KR20070023763A - Imidazole variants as modulators of gaba receptor for the treatment of gi disorders - Google Patents

Abstract

The present invention provides a novel compound having a positive allosteric GABA _B receptor (GBR) modulator effect, a method for preparing the compound, and inhibition of transient lower esophageal sphincter relaxation, treatment of gastroesophageal reflux disease, and functional gastrointestinal disorders and irritable bowel syndrome. And their use optionally in combination with GABA _B agonists for the treatment of (IBS).

Positive allosteric BAA receptor, transient lower esophageal sphincter relaxation, gastroesophageal reflux disease, functional gastrointestinal disorder

Description

IMIDAZOLE VARIANTS OF GABA RECEPTOR FOR THE TREATMENT OF GI DISORDERS}

The present invention provides a novel compound having a positive allosteric GABA _B receptor (GBR) modulator effect, a method for preparing the compound, and inhibition of transient lower esophageal sphincter relaxation, treatment of gastroesophageal reflux disease, and functional gastrointestinal disorders and And their use for the treatment of irritable bowel syndrome (IBS).

The lower esophageal sphincter (LES) is prone to intermittent relaxation. As a result, if at any point the mechanical barrier temporarily collapses, fluid from the stomach may pass over to the esophagus, hereinafter referred to as "reflux".

Gastroesophageal reflux disease (GERD) is the most common upper gastrointestinal tract disease. Current pharmacotherapy aims to reduce gastric acid secretion or to neutralize acid in the esophagus. The main mechanism after reflux has been considered to be dependent on the hypotonic lower esophageal sphincter. However, recent studies (e.g., Holloway & Dent (1990) Gastroenterol. Clin. N. Amer. 19, pp. 517-535) show that most reflux events occur during transient lower esophageal sphincter relaxation (TLESR), ie It has been found to occur during relaxation and not caused by swallowing. Gastric acid secretion is also generally known to be typical in patients with GERD.

In conclusion, there is a need for a therapy that prevents reflux by reducing the incidence of TLESR.

GABA _B -receptor agonists have been shown to inhibit TLESR and are disclosed in WO 98/11885 A1.

Functional gastrointestinal disorders, such as functional dyspepsia, are described in Thampson 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: 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].

Irritable bowel syndrome (IBS) is described by Thampson 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: 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].

GABA _B  Receptor Agonists

GABA (4-aminobutanoic acid) is an endogenous neurotransmitter of the central and peripheral nervous systems. Receptors for GABA are generally divided into GABA _A and GABA _B receptor subtypes. GABA _B receptors belong to the superfamily of G-protein coupled receptors (GPCRs).

Baclofen (4-amino-3- (p-chlorophenyl) butanoic acid; disclosed in CH 449046), the best studied GABA _B receptor agonist, is useful as an anticonvulsant. EP 356128 A2 describes the use of (3-aminopropyl) methylphosphinic acid, a GABA _B receptor agonist, for use in therapy, in particular in the treatment of central nervous system disorders.

EP 463969 A1 and FR 2722192 A1 disclose 4-aminobutanoic acid derivatives having different heterocyclic substituents on the 3-carbon of the butyl chain. EP 181833 A1 discloses substituted 3-aminopropylphosphinic acid with high affinity for the GABA _B receptor site. EP 399949 A1 discloses derivatives of (3-aminopropyl) methyl phosphinic acid described as potent GABA _B receptor agonists. In addition, other (3-aminopropyl) methylphosphinic acid and (3-aminopropyl) phosphinic acid are described in WO 01/41743 A1 and WO 01/42252 A1, respectively. Regarding their affinity for the GABA _B receptor, the relationship of structure-activity of several phosphinic acid analogs is described in [J. Med. Chem. (1995), 38, 3297-3312. Sulfinic acid analogs and their GABA _B receptor activity are described in Bioorg. & Med. Chem. Lett. (1998), 8, 3059-3064. A more general review of GABA _B ligands is given in Curr. Med. Chem.-Central Nervous System Agents (2001), 1, 27-42.

GABA _B  Positive of receptor Allosteric  control

2,6-di-tert-butyl-4- (3-hydroxy-2,2-dimethylpropyl) phenol (CGP7930) and 3- (3,5-di-tert-butyl-4-hydroxyphenyl)- 2,2-dimethylpropanal (disclosed in US 5,304,685) has been described as positive allosteric regulation of natural and recombinant GABA _B receptor activity (Society for Neuroscience, 30th Annual Meeting, New Orleans, La., Nov. 4-9, 2000: Positive allosteric regulation of natural and recombinant GABA _B receptor activity, S. Urwyler et al .; Molecular Pharmacol. (2001), 60, 963-971).

N, N-dicyclopentyl-2-methylsulfanyl-5-nitro-pyrimidine-4,6-diamine has been described as positive allosteric regulation of the GABA _B receptor (The Journal of Pharmacology and Experimental Therapeutics, 307 (2003), 322-330).

1H- Imidazole -5- Carboxylic acid  derivative

A few 4-amino-1H-imidazole-5-carboxylic acid ethyl esters are intermediates for purine synthesis (Tetrahedron Lett. (1966), 1885-1889) or imidazo [4,5-d] pyrimidone and imide As intermediates for the synthesis of multizo [4,5, -b] pyridine (Monatshefte fuer Chemie (1976), 107: 1413-1421). In addition, 1,7-dihydro-6H-purin-6-one is prepared from 4-acylamino-1H-imidazole-5-carboxylic acid ethyl ester (Tetrahedron (1982), 38: 1435-1441). However, such compounds are not known as positive allosteric modulators of the GABA _B receptor and have not been described as useful in the treatment of GERD or functional gastrointestinal disorders.

A recent review of allosteric regulation of GPCRs, see Expert Opin. Ther. Patents (2001), 11, 1889-1904.

<Overview of invention>

The present invention

1H-imidazole-5-carboxylic acid, 4- (acetylamino) -1-methyl-2- (methylthio)-, ethyl ester;

1H-imidazole-5-carboxylic acid, 4- (acetylamino) -2- (methylthio) -1-phenyl-, ethyl ester;

1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -1- (2-furanylmethyl) -2- (methylthio)-, ethyl ester;

1H-imidazole-5-carboxylic acid, 4- (acetylamino) -1- (2-furanylmethyl) -2- (methylthio)-, ethyl ester;

1H-imidazole-5-carboxylic acid, 4- (acetylamino) -2- (methylthio) -1- (2-thienylmethyl)-, ethyl ester;

* 1H-imidazole-5-carboxylic acid, 2- (methylthio) -4-[[(5-nitro-2-furanyl) carbonyl] amino] -1- (2-thienylmethyl)-, Ethyl esters;

* 1H-imidazole-5-carboxylic acid, 4-[[4- (1,1-dimethylethyl) benzoyl] amino] -1- (2-methoxyphenyl) -2- (methylthio)-, ethyl ester;

* 1H-imidazole-5-carboxylic acid, 4-[(2,4-dichlorobenzoyl) amino] -1- [4- (1-methylethyl) phenyl] -2- (methylthio)-, ethyl ester ;

* 1H-imidazole-5-carboxylic acid, 1- [4- (1-methylethyl) phenyl] -4-[(2-methyl-1-oxopropyl) amino] -2- (methylthio)-, Ethyl esters;

1H-imidazole-5-carboxylic acid, 1- [2-thienylmethyl] -4-[(chloro-acetyl) amino] -2- (methylthio)-, ethyl ester;

1H-imidazole-5-carboxylic acid, 1- [2-thienylmethyl] -4-[(dichloro-acetyl) amino] -2- (methylthio)-, ethyl ester; And

* 1H-imidazole-5-carboxylic acid, 1- [2-methoxyphenyl] -4-[(trichloro-acetyl) amino] -2- (methylthio)-, ethyl ester

Except for providing novel compounds of formula (I)

In the above formula,

R ¹ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups) ; Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups;

R ² is C ₁ -C ₁₀ alkoxy or C ₁ -C ₁₀ thioalkoxy (each optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carbo) Acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups;

R ³ is C ₁ -C ₁₀ alkoxy (optionally C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide , Nitrile or substituted with 1 or 2 aryl or heteroaryl groups); C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ Cycloalkyl, keto, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C Substituted with _1- C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or heteroaryl groups; Or amino (optionally mono- or di-substituted with C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl or C ₃ -C ₁₀ cycloalkyl);

R ⁴ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₁ -C ₁₀ alkoxy, or C ₃ -C ₁₀ cycloalkyl, each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or 1 or 2 aryl Or substituted with a heteroaryl group); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with one or two aryl or heteroaryl groups.

