WO1994024151A1

WO1994024151A1 - 1,5 benzodiazepine derivatives having cck and/or gastrin antagonistic activity

Info

Publication number: WO1994024151A1
Application number: PCT/EP1994/001130
Authority: WO
Inventors: Harry Finch; Pritom Shah; Robin Arthur Carr
Original assignee: Glaxo Inc.
Priority date: 1993-04-15
Filing date: 1994-04-13
Publication date: 1994-10-27
Also published as: HU9502977D0; AU6567594A; CZ267695A3; FI954852A; NO954091L; JPH08508743A; CN1120845A; AU688316B2; NZ265271A; CA2158972A1; HUT74091A; NO954091D0; SK125495A3; FI954852A0; EP0694040A1; PL311083A1; OA10236A

Abstract

Compounds of general formula (I) and physiologically acceptable salts thereof wherein the group NR1R2 represents a 5-7 membered saturated heterocyclic ring which may be substituted by one or two methyl groups; R3 is C1-6alkyl, C3-6cycloalkyl or phenyl optionally substituted by 1 or 2 halogen atoms; R4 is phenyl or phenyl substituted by one or two groups selected from halogen, C1-4alkyl, trifluoromethyl, trifluoromethoxy or (CH2)nR5 wherein n is zero or 1 and R5 represents C1-4alkoxy, hydroxy, nitro, cyano, CO2R6, S(O)pCH3, NR7R8, CONR7R8, SO2NR7CO(C1-4alkyl), tetrazolyl, carboxamidotetrazolyl, or a 3-trifluoromethyl 1,2,4-triazolyl; R6 is hydrogen, C1-4alkyl or benzyl; R7 is hydrogen or C1-4alkyl, R8 is hydrogen, C1-4alkyl, SO2CH3 or SO2CF3, X represents hydrogen, C1-4alkyl or halogen; m is zero, 1 or 2, and p is zero, 1 or 2; processes for their preparation and their use in medicine as antagonists of gastrin and CCK.

Description

ENZODIAZEPINE DERIVATIVES HAVING CCK AND/OR GASTRIN ANTAGONISTIC ACTIVITY

This invention relates to novel 1 ,5-benzodiazepine derivatives, to processes for their preparation, to pharmaceutical compositions containing them and to their use in medicine.

Cholecystokinins (CCK) and gastrin are structurally related peptides which exist in gastrointestinal tissue and in the central nervous system. Cholecystokinins include CCK-33, a neuropeptide of thirty-three amino acids in its originally isolated form, its carboxy terminal octapeptide, CCK-8 (also a naturally- occurring neuropeptide), and 39- and 12-amino acid forms. Gastrin occurs in 34-, 17- and 14- amino acid forms, with the miniumum active sequence being the C-terminal tetrapeptide, Trp-Met-Asp-Phe-NH₂(CCK-4), which is the common structual element shared by both CCK and gastrin.

CCK and gastrin are gastrointestinal hormones and neurotransmitters in the neural and peripheral systems and perform their respective biological roles by binding to particular receptors located at various sites throughout the body.

There are at least two subtypes of cholecystokinin receptors termed CCK-A and CCK-B and both are found in the periphery and in the central nervous system. CCK and gastrin receptor antagonists have been disclosed for preventing and treating CCK-related and/or gastrin related disorders of the gastrointestinal and central nervous systems of animals, and more particularly humans.

US Patent No. 4,988,692 describes a group of 3-acyiamino 1 -alkyl-5-phenyl 1 ,5- benzodiazepine derivatives as cholecystokinin antagonists. Further the specification teaches that the compounds have a significantly greater affinity for the CCK-A receptor over the CCK-B receptor.

We have now found a novel group of 3-ureido 1 ,5-benzodiazepine compounds which are potent and specific antagonists of gastrin and/or CCK and in particular antagonists of gastrin and /or CCK at the CCK-B receptor. The present invention thus provides compounds of the general formula (I)

and physiologically acceptable salts thereof wherein the group NR-_| R₂ represents a 5-7 membered saturated heterocylic ring which may be substituted by one or two methyl groups; R₃ is C_hal y I, C₃.₆cycloalkyl or phenyl optionally substituted by 1 or 2 halogen atoms;

R is phenyl or phenyl substituted by one or two groups selected from halogen, Cι_₄alkyl, trifluoromethyl, trifluoromethoxy or (CH₂)_nR₅ wherein n is zero or 1 and R₅ represents C^alkoxy, hydroxy, nitro, cyano, C0₂R₆, S(0)_pCH₃, NR₇R₈, CONR₇R₈ , S0₂NR₇CO(C₁.₄alkyl), tetrazolyl, carboxamidotetrazolyl, or a 3- trifluoromethyl 1 ,2,4-triazolyl;

R₆ is hydrogen, C₁.₄alkyl or benzyl;

R₇ is hydrogen or C-^alkyl,

R₈ is hydrogen, C₁.₄alkyl, S0₂CH₃ or S0₂CF₃

X represents hydrogen, C-,_₄alkyl or halogen; m is zero, 1 or 2, and p is zero, 1 or 2.

The compounds of the invention possess at least one asymmetric carbon atom and the compounds of the invention include both enantiomers and mixtures thereof including the racemate.

The term alkyl as used herein refers to both straight chain and branched chain alkyl groups. For example Chalky! includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl or hexyl. The term halogen includes fluorine, chlorine, bromine or iodine.

When R₅ represents a tetrazolyl, carboxamidotetrazolyl or 3-trifluoromethyl 1 ,2,4-triazolyl group these are linked to the rest of the molecule via a carbon atom therein and the invention includes all tautomers thereof and the C-,_₄alkyl -N substituted derivatives thereof. Examples of such groups include (1 H) tetrazol-5-yl, carboxamido-1 H-tetrazol-5-yl, 2-methyltetrazol-5-yl and (3- trifluoromethyl-1 ,2,4-triazol-5-yl.

The group NR₁ R₂ is linked to the rest of the molecule via the nitrogen thereof examples of such groups include pyrrolidino, piperidino, hexamethylenimino-, 2,5-dimethylpyrrolidino, 3,3-dimethylpiperidino, 2,6-dimethylpiperidino or 4,4- dimethylpiperidino.

When R₃ represents phenyl optionally substituted by halogen examples of such groups include phenyl optionally substituted by fluorine e.g. phenyl or 2- fluorophenyl, or 4-fluorophenyl.

When R₃ represents C₃.₆cycloalkyl examples of such groups include cyclopropyl, cyclopentyl or cyclohexyl.

When R₃ represents C^alkyl examples of such groups include methyl, ethyl, propyl, butyl, 3-methylbutyl or 3,3-dimethylbutyl.

Examples of suitable groups R₄ include phenyl optionally substituted by halogen e.g. fluorine, alkyl e.g. methyl, alkoxy e.g. methoxy, nitro, cyano, thiomethyl, carboxamido, carboxyl, dimethylamino, cyanomethyl, 1 H-tetrazol-5-yl, carboxy methyl, or N-methanesulphonylcarboxamido.

