors are plated in black walled/clear bottomed 96-wεll plates (Costar 3904) at 3.5xlO ⁴ cells per well and grown for approximately 24h in normal growth media in a 37°C CO ₂ -incubator. Before the FLIPR assay the cells of each 96-well plate are loaded with the Ca ²⁺ sensitive dye Fluo-3 (TEFLABS 0116) at 4μM in a loading media consisting of Nut Mix F12

(HAM) with Glutamax I, 22mM HEPES, 2.5mM Probenicid (Sigma P-8761) and 0.04% Pluronic F-127 (Sigma P-2443) for 1 h kept dark in a 37 ⁰ C CO ₂ -incubator. The cells are then washed three times in assay buffer (Hanks balanced salt solution (HBSS) containing 2OmM HEPES, 2.5mM Probenicid and 0.1% BSA) using a multi-channel pipette leaving them in 150μl at the end of the last wash. Serial dilutions of a test compound in assay buffer (final DMSO concentration kept below 1%) are automatically pipetted by FLIPR (Fluorometric Imaging Plate Reader) into each test well and the fluorescence intensity is recorded (excitation 488 nm and emission 530 nm) by the FLIPR CCD camera for a 2 min pre-incubation period. 50μl of the Substance P (NKi specific) or NKA (NK ₂ specific) agonist solution (final concentration equivalent to an approximate EC ₆₀ concentration) is then added by FLIPR into each well already containing 200μl assay buffer (containing the test compound or vehicle) and the fluorescence is continuously monitored for another 2 min. The response is measured as the peak relative fluorescence after agonist addition and IC ₅ os are calculated from ten-point concentration-response curves for each compound. The IC ₅ os are then converted to pKβ values with the following formula:

KB = IC ₅₀ / 1+ (EC ₆₀ cone, of agonist used in assay / EC50 agonist) pK _B = - log KB

Determining the Dissociation Constant (Ki) of compounds for Human NK ₁ /NK ₂ Receptors (Binding Assay)

Membranes are prepared from CHO cells stably transfected with human NK ₁ or NK ₂ receptors according to the following method.

Cells are detached with Accutase® solution, harvested in PBS containing 5% FBS by centrifugation, washed twice in PBS and resuspended to a concentration of 1x10 ⁸ cells/ml in Tris-HCl 50 mM, KCl 300 mM, EDTA-N ₂ 10 mM pH 7.4 (4°C). Cell suspensions are homogenized with an UltraTurrax 30 s 12.000 rpm. The homogenates are centrifuged at 38.000 x g (4 ⁰ C) and the pellet resuspended in Tris-HCl 50 mM pH 7.4. The homogenization is repeated once and the homogenates are incubated on ice for 45 min. The homogenates are again centrifuged as described above and resuspended in Tris-HCl 5OmM pH 7.4. This centrifugation step is repeated 3 times in total. After the last centrifugation step the pellet is resuspended in Tris-HCl 5OmM and homogenized with Dual Potter, 10 strokes to a homogenous solution, an aliquot is removed for protein determination. Membranes are aliquoted and frozen at -8O ⁰ C until use. The radioligand binding assay is performed at room temperature in 96-well microtiter plates (No-binding Surface Plates, Corning 3600) with a final assay volume of 200μl/well in incubation buffer (5OmM Tris buffer (pH 7.4 RT) containing 0.1 % BSA, 40 mg/L Bacitracin, complete EDTA-free protease inhibitor cocktail tablets 20 pills/L (Roche) and 3mM MnCl ₂ ). Competition binding curves are done by adding increasing amounts of the test compound. Test compounds are dissolved and serially diluted in DMSO, final DMSO concentration 1.5 % in the assay. 50μl Non-labelled ZD 6021 (a non selective NK- antagonist, lOμM final cone) is added for measurement of non-specific binding. For total binding, 50μl of 1.5% DMSO (final cone) in incubation buffer is used. [ ³ H-Sar,Met(O ₂ )- Substance P] (4nM final cone) is used in binding experiments on hNKir, [ ³ H-SR48968] (3nM final cone.) for hNK ₂ r. 50μl radioligand, 3μl test compound diluted in DMSO and

47μl incubation buffer are mixed with 5-10μg cell membranes in lOOμl incubation buffer and incubated for 30 min at room temperature on a microplate shaker. The membranes are then collected by rapid filtration on Filtermat B(Wallac), presoaked in 0.1% BSA and 0.3% Polyethyleneimine (Sigma P-3143), using a Micro 96 Harvester