In one embodiment of the invention, R ² is C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or hetero C ₁ -C ₁₀ alkoxy optionally substituted with an aryl group.

In a further embodiment of the invention, R ² is C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or hetero C ₁ -C ₁₀ thioalkoxy optionally substituted with an aryl group.

A new class of compounds of formula I, 1H-imidazole-5-carboxylic acid ethyl ester or amide, is useful as a positive allosteric GABA _B receptor modulator.

The general terms used in the definition of formula (I) have the following meanings.

C ₁ -C ₁₀ alkyl is a straight or branched alkyl group having 1 to 10 carbon atoms, for example methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl , Pentyl, isopentyl, hexyl or heptyl. The alkyl is at least one C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or one or two It may be substituted with an aryl or heteroaryl group. The alkyl group may contain one or more heteroatoms selected from O, N and S, ie one or more carbon atoms may be substituted by such heteroatoms. Examples of such groups are methyl-ethylether, methyl-ethylamine and methyl-thiomethyl. The alkyl group may form part of a ring. One or more hydrogen atoms of the alkyl may be substituted with fluorine atoms.

C ₂ -C ₁₀ alkenyl is a straight or branched alkenyl group having 2 to 10 carbon atoms, for example vinyl, isopropenyl and 1-butenyl. The alkenyl is at least one C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or one or two It may be substituted with an aryl or heteroaryl group. The alkenyl group may contain one or more heteroatoms selected from O, N and S, ie one or more carbon atoms may be substituted by such heteroatoms. One or more hydrogen atoms of the alkenyl group may be substituted with fluorine atoms.

C ₂ -C ₁₀ alkynyl is a straight or branched alkynyl group having 2 to 10 carbon atoms, for example ethynyl, 2-propynyl and but-2-ynyl. The alkynyl is at least one C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or one or two It may be substituted with an aryl or heteroaryl group. The alkynyl group may contain one or more heteroatoms selected from O, N and S, ie one or more carbon atoms may be substituted by such heteroatoms. One or more hydrogen atoms of the alkynyl group may be substituted with fluorine atoms.

C ₃ -C ₁₀ cycloalkyl is cyclic alkyl having 3 to 10 carbon atoms, for example cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. The cycloalkyl may also be unsaturated. The cycloalkyl is one or more C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or one or two It may be substituted with an aryl or heteroaryl group. The cycloalkyl group may contain one or more heteroatoms selected from O, N and S, ie one or more carbon atoms may be substituted by such heteroatoms. One or more hydrogen atoms of the cycloalkyl group may be substituted with a fluorine atom.

C ₁ -C ₁₀ alkoxy is an alkoxy group having 1 to 10 carbon atoms, for example methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy, isobutoxy, sec-butoxy, tert -Butoxy, pentoxy, hexoxy or heptoxy group. The alkoxy may be cyclic, partially unsaturated or unsaturated as in propeneoxy or cyclopentoxy. The alkoxy may be aromatic, such as in benzyloxy or phenoxy. The alkoxy group may contain one or more heteroatoms selected from O, N and S, ie one or more carbon atoms may be substituted by such heteroatoms. The alkoxy is one or more C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or one or two aryl Or a heteroaryl group.

C ₁ -C ₁₀ thioalkoxy is a thioalkoxy group having 1 to 10 carbon atoms, for example thiomethoxy, thioethoxy, n-thiopropoxy, n-thiobutoxy, thioisopropoxy, thioiso moiety Methoxy, sec-thiobutoxy, tert-thiobutoxy, thiopentoxy, thiohexoxy or thioheptoxy group. The thioalkoxy can be unsaturated, such as in thiopropenoxy, or can be aromatic, such as in thiobenzyloxy or thiophenoxy. The thioalkoxy is at least one C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or one or two It may be substituted with an aryl or heteroaryl group.

The term aryl is defined herein as an aromatic ring having 6 to 14 carbon atoms, including both single rings and polycyclic compounds, for example phenyl, benzyl or naphthyl, optionally as C ₁ as in biphenyl. -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mer Substituted with one or more substituents such as capto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile, or one or two aryl or heteroaryl groups. Polycyclic rings are saturated, partially unsaturated or saturated.

The term heteroaryl is defined herein as an aromatic ring having 3 to 14 carbon atoms, including both single rings and polycyclic compounds, wherein one or several ring atoms are oxygen, nitrogen or sulfur, for example Furanyl, thiophenyl or imidazopyridine. The heteroaryl is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -as in biphenyl. Optionally substituted with one or more substituents such as C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or one or two aryl or heteroaryl groups do. Polycyclic rings are saturated, partially unsaturated or saturated.

Halogen as used herein is selected from chlorine, fluorine, bromine or iodine.

Compounds of formula (I) have one or more asymmetric carbon atoms, which may exist in several stereochemical forms. The present invention includes mixtures of isomers, and individual stereoisomers. The present invention also includes geometric isomers, rotamers, enantiomers, racemates and diastereomers.

If appropriate, the compounds of formula (I) are in their natural state, for example as carboxylic acids, or as salts of the compounds discussed, preferably pharmaceutically acceptable salts, for example sodium, potassium, ammonium, calcium or magnesium salts. It can be used in the form.

Compounds of formula I are useful as positive allosteric GBR (GABA _B receptor) modulators. Positive allosteric modulators of GABA _B receptor is defined as to make so as to be intrinsic by ligand binding to GABA _B receptor protein at a site different from the site for binding, and more sensitive to the GABA _B receptor for GABA and GABA _B receptor agonist compound do. Positive allosteric GBR modulators work synergistically with agonists and increase the potency and / or inherent efficiency of GABA _B receptor agonists. It has also been shown that positive allosteric modulators that act on GABA _B receptors can produce agonist effects. Thus, the compounds of formula (I) may be effective as full or partial agonists.

A further aspect of the invention is a compound of formula (I) for use in therapy.

When administering a positive allosteric modulator, according to the conclusion that GABA _B receptors, it becomes more sensitive to the GABA _B receptor agonists, GABA _B I potency is observed the increase of the inhibition of transient lower esophageal sphincter relaxation (TLESR). In conclusion, the present invention relates to the use of a positive allosteric GABA _B receptor modulator according to formula I, optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for inhibiting transient lower esophageal sphincter relaxation (TLESR).

A further aspect of the invention is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for preventing reflux.

A further aspect of the invention is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment of gastroesophageal reflux disease (GERD).

Effective management of infant obstructive dysfunction will be an important method for the prevention and cure of lung diseases due to aspiration of refluxed gastric contents and for the management of growth failure, especially growth failure due to excessive loss of digested nutrients. Thus, a further aspect of the present invention is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment of lung disease.

Another aspect of the invention is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the management of growth failure.

Another aspect of the invention is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment or prevention of asthma, eg reflux-related asthma.

A further aspect of the invention is a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment or prevention of laryngitis or chronic laryngitis.

A further aspect of the invention provides a pharmaceutically and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, to a subject in need of inhibition of transient lower esophageal sphincter relaxation (TLESR). It is a method of suppressing sphincter relaxation (TLESR).

Another aspect of the invention is a method of preventing reflux, wherein a pharmaceutical and pharmacologically effective amount of a compound of formula I, optionally combined with a GABA _B receptor agonist, is administered to a subject in need of prevention of reflux.

A further aspect of the present invention is also directed to administering a gastroesophageal reflux, in which a pharmaceutical and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need of treatment of gastroesophageal reflux disease (GERD). Treatment of disease (GERD).

Another aspect of the invention is a method of treating or preventing obstructive dysfunction in which a pharmaceutically and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need thereof. .

Yet another aspect of the invention provides a method for treating infant obstruction failure, wherein a pharmaceutical and pharmacologically effective amount of a compound of Formula I, optionally in combination with a GABA _B receptor agonist, is administered to a subject in need thereof. It is a prevention method.

A further aspect of the present invention is also the treatment, prevention or inhibition of pulmonary disease, wherein a pharmaceutical and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need thereof. Way. In particular, lung diseases caused by the aspiration of refluxed gastric contents can be treated.

A further aspect of the present invention is also a method of managing growth failure, wherein a pharmaceutically and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need of treatment for growth failure.

A further aspect of the invention is administering a pharmaceutical and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, to a subject in need of treatment of asthma, eg reflux-related asthma. , For example methods of treating or inhibiting reflux-related asthma.

