OA10236A

OA10236A - 1,5 Benzodiazepine derivatives having cck and/or gastrin antagonistic activity

Info

Publication number: OA10236A
Application number: OA60721A
Authority: OA
Inventors: Finch Harry; Shah Pritom; Carr Robin Arthur Ellis
Original assignee: Glaxo Wellcome Inc
Priority date: 1993-04-15
Filing date: 1995-10-21
Publication date: 1997-10-07
Also published as: FI954852A0; WO1994024151A1; HUT74091A; CN1120845A; HU9502977D0; AU688316B2; NZ265271A; FI954852A7; EP0694040A1; CA2158972A1; PL311083A1; AU6567594A; FI954852L; NO954091D0; JPH08508743A; CZ267695A3; SK125495A3; NO954091L

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

BENZODIAZEPINE DERIVATIVES HAVING CCK AND/OR GASTRIN ANTAGONISTIC ACTIVITY 0 1 0236

This invention relates to novel 1,5-benzodiazepine dérivatives, to processes fortheir préparation, to pharmaceutical compositions containing them and to theiruse in medicine.

Cholecystokinins (CCK) and gastrin are structurally related peptides which existin gastrointestinal tissue and in the central nervous System. Cholecystokininsinclude CCK-33, a neuropeptide of thirty-three amino acids in its originallyisolated form, its carboxy terminal octapeptide, CCK-8 (also a naturally-occurring neuropeptide), and 39- and 12-amino acid forms. Gastrin occurs in34-, 17- and 14- amino acid forms, with the miniumum active sequence beingthe C-terminal tetrapeptide, Trp-Met-Asp-Phe-NH2(CCK-4), which is thecommon structual element shared by both CCK and gastrin. CCK and gastrin are gastrointestinal hormones and neurotransmitters in theneural and peripheral Systems and perform their respective biologicai rôles bybinding to particular receptors located at various sites throughout the body.

There are at least two subtypes of cholecystokinin receptors termed CCK-A andCCK-B and both are found in the periphery and in the central nervous System.CCK and gastrin receptor antagonists hâve been disclosed for preventing andtreating CCK-related and/or gastrin related disorders of the gastrointestinal andcentral nervous Systems of animais, and more particularly humans. US Patent No. 4,988,692 describes a group of 3-acylamino 1-alkyl-5-phenyl 1,5-benzodiazepine dérivatives as cholecystokinin antagonists. Further thespécification teaches that the compounds hâve a significantly greater affinity forthe CCK-A receptor over the CCK-B receptor.

We hâve now found a novel group of 3-ureido 1,5-benzodiazepine compounds which are potent and spécifie antagonists of gastrin and/or CCK and in particular antagonists of gastrin and /or CCK at the CCK-B receptor. Oî 0236

The présent invention thus provides compounds of the general formula (I)

and physiologically acceptable salts thereof wherein the group NR-|R2represehts a 5-7 membered saturated heterocylic ring which may be substitutedby one or two methyl groups; R3 is Cvgalkyl, C3.6cycloalkyl or phenyl optionally substituted by 1 or 2 halogenatoms; R4 is phenyl or phenyl substituted by one or two groups selected from halogen,C^alkyl, trifluoromethyl, trifluoromethoxy or (CH2)nR5 wherein n is zéro or 1and R5 représente C^alkoxy, 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, C-,_4aikyl or benzyi; R7 is hydrogen or C^alkyl, R8 is hydrogen, C^alkyl, SO2CH3 or SO2CF3 X represents hydrogen, Cwalkyl or halogen; m is zéro, 1 or 2, and p is zéro, 1 or 2.

The compounds of the invention possess at least one asymmetric carbon atomand the compounds of the invention include both enantiomers and mixturesthereof including the racemate.

The term alkyl as used herein refers to both straight Chain and branched chain alkyl groups. For example Cv6alkyl includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl or hexyl. 010236

The term halogen includes fluorine, chlorine, bromine or iodine.

When Rs represents a tetrazolyl, carboxamidotetrazolyl or 3-trifluoromethyl1,2,4-triazolyl group these are linked to the rest of the molécule via a carbon 5 atom therein and the invention includes ail tautomers thereof and the C-, _4alkyl-N substituted dérivatives thereof. Examples of such groups include (1H)tetrazol-5-yl, carboxamido-1 H-tetrazol-5-yl, 2-methyltetrazol-5-yl and (3-trifluoromethyl-1,2,4-triazol-5-yl. 10 The group NR-,R2 is linked to the rest of the molécule via the nitrogen thereofexamples of such groups include pyrrolidino, piperidino, hexamethylenimino-,2,5-dimethylpyrrolidino, 3,3-dimethylpiperidino, 2,6-dimethylpiperidino or 4,4-dimethylpiperidino. 15 When R3 represents phenyl optionally substituted by halogen exampies of suchgroups 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 20 cyclopropyl, cyclopentyl or cyclohexyl.

When R3 represents C-,.6alkyl examples of such groups include methyl, ethyl,propyl, butyl, 3-methylbutyl or 3,3-dimethylbutyl. 25 Examples of suitable groups R4 include phenyl optionally substituted by halogene.g. fluorine, alkyl e.g. methyl, alkoxy e.g. methoxy, nitro, cyano, thiomethyl,carboxamido, carboxyl, dimethylamino, cyanomethyl, 1 H-tetrazol-5-yl,carboxymethyl, or N-methanesulphonylcarboxamido. 30 A preferred class of compound according to the invention are those wherein thegroup NR1R2 represent pyrrolidino, piperidino, 3,3-dimethylpiperidino 4,4-dimethylpiperidino, 2,6-dimethylpiperidino or 2,5-dimethylpyrrolidino. Within thisclass the group NR^ is conveniently pyrrolidino, piperidino, or 3,3-dimethylpiperidino. 35 01 0236 4

The group X is conveniently halogsn e.g. bromine, fluorine or fluorine or moreparticularly hydrogen. A further preferred class of compounds of formula (I) are those wherein R3 isphenyl, 2-fluorophenyl or cyclohexyl and more particularly 2-fluorophenyl orcyclohexyl.