5 (Skatron Instruments, Norway), Filters are washed by the harvester with ice-cold wash buffer (5OmM Tris-HCl, pH 7.4 at 4°C, containing 3mM MnCl ₂ ) and dried at 50 ⁰ C for 30- 60 min. Meltilex scintillator sheets are melted on to filters using a Microsealer (Wallac, Finland) and the filters are counted in a β-Liquid Scintillation Counter (1450 Microbeta, Wallac, Finland). Q The Kj value for the unlabeled ligand is calculated using the Cheng-Prusoff equation (Biochem. Pharmacol. 22:3099-3108, 1973): where L is the concentration of the radioactive ligand used and Kd is the affinity of the radioactive ligand for the receptor, determined by saturation binding. Data is fitted to a four-parameter equation using Excel Fit. )

Results

In general, the compounds useful in accordance with the present invention demonstrated statistically significant antagonistic activity at the NKj receptor within the interval 7-9 for Q the pKβ. For the NK2 receptor the interval for the pKg was 7-9.

Thus, the tested compounds useful in accordance with the present invention have been shown to be dual NKj/ NK2 receptor antagonists.

5

Biological evalution

Gerbil Foot Tap (NKi specific test model)

Male Mongolian gerbils (60-80g) are purchased from Charles River, Germany. On arrival, they are housed in groups often, with food and water ad libitium in temperature and ₀ humidity-controlled holding rooms. The animals are allowed at least 7 days to acclimatize

to the housing conditions before experiments. Each animal is used only once and killed immediately after the experiment by heart punctuation or a lethal overdose of penthobarbital sodium.

Gerbils are anaesthetized with isoflurane. Potential CNS-peπneable NKl receptor antagonists are administered intraperitoneally, intravenously or subcutaneously. The compounds are given at various time points (typically 30-120 minutes) prior to stimulation with agonist.

The gerbils are lightly anaesthetized using isofluoranε and a small incision is made in the skin over bregma. 10 pmol of ASMSP, a selective NKi receptor agonist, is administered icv in a volume of 5 μl using a Hamilton syringe with a needle 4 mm long. The wound is clamped shut and the animal is placed in a small plastic cage and allowed to wake up. The cage is placed on a piece of plastic tubing filled with water and connected to a computer via a pressure transducer. The number of hind feet taps is recorded.

Fecal pellet output (NK ₂ specific test model)

The in vivo effect (NK ₂ ) of the compounds of formula I can be determined by measuring NK2 receptor agonist-induced fecal pellet output using gerbil as described in e.g. The Journal of Pharmacology and Experimental Therapeutics (2001) 559-564.

Biological evaluation Methods An in vivo gastric distension model is used as a model for functional gastrointestinal disorders, in particular for functional dyspepsia (Bayati A, Astin M, Ekman C, Mattsson H. Wistar Kyoto rats have impaired gastric adaptive accommodation in response to gastric distension. Gastroenterology 2003; 124 (4, suppl 1): W1471 (abstract)).

The gastric distension model enables detailed analysis of the physico-mechanical properties of the stomach, e.g. basal gastric tone, threshold for accommodation,

accommodation rate, accommodation volume, and maximal gastric volume. By using the same model in both rats and humans it has been found that the gastric volume responses is very similar in the rat glandular stomach to that in human proximal stomach. Furthermore, it has been shown that patients with Functional Dyspepsia as well as Wistar Kyoto (WKY) rats have an impaired gastric adaptive response and also a lower total gastric volume as compared to healthy subjects and Sprague Dawley (SD) rats, respectively. In addition, the method has shown to be reproducible and reliable. Moreover, the advantage of the presently used barostat technique compared to other barostat techniques normally used in experimental clinical studies is that it is possible to discriminate between if a compound exerts its effect directly on gastric smooth muscles or if the effect involves the vagal reflex mechanism.

The rats are equipped with fistulas chronically implanted into the stomach. During gastric experiments, a small inflatable plastic bag with a spherical shape is inserted through fistula into the glandular part of the stomach (middle to distal part in the rat). The experiments are performed in conscious rats. For detailed analysis of the physico-mechanical properties of the stomach, a combination of ramp and tonic distension paradigm is used. Pressure and volume data collected during experiments are saved for and further analysis.