A further aspect of the invention is the treatment of laryngitis or chronic laryngitis, wherein a pharmaceutical and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need of treatment of laryngitis or chronic laryngitis, or It is a prevention method.

A further embodiment is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment of functional gastrointestinal disorders (FGD). Another aspect of the invention is a method of treating a functional gastrointestinal disorder, wherein an effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject with a functional gastrointestinal disorder.

A further embodiment is the use of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment of functional dyspepsia. Another aspect of the invention is a method of treating functional dyspepsia, wherein an effective amount of a compound of Formula I, optionally in combination with a GABA _B receptor agonist, is administered to a subject with a functional dyspepsia.

Functional dyspepsia refers to pain or discomfort around the upper abdomen. The discomfort may be characterized by or combined with satiety of the upper abdomen, early overeating, bloating or nausea. According to etiology, patients with functional dyspepsia can be divided into two groups.

1- Patients exhibiting identifiable pathophysiological or microbiological abnormalities of unspecified clinical relevance (e.g. Helicobacter pylori ) gastritis, histological duodenitis, gallstones, visceral hypersensitivity, gastroduodenal dyskinesia)

2- Patients with no definitive explanation of symptoms.

Functional dyspepsia can be diagnosed as follows.

(More than 12 weeks, not necessarily consistent within the previous 12 months)

1- persistent or recurrent indigestion (pain or discomfort centered around the upper abdomen) and

2- no evidence of organ disease (including in upper gastrointestinal endoscopy) that can explain symptoms; and

3- indigestion without evidence, alleviated only by defecation or associated with the onset of change in fecal frequency or form

Functional dyspepsia can be divided into subsets such as ulcer-type dyspepsia, dyskinesia-type dyspepsia and nonspecific (nonspecific) dyspepsia, depending on the distinct symptom pattern.

Current treatments for functional dyspepsia are associated with the relief of very empirical and significant symptoms. The most commonly used therapies still include antidepressants.

A further aspect of the present invention is optionally in combination with a GABA _B receptor agonist for the manufacture of a medicament for the treatment or prophylaxis of irritable bowel syndrome (IBS), eg, constipation predominant IBS, diarrhea predominant IBS or alternating intestinal exercise predominant IBS. The use of compounds according to formula (I).

A further aspect of the present invention is directed to administering said irritable bowel syndrome (IBS), wherein a pharmaceutical and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need of treatment of irritable bowel syndrome (IBS). ) Is a method of treatment or prevention.

IBS is defined herein as a chronic dysfunction with specific symptoms, often abdominal bloating and abdominal distension, including continuous or recurrent abdominal pain and discomfort accompanied by alternating bowel function. In general, it is divided into three subgroups according to the dominant field pattern.

1- diarrhea predominant

2- constipation predominant

3- shift bowel movement type.

Abdominal pain or discomfort is a hallmark of IBS and is present in these three subgroups.

IBS symptoms are classified according to the Rome criteria and then adjusted for the Rome II criteria. This suitability for describing IBS symptoms helps to achieve consensus in designing and evaluating IBS clinical studies.

ROM II diagnostic criteria are as follows.

1- Abdominal pain or discomfort for more than 12 weeks (not necessarily continuous) last year

2- Two or more of the following symptoms:

a) relief from bowel movements

b) initiation associated with changes in stool frequency

c) initiation associated with changes in fecal hardness

A further aspect of the present invention is the use of a compound according to formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for the treatment or prevention of CNS disorders such as anxiety.

A further aspect of the present invention provides a pharmaceutical and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, to a subject in need of treatment for a CNS disorder, eg anxiety, such a CNS disorder, eg For example is the treatment or prevention of anxiety.

A further aspect of the invention is the use of a compound according to formula (I), optionally in combination with a GABA _B receptor agonist, for the manufacture of a medicament for treating or preventing depression.

A further aspect of the invention is a method of treating or preventing depression, wherein a pharmaceutically and pharmacologically effective amount of a compound of formula (I), optionally in combination with a GABA _B receptor agonist, is administered to a subject in need thereof.

For the purposes of the present invention, the term "agonist" is understood to include full agonists and partial agonists, whereby "partial agonists" are understood as compounds capable of partially but partially activating GABA _B receptors. Can be.

The term “TLESR”, transient lower esophageal sphincter relaxation, is described herein in Mittal, R.K., Holloway, R.H., Penagini, R., Blackshaw, L.A., Dent, J., 1995; Transient lower esophageal sphincter relaxation. Gastroenterology 109, pp. 601-610.

The term “reflux” is defined as the mechanical barrier being temporarily collapsed at some point so that fluid from the stomach can pass into the esophagus.

The term "GERD", gastroesophageal reflux disease is described by van Heerwarden, M.A., Smout A.J.P.M., 2000; Diagnosis of reflux disease. Bailliere's Clin. Gastroenterol. 14, pp. 759-774.

"Formulations" according to the invention may exist as "fix formulations" or "partial formulation kits".

"Fixed formulation" includes (i) a compound of Formula (I); And (ii) a combination in which the GABA _B receptor agonist is present in one unit. One example of a “fixed formulation” is a pharmaceutical composition wherein (i) the compound of Formula I and (ii) the GABA _B receptor agonist are present in a mixture. Another example of a “fixed formulation” is a pharmaceutical composition wherein (i) the compound of Formula I and (ii) the GABA _B receptor agonist are present in one unit without mixing.

A "partial combination kit" is defined as a combination in which (i) a compound of Formula (I) and (ii) a GABA _B receptor agonist is present in one or more units. One example of a "partial combination kit" is a combination in which (i) a compound of Formula (I) and (ii) a GABA _B receptor agonist are present separately. The components of the "partial combination kit" can be administered simultaneously, successively or separately, ie individually or together.

The term "positive allosteric modulator" is defined as a compound that binds to a receptor protein at a site different from the site to which the endogenous ligand binds, thereby making the receptor more sensitive to receptor agonists.

The terms "treatment" and the term "treatment" also include "prevention" and / or prophylaxis, unless stated otherwise. The terms "therapeutic" and "therapeutically" can be interpreted accordingly.

Pharmaceutical Formulation

Compounds of formula (I) may be formulated alone or in combination with GABA _B receptor agonists.

For clinical purposes, it is suitable that the compound of formula (I), optionally in combination with a GABA _B receptor agonist, is formulated in a pharmaceutical formulation for oral administration in accordance with the present invention. In addition, rectal, parenteral or other routes of administration can also be expected for formulators. Thus, compounds of formula (I), optionally in combination with GABA _B receptor agonists, may be formulated with pharmaceutically and pharmaceutically acceptable carriers or adjuvants. The carrier may be in the form of a solid, semisolid or liquid diluent.

In the preparation of oral pharmaceutical preparations according to the invention, a compound of formula (I) to be formulated, optionally in combination with a GABA _B receptor agonist, comprises a solid powdered component such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin , Cellulose derivatives, gelatin, or other suitable ingredients, and with disintegrants and lubricants such as magnesium stearate, calcium stearate, sodium stearyl formate and polyethylene glycol wax. This mixture is then processed into granules or compressed into tablets.

Soft gelatin capsules may be prepared as capsules containing a compound of formula (I), optionally with a mixture of GABA _B receptor agonists, and other suitable vehicles for vegetable oils, fats, or soft gelatin capsules. Hard gelatine capsules are formulated with a compound of formula (I), optionally with a GABA _B receptor agonist, and solid powder components such as lactose, saccharose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives or gelatin It may contain.

Dosage units for rectal administration are in the form of (i) suppositories containing the active ingredient (s) mixed with triglyceride substrates; (ii) in the form of a gelatin rectal capsule containing a compound of formula (I), optionally in combination with a GABA _B receptor agonist and other suitable vehicle for vegetable oil, paraffin oil, or gelatin rectal capsules; (iii) in the form of a ready-made micro enema; Or (iv) in the form of a dry micro enema formulation which is reconstituted in a suitable solvent immediately prior to administration.

Liquid preparations for oral administration are syrups containing a compound of formula (I), optionally a GABA _B receptor agonist, and a remainder of the formulation consisting of a sugar or sugar alcohol and a mixture of ethanol, water, glycerol, propylene glycol and polyethylene glycol, or It may be prepared in the form of a suspension, for example a solution or a suspension. If desired, such liquid preparations may contain coloring agents, flavoring agents, saccharin and carboxymethylcellulose or other thickeners. Liquid preparations for oral administration may also be prepared in the form of a dry powder which is reconstituted with a suitable solvent immediately before use.