Another preferred class of compounds of formula (I) are those wherein R4 isphenyl 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-nitrophenyI, 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 arethose wherein NRiR2 représente pyrrolidino, piperidine or 3,3-dimethyl-,piperidino, R3 represents 2-fluorophenyl or cyclohexyl, and X represents ahydrogen atom. Within this group particularly preferred compounds includethose 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-phenyi)-2,4-dioxo-5-(2-oxo-2-pyrrofidin-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 acidand enantiomers thereof. 01 0236

The physiologically acceptable salts of the compounds of formula (I) înclude conventional satts 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, 5 phosphoric, nitric, perchloric, fumaric, acetic, propionic, succinic, glycolic, formic,lactic, maleic, tartane, citric, pamoic, malonic, hydroxymaleic, phenylacetic,glutamic, benzoic, salicylic, fumaric, toluenesulphonic, methanesuiphonic,naphthalene-2-sulphonic, benzenesulphonic and the like. Other acids such asoxalic, while not in themselves pharmaceutically acceptable, may be useful in 10 the préparation of salts useful as intermediates in obtaining the compounds ofthe invention and their pharmaceutically acceptable salts. Référencés hereinafter to a compound according to the invention includes bothcompounds of formula (I) and their pharmaceutically acceptable salts and 15 solvatesi

Compounds of the invention modulate the effect of gastrin and/or CCK inmammals. In particular compounds of the invention are antagonists of gastrinand/or CCK. 20

Compounds of the invention hâve been shown to be antagonists of CCK,particularly at CCK-B receptors as demonstrated for example by thecompound's ability to inhibit the contractile actions of CCK-4 in the presence ofa CCK-A antagonist, in the guinea-pig isolated ileum longitudinal muscle- 25 myenteric plexus.

The préparation 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 30 al in J. Pharmacol. Exp. Ther., (1991) 256, 695-703.

Compounds of the invention hâve also been shown to hâve a greater affinity forthe CCK-B receptorthan for the CCK-A receptor. This may be determined usingthe CCK receptor binding assays described by Fornos et al J. Pharmacol Exp. 35 Ther., 261 1056-1063, 1992. 010236 6

Afternativeiy 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 yvith the human CCK-B receptor or, COS-M6 cell membranes that are transiently transfected with the human CCK-A receptor. 5

Compounds of the invention hâve also been shown to be antagonists of gastrinas demonstrated by their ability to înhibit pentagastrin-stimulated acid sécrétionfrom rat isolated gastric mucosa using the procedure described by J.J. Reevesand R. Stables in Br, J. Pharmac.. 1985 86. p.677-684. 10

Compounds of the invention hâve also been shown to inhibit pentagastrinstimulated acid sécrétion in conscious gastric fistula rats using the methodsdescribed by Hedges and Parsons Journal of Physiology 1977, 267.181-194. 15 The compounds of the invention are therefore useful for the treatment and/orprévention of disorders in mammals, especially humans, where modification ofthe effects of gastrin or CCK is of therapeutic benefit. Thus the compounds ofthe invention are useful for the treatment of gastrointestinal disorders especiallythose where there is an advantage in lowering gastric acidity. Such disorders 20 include peptic ulcération, reflux oesophagitis and Zollinger Ellison syndrome.They may also be useful for the treatment of gastrointestinal disorders such asirritable bowel syndrome, excess pancreatic sécrétion, acute pancreatitis,motility disorders, antral G cell hyperplasia, fundic mucosal hyperplasia orgastrointestinal neoplasms. The compounds of the invention are also useful for 25 the treatment of central nervous System disorders where CCK and/or gastrin areinvolved. For example anxiety disorders (including panic disorder, agoraphobia,social phobia, simple phobia, obsessive compulsive disorders, post traumaticstress disorder, and general anxiety disorder), dépréssion, tardive dyskinesia,Parkinson's disease or psychosis. They may also be useful for the treatment of 30 dependency on drugs or substances of abuse and withdrawal, Gilles de taTourette syndrome, or dysfunction of appetite regulatory Systems; as well as thetreatment of certain tumours of the lower oesophagus, stomach, intestines andcolon. Compounds of the invention are also useful for directly inducinganalgesia, or enhancing opiate or non-opiate mediated analgesia, as well as 35 anaesthesia or loss of the sensation of pain. 0 1 0236

The invention therefore provides a compound of formula (I) or a pharmaceutically acceptable sait or solvaté thereof for use in therapy, in particular in human medicine. 5

According to another aspect the invention provides the use of a compound offormula (I) or a pharmaceutically acceptable sait or solvaté thereof for the · manufacture of a médicament for the treatment of conditions where modificationof the effects of gastrin and/or CCK is of therapeutic benefit. 10

According to a further aspect of the invention we provide a method for thetreatment of a mammal, including man, in particular in the treatment ofconditions where modification of the effects of gastrin and/or CCK is oftherapeutic benefit which method comprises administering an effective amount 15 of a corhpound of formula (I) or a pharmaceutically acceptable sait or solvatéthereof to the patient.

It will be appreciated by those skilled in the art that reference herein totreatment extends to prophylaxis as well as the treatment of established 20 diseases or symptoms.

It will further be appreciated that the amount of a compound of the inventionrequired for use in treatment will vary with the nature of the condition beingtreated and the âge and the condition of the patient and will be ultimately at the

25 discrétion of the attendant physician or veterinarian. In general however doses I employed for adutt human treatment will typically be in the range of 1-2000mg | per day e.g l0-500mg per day. |

The desired dose may conveniently be presented in a single dose or as divided | 30 doses administered at appropriate intervals, for example as two, three, four or | more sub-doses per day. i

Compounds of the invention which antagonise the fonction of CCK in animais, may also be used as feed additives to increase the food intake in animais in i 35 daily dosages of around 1 mg/kg to 10mg/kg. 010236

While it is possible that, for use in therapy, a compound of the invention may beadministered as the raw Chemical it is preferabie to présent the active ingrédientas a pharmaceutical formulation. 5

The invention thus further provides a pharmaceutical formulation comprising acompound of formula (I) or a pharmaceutically acceptable sait thereof togetherwith one or more pharmaceutically acceptable carriers therefor and, optionally,other therapeutic and/or prophylactic ingrédients. The carrier(s) must be 10 'acceptable' in the sense of being compatible with the other ingrédients of theformulation and not deleterious to the récipient thereof. r

The compositions of the invention include those in a form especially formulatedfor oral, buccal, parentéral, implant, or rectal administration. Oral administration 15 is preferred.