A preferred class of compound according to the invention are those wherein the group NR-_|R₂ represent pyrrolidino, piperidino, 3,3-dimethylpiperidino 4,4- dimethylpiperidino, 2,6-dimethylpiperidino or 2,5-dimethylpyrrolidino. Within this class the group NR-,R₂ is conveniently pyrrolidino, piperidino, or 3,3- dimethylpiperidino. The group X is conveniently halogen e.g. bromine, fluorine or fluorine or more particularly hydrogen.

A further preferred class of compounds of formula (I) are those wherein R₃ is phenyl, 2-fluorophenyl or cyclohexyl and more particularly 2-fluorophenyl or cyclohexyl.

Another preferred class of compounds of formula (I) are those wherein R₄ is phenyl or phenyl substituted by methyl e.g. 3-methylphenyl or 3,5- dimethylphenyl, 3-dimethylaminophenyl, phenyl substituted by fluorine e.g. 4- fluorophenyl, phenyl substituted by methoxy e.g. 3-methoxyphenyl or 4- methoxyphenyl, 3-nitrophenyl, 3-cyanomethylphenyl, 3-carboxamidophenyl, 3- carboxyphenyl, 3-carboxymethylphenyl, or 3-(1 H)-tetrazol-5-ylphenyl.

A particularly preferred group of compounds according to the invention are those wherein NR-|R₂ represents pyrrolidino, piperidine or 3,3-dimethyl-, piperidino, R₃ represents 2-fluorophenyl or cyclohexyl, and X represents a hydrogen atom. Within this group particularly preferred compounds include those wherein R₄ is phenyl, 4-fluorophenyl, 3-dimethylaminophenyl, 3-carboxyphenyl, 3-carboxymethylphenyl or 3-(1 H)-tetrazolyl-5-yl-phenyl.

A particularly preferred compound of the invention is:-

1 -[1 -Cyclohexyl-2,4-dioxo-5-(2-oxo-2-pyrrolidin-1 -yl-ethyl)-2,3,4,5-tetrahydro-

1 H-benzo[b][1 ,4]diazepin-3-yl]-3-(4-fluoro-phenyl)urea and enantiomers thereof.

Further preferred compounds of the invention include

3-{3-[1-Cyclohexyl-2,4-dioxo-5-(2-oxo-2-pyrrolidin-1-yl-ethyl)-2,3,4,5-tetrahydro-

1 H-benzo[b][1 ,4]diazepin-3-yl]-ureido}-benzoic acid;

(3-{3-[1 -(2-Fluoro-phenyl)-2,4-dioxo-5-(2-oxo-2-pyrrolidin-1-yl-ethyl)-2,3,4,5- tetrahydro-1 H-benzo[b][1 ,4]diazepin-3-yl]-ureido}-phenyl)-acetic acid;

3-{3-[1-[2-(3,3-Dimethyl-piperidin-1 -yl)-2-oxo-ethyl]-5-(2-fluoro-phenyl)-2,4- dioxo-2,3,4,5-tetrahydro-1 H-benzo[b][1 ,4]diazepin-3-yl]-ureido}-benzoic acid , and enantiomers thereof. The physiologically acceptable salts of the compounds of formula (I) include conventional salts formed for example from pharmaceutically acceptable inorganic or organic acids as well as quaternary ammonium acid addition salts. Examples of suitable salts include hydrochloric, hydrobromic, sulfuric, phosphoric, nitric, perchloric, fumaric, acetic, propionic, succinic, glycolic, formic, lactic, maleic, tartaric, citric, pamoic, malonic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, fumaric, toluenesulphonic, methanesulphonic, naphthalene-2-sulphonic, benzenesulphonic and the like. Other acids such as oxalic, while not in themselves pharmaceutically acceptable, may be useful in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable salts.

References hereinafter to a compound according to the invention includes both compounds of formula (I) and their pharmaceutically acceptable salts and solvates.

Compounds of the invention modulate the effect of gastrin and/or CCK in mammals. In particular compounds of the invention are antagonists of gastrin and/or CCK.

Compounds of the invention have been shown to be antagonists of CCK, particularly at CCK-B receptors as demonstrated for example by the compound's ability to inhibit the contractile actions of CCK-4 in the presence of a CCK-A antagonist, in the guinea-pig isolated ileum longitudinal muscle- myenteric plexus.

The preparation and use of guinea-pig isolated ileum longitudinal muscle- myenteric plexus has been described by K-H Buchheit et al in Nauyn- Schmeideberg's Arch. Pharmacol., (1985), 329, p36-41 and by V.L. Lucaites et al in J. Pharmacol. Exp. Ther., (1991 ) 256, 695-703.

Compounds of the invention have also been shown to have a greater affinity for the CCK-B receptor than for the CCK-A receptor. This may be determined using the CCK receptor binding assays described by Fornos et al J. Pharmacol Exp. Ther., 201 1056-1063, 1992. Alternatively the binding affinity of compounds of the invention for CCK-A and CCK-B receptors may be determined using HeLa cell membranes that are transfected with the human CCK-B receptor or, COS-M6 cell membranes that are transiently transfected with the human CCK-A receptor.

Compounds of the invention have also been shown to be antagonists of gastrin as demonstrated by their ability to inhibit pentagasthn-stimulated acid secretion from rat isolated gastric mucosa using the procedure described by J.J. Reeves and R. Stables in Br. J. Pharmac. 1985 8_£, p.677-684.

Compounds of the invention have also been shown to inhibit pentagastrin stimulated acid secretion in conscious gastric fistula rats using the methods described by Hedges and Parsons Journal of Physiology 1977, 267. 181-194.

The compounds of the invention are therefore useful for the treatment and/or prevention of disorders in mammals, especially humans, where modification of the effects of gastrin or CCK is of therapeutic benefit. Thus the compounds of the invention are useful for the treatment of gastrointestinal disorders especially those where there is an advantage in lowering gastric acidity. Such disorders include peptic ulceration, reflux oesophagitis and Zollinger Ellison syndrome. They may also be useful for the treatment of gastrointestinal disorders such as irritable bowel syndrome, excess pancreatic secretion, acute pancreatitis, motility disorders, antral G cell hyperplasia, fundic mucosal hyperplasia or gastrointestinal neoplasms. The compounds of the invention are also useful for the treatment of central nervous system disorders where CCK and/or gastrin are involved. For example anxiety disorders (including panic disorder, agoraphobia, social phobia, simple phobia, obsessive compulsive disorders, post traumatic stress disorder, and general anxiety disorder), depression, tardive dyskinesia, Parkinson's disease or psychosis. They may also be useful for the treatment of dependency on drugs or substances of abuse and withdrawal, Gilles de la Tourette syndrome, or dysfunction of appetite regulatory systems; as well as the treatment of certain tumours of the lower oesophagus, stomach, intestines and colon. Compounds of the invention are also useful for directly inducing analgesia, or enhancing opiate or non-opiate mediated analgesia, as well as anaesthesia or loss of the sensation of pain. The invention therefore provides a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof for use in therapy, in particular in human medicine.

According to another aspect the invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof for the manufacture of a medicament for the treatment of conditions where modification of the effects of gastrin and/or CCK is of therapeutic benefit.

According to a further aspect of the invention we provide a method for the treatment of a mammal, including man, in particular in the treatment of conditions where modification of the effects of gastrin and/or CCK is of therapeutic benefit which method comprises administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof to the patient.