Solution for parenteral administration may be prepared as a solution of a compound of formula (I), optionally in combination with a GABA _B receptor agonist in a pharmaceutically acceptable solvent. Such solutions may also contain stabilizing and / or buffering components and are dispensed in unit doses in the form of ampoules or vials. Solutions for parenteral administration can also be prepared as dry preparations which are reconstituted with a suitable solvent immediately before use.

In one aspect of the invention, the compound of formula (I), optionally in combination with a GABA _B receptor agonist, may be administered once or twice daily, depending on the severity of the patient's condition. Typical daily dosages of compounds of formula (I) are 0.1-100 mg / kg body weight of the subject to be treated, but this will depend on various factors such as the route of administration, the age and weight of the patient, and the severity of the patient's condition.

Manufacturing method

Compounds of formula I according to the invention, wherein R ¹ , R ² , R ³ and R ⁴ are each and independently as defined above, can be prepared according to the following general methods (Scheme 1; related literature). Tetrahedron (1982), 38: 1435-1441).

Here, aminoimidazole II is efficiently acylated with a compound of formula I using acyl chloride (typically 1.5 to 2.5 equivalents) in an organic solvent such as THF or the like. The reaction is carried out at ambient temperature to 50 ° C. with stirring over 4 to 18 hours in the presence of polymer-supported diisopropylethylamine (PS-DIPEA; 1.5 to 3 equivalents). The reaction mixture is filtered over the nucleophilic anion exchange resin Isolute-NH ₂ eluting with THF and evaporated in vacuo to afford the desired product as an oil or amorphous solid.

Aminoimidazole II is prepared from intermediate III or IV by heating the reagent with an alpha halo carbonyl compound under base conditions (Scheme 2; Tetrahedron Lett. (1966), 1885-1889 and Monatshefte fuer Chemie (1976), 107). : 1413-1421]).

Intermediate III, wherein R ² is a C ₁ -C ₇ alkoxy group, is prepared by replacing the thiomethoxy group in intermediate IV with the corresponding C ₁ -C ₇ alkoxy group according to Scheme 3.

Intermediate IV is prepared by treating dimethylcyanodithioimidocarbonate with 1-2 equivalents of primary amine in ethanol and refluxing for 3-5 hours (see Scheme 4). The desired compound is collected by cooling the reaction mixture, evaporating in vacuo and then filtering directly or by adding water to precipitate the product and subsequently filtering.

An alternative route to intermediate II consists in treating intermediate IV with an alpha halo carbonyl compound in the presence of a base such as potassium carbonate to provide intermediate V. Subsequent treatment of intermediate V with a nucleophile such as an alkoxy or thioalkoxy derivative (eg, NaOMe, NaOEt) provides intermediate II via thiomethyl group substitution and ring closure.

Additionally, Intermediate II can be prepared in a similar manner as described above by using diphenylcyanocarbonimidate as starting material instead of dimethylcyanodithioimidocarbonate (Scheme 6; Bioorg. Med. Chem Lett. 2001, 11, 2225-2228 and Bioorg. Med. Chem. Lett. 2004, 14, 4225-4229.

4- Acylamino -1H- Imidazole -5- Carboxylic acid  Multiple Parallel Synthesis of Ethyl Ester

Polymer-supported diisopropylethylamine (35 mg, 3.5 mmol / g) using Titan Resin Loader ™ in 80 wells of Robbins Flex Chem teflon 96 well block ) The 80 wells were then followed by 1H-imidazole-5-carboxylic acid, 4-amino-2- (methylthio) -1- (phenylmethyl)-, ethyl ester in THF (600 μl, 0.05 mmol) 1 (2 to 2.5 equivalents) of acyl chloride 80 dissolved in THF (600 μl) is added. The block is sealed and spun at room temperature for 18 hours, after which each well is filtered while eluting with THF (1.5 ml) on isolute-NH ₂ (200 mg).

THF is evaporated in vacuo to give the acylated product as an amorphous solid or oil.

4- Acylamino -1H- Imidazole -5- Carboxylic acid  Multiple Parallel Synthesis of Ethyl Ester

To 24 wells of Ribbon Flex Chem Teflon 96 well blocks are added polymer-supported diisopropylethylamine (70 mg, 3.5 mmol / g) using a titanium resin loader ™. Thereafter, 1 well of aminoimidazole 8 in THF (600 μl, 0.1 mmol) per column was added to the well, and then 1 (2 to 2.5 equivalents) of acyl chloride 3 dissolved in THF (600 μl) per row. Add). The block is sealed and spun at room temperature for 18 hours, after which each well is filtered while eluting with THF (2.5 ml) on isolute-NH ₂ (400 mg).

THF is evaporated in vacuo to give the acylated product as an amorphous solid or oil.

Example  1: 1H- Imidazole -5- Carboxylic acid , 4-[(3,4-dichlorobenzoyl) amino] -2- (methylthio) -1- (phenylmethyl)-, synthesis of ethyl ester

1H-imidazole-5-carboxylic acid, 4-amino-2- (methylthio) -1- (phenylmethyl)-, ethyl ester (29 mg, 0.1 mmol) in 1 ml vial in THF (700 μl) Dissolved. 50 mg (3.5 mmol / g) of polymer-supported diisopropylethylamine and then 3,4-dichlorobenzoyl chloride (31 mg, 0.15 mmol) were added. The reaction mixture was stirred overnight at room temperature and then filtered over an Isolute-NH ₂ column (200 mg), washing through THF (1 ml). THF was evaporated in vacuo to yield the product (35 mg, 75%).

Example  2: 1H- Imidazole -5- Carboxylic acid , 4-amino-2- (methylthio) -1- (phenylmethyl)-, ethyl ester (used as intermediate)

Carbamimidothioic acid, N'-cyano-N- (phenylmethyl)-, methyl ester (2 g, 9.7 mmol), bromo ethyl acetate (1.79 g, 10.7 mmol) and potassium carbonate (1.48 g, 10.7 mmol ) Was dissolved / suspended in DMF (20 ml) and stirred at 60 ° C. for 1 hour. During the same time, 0.56 g of sodium metal was dissolved in 12 ml of ethanol (99.9%). The reaction was cooled and sodium ethoxide solution was added dropwise over 5 minutes. The reaction was then dissolved for 5 minutes to 90 ° C., cooled to room temperature and water was added until the product precipitated. The product was filtered off and washed with ethanol / water (1: 1) to give 1.9 g of 1H-imidazole-5-carboxylic acid, 4-amino-2- (methylthio) -1- (phenylmethyl)-, ethyl ester (67%) was obtained. 0.5 g was recrystallized from ethanol / water to give> 95% purity 1H-imidazole-5-carboxylic acid, 4-amino-2- (methylthio) -1- (phenylmethyl)-, ethyl ester (410 mg ) Was obtained.

Example  3: Carbamimidothioic acid , N ' - Cyano -N- ( Phenylmethyl )-, methyl  Synthesis of Esters (Used as Intermediates)

Benzylamine (10 g, 93 mmol) was added to dimethylcyanodithioimidocarbonate (8.2 g, 56 mmol) dissolved in ethanol (150 ml, 99.9%). A thick white precipitate formed after about 10 seconds. The mixture was heated at reflux for 3 hours. The reaction was cooled to room temperature and the precipitate formed was collected by filtration and washed with ethanol (6.43 g obtained). The filtrate was evaporated and ethanol (50 ml) was added. The resulting white precipitate was filtered and washed with ethanol to give a second crop (2.66 g was obtained). The total yield of carbamimidiothioic acid, N'-cyano-N- (phenylmethyl)-, and methyl ester was 79%.

The following compounds were synthesized in a similar manner to Example 1 (RT = retention time, TOF ES + = Electrospray MS, ie molecular weight + 1):

Intermediate:

Example  4

methyl  N- Cyano - N ' - Phenylimidothiocarbamate

Aniline (0,093 mol) was added to a solution of dimethylcyanodithioimidocarbonate (0.146 mol) dissolved in 250 mL of ethanol (99.9%). The suspension was heated for 3 hours. The reaction was allowed to reach room temperature and the resulting precipitate was removed by filtration. The solid was washed with cold ethanol and dried in vacuo to afford the product.