Tablets and capsules for oral administration may contain conventionalexcipients such as binding agents, for example, syrup, acacia, gelatin, sorbitol,tragacanth, mucilage of starch or poiyvinylpyrrolidone; fillers, for example, 20 lactose, sugar, microcrystalline celluiose, maize-starch, calcium phosphate orsorbitol; lubricants, for example, magnésium stéarate, stearic acid, talc,polyethylene glycol or silica; disintegrants, for example, potato starch or sodiumstarch glycollate, or wetting agents such as sodium lauryl sulphate. The tabletsmay be coated according to methods well known in the art. Oral liquid 25 préparations may be in the form of, for example, aqueous or oily suspensions,solutions émulsions, syrups or élixirs, or may be presented as a dry product forconstitution with water or other suitable vehicle before use. Such liquidpréparations may contain conventional additives such as suspending agents, forexample, sorbitol syrup, methyl cellulose, glucose/sugar syrup, gelatin, 30 hydroxyethylcellulose, carboxymethyl cellulose, aluminium stéarate gel orhydrogenated edible fats; emuisifying agents, for example, lecithin, sorbitanmono-oleate or acacia; non-aqueous vehicles (which may include edible oils),for exampie, almond oil, fractionated coconut oil, oily esters, propylene glycol orethyl alcohol; and preservatives, for example, methyl or propyl β- 35 hydroxybenzoates or ascorbic acid. The compositions may also be formulated 010236 as suppositories, e.g. containing conventional suppository bases such as cocoabutter or other glycerides.

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

The composition according to the invention may be formulated for parentéraladministration by injection or continuous infusion. Formulations for injectionmay be presented in unit dose form in ampoules, or in multi-dose containerswith an added preservative. The compositions may take such forms assuspensions, solutions, or émulsions in oily or aqueous vehicles, and maycontain formulatory agents such as suspending, stabilising and/or dispersingagents. Alternatively the active ingrédient may be in powder form forconstitution with a suitable vehicle, e.g. stérile, pyrogen-free water, before use.

The composition according to the invention may also be formulated as a depotpréparation. Such long acting formulations may be administered byimplantation (for example subcutaneously or intramuscularly) or byintramuscular injection. Thus for example, the compounds of the invention maybe formulated with suitable polymeric or hydrophobie materials (for example asan émulsion in an acceptable oil) or ion exchange resins, or as sparingly solubledérivatives, for example, as a sparingly soluble sait.

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

Compounds of general formula (I) and salts thereof may be prepared by thegeneral methods outlined hereinafter. In the following description, the groupsR-pRg and X are as defined for the compounds of formula (I) unless otherwisestated.

According to a first general process A, compounds of formula (I) may be prepared by the reaction of an amine of formula (II) wherein R1t R2, r3 , χ and m hâve the meanings defined in formula (I). 010236

with a compound R4Y, wherein R4Y is an isocyanate of formula (lll), carbamoyl 5 chloride of formula (IV), imidazolide of formula (V), or an optionally substitutedphenyl çarbamate of formula (VI) wherein Ra is an optionally phenxoy group. o zxÂ- O=C = N—R C1CONHR. N ' NZ N R,

' \=J H (ΠΙ) (IV) (V) RCONHR, (VI) 10 The reaction conveniently takes place in the presence of a suitable solvent suchas a halohydrocarbon (e.g. dichloromethane), an ether (e.g. tetrahydrofuran) ornitrile (e.g. acetronitrile) or a mixture thereof at a température in the range of0°-80°C. 15 Isocyanates of formula (lll) may be purchased or prepared by the reaction ofamines H2N-R4 with phosgene or triphosgene in a suitable solvent such asmethylene chloride. Carbamoyl chlorides of formula (IV) are also prepared bythe reaction of amines H2NR4 with phosgene or triphosgene in a suitablesolvent such as methylene chloride. Imidazolides of formula (VI) are prepared 20 by treatment of amines H2N-R4 with carbonyl diimidazole in a suitable solvent(dichloromethane, ether, tetrahydrofuran) at a température ranging from 0-80° C(conveniently at room température). Optionally substituted phenyl carbamates offormula (VI) are prepared by the reaction of amines H2N-R4 with the appropriate 010236 11 chloroformate RaCOCI in the presence of a base (pyridine, triethylamine) in asuitable solvent (dichloromethane) and at a température of 0 - 50° C.

According to a further general process B, compounds of formula (I) may be5 prepared by reaction of an intermediate of formula (VU).

wherein Y is the group -NCO, -NHCOCI or NHCORa wherein Ra is an optionallysubstituted phenoxy group or a 1-imidazole group with an amine (VIII) 10 H2N-R4 (VIII) and optionally in the the presence of a base such as a tertiary amine (e.g.triethylamine). 15

The reaction conveniently takes place in a suitable solvent such as ahalogenated hydrocarbon (e.g. dichloromethane) or an ether (e.g.tetrahydrofuran) or an amide (e.g. Ν,Ν-dimethyl formamide) optionally at atempérature ranging from room température to the reflux température of the 20 solvent.

Conveniently the compound of formula (VII) are prepared in situ from the amine(II). 25 In a particular aspect of the process (B) when Y is the group NHCORa and Ra 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) 010236

in the presence of carbonyldiimidazole under the aforementioned conditions.

For process B when Y is the group NHCORa and Ra is optionally substituted 5 phenoxy group the reaction with the primary amine (VIH) is preferably carriedout 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=O the reaction with theprimary amine (VIII) is preferably carried out in an aprotic solvent such as a 10 halohydrocarbon e.g. methylene chloride. Conveniently the isocyanate isqenerated in situ prior to the addition of the primary amine (VIII).

The compounds of formula (VII) wherein Ra is an optionally substituted phenoxygroup may be prepared from the primary amine (II) by reaction with the 15 corresponding optionally substituted phenyl chloroformate in the presence of abase such as pyridine. The reaction may be carried out in a solvent such as ahalohydrocabon e.g. dichloromethane and at a température from 0-50°.