It will be appreciated by those skilled in the art that reference herein to treatment extends to prophylaxis as well as the treatment of established diseases or symptoms.

It will further be appreciated that the amount of a compound of the invention required for use in treatment will vary with the nature of the condition being treated and the age and the condition of the patient and will be ultimately at the discretion of the attendant physician or veterinarian. In general however doses employed for adult human treatment will typically be in the range of 1-2000mg per day e.g 10-500mg per day.

The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example as two, three, four or more sub-doses per day.

Compounds of the invention which antagonise the function of CCK in animals, may also be used as feed additives to increase the food intake in animals in daily dosages of around 1 mg/kg to 10mg/kg. While it is possible that, for use in therapy, a compound of the invention may be administered as the raw chemical it is preferable to present the active ingredient as a pharmaceutical formulation.

The invention thus further provides a pharmaceutical formulation comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with one or more pharmaceutically acceptable carriers therefor and, optionally, other therapeutic and/or prophylactic ingredients. The carrier(s) must be 'acceptable' in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The compositions of the invention include those in a form especially formulated for oral, buccal, parenteral, implant, or rectal administration. Oral administration is preferred.

Tablets and capsules for oral administration may contain conventional excipients such as binding agents, for example, syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch or polyvinylpyrrolidone; fillers, for example, lactose, sugar, microcrystalline celluiose, maize-starch, calcium phosphate or sorbitol; lubricants, for example, magnesium stearate, stearic acid, talc, polyethylene glycol or silica; disintegrants, for example, potato starch or sodium starch glycollate, or wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in the art. Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions emulsions, syrups or elixirs, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example, sorbitol syrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenated edible fats; emulsifying agents, for example, lecithin, sorbitan mono-oleate or acacia; non-aqueous vehicles (which may include edible oils), for example, almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; and preservatives, for example, methyl or propyl p.- hydroxybenzoates or ascorbic acid. The compositions may also be formulated as suppositories, e.g. containing conventional suppository bases such as cocoa butter or other glycerides.

For buccal administration the composition may take the form of tablets or lozenges formulated in conventional manner.

The composition according to the invention may be formulated for parenteral administration by injection or continuous infusion. Formulations for injection may be presented in unit dose form in ampoules, or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilising and/or dispersing agents. Alternatively the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use.

The composition according to the invention may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus for example, the compounds of the invention may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

The compositions according to the invention may contain between 0.1 - 99% of the active ingredient, conveniently from 30-95% for tablets and capsules and 3- 50% for liquid preparations.

Compounds of general formula (I) and salts thereof may be prepared by the general methods outlined hereinafter. In the following description, the groups R .-R₈ ^ancl * ^{are as} defined for the compounds of formula (I) unless otherwise stated.

According to a first general process A, compounds of formula (I) may be prepared by the reaction of an amine of formula (II) wherein R_{1 f} R₂, R₃ , X and m have the meanings defined in formula (I).

with a compound R₄Y, wherein R Y is an isocyanate of formula (III), carbamoyi chloride of formula (IV), imidazolide of formula (V), or an optionally substituted phenyl carbamate of formula (VI) wherein R_a is an optionally phenxoy group.

R a CONHR 4_Λ

(VI)

The reaction conveniently takes place in the presence of a suitable solvent such as a halohydrocarbon (e.g. dichloromethane), an ether (e.g. tetrahydrofuran) or nitrile (e.g. acetronitrile) or a mixture thereof at a temperature in the range of 0°-80°C.

Isocyanates of formula (III) may be purchased or prepared by the reaction of amines H₂N-R₄ with phosgene or triphosgene in a suitable solvent such as methylene chloride. Carbamoyi chlorides of formula (IV) are also prepared by the reaction of amines H₂NR₄ with phosgene or triphosgene in a suitable solvent such as methylene chloride. Imidazolides of formula (VI) are prepared by treatment of amines H₂N-R₄ with carbonyl diimidazole in a suitable solvent (dichloromethane, ether, tetrahydrofuran) at a temperature ranging from 0-80° C (conveniently at room temperature). Optionally substituted phenyl carbamates of formula (VI) are prepared by the reaction of amines H₂N-R₄ with the appropriate chloroformate R_aCOCI in the presence of a base (pyridine, triethylamine) in a suitable solvent (dichloromethane) and at a temperature of 0 - 50° C.

According to a further general process B, compounds of formula (I) may be prepared by reaction of an intermediate of formula (VII).

wherein Y is the group -NCO, -NHCOCI or NHCOR_a wherein R_a is an optionally substituted phenoxy group or a 1-imidazole group with an amine (VIII)

H₂N-R₄ (VIII)

and optionally in the the presence of a base such as a tertiary amine (e.g. triethylamine).

The reaction conveniently takes place in a suitable solvent such as a halogenated hydrocarbon (e.g. dichloromethane) or an ether (e.g. tetrahydrofuran) or an amide (e.g. N,N-dimethyl formamide) optionally at a temperature ranging from room temperature to the reflux temperature of the solvent.

Conveniently the compound of formula (VII) are prepared in situ from the amine (II).

In a particular aspect of the process (B) when Y is the group NHCOR_a and R_a is a 1-imidazole group, the imidazolide (VII) may be formed in situ in which case the amine of formula (VIII) will be mixed with the compound of formula (II)

in the presence of carbonyidiimidazole under the aforementioned conditions.

For process B when Y is the group NHCOR_a and R_a is optionally substituted phenoxy group the reaction with the primary amine (VIII) is preferably carried out in the presence of a base such as a tertiary amine e.g. triethylamine.

For process B when Y is the isocyanate group -N=C=0 the reaction with the primary amine (VIII) is preferably carried out in an aprotic solvent such as a halohydrocarbon e.g. methylene chloride. Conveniently the isocyanate is generated in situ prior to the addition of the primary amine (VIII).

The compounds of formula (VII) wherein R_a is an optionally substituted phenoxy group may be prepared from the primary amine (II) by reaction with the corresponding optionally substituted phenyl chloroformate in the presence of a base such as pyridine. The reaction may be carried out in a solvent such as a halohydrocabon e.g. dichloromethane and at a temperature from 0-50°.

Compounds of formula (VII) wherein R_a is a 1-imidazole group may be prepared by reacting a compound of formula (II) with carbonyidiimidazole in the presence of a suitable solvent such as a halogenated hydrocarbon (e.g. dichloromethane) or an ether (e.g. tetrahydrofuran) at a temperature ranging from 0° to 80° (conveniently at room temperature).

Compounds of formula (VII) wherein Y is the isocyanate grouping -N=C=0 or carbamoyi chloride -NHCOCI may be prepared from the primary amine (II) by reaction with phosgene (COCI2) or triphosgene in a suitable solvent such as methylene chloride. According to a further general process C compounds of formula (I) may also be prepared by a reaction of the compound of formula (IX)

with the halide R₂R-_|NCOCH₂Z wherein Z = a leaving group e.g. bromine, chlorine or iodine.

The reaction is conveniently carried out by treating the compound of formula (IX) with a strong base such as sodium hydride in a polar aprotic solvent such as N,N-dimethylformamide followed by reaction with the alkylating agent F^ RgNCOCHgZ.

Compounds of formula (II) may be prepared by reduction of compounds of formula (X)

wherein W is CH-N3 or C=N-NHPh.