Example  5

2- Methylthio -Substituted intermediate ( II General procedure of)

All reactions were carried out under an inert gas atmosphere using dry glassware. The corresponding bromoacetate (0.023 mol) was added dropwise to a suspension of methyl N-cyano-N'-phenylimidothiocarbamate (0.019 mol) and potassium carbonate (0,023 mol). The reaction mixture was heated at 85-90 ° C. for 2 hours. After cooling to room temperature, 12 mL of 4 M NaOH (0.048 mol) solution was added to the reaction mixture. The reaction was terminated after 15 minutes by adding EtOAc. The organic phase was washed with brine, dried over NaSO ₄ , filtered and concentrated by evaporation. The desired product was obtained by precipitating the crude product from EtOAc and small amount of heptane.

The following compounds were prepared according to Example 5.

tert -Butyl 4-amino-2- ( Methylthio )-One- Phenyl -1H- Imidazole -5-carboxylate

tert -Butyl 4-amino-2- ( Methylthio )-One- Phenyl -1H- Imidazole -5- Carboxylate

Cyclopentyl 4 -Amino-2- ( Methylthio )-One- Phenyl -1H- Imidazole -5- Carboxylate

Isopropyl 4-amino-2- ( Methylthio )-One- Phenyl -1H- Imidazole -5- Carboxylate

2,2-dimethylpropyl 4-amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

One- Cyano -One- Methylethyl 4 -Amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

1,1- Dimethylpentyl 4 -Amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

Tetrahydrofuran-3-yl 4-amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

2- Methoxy -1,1-dimethylethyl 4-amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

1,1-dimethyl-2- Oxopropyl  4-amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

One-( Methoxymethyl ) Propyl 4-amino-2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

Example  6

2- Methoxy -Substituted intermediate ( II General procedure for)

All reactions were carried out under an inert gas atmosphere using dry glassware. The corresponding bromoacetate (0.014 mol) was added dropwise to a suspension of methyl N-cyano-N'-phenylimidothiocarbamate (0.012 mol) and potassium carbonate (0.014 mol) in 23 mL of dry DMF. The reaction mixture was heated at 60 ° C. for 1.5 h. After cooling to room temperature, NaOMe (0.060 mol) dissolved in 30 mL of dry MeOH was added slowly to deform. After addition, the reaction was cooled in an ice bath and water was added to give a precipitate which was filtered and dried in vacuo. The crude was purified by high performance chromatography (MeCN: NH ₄ OAc-buffer gradient 5:95 to 95: 5%) to afford the desired product.

The following compounds were prepared according to Example 6:

2- Methoxy -1,1-dimethylethyl 4-amino-2- Methoxy -One- Phenyl -1H- Imidazole -5- Carboxylate

The following compounds were synthesized according to Example 1 and analyzed at 400 MHz by NMR:

tert -Butyl 4-[(4-fluorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

2- Methoxy -One- Methylethyl 4 -[(4- Chlorobenzoyl ) Amino] -2- ( Methylthio )-One- Phenyl -1H- Imidazole -5- Carboxylate

Cyclopentyl 4 -[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

tert -Butyl 4-[(methoxyacetyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

Isopropyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate

2,2-dimethylpropyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imi Dazole -5-carboxylate

One- Cyano -One- Methylethyl 4 -[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H- Imidazole -5-carboxylate

1,1- Dimethylpentyl 4 -[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imida Sol -5-carboxylate

Tetrahydrofuran-3-yl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H- Imidazole -5-carboxylate

2- Methoxy -1,1-dimethylethyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H- Imidazole -5-carboxylate

1,1-dimethyl-2- Oxopropyl  4-[(4- Chlorobenzoyl ) Amino] -2- ( Methylthio )-One- Phenyl -1H- Imidazole -5- Carboxylate

One-( Methoxymethyl ) Propyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H- Imidazole -5-carboxylate

2- Methoxy -1,1-dimethylethyl 4-[(2,3-dihydro-1,4-benzodioxin-2-ylcarbonyl) amino] -2- Methoxy -One- Phenyl -1H- Imidazole -5-carboxylate

2- Methoxy -1,1-dimethylethyl 4-{[(3-chlorophenoxy) acetyl] amino} -2-methoxy-1- Phenyl -1H- Imidazole -5-carboxylate

2- Methoxy -1,1-dimethylethyl 4-[(4-chlorobenzoyl) amino] -2-methoxy-1-phenyl-1H- Imidazole -5-carboxylate

2- Methoxy -1,1-dimethylethyl 4-[(2,4-dichlorobenzoyl) amino] -2-methoxy-1-phenyl-1H- Imidazole -5-carboxylate

analysis

LC-MS analysis was performed using a Micromass 8 probe MUX-LTC ESP + system and purity was measured by single wavelength (254 nm) UV detection. Chromatography was performed on eight parallel Xterra ™ MS C8 3.5 um, 4.6 × 30 mm columns. The flow of 15 ml / min was divided into eight columns to obtain a flow rate of 1.9 ml / min. The 10-minute chromatography gradient was as follows.

Mobile phase A: 95% ACN + 5% 0,010 M NH ₄ OAc

Mobile phase B: 5% ACN + 95% 0,010 M NH ₄ OAc

10 minutes 0,0 minutes 0% A

8,0 min 100% A

9,0 min 100% A

9,1 min 0% A

NMR analysis was performed at 400 MHz.

Biological assessment

In vitro  Positive in functional assay Allosteric GABA _B  Effect of Receptor Modulators

The effect of GABA and baclofen on intracellular calcium release in CHO cells expressing GABA _{B (1A, 2)} receptor heterodimer was studied in the presence or absence of a positive allosteric modulator. Positive allosteric modulators according to the present invention increased both the efficacy and efficacy of GABA.

Ability of the compounds to reduce the compound potency, i.e., EC ₅₀ of GABA was revealed by the concentration of reducing the GABA EC ₅₀ to 50%. This efficacy was similar to the efficacy reported for CGP7930 by Urwyler et al. (Available from Tocris, Northwest Point, Ms. 11 8T Bristol, Avonmouth, Bristol, UK). CGP7930 increases GABA potency from EC ₅₀ about 170 to 180 nM to EC ₅₀ about 35 to 50 nM.

Experimental procedure

material

Nut mix F-12 (Ham) cell culture medium, serum reduced OPTI-MEM I medium, fetal bovine serum (FBS), penicillin / streptomycin solution (PEST), geneticin, HEPES ( 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (buffer), 1 M solution), Hank Balanced Salt Solution (HBSS) and Zeocin Life Technologies (Scotland Paisley) From); Polyethylenimine, Provenidide, Barclofen and γ-aminobutyric acid (GABA) are obtained from Sigma (St. Louis, USA); Fluo-3 AM was obtained from Molecular Probes (Oregon, USA). 4-amino-n- [2,3- ³ H] butyric acid ([ ³ H] GABA) was obtained from Amersham Pharmacia Biotech (Wussala, Sweden).

GABA _B  Generation of Cell Lines Expressing Receptors

GABA _B Rla and GABA _B R2 were cloned from human brain cDNA and subcloned into pCI-Neo (Promega) and pALTER-1 (Promega), respectively. GABA _B R1a-G _{α qi5} fusion protein expression vector was constructed using the pCI-Neo-GABA _B R1a cDNA plasmid and pLEC1-G _{α qi5} (Molecular Devices ISIS (Molecular Devices), CA). Cys356 was mutated to Gly using standard PCR methods using primers 5′-GGATCCATGGCATGCTGCCTGAGCGA-3 ′ (forward) and 5′-GCGGCCGCTCAGAAGAGGCCGCCGTCCTT-3 ′ (reverse) to make G _{α qi5} pertussis toxic insensitive. G _{α qi5mut} cDNA was ligated to the BamHI and NotI sites of pcDNA3.0 (Invitrogen). GABA _B R1a coding sequence was amplified by PCR from pCI-Neo-GABA _B R1a using primers, namely 5'-GGATCCCCGGGGAGCCGGGCCC-3 '(forward) and 5'-GGATCCCTTATAAAGCAAATGCACTCGA-3' (reverse), and pcDNA3.0- _Subcloning into the BamHI site of G _{α qi5mut} .

To optimize the Kozak conserved sequence of GABA _B R2, in situ mutagenesis was performed using the alteration site mutagenesis kit with primer 5'-GAATTCGCACCATGGCTTCCC-3 'according to the manufacturer's instructions (Promega). The optimized GABA _B R2 is then restricted from pALTER-1 to Xho I + Kpn I and subcloned into the mammalian expression vector pcDNA3.1 (−) / Zeo (Invitrogen) to yield the final construct pcDNA3.1 (−) / Zeo-GABA _B R2 was generated.