Compounds of formula (VII) wherein Ra is a 1-imidazole group may be prepared 20 by reacting a compound of formula (II) with carbonyldiimidazole in the presenceof a suitable solvent such as a halogenated hydrocarbon (e.g. dichloromethane)or an ether (e.g. tetrahydrofuran) at a température ranging from 0° to 80°(conveniently at room température). 25 Compounds of formula (Vil) wherein Y is the isocyanate grouping -N=C=O orcarbamoyl chloride -NHCOCI may be prepared from the primary amine (II) byreaction with phosgene (COCI2) or triphosgene in a suitable solvent such asmethylene chloride. 010236 13

According to a further general process C compounds of formula (I) may also beprepared 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 formula10 (IX) with a strong base such as sodium hydride in a polar aprotic solvent suchas Ν,Ν-dimethylformamide followed by reaction with the alkylating agent R^NCOCHgZ. 15

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

wherein W is CH-N3 or C=N-NHPh. 20 Compounds of formula (X) wherein W is CH-N3 may be reduced to a compoundof formula (II) by hydrogénation in the presence of a suitable catalyst such aspalladium, on a support such as carbon or calcium carbonate, or platinum (IV)oxide. The reaction conveniently takes place in the presence of a solvent suchas an alkanol (e.g. éthanol) an ester (e.g. ethyl acetate) or acetic acid. 25 010236 14

Compounds of formula (X) wherein W is C=N-NHPh may be reduced to acompound of formula (II) by reaction with zinc and acetic acid. This reactionmay be carried out a température with the range 0-50°. 5 Compounds of formula (X) wherein W is CHN3 may be prepared from acompound of formula (X) wherein W is CH2 by treatment with a strong basesuch as sodium hydride or potassium tert-butoxide followed by tri-isopropylbenzenesulphonyl azide. The reaction conveniently takes place in a solventsuch as an ether (e.g. tetrahydrofuran) at a température in the range of -78° to 10 20°.

Compoünds of formula (X) in which W is C=NNHPh may be prepared byreaction of the ortho-phenylenediamine (XI) with the diacid chioride (XII), in asuitable solvent such as an ether e.g. tetrahydrofuran 15 CONR^

(XI)

Compounds of formula (X) wherein W is CH2 may be prepared by reaction ofthe compound of formula (XI) with the diacid chioride (XIII) 20 25 cioq^ .CH,

CIOC (ΧΙΠ)

Compounds of formula (XI) are either known compounds or may be prepared byanalogous methods. Thus for example a compound of formula (XI) may beprepared by alkylation of the amine (XIV). 010236 \·

(XIV)

Thus the amine (XIV) may be reacted with the compound R1R2NCOCH2Z , inwhiçh Z is chlorine or bromine, optionally in the presence of sodium iodide in a 5 solvent such as N,N-dimethyiformamide.

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

According to a further process (D) a compound of formula (I) may be preparedfrom the compound of formula (XV) wherein Rb is hydrogen

15 20 25 by reaction of an activated dérivative thereof with the amine RiR2NH.Conveniently the reaction is carried out using the acid in the presence of adiimide such as dicyciohexyl carbodimide and hydroxybenzotriazole in a solventsuch 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 Rb is a hydrogen may be prepared byhydrolysis of the corresponding compound of formula (XV) wherein Rb is a t-butyl group, for example by reaction with trifluoroacetic acid. The compound offormula (XV) wherein Rb is t-butyi may be prepared by alkylation of thecorresponding compound of formula (IX) with the halo ester Z-CH2CO2Rbwherein Z is halogen. Alternatively the compound (XV) wherein Rb is t-butyl maybe prepared using the general processes A and B described above for 01 0236 16 preparing the compounds of formula (I) but starting with the appropriate N- substituted benzodiazépine dérivative.

According to a further process (E) a compound of formula (I) may be converted into another compound of formula (I) using conventional techniques. 5

Thus compounds of formula (I) wherein R4 is a phenyl group substituted by acarboxyl or carboxymethyl group may be prepared by hydrolysis of thecorresponding compound of formula (I) wherein R4 is a phenyl groupsubstituted by an alkoxycarbonyl or alkoxycarbonylmethyl group. 10

Compounds of formula (IX) may be prepared from the diamine (XIV), in whichthe prirpary amino group is protected as an p-methoxybenzyl dérivative thereof,using the general procedures described above for preparing the compounds offormula (I) from the corresponding orthophenylenediamine (XI) followed by 15 removal of the N-protecting group p-methoxybenzyl using conventionalprocedures.

Compounds of formula (I) contain at least one asymmetric carbon atom, namelythe carbon atom of the diazepine ring to which the substituted urea grouping is 20 attached. Spécifie enantiomers of the compounds of formula (I) may be obtainedby resolution of the racemic compound using conventional procedures such aschiral HPLC. Alternatively the required enantiomer may be prepared from thecorresponding enantiomeric amine of formula (II) using any of the processesdescribed above for preparing compounds of formula (I) from the amine (II). The 25 enantiomers of the amine (11) may be prepared from the racemic amine (11)using conventional procedures such as sait formation with a suitably opticallyactive acid such as R- camphorsulphonic acid or by préparative chiral HPLC.

In order that the invention may be more fully understood the following examples 30 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. Ailtempératures refers to °C. Coiumn chromatography was carried out over silica 35 gel. T.l.c. refers to thin layer chromatography on silica plates. Dried refers to 010236 17 solutions dried cver anhydrous sodium sulphate. The following abbreviation arealso 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, RT = rétention time. 5

Intermediate 1

Cvclohexvl-(2-nitro-Dhenyn-amine· A mixture of 2-chloronitrobenzene (20g), potassium carbonate (35g) and copper(I) iodide (1.21g) in cyclohexylamine (43.6ml) was heated at 150° under 10 nitrogen for 18h. The mixture was allowed to cool to room température and wasadsorbed onto silica. This was chromatographed with hexane-EA (98:2) aseluent to give the title compound (22.94g) as an orange solid. T.l.c. (98:2 hexane-EA) Rf 0.38 15 Intermediate 2 N2C.vclohexvl-benzene-1.2-diamine. A solution of Intermediate 1 (10g) in ethyl acetate (400ml) was hydrogenated at23° and 1 atm. pressure over 10% palladium on carbon (1g) for 4h. The catalystwas removed by filtration through hyflo and the filtrate evaporated to give the 20 title compound (8.5g) as an orange oil. T.Lc. (9:1 hexane-EA) Rf 0.36