Compounds of formula (X) wherein W is CH-N3 may be reduced to a compound of formula (II) by hydrogenation in the presence of a suitable catalyst such as palladium, on a support such as carbon or calcium carbonate, or platinum (IV) oxide. The reaction conveniently takes place in the presence of a solvent such as an alkanol (e.g. ethanol) an ester (e.g. ethyl acetate) or acetic acid. Compounds of formula (X) wherein W is C=N-NHPh may be reduced to a compound of formula (II) by reaction with zinc and acetic acid. This reaction may be carried out a temperature with the range 0-50°.

Compounds of formula (X) wherein W is CHN3 may be prepared from a compound of formula (X) wherein W is CH2 by treatment with a strong base such as sodium hydride or potassium tert-butoxide followed by tri-isopropyl benzenesulphonyl azide. The reaction conveniently takes place in a solvent such as an ether (e.g. tetrahydrofuran) at a temperature in the range of -78° to 20°.

Compounds of formula (X) in which W is C=NNHPh may be prepared by reaction of the ortho-phenylenediamine (XI) with the diacid chloride (XII), in a suitable solvent such as an ether e.g. tetrahydrofuran

CONR.R-

I ^Λ

CH.

(XI)

Compounds of formula (X) wherein W is CH2 may be prepared by reaction of the compound of formula (XI) with the diacid chloride (XIII)

ClOC

(XIII)

Compounds of formula (XI) are either known compounds or may be prepared by analogous methods. Thus for example a compound of formula (XI) may be prepared by alkylation of the amine (XIV).

Thus the amine (XIV) may be reacted with the compound R-_| R₂NCOCH₂Z , in which Z is chlorine or bromine, optionally in the presence of sodium iodide in a solvent such as N,N-dimethylformamide.

In general, the compounds of formula (III), (IV), (V) and (VI) are either known compounds or may be prepared according to methods simlar to those used for the preparation of known compounds,

According to a further process (D) a compound of formula (I) may be prepared from the compound of formula (XV) wherein R is hydrogen

by reaction of an activated derivative thereof with the amine R₁R₂NH. Conveniently the reaction is carried out using the acid in the presence of a diimide such as dicyclohexyl carbodimide and hydroxybenzotriazole in a solvent such as dichloromethane or in the presence 2-ethoxy-1-ethoxycarbonyl-1 ,2- dihydroquinoline in a solvent such as dimethoxyethane.

The compounds of formula (XV) wherein R_b is a hydrogen may be prepared by hydrolysis of the corresponding compound of formula (XV) wherein R_b is a t- butyl group, for example by reaction with trifluoroacetic acid. The compound of formula (XV) wherein R_b is t-butyl may be prepared by alkylation of the corresponding compound of formula (IX) with the halo ester Z-CH₂C0₂R_b wherein Z is halogen. Alternatively the compound (XV) wherein R_b is t-butyl may be prepared using the general processes A and B described above for preparing the compounds of formula (I) but starting with the appropriate N- substituted benzodiazepine derivative.

According to a further process (E) a compound of formula (I) may be converted into another compound of formula (I) using conventional techniques.

Thus compounds of formula (I) wherein R is a phenyl group substituted by a carboxyl or carboxymethyl group may be prepared by hydrolysis of the corresponding compound of formula (I) wherein R₄ is a phenyl group substituted by an alkoxycarbonyl or alkoxycarbonylmethyl group.

Compounds of formula (IX) may be prepared from the diamine (XIV), in which the primary amino group is protected as an p-methoxybenzyl derivative thereof, using the general procedures described above for preparing the compounds of formula (I) from the corresponding orthophenylenediamine (XI) followed by removal of the N-protecting group p-methoxybenzyl using conventional procedures.

Compounds of formula (I) contain at least one asymmetric carbon atom, namely the carbon atom of the diazepine ring to which the substituted urea grouping is attached. Specific enantiomers of the compounds of formula (I) may be obtained by resolution of the racemic compound using conventional procedures such as chiral HPLC. Alternatively the required enantiomer may be prepared from the corresponding enantiomeric amine of formula (II) using any of the processes described above for preparing compounds of formula (I) from the amine (II). The enantiomers of the amine (II) may be prepared from the racemic amine (II) using conventional procedures such as salt formation with a suitably optically active acid such as R- camphorsulphonic acid or by preparative chiral HPLC.

In order that the invention may be more fully understood the following examples are given by way of illustration only.

In the Intermediates and Examples unless otherwise stated. Melting points (mp) are determined on a Gallenkamp mp apparatus and are uncorrected. All temperatures refers to °C. Column chromatography was carried out over silica gel. T.l.c. refers to thin layer chromatography on silica plates. Dried refers to solutions dried ever anhydrous sodium sulphate. The following abbreviation are also used in the Examples. EA = ethyl acetate, DMF = N,N-dimethylformamide, THF = tetrahydrofuran, DE = diethyl ether, DCM = dichloromethane, MeOH = methanol, AcOH = acetic acid, ee = enantiomeric excess, R_τ = retention time.

Intermediate 1

Cvclohexvl-.2-nitro-phenvn-amine.

A mixture of 2-chloronitrobenzene (20g), potassium carbonate (35g) and copper (I) iodide (1 -21 g) in cyclohexylamine (43.6ml) was heated at 150° under nitrogen for 18h. The mixture was allowed to cool to room temperature and was adsorbed onto silica. This was chromatographed with hexane-EA (98:2) as eluent to give the title compound (22.94g) as an orange solid. T.l.c. (98:2 hexane-EA) Rf 0.38

Intermediate 2

N-Cyclohexvl-benzene-1.2-diamine.

A solution of Intermediate 1 (10g) in ethyl acetate (400ml) was hydrogenated at 23° and 1 atm. pressure over 10% palladium on carbon (1g) for 4h. The catalyst was removed by filtration through hyflo and the filtrate evaporated to give the title compound (8.5g) as an orange oil. T.l.c.^'(9:1 hexane-EA) Rf 0.36

Intermediate 3

2-(2-Cyclohexylamino-phenylaminoV1-pyrrolidin-1-yl-ethanone. A mixture of Intermediate 2 (8.5g), 2-oxo-2-(pyrrolidin-1-yl)ethylbromide (9.4g) and potassium carbonate (18.5g) in dry DMF (250ml) was stirred at 23° under nitrogen for 18h. The mixture was poured into water (600ml) and extracted with ethyl acetate (3x300ml). The combined extracts were washed with water (3x300ml) and saturated brine (200ml), dried and evaporated to give a brown oil. This was chromatographed with hexane-EA (1 :1) as eluent to give the title compound (9.8g) as a cream solid, m.p.108-110°. T.l.c. (1 :1 hexane-EA) Rf 0.42 Intermediate 4

1 -Cvclohexyl-5-(2-oxo-2-pyrrolidin-1 -vl-ethyl.-3-.phenyl-hvdrazono V1.5-dihvdro- benzo[b]ri .4]diazepine-2.4-dione.