To produce stable cell lines, CHO-K1 cells were immersed in a wet CO ₂ -incubator at 37 ° C. in nut mix F-12 (containing) medium supplemented with 10% FBS, 100 U / ml penicillin and 100 μg / ml streptomycin. Grown. The cells were detached with 1 mM EDTA in PBS and 1 million cells were seeded in 100 mm Petri dishes. After 24 hours, the culture medium was replaced with OptiMEM and incubated in CO ₂ -incubator for 1 hour.

To generate cell lines expressing GABA _B R1a / GABA _B R2 heterodimer, GABA _B R1a plasmid DNA (4 μg), GABA _B R2 plasmid DNA (4 μg) and lipofectamine (24 μl) were added in 5 ml OptiMEM. Mix and incubate for 45 minutes at room temperature. Cells were exposed to transfection medium for 5 hours and then replaced with culture medium. After the cells were incubated for an additional 10 days, optional agents (300 μg / ml hygromycin and 400 μg / ml geneticin) were added. After 24 days of transfection, single cells were separated into 96-well plates by flow cytometry using FACS Vantage SE (Palo Alto Becton Dickinson, CA). After expansion, GABA _B receptor functional responses were tested by the FLIPR assay described below. Clones with the highest functional response were collected, expanded, and subcloned by single cell isolation. Clonal cell lines showing the highest peak response in FLIPR were used in this study.

GABA _B R1a-G _{α qi5} to generate stable cell lines expressing the fusion protein and a GABA _B R2, GABA _B R1a-G _{α qi5mut} plasmid _{DNA (8 ㎍), GABA B} R2 plasmid DNA (8 ㎍) and lipofectamine (24 μl) was mixed in 5 ml OptiMEM and incubated for 45 minutes at room temperature. Cells were exposed to transfection medium for 5 hours and then replaced with culture medium. After 48 hours, cells were detached, seeded in 6 well plates (2000 cells / well), and grown in culture medium supplemented with Geneticin (400 μg / ml) and Zeosin (250 μg / ml). After 4 days, cells were collected from single colonies and transferred to 24-well plates. After 10 days, cell clones were seeded in T-25 flasks and grown for an additional 16 days, after which they were tested for GABA _B receptor mediated functional responses. Clones showing the highest peak response were collected, subcloned by seeding the cells in 6-well plates (1000 cells / well) and the steps described above were repeated. Clonal cells showing the highest peak response in FLIPR were used in this study.

FLIPR in Intracellular  Calcium GABA _B  Measurement of Receptor-Dependent Release

Coward et al. Anal. Biochem. (1999) 270, 242-248, with some modifications to determine GABA _B receptor dependent release of intracellular calcium in a fluorescent image plate reader (FLIPR). The transfected CHO cells were nut mix with Glutamax-I and supplemented with 10%, 100 U / ml penicillin and 100 μg / ml streptomycin, 250 μg / ml zeocin and 400 μg / ml geneticin Incubated in F-12 (with). Twenty four hours prior to the experiment, cells (35,000 cells / well) were seeded in a culture medium without a selection agent in 96-well poly-D-lysine coated black-wall plates (Becton Dickinson, Bedford, UK). It was. The cell culture medium was aspirated and 100 μl of fluor-3 loading solution (4 μM fluor-3, 2.5 mM probeneside and 20 mM Hepes in nut mix F-12) was added. After incubation in a 5% CO ₂ incubator at 37 ° C. for 1 hour, the dye-solution is aspirated and the cells are washed twice with 150 μl of wash solution (2.5 mM probeneside and 20 mM Hepes in HBSS) and then washed The solution was washed with an additional 150 μl. Cells were then analyzed in a fluorescent image plate reader (Molecular Devices Corp., Calif.). Test compounds were diluted to 50 μΜ concentration in HBSS containing 20 mM Hepes and 5% DMSO and added to 50 μΐ volume. Fluorescence was sampled every second for 60 seconds (15 seconds before and 50 seconds after addition of test compound), then GABA (50 μl 7.6 nM-150 μM) was added and sampled every 60 seconds for an additional 120 seconds Continued.

GTPgS

[ ³⁵ S] -GTPγS binding assay was performed with 0.025 μg / μl of membrane protein (prepared from the cell line described above) with 0.01% bovine serum albumin (without fatty acids), 10 μM GDP, 100 μM DTT and 0.53 nM [ ³⁵ S ] -GTPγS (Amersham-Pharmacia Biotech) was performed for 45 minutes at 30 ° C. in membrane buffer (100 mM NaCl, 5 mM, 1 mM EDTA, 50 mM HEPES, pH 7.4) (final volume 200 μl). . Nonspecific binding was measured in the presence of 20 μM GTPγS. The reaction was initiated by adding GABA at a concentration of 1 mM to 0.1 nM in the presence or absence of the required concentration of PAM. The reaction was terminated by the addition of ice cold wash buffer (50 mM Tris-HCl, 5 mM MgCl ₂ , 50 mM NaCl, pH 7.4) and then printed filtermate using a micro 96 harvester (Skatron Instruments). It was rapidly filtered under vacuum through a Printed Filtermat A glass fiber filter (Wallac) (0.05% PEI treated). The filter was dried at 50 ° C. for 30 minutes, then the paraffin scintillant pad was melted on the filter and binding radioactivity was measured using a 1450 Microbeta Trilux (Wallac) scintillation counter.

conclusion

GABA doses in the presence and absence of test compound - response curve is a 4-parameter log equation _{y = y max + ((y} min y _max ) / 1 + (x / C) ^D ), where C = EC ₅₀ and D = slope factor.

The efficacy of PAM in the GTPγS assay was determined by plotting log EC ₅₀ against GABA against the log concentration of the positive allosteric modulator, taking measurements in the presence of the positive allosteric modulator.

In general, the efficacy of the compounds of formula (I) is EC ₅₀ 20 μM to 0.001 μM. The individual EC ₅₀ values of the examples are as follows.

compound EC ₅₀ 1H-imidazole-5-carboxylic acid, 4-[(3,4-dichlorobenzoyl) amino] -2- (methylthio) -1- (phenylmethyl)-, ethyl ester 2.3 μM 1H-imidazole-5-carboxylic acid, 2- (methylthio) -4-[(1-oxo-2-phenylbutyl) amino] -1-phenyl-, ethyl ester 0.6 μM 1,1-dimethylpentyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate 0.3 μM 2-methoxy-1,1-dimethylethyl 4-[(2,3-dihydro-1,4-benzodioxin-2-ylcarbonyl) amino] -2-methoxy-1-phenyl-1H- Imidazole-5-carboxylate 0.3 μM

IBS  Effect of Compounds in Model (Colonic Bloating)

Colorectal distension ( CRD )

For CRD, a balloon bottom with a 3 cm polyethylene balloon (self-made) with a catheter for connection during optical isopluran anesthesia (Forene ^® , Abbott Scandinavia AB, Sweden) From distal colon to 2 cm from to anus. The catheter was fixed to the bottom of the tail with tape. At the same time, an intravenous catheter (Neople Ron (Neoflon) ^®, Becton Dickinson Abbe Swedish material) was inserted into the tail for compound administration. The rats were then placed in a Bolman cage and the experiments started after recovery from sedatives for at least 15 minutes.

During the CRD procedure, the balloon was connected to a pressure transducer (P-602, CFM-k33, 100 mmHg; Bronckhost Hi-Tec, Paynendal, The Netherlands). A custom Barostat (AstraZeneca, Nuremberg, Sweden) was used to control air expansion and pressure in the balloon. Custom computer software running on a standard PC (PharmLab on-line 4.0.1) was used to control the immediate start and perform data collection and storage. The bloating paradigm caused by Brostart was achieved by generating a pulse pattern on the analog output channel. The CRD paradigm used consists of repetitive phase distension (12 times at 80 mmHg, pulse duration 30 seconds at 5 minute intervals).

Response to CRD was assessed by recording and quantifying the phase change in pressure in the balloon for a wide pulse. Pressure fluctuations during isostatic expansion of the colonic colon reflect the abdominal muscle contractions associated with the distension procedure, which is therefore considered an effective assessment of the visceral motor response (VMR) associated with the presence of pain in the visceral organs.