Intermediate 3 2τ(2-ΟνοΙοΙΐ6χνΐ3Γηΐηο-ρΙΐ6ηνΐ3Γηΐηο)-1-ρνπΌΐΐΰιη-1-ν[Γ6^3ηοη6, 25 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° undernitrogen for 18h. The mixture was poured into water (600ml) and extracted withethyl acetate (3x300ml). The combined extracts were washed with water(3x300ml) and saturated brine (200ml), dried and evaporated to give a brown 30 oil. This was chromatographed with hexane-EA (1:1) as eluent to give the titlecompound (9.8g) as a cream solid, m.p.108-110°. T.l.c. (1:1 hexane-EA) Rf 0.42 010236 18 tntermediate 4 1 -Cyclohexvl-5-(2-oxo-2-pvrrolidin-1 -vl-ethyl)-3-(phenyt-hvdrazono)-1.5rdihvdror b.g,QZQfbK1,41diazepine-2.4-dione·

Solutions of Intermediate 3 (9.8g) and 2-(phenyl-hydrazono)-propanedioyl 5 dichloride (8.36g) in dry THF (75ml) were added at equai rates to dry THF(75ml) cooled to -10° under nitrogen. When the addition was complété themixture was allowed to warm to room température and stir for 3h. The solventwas removed by évaporation to give the crude title compound (19.5g) as ayellow crunchy foam. 10 T.l.c. (DE) Rf 0.23

Intermediate 5 3zAmino-1 -cvclohexvl-5-(2-oxo-2-Dyrrolidin-1 -vl-ethvn-1,5-dihvdror benzofbïïl .41diazepine-2.4-dione. 15 A solution of Intermediate 4 (19g) in glacial acetic acid (200ml) was addeddropwise to a stirring suspension of zinc dust (15g) in glacial acetic acid. Themixture was stirred at 23° for 1.5h whereupon the zinc was removed by filtrationthrough hyflo and the filtrate evaporated to give a red oil. This was partitionedbetween water (200ml) and ethyl acetate (75ml). The aqueous portion was 20 adjusted to pH9 with solid NagCOg and extracted with ethyl acetate (4x100ml).The combined organic extracts were dried and evaporated to give a brown oilwhich was chromatographed with DCM-MeOH (95:5) as eluent to give the titlecompound (7.4g) as a pink crunchy foam. T.l.c. (95:5 DCM-MeOH) Rf 0.26 25

Intermediate 6 3-(3-f1-Cyclohexyl-2.4-dioxo-5-(2-oxo-2-pyrrQlidin-1-yl-ethyl)-2.3.4.5-tetrahydro- 1 H-benzofb1î1.4)diazepin-3-vri-ureido)-benzoic acid.benzvl ester,

Triethylamine (108μΙ) and triphosgene (77mg) were added sequentially to a 30 solution of 3-amino-benzoic acid benzyl ester (177mg) in dry THF (10ml) at 0°under nitrogen. More triethylamine (108μΙ) was added and stirring continued at0° for 30min. A solution of Intermediate 5 (250mg) in dry THF (10ml) was addedand stirring continued at 23° for 18h. The mixture was then partitioned between2N hydrochioric acid (50ml) and ethyl acetate (50ml). The dried organic extract 010236 19 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.I.c. (2.Ί EA-hexane) Rf 0.17 l.r. (Solution in CHCI3) 3384;2939;1690;1658;1423cm’1 5

Intermediate 7 3-Nitro-benzoic acid tert-butvl ester. A solution of 3-nitrobenzoic acid (1.4g) in dry toluene (10ml) was treated withΝ,Ν-dimethylformamide di-tertbutyl acetal (10ml). The mixture was heated under 10 reflux for 18h then cooled, diluted with ethyl acetate and washed consecutivelywith water, 8% sodium bicarbonate solution and saturated brine. The driedorganic phase was evaporated to give the title compound (1.02g) as a yellowoil. T.I.c. (97:3 DCM-MeOH) Rf 0.66 15

Intermediate 8 . 3rAmino-benzoic acid tert-butvl ester. A solution of Intermediate 7 (1g) in éthanol (20ml) was hydrogenated at 23° and1atm pressure in the presence of 10% palladium on carbon as catatyst (200mg). 20 After 2h the mixture was filtered through hyflo and the filtrate evaporated. Theresidue was chromatographed with DCM-MeOH (97:3) as eluent to give the titlecompound (727mg) as a colourless oil. T.I.c. (97:3 DCM-MeOH) Rf 0.35 25 Intermediate 9 (3- Nitro-ohenvB-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 with1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (7.4g). The resulting yellow 30 mixture was stirred at 23° for 3days whereupon the solvent was removed invacuo. The residue was partitioned between water (100ml) and ethyl actate(100ml) and the organic phase washed with water (2x100ml) and saturatedbrine (100ml) then dried and evaporated. The residue was chromatographedwith hexane-EA (4:1) as eluent then again with isohexane-DCM (1:1) as eluent 35 to give the title compound (2.05g) as a pale yellow oil. 010236 20 T.l.c. (4:1 hexane-EA) fil 0.35

Intermediate 10 i3jLAmino-phgnvl)-acetic acid benzvl ester. 5 A solution of Intermediate 9 (2.05g) in ethyl acetate (100ml) was hydrogenatedat 23° and 1atm pressure in the presence of 5% platinum on carbon (200mg) ascatalyst. After 90min the mixture was filtered through hyflo and the filtrateevaporated to give the title compound (1,77g) as a pale yellow oil T.l.c. (DCM) Rf 0.35 10

Intermediate 11 i2rEluoro-DhenylW2-nitro-phenyn-amine. A mixture of 2-fluoroaniline (5.0g), potassium carbonate (2.5g) andcopper(,)iodide (414mg) in 2-fluoronitrobenzene (16.9ml) was heated to 180° 15 under nitrogen for 18h. The cooled mixture was poured into water (300ml) andextracted with ethyl acetate (2x250ml) then the combined extracts were washedwith saturated brine and evaporated. The residual brown oil was azeotropedwith ethanol/water then toluene. The residue was chromatographed withhexane-DE (100:0 to 95:5) to give the title comoound (3.25g) as a bright orange 20 solid, m.p.58-9°. T.l.c.(95:5 hexane-DE) Rf 0.45 lüteimediate 12 bLf2-Fluoro-phenvn-benzene-1.2-diamine, 25 A solution of Intermediate 11 (15.6g) in ethyl acetate (400ml) was hydrogenatedat 23° and 1atm pressure in the presence of 5% platinum on carbon (2g) ascatalyst. After 1h the mixture was filtered through hyflo and the fittrateevaporated to give the title compound (13.45g) as a yellow solid. T.l.c. (9:1 hexane-DE) Rf 0.25 30