Solutions of Intermediate 3 (9.8g) and 2-(phenyl-hydrazono)-propanedioyl dichloride (8.36g) in dry THF (75ml) were added at equal rates to dry THF (75ml) cooled to -10° under nitrogen. When the addition was complete the mixture was allowed to warm to room temperature and stir for 3h. The solvent was removed by evaporation to give the crude title compound (19.5g) as a yellow crunchy foam. T.l.c. (DE) Rf 0.23

Intermediate 5

3-Amino-1 -cvclohexyl-5-(2-oxo-2-pyrrolidin-1 -yl-ethyl V1.5-dihydro- benzo[b][1.4]diazepine-2.4-dione. A solution of Intermediate 4 (19g) in glacial acetic acid (200ml) was added dropwise to a stirring suspension of zinc dust (15g) in glacial acetic acid. The mixture was stirred at 23° for 1.5h whereupon the zinc was removed by filtration through hyflo and the filtrate evaporated to give a red oil. This was partitioned between water (200ml) and ethyl acetate (75ml). The aqueous portion was adjusted to pH9 with solid Na2Cθ3 and extracted with ethyl acetate (4x100ml). The combined organic extracts were dried and evaporated to give a brown oil which was chromatographed with DCM-MeOH (95:5) as eluent to give the title compound (7.4g) as a pink crunchy foam. T.l.c. (95:5 DCM-MeOH) Rf 0.26

Intermediate 6

3-{3-[1 -Cvclohexyl-2.4-dioxo-5-.2-oxo-2-pyrrolidin-1-yl-ethyl.-2.3.4.5-tetrahvdro- 1 H-benzo[b][1.4]diazepin-3-vl]-ureido}-benzoic acid.benzyl ester. Triethylamine (108μl) and triphosgene (77mg) were added sequentially to a solution of 3-amino-benzoic acid benzyl ester (177mg) in dry THF (10ml) at 0° under nitrogen. More triethylamine (108μl) was added and stirring continued at 0° for 30min. A solution of Intermediate 5 (250mg) in dry THF (10ml) was added and stirring continued at 23° for 18h. The mixture was then partitioned between 2N hydrochloric acid (50ml) and ethyl acetate (50ml). The dried organic extract was evaporated and the residue chromatographed with EA-hexane (2:1 ) as eluent to give the title compound (320mg) as a paie yellow solid.

T.l.c. (2:1 EA-hexane) Rf 0.17 l.r. (Solution in CHCI₃) 3384;2939;1690;1658;1423cm^"1

Intermediate 7

3-Nitro-benzoic acid tert-butyl ester.

A solution of 3-nitrobenzoic acid (1.4g) in dry toluene (10ml) was treated with

N,N-dimethylformamide di-tertbutyl acetal (10ml). The mixture was heated under reflux for 18h then cooled, diluted with ethyl acetate and washed consecutively with water, 8% sodium bicarbonate solution and saturated brine. The dried organic phase was evaporated to give the Jtili≤. compound (1.02g) as a yellow oil.

T.l.c. (97:3 DCM-MeOH) Rf 0.66

Intermediate 8 3-Amino-benzoic acid tert-butyl ester.

A solution of Intermediate 7 (1g) in ethanol (20ml) was hydrogenated at 23° and

1 atm pressure in the presence of 10% palladium on carbon as catalyst (200mg). After 2h the mixture was filtered through hyflo and the filtrate evaporated. The residue was chromatographed with DCM-MeOH (97:3) as eluent to give the title compound (727mg) as a colourless oil.

T.l.c. (97:3 DCM-MeOH) Rf 0.35

Intermediate 9

.3- Nitro-phenvl.-acetic acid benzvl ester.

A mixture of 3-nitrophenylacetic acid (5g), benzyl alcohol (2.9ml) and 4,4- dimethylaminopyridine (300mg) in dry dichloromethane (50ml) was treated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (7.4g). The resulting yellow mixture was stirred at 23° for 3days whereupon the solvent was removed in vacuo. The residue was partitioned between water (100ml) and ethyl actate (100ml) and the organic phase washed with water (2x100ml) and saturated brine (100ml) then dried and evaporated. The residue was chromatographed with hexane-EA (4:1 ) as eluent then again with isohexane-DCM (1 :1 ) as eluent to give the title compound (2.05g) as a pale yellow oil. T.l.c. (4:1 hexane-EA) Rf 0.35

Intermediate 10

(3- Amino-phenyl.-acetic acid benzyl ester.

A solution of Intermediate 9 (2.05g) in ethyl acetate (100ml) was hydrogenated at 23° and 1atm pressure in the presence of 5% platinum on carbon (200mg) as catalyst. After 90min the mixture was filtered through hyflo and the filtrate evaporated to give the title compound (1.77g) as a pale yellow oil

T.l.c. (DCM) Rf 0.35

Intermediate 11

(2-Fluoro-phenylW2-nitro-phenyl.-amine.

A mixture of 2-fluoroaniline (5.0g), potassium carbonate (2.5g) and copper(l)iodide (414mg) in 2-fluoronitrobenzene (16.9ml) was heated to 180° under nitrogen for 18h. The cooled mixture was poured into water (300ml) and extracted with ethyl acetate (2x250ml) then the combined extracts were washed with saturated brine and evaporated. The residual brown oil was azeotroped with ethanol/water then toluene. The residue was chromatographed with hexane-DE (100:0 to 95:5) to give the title compound (3.25g) as a bright orange solid, m.p.58-9°.

T.l.c.(95:5 hexane-DE) Rf 0.45

Intermediate d

N-.2-Fluoro-phenvl.-benzene-1.2-diamine. A solution of Intermediate 11 (15.6g) in ethyl acetate (400ml) was hydrogenated at 23° and 1atm pressure in the presence of 5% platinum on carbon (2g) as catalyst. After 1 h the mixture was filtered through hyflo and the filtrate evaporated to give the title compound (13.45g) as a yellow solid.

T.l.c. (9:1 hexane-DE) Rf 0.25

Intermediate 13

2-[2-.2-Fluoro-phenvlamino.-phenvlamino1-1-pvrrolidin-1-vl-ethanone.

A solution of 2-oxo-2-(pyrrolidin-1-yl)ethylbromide (12.8g) in dry DMF (60ml) was added dropwise to a mixture of Intermediate 12 (13.45g) and potassium carbonate (27.5g) in dry DMF (100ml) at 23° under nitrogen. The mixture was stirred at 60° for 4h then poured into 2N sodium carbonate solution (500ml) and extracted with ethyl acetate (400ml). The organic extract was washed with water (2x250ml) and saturated brine (250ml), dried and evaporated. The residue was chromatographed with hexane-DE (50:50 to 0:100) as eluent to give the s. compound (14.12g) as a pale brown solid. T.l.c. (DE) Rf 0.53

Intermediate 14

1 -te-Fluoro-phenvn-5-.2-oxo-2-pvrrolidin-1 -yl-ethyl .-3-. phenvl-hvdrazono )-1.5- dihvdro-benzofbl[1.4]diazepine-2.4-dione.

Solutions of Intermediate 13 (14.12g) and 2-(phenyl-hydrazono)-propanedioyl dichloride (11.04g) in dry THF (100ml) were added simultaneously and dropwise at equal rates to dry THF (100ml) cooled to -10° under nitrogen.

When the addition was complete the mixture was allowed to warm to room temperature and stir for 3.5h. The solvent was removed by evaporation to give the crude title compound (23g) as a yellow crunchy foam.