In order to determine an animal's maximum gastric accommodation capacity, a balloon is inserted into the stomach of the animal and a four phase protocol which includes a start phase, a ramp phase, a tonic phase and an end phase is performed. The pressure applied to the balloon and the corresponding changes to the volume of the balloon are monitored throughout, e.g., using any barostat system known in the art (e.g., see Toma et al, Neurogastroenterol. Mot, 8, 19-28, 1996).

During the start phase a minimum distension pressure, e.g., 1 rnmHg, is applied to the balloon until base line values are obtained. This is followed by a Ramp Phase. During this phase the pressure applied to the balloon is increased linearly with a constant increase in pressure. The pressure delivered to the balloon can be between 2-20 mmHg. This phase is then followed by the Tonic Phase. During the tonic phase the pressure is kept constant at

the maximum pressure. Finally the pressure is dropped to the starting minimum distension pressure and this period is known as the End Phase.

To determine if an agent, e.g., a compound is useful in the treatment of FD, the maximum gastric accommodation capacity in the animal following administration of the compound is calculated. A compound of interest will be a compound that alters the maximum gastric accommodation capacity in the animal and this is calculated by determining a difference in the maximum gastric accommodation capacity before and after administration of the compound.

The Wistar Kyoto rats (WKY; M&B Denmark) are starved about 8 or 18 hours before each experiment depending on if the experiments are performed in the morning or in the afternoon. A small, inflatable balloon is inserted through the central hole of the fistula into the distal part of stomach under isoflurane anaesthesia (Forene ^® , Abbott Scandinavia AB) and fixed in its position through the tightening of the fistula. The balloon has a spherical shape with a wall thickness of about 15 μm, a non-distensible max diameter of 25 mm and a max volume of about 7 ml. The balloon is connected to a double-lumen polyethylene catheter with an outer diameter of 1.40 mm and a length of about 20 cm. The inner lumen diameter of the catheter was about 0.58 mm. The animals are placed in a specially designed Bollmann cage, with an inner diameter of 60 mm for females and 70 mm for males. The catheter is then, via a pressure transducer, connected to a barostat system.

A barostat system maintains the pressure by pumping air into and out of the balloon. After the experiment the balloon and the connecting cable are removed under isoflurane anaesthesia and the animals are returned to their normal cages.

A combination of ramp and tonic distension is used in all the experiments. The protocol starts with a minimum distension pressure of 1 mrnHg and continues for 20 min in order to collect base line values. The pressure is then increased by a velocity of 1-4 mmHg/min for 10 min to a maximum pressure of 10-20 mmHg (ramp phase). The barostat then keeps the pressure at the maximum pressure for 10 more min (tonic phase). After the tonic phase the

pressure drops to the minimum distension pressure of 1 mmHg in about Is. The pressure is then kept at this level for another 20-minute period.

Results

The results shown in Fig 1 show that (S)-N-methyl-N-(4-(4-acetylamino-4-phenyl piperidino)-2-(3,4-dichlorophenyl)butyl benzamide) (SR48968), a NK ₂ receptor antagonist (can be synthesized as described in Bioorganic & Medicinal Chemistry Letters 1993, 3:5, 925-930) in a dose of 10 μmol/kg in WKY rats induced an increased gastric volume both during the tonic phase and the ramp phase in addition to an increased maximum gastric volume compared to the control situation. The increased maximum gastric volume is probably due to the increased accommodation rate seen (the slope of volume curve during the tonic phase). These results thus conclude that (S)-N-methyl-N-(4-(4-acetylamino-4- phenyl piperidino)-2-(3,4-dichlorophenyl)butyl benzamide) increases the accommodation capacity in WKY rats.

The results shown in Fig 2 indicate that (3aR,7aR)-2-[2-(2-methoxyphenyl)ethanimidoyl]~ 7,7-diphenyloctahydro-4H-isoindol-4-one (RP67580), a NK ₁ receptor antagonist (commercially available from Tocris) in a dose of 3 μmol/kg in WKY rats induced an increased gastric volume both during the tonic phase and the ramp phase in addition to an increased maximum gastric volume compared to the control situation. The increased maximum gastric volume is probably due to the increased accommodation rate seen (the slope of volume curve during the tonic phase). The conclusion is thus, that (3aR,7aR)-2-[2- (2-methoxyphenyl)ethanimidoyl]-7,7-diphenyloctahydro-4H-isoi ndol-4-one increases the accommodation capacity in WKY rats.