Intermediate 13 2T2-(2-Fluoro-phenvlaminoVphenvlaminoM-pyrrolidiri-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 35 carbonate (27.5g) in dry DMF (100ml) at 23° under nitrogen. The mixture was 01 0236

21 stirred at 60° for 4h then poured into 2N sodium carbonate solution (500ml) andextracted with ethyl acetate (400ml). The organic extract was washed with water(2x250ml) and saturated brine (250ml), dried and evaporated. The residue waschromatographed with hexane-DE (50:50 to 0:100) as eluent to give the title 5 compound (14.12g) as a pale brown solid. T.I.C. (DE) Rf 0.53

Jntermediate 14

Jzi2:Fluoro-phenvn-5-(2-oxo-2-pynOlidin-1-yl-ethvl)-3-(phenyl-hydrazono)-1,5:-_ 10 .dihvdrQ-benzQlblfl .41diazepine-2.4-dione.

Solutions of Intermediate 13 (14.12g) and 2-(phenyl-hydrazono)-propanedioyldichloride (11.04g) in dry THF (100ml) were added simultaneously anddropwise at equal rates to dry THF (100ml) cooled to -10° under nitrogen.When the addition was complété the mixture was allowed to warm to room 15 température and stir for 3.5h. The solvent was removed by évaporation to givethe crude title compound (23g) as a yellow crunchy foam. T.l.c. (EA) Rf 0.5 jolermediate 15 20 3zAmino-1-f2-fluoro-phenvlV5-f2-oxo-2-pvrrolidin-1-vl-ethvn-1.5-dihvdro- benzofblH .41diazepine-2.4-dione. A solution of Intermediate 14 (23g) in glacial acetic acid (200ml) was addeddropwise 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 25 the zinc was removed by filtration through hyflo and the filtrate evaporated. Theresidue was partitioned between water (150ml) and ethyl acetate (100ml) andthe aqueous portion basified with solid Na2CO3- The layers were separated andthe aqueous phase further extracted with ethyl acetate then the combinedorganic extracts were washed with saturated brine, dried and evaporated. The 30 residue was chromatographed with EA-MeOH (100:0 to 95:5) as initial eluentfollowed by DCM-MeOH (80:20) to give the title compound (11.07g) as a fawnpowder. T.l.c. (95:5 DCM MeOH) Rf 0.23 01 0236 22 r f J.·

Intermediate 16 i3-f341-(2-Fluoro-Dhenvn-2.4-dioxo-5-(2-oxo-2-pvrrolidin-1-vl-ethyl)-2.3.4.5- tetrahvdro-1 H-benzofb1f1 .41diazepin-3-vn-ureido)-phenvl)-acetic acid benzyl astei 5 A solution of Intermediate 15 (3g) in dry dichloromethane (40ml) was addeddropwise to a stirred solution of phosgene in toluene (60ml) under nitrogen. Theresulting mixture was stirred at 23° for 4h then nitrogen was bubbled throughthe mixture into ammonia solution for 18h. The mixture was then evaporatedand the residue dried at 60° in vacuo for 3h to give the intermediate isocyanate 10 (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 toa stirred, solution of (3- amino-phenyl)-acetic acid benzyl ester (241 mg) in drydichloromethane (2ml) at 23° under nitrogen. After 60min the solvent wasremoved in vacuo and the residue triturated with diethyl ether. The solid was 15 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 17 20 2z.B.romo-1 -(3.3-dimethvl-pioeridin-1 -vQ-ethanone, A mixture of 3,3-dimethylpiperidine (10g) and triethylamine (12.3ml) in drydichloromethane (50ml) was added dropwise to an ice-cold solution ofbromoacety, bromide (7.7ml) in dry dichloromethane (100ml). The mixture wasstirred at 23° for 18h then recooled to 0°. Iced water (200ml) was added and the 25 layers separated. The aqueous layer was further extracted withdichloromethane (2x200ml) then the combined extracts were washed with 2NHCl (200ml) and saturated brine (200ml), dried and evaporated to give the titlecompound (17.28g) as a brown oil. T.l.c. (DE) Rf 0.49 30

Intermediate 18 1-(3.3-Dimethvl-Dioeridin-1-vn-2-f2-i2-fluoro-phenvlamino)-phenvlamino]- .ethanene,

A mixture of Intermediate 12 (3g)) and potassium carbonate (6.15g) in dry DMF 35 (10ml) was treated with a solution of 2-bromo-1-(3,3-dimethyl-piperidin-1-yl)- 010236 23

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. Thecombined extracts were washed with water and saturated brine, dried andevaporated to give a brown oil. This was chromatographed with DCM-MeOH 5 (100:0 to 97:3) as eluent to give the title comoound (3.17g) as a pale brown foam. T.l.c. (DCM) Rf 0.23

Intermediate 19 10 .1-f2-(3x3-Dirnethvl-PiDeridin-1-yl)-2-oxo-ethvl]-5-f2-fluoro-phenyl)-3-(phe.ayl7 hvdrazonoM ,5-dihvdro-benzo(biri ,4)diazeDine-2.4-dione.