T.l.c. (EA) Rf 0.5

Intermediate 15 3-Amino-1 - -fluoro-phenvl .-5-.2-oxo-2-pvrrolidin-1 -vl-ethvl.-1.5-dihvdro- benzo[b][1.4]diazepine-2.4-dione.

A solution of Intermediate 14 (23g) in glacial acetic acid (200ml) was added dropwise to a stirred suspension of zinc dust (22.2g) in glacial acetic acid (100ml) in a cold water-bath. The mixture was stirred at 23° for 2.5h whereupon the zinc was removed by filtration through hyflo and the filtrate evaporated. The residue was partitioned between water (150ml) and ethyl acetate (100ml) and the aqueous portion basified with solid Na2Cθ3- The layers were separated and the aqueous phase further extracted with ethyl acetate then the combined organic extracts were washed with saturated brine, dried and evaporated. The residue was chromatographed with EA-MeOH (100:0 to 95:5) as initial eluent followed by DCM-MeOH (80:20) to give the title compound (11.07g) as a fawn powder. T.l.c. (95:5 DCM MeOH) Rf 0.23 Intermediate 16 r3-{3-[1-.2-Fluoro-phenyl.-2.4-dioxo-5-f2-oxo-2-pyrrolidin-1-yl-ethyl.-2.3.4.5- tetrahydro-1 H-benzo[b1f1.4]diazepin-3-vl1-ureido}-phenvn-acetic acid benzyl ester. A solution of Intermediate 15 (3g) in dry dichloromethane (40ml) was added dropwise to a stirred solution of phosgene in toluene (60ml) under nitrogen. The resulting mixture was stirred at 23° for 4h then nitrogen was bubbled through the mixture into ammonia solution for 18h. The mixture was then evaporated and the residue dried at 60° in vacuo for 3h to give the intermediate isocyanate (3.3g) as a buff solid which was used without further characterisation. A solution of this isocyanate (400mg) in dry dichloromethane (3ml) was added dropwise to a stirred solution of (3- amino-phenyl)-acetic acid benzyl ester (241 mg) in dry dichloromethane (2ml) at 23° under nitrogen. After 60min the solvent was removed in vacuo and the residue triturated with diethyl ether. The solid was filtered off and dried to give the title compound (263mg) as an off-white solid, m.p.104-6°. T.l.c. (EA) Rf 0.54

Intermediate d 2-Bromo-1 -(3.3-dimethyl-piperidin-1 -yl ,-ethanone.

A mixture of 3,3-dimethylpiperidine (10g) and triethylamine (12.3ml) in dry dichloromethane (50ml) was added dropwise to an ice-cold solution of bromoacetyl bromide (7.7ml) in dry dichloromethane (100ml). The mixture was stirred at 23° for 18h then recooled to 0°. Iced water (200ml) was added and the layers separated. The aqueous layer was further extracted with dichloromethane (2x200ml) then the combined extracts were washed with 2N

HCI (200ml) and saturated brine (200ml), dried and evaporated to give the title compound (17.28g) as a brown oil.

T.l.c. (DE) Rf 0.49

Intermediate 18

1-f3.3-Dimethyl-piperidin-1-yl.-2-[2-^2-fluoro-phenylamino.-phenylamino]- ethanone.

A mixture of Intermediate 12 (3g)) and potassium carbonate (6.15g) in dry DMF (10ml) was treated with a solution of 2-bromo-1-(3,3-dimethyl-piperidin-1-yl)- ethanone (3.5g) in dry DMF (10ml) and stirred at 23° under nitrogen for 3 days. The mixture was poured into water and extracted with ethyl acetate. The combined extracts were washed with water and saturated brine, dried and evaporated to give a brown oil. This was chromatographed with DCM-MeOH (100:0 to 97:3) as eluent to give the title compound (3.17g) as a pale brown foam. T.l.c. (DCM) Rf 0.23

Intermediate 19 1-[2-r3.3-Dimethyl-piperidin-1-ylV2-oxo-ethyl]-5-r2-fluoro-phenyl.-3- henyl- hvdrazonoVI .5-dihvdro-benzo[b][1.4]diazepine-2.4-dione.

Solutions of Intermediate 18 (3.08g) and 2-(phenyl-hydrazono)-propanedioyl dichloride (2.12g) in dry THF (100ml) were added at equal rates to dry THF

(50ml) cooled to -10° under nitrogen. When the addition was complete the mixture was allowed to warm to room temperature and stirred for 2h. The solvent was removed by evaporation to give the crude title compound (5.25g) as a yellow crunchy foam.

T.l.c. (95:5 DCM-MeOH) Rf 0.68

Intermediate 20

3-Amino-1-r2-(3.3-dimethvl-piperidin-1-vl.-2-oxo-ethvll-5-(2-fluoro-phenvl.-1.5- dihvdro-benzo[b]f1.4]diazepine-2.4-dione.

A solution of Intermediate 19 (5.15g) in glacial acetic acid (75ml) was added dropwise to a stirring suspension of zinc dust (4.47g) in glacial acetic acid (50ml) in a cold water-bath. The mixture was stirred at 23° for 6h whereupon the zinc was removed by filtration through hyflo and the filtrate evaporated. The residue was partitioned between water and ethyl actate and the aqueous portion basified with solid Na2Cθ3. The organic extract was washed with saturated brine, dried and evaporated. The residue was chromatographed with DCM-MeOH (95:5) as eluent to give the title compound (2.38g) as a pale brown solid. T.l.c. (95:5 DCM-MeOH) Rf 0.18 Intermediate 21

3-{3-[1-[2-.3.3-Dimethyl-piperidin-1-yl.-2-oxo-ethyl]-5-f2-fluoro-phenylV2.4- dioxo-2.3.4.5-tetrahvdro-1 H-benzo[b][1.4]diazepin-3-yl]-ureido}-benzoic acid tert-butyl ester. Triethylamine (0.31 ml) and triphosgene (217mg) were added sequentially to a solution of 3-amino-benzoic acid tert-butyl ester (423mg) in dry THF (10ml) at 0° under nitrogen. More triethylamine (0.31 ml) was added and stirring continued at 0° for 30min. A solution of Intermediate 20 (800mg) in dry THF (10ml) was added and stirring continued at 23° for 18h. The mixture was then partitioned between phosphate buffer solution (pH6.5) and dichloromethane. The combined organic extracts were washed with saturated brine, dried and evaporated and the residue chromatographed with DCM-MeOH (97:3) as eluent to give a pale green solid which was triturated with DE-hexane (1 :1 ) to give the title compound (1.07g) as a white solid, m.p.170°. T.l.c. (97:3 DCM-MeOH) Rf 0.15

Example 1

1 -[1 -Cvclohexyl-2.4-dioxo-5-(2-oxo-2-pyrrolidin-1 -yl-ethyl ,-2.3.4.5-tetrahvdro-

1 H-benzo[b][1.4]diazepin-3-yl]-3-.4-fluoro-phenyl .urea. 4-Fluorophenyl isocyanate (650μl) was added to a solution of Intermediate 5

(2g) in dry dichloromethane (50ml) at 23° under nitrogen. The resulting mixture was stirred for 1 h then adsorbed onto silica and chromatographed with EA-

MeOH (100:0 to 95:5) as eluent to give the title compound (1.92g) as a white solid, m.p. 181-3°. T.l.c. (EA) Rf 0.42

The compound of Example 1 (1.7g) was separated into its 2 enantiomers (Isomer 1 and Isomer 2) by chiral HPLC.