Data  Collect and Analyze

The balloon pressure signal was sampled at 50 Hz and then subjected to digital filtering. Using a highpass filter at 1 Hz, the shrinkage-induced pressure change was distinguished from the slowly changing pressure generated by Barostart. Resistance in the airflow between the pressure generator and the pressure transducer further enhanced the pressure fluctuations induced by abdominal contraction in the animal. In addition, line frequency interference was eliminated using a band-stop filter at 49-51 Hz. Custom computer software (PalmLab off-line 4.0.1) was used to quantify the phase change of the balloon pressure signal. The average rectified value (ARV) of the balloon pressure signal was calculated for 30 seconds before the pulse (baseline activity) and during the pulse period (as a measure of VMR for bloating). When performing pulse analysis, the first and last seconds of each pulse reflect the artificial signal produced by the Barostart during balloon expansion and release and are not of animal origin, so the first and last seconds of each pulse are Seconds were excluded.

result

The effects of positive allosteric modulators were examined for VMR against isostatic CRD in rats. A paradigm of 12 folds was used at 80 mmHg. Compounds were administered at a dose of 1-50 μmol / kg and the VMR response to CRD was compared with the vehicle control. Compounds are effective at reducing VMR against CRD (inhibiting at least 20% compared to vehicle use).

Claims (36)

1H-imidazole-5-carboxylic acid, 4- (acetylamino) -1-methyl-2- (methylthio)-, ethyl ester; 1H-imidazole-5-carboxylic acid, 4- (acetylamino) -2- (methylthio) -1-phenyl-, ethyl ester; 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -1- (2-furanylmethyl) -2- (methylthio)-, ethyl ester; 1H-imidazole-5-carboxylic acid, 4- (acetylamino) -1- (2-furanylmethyl) -2- (methylthio)-, ethyl ester; 1H-imidazole-5-carboxylic acid, 4- (acetylamino) -2- (methylthio) -1- (2-thienylmethyl)-, ethyl ester; * 1H-imidazole-5-carboxylic acid, 2- (methylthio) -4-[[(5-nitro-2-furanyl) carbonyl] amino] -1- (2-thienylmethyl)-, Ethyl esters; * 1H-imidazole-5-carboxylic acid, 4-[[4- (1,1-dimethylethyl) benzoyl] amino] -1- (2-methoxyphenyl) -2- (methylthio)-, ethyl ester; * 1H-imidazole-5-carboxylic acid, 4-[(2,4-dichlorobenzoyl) amino] -1- [4- (1-methylethyl) phenyl] -2- (methylthio)-, ethyl ester ; * 1H-imidazole-5-carboxylic acid, 1- [4- (1-methylethyl) phenyl] -4-[(2-methyl-1-oxopropyl) amino] -2- (methylthio)-, Ethyl esters; 1H-imidazole-5-carboxylic acid, 1- [2-thienylmethyl] -4-[(chloro-acetyl) amino] -2- (methylthio)-, ethyl ester; 1H-imidazole-5-carboxylic acid, 1- [2-thienylmethyl] -4-[(dichloro-acetyl) amino] -2- (methylthio)-, ethyl ester; And * 1H-imidazole-5-carboxylic acid, 1- [2-methoxyphenyl] -4-[(trichloro-acetyl) amino] -2- (methylthio)-, ethyl ester Except for the compounds of formula (I). <Formula I>

In the above formula, R ¹ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups) ; Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ² is C ₁ -C ₁₀ alkoxy or C ₁ -C ₁₀ thioalkoxy (each optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carbo) Acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ³ is C ₁ -C ₁₀ alkoxy (optionally C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide , Nitrile or substituted with 1 or 2 aryl or heteroaryl groups); C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ Thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C Substituted with _1- C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or heteroaryl groups; Or amino (optionally mono- or di-substituted with C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl or C ₃ -C ₁₀ cycloalkyl); R ⁴ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₁ -C ₁₀ alkoxy, or C ₃ -C ₁₀ cycloalkyl, each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or 1 or 2 aryl Or substituted with a heteroaryl group); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; Here, each alkyl, alkenyl, alkynyl and cycloalkyl may independently have one or more carbon atom (s) substituted by O, N or S. The compound of claim 1, wherein R ² is optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl Or C ₁ -C ₁₀ alkoxy substituted with a heteroaryl group. The compound of claim 2, wherein R ² is optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or 1 or 2 aryl Or C ₁ -C ₅ alkoxy substituted with a heteroaryl group. The compound of claim 1, wherein R ² is optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl Or C ₁ -C ₁₀ thioalkoxy substituted with a heteroaryl group. 5. The compound of claim 4, wherein R ² is optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or 1 or 2 aryls. Or C ₁ -C ₅ thioalkoxy substituted with a heteroaryl group. 6. The compound of claim 1, wherein R ¹ is C ₁ -C ₇ alkyl or C ₃ -C ₇ cycloalkyl, each optionally and independently C ₁ -C ₇ alkoxy, C ₃ -C ₁₀ cycloalkyl, hydroxy, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with one aryl or heteroaryl group); Aryl or heteroaryl (each optionally and independently C ₁ -C ₇ alkyl, C ₂ -C ₇ alkenyl, C ₂ -C ₇ alkynyl, C ₃ -C ₇ cycloalkyl, C ₁ -C ₇ alkoxy, C ₁ -C ₇ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups) Phosphorus compounds. 7. Compounds according to claim 6, wherein R ¹ represents C ₁ -C ₄ alkyl optionally and independently substituted with one aryl or heteroaryl group. 8. The compound of claim 1, wherein R ³ is optionally and independently C ₁ -C ₁₀ thioalkoxy, keto, C ₃ -C ₁₀ cycloalkyl, halogen, hydroxy, mercapto, carboxylic acid. , Carboxylic ester, carboxylic acid amide, nitrile or C ₁ -C ₇ alkoxy substituted with one or two aryl or heteroaryl groups. 8. The compound of claim 1, wherein R ³ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each Is optionally and independently C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid Amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups. The compound of claim 9, wherein R ³ is optionally and independently C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, Carboxylic ester, carboxylic acid amide, nitrile or C ₁ -C ₁₀ alkyl substituted with one or two aryl or heteroaryl groups. 11. The compound of claim 1, wherein R ⁴ is C ₁ -C ₇ alkyl, C ₂ -C ₇ alkenyl, C ₂ -C ₇ alkynyl or C ₃ -C ₇ cycloalkyl, Optionally and independently C ₁ -C ₇ alkoxy, C ₁ -C ₇ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or hetero Substituted with an aryl group). 11. The compound of claim 1, wherein R ⁴ is optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cyclo Alkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or 1 or 2 aryl Or aryl or heteroaryl substituted with a heteroaryl group. The method of claim 1, * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(3,4-dichlorobenzoyl) amino] -2- (methylthio) -1- (2-thienylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-bromobenzoyl) amino] -2- (methylthio) -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1- (2-thienylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 2- (methylthio) -4-[(1-oxo-2-phenylbutyl) amino] -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-methoxybenzoyl) amino] -2- (methylthio) -1- (2-thienylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-methoxybenzoyl) amino] -2- (methylthio) -1-phenyl-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[[4- (1,1-dimethylethyl) benzoyl] amino] -2- (methylthio) -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(3,4-dichlorobenzoyl) amino] -2- (methylthio) -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(3,4-dichlorobenzoyl) amino] -2- (methylthio) -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -1- (3-methylbutyl) -2- (methylthio)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(3,4-dichlorobenzoyl) amino] -1- (3-methylbutyl) -2- (methylthio)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 2- (methylthio) -4-[(1-oxo-2-phenoxypropyl) amino] -1- (phenylmethyl)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 2- (methylthio) -4-[(1-oxo-2-phenylbutyl) amino] -1-phenyl-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-iodobenzoyl) amino] -2- (methylthio) -1-phenyl-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(3,4-dichlorobenzoyl) amino] -1- (2-furanylmethyl) -2- (methylthio)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -1- (2-methoxyphenyl) -2- (methylthio)-, ethyl ester * 1H-imidazole-5-carboxylic acid, 4-[(4-chlorobenzoyl) amino] -1- [4- (1-methylethyl) phenyl] -2- (methylthio)-, ethyl ester tert-butyl 4-[(4-fluorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 2-methoxy-1-methylethyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * Cyclopentyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate tert-butyl 4-[(methoxyacetyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate Isopropyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 2,2-dimethylpropyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 1-cyano-1-methylethyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 1,1-dimethylpentyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate Tetrahydrofuran-3-yl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate 2-methoxy-1,1-dimethylethyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 1,1-dimethyl-2-oxopropyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 1- (methoxymethyl) propyl 4-[(4-chlorobenzoyl) amino] -2- (methylthio) -1-phenyl-1H-imidazole-5-carboxylate * 2-methoxy-1,1-dimethylethyl 4-[(2,3-dihydro-1,4-benzodioxin-2-ylcarbonyl) amino] -2-methoxy-1-phenyl-1H -Imidazole-5-carboxylate * 2-methoxy-1,1-dimethylethyl 4-{[(3-chlorophenoxy) acetyl] amino} -2-methoxy-1-phenyl-1H-imidazole-5-carboxylate * 2-methoxy-1,1-dimethylethyl 4-[(4-chlorobenzoyl) amino] -2-methoxy-1-phenyl-1H-imidazole-5-carboxylate * 2-methoxy-1,1-dimethylethyl 4-[(2,4-dichlorobenzoyl) amino] -2-methoxy-1-phenyl-1H-imidazole-5-carboxylate Compound selected from. Compounds of formula (I) for therapeutic use. <Formula I>