Solutions of Intermediate 18 (3.08g) and 2-(phenyl-hydrazono)-propanedioyldichloride (2.12g) in dry THF (100ml) were added at equal rates to dry THF(50ml) cooled to -10° under nitrogen. When the addition was complété the 15 mixture was allowed to warm to room température and stirred for 2h. Thesolvent was removed by évaporation to give the crude title compound (5.25g) asa yellow crunchy foam. T.l.c. (95:5 DCM-MeOH) Rf 0.68 20 lntsnnedlate-20 3rAmiaO--1-r2-(3.3-dimethvl-pineridin-1-vn-2-oxo-ethvll-5-i2-fluoro-Dhenvn-1.5- dihvdro-benzofblfl .41diazepine-2.4-dione. A solution of Intermediate 19 (5.15g) in glacial acetic acid (75ml) was addeddropwise to a stirring suspension of zinc dust (4.47g) in glacial acetic acid 25 (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. Theresidue was partitioned between water and ethyl actate and the aqueousportion basified with solid Na2CO3- The organic extract was washed withsaturated brine, dried and evaporated. The residue was chromatographed with 30 DCM-MeOH (95:5) as eluent to give the title compound (2.38g) as a pale brownsolid. T.l.c. (95:5 DCM-MeOH) Rf 0.18 01 0236 24

Inîgrmstf ate 21 3-f3-i1-f2-f3.3-Dimethvl-Diperidin-1-vn-2-oxo-ethvl]-5-f2-fluQro-Dhenvl)-2Ædioxo^2.3.4.5-tetrahvdrQ-1H-benzo[b1f1.41diazepin-3-vH-ureido}-bgQZQk-3£id tert-butvl ester. 5 Triethylamine (0.31ml) and triphosgene (217mg) were added sequentially to asolution of 3-amîno-benzoic acid tert-butyl ester (423mg) in dry THF (10ml) at0° under nitrogen. More triethylamine (0.31ml) was added and stirringcontinued 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 10 partitioned between phosphate buffer solution (pH6.5) and dichloromethane.The combined organic extracts were washed with saturated brine, dried andevaporated and the residue chromatographed with DCM-MeOH (97:3) as eluentto give a pale green solid which was triturated with DE-hexane (1:1 ) to give thetitle compound (1.07g) as a white solid, m.p.170°. 15 T.l.c. (97:3 DCM-MeOH) Rf 0.15

Exemple .1 1-ri-Cvclohexvl-2.4-dioxo-5-i2-oxo-2-Dvrrolidin-1-yl-ethyn-2.3.4.5.-tetrahvdrQz 1 H-benzofblf1 .4]diazepin-3-vl]-3-(4-fluoro-phenvnurea. 20 4-Fluorophenyl isocyanate (650μΙ) was added to a solution of Intermediate 5(2g) in dry dichloromethane (50ml) at 23° under nitrogen. The resulting mixturewas stirred for 1h 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 whitesolid, m.p. 181-3°. 25 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. i 30 Column: Chiralcel OD 25cm x 20mm id

Eluent: Hexane-EtOH (70:30)

Flow rate: 20 ml min*1Détection: uv @ 254nm

Isomer 1, (452mg) as a white solid - Ry 8.6 min. H.p.l.c. >99.5% ee 35 01 0236 25 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- RT 15.1 min. K.p.l.c. 98.8%ee T.l.c. (EA) Rf 0.42 l.r. (KBr dise) 3366;2935;1695;1657;1558;1510;1422;1206;833;763cm·1

Examp.le.2 3-{3-[1-Cyclohexvl-2.4-dioxo-5-(2-oxo-2-Dvrrolidin-1-vl-ethvl)-2.3.4.5-tetrahydro- 1 H-benzo(b)(1,41diazepin-3-vî]-ureido)-benzoic acid. A solution of Intermediate 6 (248mg) in ethyl acetate (10ml) and THF (5ml) washydrogenated at 23° and 1 atm pressure overnight in the presence of 10%palladium on carbon (25mg) as catalyst. More catalyst (25mg) was added andhydrogénation continued for 2h whereupon the mixture was filtered throughhyflo and the filtrate evaporated to give the title compound (216mg) as a whitesolid, m.p.275-6°. T.l.c. (100:2 EA-AcOH) Rf 0.25 l.r. (KBr dise) 3369;2935;1700;1659;1557;1499;1431;1233;760cm·1 fxampls_2 f3-{3-T1-(2-Fluoro-Dhenvn-2.4-dioxo-5-(2-oxo-2-Pvrrolidin-1-vl-ethvD-2.3.4.5- tetrahvdro-1 H-benzoibiri .41diazeDin-3-vn-ureidoÎ-phenvn-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) ascatalyst. After 90min the mixture was filtered through hyflo and the filtrateevaporated to give the title compound (67mq) as a white solid, m.p.197-9°. T.l.c. (EA) Rf0.17 l.r. (KBr dise) 3331 ;1708;1651 ;1562;1499;1454;1403;1238;761cm·1

Example 4 3-f3-ri-[2-f3.3-Dimethvl-piperidind-vn-2-oxo-ethvn-5-(2-fluoro-Dhenvn-2.4- dioxo-2.3.4.5-tetrahvdro-1 H-benzofblf1.41diazeDin-3Aureidol-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 010236 mixture was concentrâtes 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 followingcompounds of the invention hâve also been prepared. 26

Table 1 10 con

hxJÜfi X fis ma Process 5 H 3,5-diCH3 254-5° A 6 H 4-OCHo 180-2° A 7 H 3-F 188-90° A 8 H 3-NO2 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 010236 27

Table 2

Ex. No fia fis mp Process 14 Ph 3-tetrazolyl 243° A 15 Ph 3-CH3 234-6° A 16 Ph 3-CONHSO2CH3 199-200° B 17 2-F-Ph 4-F 155-7° A 18 2F-Ph 3-CH3 195-7° . A 19 2F-Ph 3-CO2H -225-7° B

Table 3

10 010236 28 EXu-No, NH1R2

5 Pharmacv ExamplesTablets

H3Q Er.Qgg.SS

240° A

170° A

260° A

220-5°dec C

255-8° D

Active ingrédient 50mg Lactose anhydrous USP 163mg Microcrystalline Cellulose NF 69mg Pregelatinised starch Ph.Eur. 15mg Magnésium stéarate USP 3,ma Compression weight 300mg 15

The active ingrédient, microcrystalline cellulose, lactose and pregelatinisedstarch are sieved through a 500 micron sieve and biended in a suitable mixer.The magnésium stéarate is sieved through a 250 micron sieve and biended withthe active blend. The blend is compressed into tablets using suitable punches. 010236 29

Active ingrédient 50mg Lactose monohydrate USP 120mg Pregelatinised starch Ph.Eur. 20mg Crospovidone NF 8mg Magnésium stéarate USP 2ma Compression weight 200mg

The active ingrédient, lactose and pregelatinised starch are blended together10 and granulated with water. The wet mass is dried and milled. The magnésiumstéarate and Crospovidone are screened through a 250 micron sieve andblended with the granule. The résultant blend is compressed using suitable tablet punches. 15 Capsules a. Active ingrédient 50mg Pregelatinised Starch Ph.Eur. 148mg Magnésium stéarate USP 20 RII weight 200mg

The active ingrédient and pregelatinised starch are screened through a 500micron mesh sieve, blended together and lubricated with magnésium stéarate(meshed through a 250 micron sieve). The blend is filled into hard gelatin 25 capsules ot a suitable size.