Column: Chiralcel OD 25cm x 20mm id

Eluent: Hexane-EtOH (70:30)

Flow rate: 20 ml min^"1

Detection: uv <3> 254nm Isomer 1 , (452mg) as a white solid - R_τ 8.6 min. H.p.l.c. >99.5% ee T.l.c. (EA) Rf 0.42 l.r. (Solution in CHCI₃) 3622;3091 ;2938;2895;2403;1657;1515;1425;1189;

1047;929cm^"1

Isomer 2, (488mg) as a white solid - R_τ 15.1 min. H.p.l.c. 98.8%ee T.l.c. (EA) Rf 0.42 l.r. (KBr disc) 3366;2935;1695;1657;1558;1510;1422;1206;833;763cm^"1

Example 2 3-{3-ri-Cvclohexvl-2.4-dioxo-5-f2-oxo-2-pyrrolidin-1-yl-ethyl.-2.3.4.5-tetrahvdro-

1 H-benzo[b][1.4]diazepin-3-vl]-ureido}-benzoic acid.

A solution of Intermediate 6 (248mg) in ethyl acetate (10ml) and THF (5ml) was hydrogenated at 23° and 1 atm pressure overnight in the presence of 10% palladium on carbon (25mg) as catalyst. More catalyst (25mg) was added and hydrogenation continued for 2h whereupon the mixture was filtered through hyflo and the filtrate evaporated to give the title compound (216mg) as a white solid, m.p.275-6°.

T.l.c. (100:2 EA-AcOH) Rf 0.25 l.r. (KBr disc) 3369;2935;1700;1659;1557;1499;1431 ;1233;760cm^"1

Example 3

■ 3-{3-[1-(2-Fluoro-phenvlV2.4-dioxo-5-.2-oxo-2-pvrrolidin-1-yl-ethyl.-2.3.4.5- tetrahvdro-1 H-benzofb][1.4]diazepin-3-vn.-ureido}-phenvl.-acetic acid.

A solution of Intermediate 16 (177mg) in THF (10ml) was hydrogenated at 23° and 1atm pressure in the presence of 10% palladium on carbon (10mg) as catalyst. After 90min the mixture was filtered through hyflo and the filtrate evaporated to give the title compound .67mo. as a white solid, m.p.197-9°.

T.l.c. (EA) Rf 0.17 l.r. (KBrdisc) 3331;1708;1651_.1562;1499;1454;1403;1238;761cm^"1

Example 4

3-{3-[1-[2-.3.3-Dimethyl-piperidin-1-yl.-2-oxo-ethyl]-5-.2-fluoro-phenvh-2.4- rtioxo-2.3.4.5-tetrahvdro-1 H-benzofb]f1.41diazeDin-3-vl1-ureido}-benzoic acid A solution of Intermediate 21 (400mg) indichloromethane (20ml) was treated with trifluoroacetic acid (2ml) and stirred at 23° under nitrogen for 1h. The mixture was concentrated in vacuo and the residue chromatographed with DCM- MeOH (95:5) as eluent to give the title compound (344mg) as a white solid, m.p.225°dec. T.l.c. (95:5 DCM-MeOH) Rf 0.26

Using the general processes A, B, C and D described above the following compounds of the invention have also been prepared.

Table 1

Ex. No Process

5 H 3,5-diCH₃ 254-5° A

6 H 4-OCH₃ 180-2° A

7 H 3-F 188-90° A

8 H 3-N0₂ 173-5° A

9 H 3-CH₂CN 210-2° A

10 H H 216-8° A

11 H 3-CONH₂ 275-7° A

12 H 3-N(CH₃)₂ 251 -3° A

13 Br 4-F ^*Rf 0.3 D

^* T.l.c. cyclohexane : E A:AcOH 1 :1 :0.0.4 Table 2

Ex. No B-3 mp Process

14 Ph 3-tetrazolyl 243° A 15 Ph 3-CH₃ 234-6° A 16 Ph 3-CONHS0₂CH₃ 199-200° B 17 2-F-Ph 4-F 155-7° A 18 2F-Ph 3-CH₃ 195-7° A 19 2F-Ph 3-C0₂H -225-7° B

Table 3

Ex. No. NB 1.-R--:2 mp Process

Pharmacy Examples

Tablets a. Active ingredient 50mg Lactose anhydrous USP 163mg Microcrystallme Cellulose NF 69mg Pregelatinised starch Ph. Eur. 15mg Magnesium stearate USP 3mα Compression weight 300mg

The active ingredient, microcrystalline cellulose, lactose and pregelatinised starch are sieved through a 500 micron sieve and blended in a suitable mixer. The magnesium stearate is sieved through a 250 micron sieve and blended with the active blend. The blend is compressed into tablets using suitable punches. Active ingredient 50mg

Lactose monohydrate USP 120mg

Pregelatinised starch Ph. Eur. 20mg

Crospovidone NF 8mg

Magnesium stearate USP 2mg

Compression weight 200mg

The active ingredient, lactose and pregelatinised starch are blended together and granulated with water. The wet mass is dried and milled. The magnesium stearate and Crospovidone are screened through a 250 micron sieve and blended with the granule. The resultant blend is compressed using suitable tablet punches.

Capsules

Active ingredient 50mg Pregelatinised Starch Ph. Eur. 148mg Magnesium stearate USP 2mg Fill weight 200mg

The active ingredient and pregelatinised starch are screened through a 500 micron mesh sieve, blended together and lubricated with magnesium stearate (meshed through a 250 micron sieve). The blend is filled into hard gelatin capsules of a suitable size.

Active ingredient 50mg

Lactose monohydrate USP 223mg

Povidone USP 12mg

Crospovidone NF 12mg

Magnesium stearate 3mg

Fill weight 300mg

The active ingredient and lactose are blended together and granulated with a solution of Povidone. The wet mass is dried and milled. The magnesium stearate and Crospovidone are screened through a 250 micron sieve and blended with the granule. The resultant blend is filled into hard gelatin capsules of a suitable size.

CCK-B. Receptor Binding Affinity

CCK-B receptor binding studies were carried out using HeLa cell membranes which had been stabily transfected with the human CCK-B receptor cloned from temporal cortex cDNA library.

Measurement of CCK-B binding affinity.

The transfected HeLa cell membranes were incubated with 30pM [¹²⁵I]BH- sCCK-8 in the presence of various concentrations of the test compound (1 pM to 1 μM). All experiments were performed in an assay volume of 250μl. Membranes were harvested onto GF/C glass fibre filter paper (Whatman, UK) by rapid filtration using a Cell Harvester (Brandel model M-24R) and washed three times with HEPES Wash buffer chilled to 4°C. The radioactivity trapped on individual filter discs was counted over 60 seconds using a gamma counter (Mini Gamma counter, LKB, Wallac, Finland) at a counting efficiency of 77%. A control experiment was also carried out in the absence of a test compound.