In the above formula, R ¹ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups) ; Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ² is C ₁ -C ₁₀ alkoxy or C ₁ -C ₁₀ alkoxy (each of which is optionally and independently C ₁ - C ₁₀ alkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic Acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ³ is C ₁ -C ₁₀ alkoxy (optionally C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide , Nitrile or substituted with 1 or 2 aryl or heteroaryl groups); C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ Thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C Substituted with _1- C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or heteroaryl groups; Or amino (optionally mono- or di-substituted with C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl or C ₃ -C ₁₀ cycloalkyl); R ⁴ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₁ -C ₁₀ alkoxy, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C) ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carboxylic acid amide, nitrile or one or two Substituted with an aryl or heteroaryl group); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; Here, each alkyl, alkenyl, alkynyl and cycloalkyl may independently have one or more carbon atom (s) substituted by O, N or S. 14. A compound according to any one of claims 1 to 13 for use as a positive allosteric GABA _B receptor modulator. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 as an active ingredient and a pharmaceutically acceptable carrier or diluent. Use of a compound of formula (I) according to claim 14 or 15 optionally in combination with a GABA _B receptor agonist for the manufacture of a medicament for the treatment of gastroesophageal reflux disease (GERD). Use of a compound of formula (I) according to claim 14 or 15 optionally in combination with a GABA _B receptor agonist for the manufacture of a medicament for preventing reflux. Use of a compound of formula (I) according to claim 14 or 15 optionally in combination with a GABA _B receptor agonist for the manufacture of a medicament for inhibiting transient lower esophageal sphincter relaxation (TLESR). Use of a compound of formula (I) according to claim 14 or 15 optionally in combination with a GABA _B receptor agonist for the manufacture of a medicament for the treatment of functional gastrointestinal disorders. The use of claim 20, wherein the functional gastrointestinal disorder is functional dyspepsia. Use of a compound of formula (I) according to claim 14 or 15 optionally in combination with a GABA _B receptor agonist for the manufacture of a medicament for the treatment of irritable bowel syndrome (IBS). 23. The use of claim 22, wherein the IBS is constipation predominant IBS. Use according to claim 22, wherein the IBS is diarrhea predominant IBS. 23. The use of claim 22 wherein the IBS is an alternating intestinal motility predominant type IBS. A gastroesophageal reflux comprising administering a pharmaceutically and pharmacologically effective amount of a compound of formula (I) according to claim 14 optionally in combination with a GABA _B receptor agonist to a subject in need of treatment of gastroesophageal reflux disease (GERD) Method of treatment of the disease. A method of preventing reflux, comprising administering to a subject in need thereof a pharmaceutically and pharmacologically effective amount of a compound of formula (I) according to claim 14 optionally in combination with a GABA _B receptor agonist. A temporary, lower dose comprising administering to the subject in need of inhibition of temporary lower esophageal sphincter relaxation (TLESR) a pharmaceutical and pharmacologically effective amount of a compound of formula (I) according to claim 14 optionally combined with a GABA _B receptor agonist. Method of suppressing esophageal sphincter relaxation. A method of treating a functional gastrointestinal disorder comprising administering to a subject in need thereof a pharmaceutical and pharmacologically effective amount of a compound of formula I according to claim 14 optionally in combination with a GABA _B receptor agonist. The method of claim 29, wherein the functional gastrointestinal disorder is functional dyspepsia. A pharmaceutical and pharmacologically effective amount of a compound of formula (I) according to claim 14, optionally in combination with a GABA _B receptor agonist, comprising administering to a subject in need of treatment of irritable bowel syndrome (IBS) Method of treatment. 32. The method of claim 31, wherein said IBS is constipation predominant IBS. 32. The method of claim 31, wherein said IBS is diarrhea predominant IBS. 32. The method of claim 31, wherein the IBS is alternating bowel movement predominant IBS. The following intermediate III is prepared by preparing the following intermediate IV from dimethylcyanodithioimidocarbonate and substituting the thiomethoxy group in the intermediate IV with the corresponding C ₁ -C ₇ alkoxy group; The following aminoimidazole II is prepared from intermediate III or IV by heating the reagent with an alpha halo carbonyl compound under basic conditions, and acyl chloride is used to acylate the aminoimidazole of formula II, Method for preparing the compound of I. <Formula II>

<Formula III>

<Formula IV>

In the above formula, R ¹ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups) ; Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ² is C ₁ -C ₁₀ alkoxy or C ₁ -C ₁₀ thioalkoxy (each optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carbo) Acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ³ is C ₁ -C ₁₀ alkoxy (optionally C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide , Nitrile or substituted with 1 or 2 aryl or heteroaryl groups); C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ Thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C Substituted with _1- C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or heteroaryl groups; Or amino (optionally mono- or di-substituted with C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl or C ₃ -C ₁₀ cycloalkyl); Here, each alkyl, alkenyl, alkynyl and cycloalkyl may independently have one or more carbon atom (s) substituted by O, N or S. 1H-imidazole-5-carboxylic acid, 4-amino-1-methyl-2- (methylthio)-, ethyl ester; Ethyl-4-amino-1- (2-furylmethyl) -2- (methylthio) -1H-imidazole-5-carboxylate; Ethyl-4-amino-1- (4-isopropylphenyl) -2- (methylthio) -1H-imidazole-5-carboxylate; Ethyl-4-amino-1- (2-methoxyphenyl) -2- (methylthio) -1H-imidazole-5-carboxylate; Metanon, [4-amino-1-methyl-2- (methylthio) -1H-imidazol-5-yl] phenyl; Metanon, [4-amino-2- (methylthio) -1-phenyl-1H-imidazol-5-yl] phenyl-; 1H-imidazole-5-carboxylic acid, 4-amino-2- (methylthio) -1-phenyl-, ethyl ester; 1H-imidazole-5-carboxylic acid, 4-amino-1-methyl-2- (methylthio)-, ethyl ester; And 1H-imidazole-5-carboxylic acid, 4-amino-2- (methylthio) -1- (2-propenyl)-, ethyl ester Except for the compound of formula II. <Formula II>

In the above formula, R ¹ is C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (each optionally and independently C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups) ; Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ² is C ₁ -C ₁₀ alkoxy or C ₁ -C ₁₀ thioalkoxy (each optionally and independently C ₁ -C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, carboxylic acid, carboxylic acid ester, carbo) Acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups; R ³ is C ₁ -C ₁₀ alkoxy (optionally C ₁ -C ₁₀ thioalkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide , Nitrile or substituted with 1 or 2 aryl or heteroaryl groups); C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, or C ₃ -C ₁₀ cycloalkyl (optionally and independently C ₁ -C ₁₀ alkoxy, C ₁ -C ₁₀ thio, respectively) Alkoxy, C ₃ -C ₁₀ cycloalkyl, keto, halogen, hydroxy, mercapto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or substituted with 1 or 2 aryl or heteroaryl groups); Aryl or heteroaryl (each optionally and independently C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C ₁₀ alkoxy, C Substituted with _1- C ₁₀ thioalkoxy, halogen, hydroxy, mercapto, nitro, keto, carboxylic acid, carboxylic ester, carboxylic acid amide, nitrile or 1 or 2 aryl or heteroaryl groups; Or amino (optionally mono- or di-substituted with C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₂ -C ₁₀ alkynyl or C ₃ -C ₁₀ cycloalkyl); Here, each alkyl, alkenyl, alkynyl and cycloalkyl may independently have one or more carbon atom (s) substituted by O, N or S.