Analysis of the data was performed by computer-assisted non-linear regression (ALLFIT programmes DeLean et al., 1978;). IC₅₀ values for the test compounds were converted to Kj values using the Cheng & Prussof equation (K_j = IC₅₀/(1 + [L]/K_D)) (Cheng and Prussof, 1973). Results obtrained with representative compounds of the invention in this test are given below and expressed as pKi values. CCK-A Receptor Binding Affinity

CCK-A receptor binding studies were carried out using COS-M6 cell membranes which have been transiently transfected with human gall bladder CCK-A receptor cDNA.

The clone was transiently transfected into COS-M6 cells grown to 80% confluency using the dimethylaminoethyl - dextran method based on the method of Seed and Araffo. Proc. Natl, Acad, Sci. USA 84, 3365 1987. The required transfected cell membranes were then isolated in a conventional manner.

The affinity of compounds of the invention for the CCK-A receptor was determined using the procedures described above for the CCK-B receptor however, in these experiments the test compound was tested at concentrations in the range 10pM to 10μM.

CCK-B Binding Studies

Example No pKi

1 8.5

1 (isomer 1 ) 8.5

1 (isomer 2) 8.1

2 8.7

3 8.4

4 8.8

7 8.3

8 8.6

10 8.4

12 9.0

14 8.5

15 8.8

20 8.7 The compounds of the invention are essential non-toxic a theraapeutically active doses. Thus no untoward effects were observed when the compound of Example 1 (isomer 1) was administered to the gastrin fistula rat of doses of 0.3 and 1 mg/kg iv at these doses gastric acid secretion was significally inhibited.

Claims

1. Compounds of the general formula (I)

and physiologically acceptable salts thereof wherein the group NR₁R₂ represents a 5-7 membered saturated heterocylic ring which may be substituted by one or two methyl groups;

R₃ is C_halky!, C₃.₆cycloalkyl or phenyl optionally substituted by 1 or 2 halogen atoms;

R₄ is phenyl or phenyl substituted by one or two groups selected from halogen,

C₁_ alkyl, trifluoromethyl, trifluoromethoxy or (CH₂)_nR₅ wherein n is zero or 1 and R₅ represents C^alkoxy, hydroxy, nitro, cyano, C0₂R₆, S(0)_pCH₃, NR₇R₈,

CONR₇R₈ , S0₂NR₇CO(C₁_₄alkyl), tetrazolyl, carboxamidotetrazolyl, or a 3- trifluoromethyl 1 ,2,4-triazolyl;

R₆ is hydrogen, Cι_₄alkyl or benzyl;

R₇ is hydrogen or C₁_₄alkyl, R₈ is hydrogen, C^alkyl, S0₂CH₃ or S0₂CF₃

X represents hydrogen, C₁.₄alkyl or halogen; m is zero, 1 or 2, and p is zero, 1 or 2.

2. Compounds as claimed in Claim 1 wherein NR₁R₂ represents pyrrolidino, 2,5-dimethylpyrrolidino, piperidino, 3,3-dimethylpiperidino, 4,4- dimethylpiperidino or 2,6-dimethylpiperidino.

3. Compounds as claimed in Claim 1 or 2 wherein NR.,R₂ represents pyrrolidino.

4. Compounds as claimed in any of Claims 1 to 3 wherein R₃ represents phenyl, 2-fluorophenyl or cyclohexyl.

5. Compounds as claimed in any of claims 1 to 4 wherein R₃ represents cyclohexyl.

6. Compounds as claimed in any of Claims 1 to 5 wherein R represents phenyl, 3-methylphenyl, 3,5-dimethylphenyl, 3-dimethylaminophenyl, 4- fluorophenyl, 3-methoxyphenyl, 4-methoxyphenyl, 3-nitrophenyl, 3- cyanomethylphenyl, 3-carboxamidophenyl, 3-carboxyphenyl, 3- carboxymethylphenyl or 3-(1 H)-tetrazol-5-yl-phenyl.

7. Compounds as claimed in any of claims 1 to 6 wherein R represent phenyl, 4-fluorophenyl, 3-dimethylaminophenyl, 3-carboxylphenyl, 3- carboxymethylphenyl or 3-(1 H)-tetrazol-5-yl-phenyl.

8. 1 -[1 -Cyclohexyl-2,4-dioxo-5-(2-oxo-2-pyrrolidin-1-yl-ethyl)-2,3,4,5- tetrahydro-1 H-benzo[b][1 ,4]diazepin-3-yl]-3-(4-fluoro-phenyl)urea and enantiomers thereof.

9. A compound selected from

3-{3-[1 -Cyclohexyl-2,4-dioxo-5-(2-oxo-2-pyrrolidin-1-yl-ethyl)-2,3,4,5-tetrahydro- 1 H-benzo[b][1 ,4]diazepin-3-yl]-ureido}-benzoic acid; (3-{3-[1 -(2-Fluoro-phenyl)-2,4-dioxo-5-(2-oxo-2-pyrrolidin-1 -yl-ethyl)-2,3,4,5- tetrahydro-1 H-benzo[b][1 ,4]diazepin-3-yl]-ureido}-phenyl)-acetic acid; 3-{3-[1-[2-(3,3-Dimethyl-piperidin-1-yl)-2-oxo-ethyl]-5-(2-fluoro-phenyl)-2,4- dioxo-2,3,4,5-tetrahydro-1 H-benzo[b][1 ,4]diazepin-3-yl]-ureido}-benzoic acid , and enantiomers thereof.

10. Compounds as defined in any of Claims 1 to 9 for use in therapy.

11. The use of a compound as defined in any of Claims 1 to 9 in the manufacture of a medicament for the treatment of conditions where modification of the effects of gastrin and/or CCK is of therapeutic benefit.

12. A pharmaceutical comparison comprising a compound is defined in any of Claims 1 to 9 in admixture with one or more physiologically acceptable carriers or excipients.

13. A method of treatment of a mammal including man for conditions where modifications of the effects of gastrin and/or CCK is of therapeutic benefit comprising administration of an effective amount of a compound as defined in any of Claims 1 to 9.

14. A process for the preparation of a compound as defined in Claim 1 which comprises.

(a) reacting a compound of formula (II),

wherein R-, R₂ R₃ X and n have the meanings defined in formula (I) with a compound R₄Y selected from an isocyante (III), carbamoyi chloride (IV), imidazolide (V) or phenyl carbamate (VI) wherein R_a is an optionally substituted phenoxy group, o

R a CONHR 4,

(VI)

and R₄ has the meanings defined in formfula (II). (b) reacting a compound of formula (VII)

wherein R₁ , R₂, R₃, X and m have the meanings defined in formula (I) and Y is a group selected from NCO, NHCOCI, or NHCOR_a wherein R_a is an optionally substituted phenoxy or imidazole group with the amine R₄NH₂ (VIII) wherein R₄ has the meaning defined in formula (I).

(c) reacting a compound of formula (IX) wherein R₃, R₄, X and m have the meanings defined in formula (I).

with the compound R₂R-,NCOCH₂Z wherein Z is a leaving group;

(d) Reacting an activated derivative of the compound of formula (XV) wherein R₃, R₄, X and m have the meanings defined in formula (I) and R represents hydrogen

with amine R-_| R₂NH; and thereafter if necessary or desired either before or after separation into its stereochemical isomers converting one compound of formula (I) into another compound of formula (I).