Active ingrédient 50mg Lactose monohydrate USP 223mg Povidone USP 12mg Crospovidone NF 12mg Magnésium stéarate RII weight 300mg

The active ingrédient and lactose are blended together and granulated with a35 solution ot Povidone. The wet mass is dried and milled. The magnésium 010236 30 stéarate and Crospovidone are screened through a 250 micron sieve andblended with the granule. The résultant blend is filled into hard gelatin capsulesof a suitable size. 5 CCK-B. Receptor Binding Affinitv CCK-B receptor bînding studies were carried out using HeLa cell membraneswhich had been stabily transfected with the human CCK-B receptor cloned fromtemporal cortex cDNA library. 10

Measurement of CCK-B bindina affinitv,

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 15 1 μΜ). Ail experiments were performed in an assay volume of 250μΙ.

Membranes were harvested onto GF/C glass fibre filter paper (Whatman, UK)by rapid filtration using a Cell Harvester (Brandel model M-24R) and washedthree times with HEPES Wash buffer chilled to 4°C. The radioactivity trapped onindividual filter dises was counted over 60 seconds using a gamma counter 20 (Mini Gamma counter, LKB, Wallac, Finland) at a counting efficiency of 77%. Acontrol experiment was also carried out in the absence of a test compound.

Analysis of the data was performed by computer-assisted non-linear régression(ALLFIT programmes DeLean et al., 1978;). IC50 values for the test compounds 25 were converted to Kj values using the Cheng & Prussof équation (K( = IC50/(1 +(L]/Kd)) (Cheng and Prussof, 1973). Results obtrained with représentativecompounds of the invention in this test are given below and expressed as pKivalues. 1 31 010236 CCK-A Receptor Binding Affinity CCK-A receptor binding studies were carried out using COS-M6 cell5 membranes which hâve been transiently transfected with human gall bladder CCK-A receptor cDNA.

The clone was transiently transfected Înto COS-M6 cells grown to 80%confluency using the dimethylaminoethyl - dextran method based on the method 10 of Seed and Araffo. Proc. Natl, Acad, Sci. USA 84, 3365 1987. The requiredtransfected cell membranes were then isolated in a conventional manner.

The affinity of compounds of the invention for the CCK-A receptor wasdetermined using the procedures described above for the CCK-B receptor 15 however, in these experiments the test compound was tested at concentrationsin the range 10pM to 10μΜ. CCK-B Binding Studies

Example No βΚΐ 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 010236 32

The compounds of the invention are essential non-toxic a theraapeuticallyactive doses. Thus no untoward effects were observed when the compound ofExample 1 (isomer 1) was administered to the gastrin fistula rat of doses of 0.3and 1mg/kg iv atthese doses gastric acid sécrétion was significally inhibited.

Claims

01 0236 33 CL Al M S 1. Compounds of the general formula (I)

NHCONHR. (D and physiologically acceptable salts thereof wherein the group NR1R2represents a 5-7 membered saturated heterocylic ring which may be substituted 10 by one ortwo methyl groups; R3 is Cv6alkyl, C3.6cycloalkyl or phenyl optionally substituted by 1 or 2 halogenatoms; R4 is phenyl or phenyl substituted by one or two groups selected from halogen,C-,_4alkyl, trifluoromethyl, trifluoromethoxy or (CH2)nRs wherein n is zéro or 1 15 and R5 represents C^alkoxy, hydroxy, nitro, cyano, CO2R6· S(O)pCH3, NR7R8,CONR7R8 , SO2NR7CO(Cwalkyl), tetrazolyl, carboxamidotetrazolyl, or a 3-trifluoromethyl 1,2,4-triazolyl; R6 is hydrogen, Cv4alky! or benzyl; R7 is hydrogen or C^alkyl, 20 R8 is hydrogen, C^alkyl, SO2CH3 or SO2CF3X represents hydrogen, C1.4alkyl or halogen;m is zéro, 1 or 2, and p is zéro, 1 or 2.
2. Compounds as claimed in Claim 1 wherein NR1R2 represents 25 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 NR1R2 representspyrrolidino. 010236 34
4. Compounds as claimed in any of Claims 1 to 3 wherein R3 representsphenyl, 2-fluorophenyl or cyclohexyl.
5. Compounds as claimed in any of claims 1 to 4 wherein R3 representscyclohexyl.
6. Compounds as claimed in any of Claims 1 to 5 wherein R4 representsphenyl, 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 R4 representphenyl, 4-fluorophenyl, 3-dimethylaminophenyl, 3-carboxylphenyl, 3-carboxymethyiphenyl 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-phenyI)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}-phenyI)-acetic acid;3-{3-[1-[2-(3,3-Dimethyl-piperidin-1-yl)-2-oxo-ethylJ-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 themanufacture of a médicament for the treatment of conditions where modificationof the effects of gastrin and/or CCK is of therapeutic benefit. V’ - .'i . M'w’i : ?.. ,<ν·.'Λ> · : i. 010236 35
12. A pharmaceutical comparison comprising a compound is defined inany of Claims 1 to 9 in admixture with one or more physiologically acceptablecarriers or excipients. 5
13. A method of treatment of a mammal including man for conditionswhere modifications of the effects of gastrin and/or CCK is of therapeutic benefitcomprising administration of an effective amount of a compound as defined inany of Claims 1 to 9. 10
14. A process for the préparation of a compound as defined in Claim 1which comprises. (a) reacting a compound of formula (II),

20 wherein R1 R2t R3< X and n hâve the meanings defined in formula (I) with acompound R4Y selected from an isocyante (III), carbamoyl chloride (IV),imidazolide (V) or phenyl carbamate (VI) wherein Ra is an optionally substitutedphenoxy group, o O=c = N — R4 ciconhr4 (III) (IV) N Nz \=J

R CONHR a 4 (VI) (V) and R4 has the meanings defined in formfula (II) 01 0236 36 (b) reacting a compound of formula (VII)

(VH) wherein R1t R2, R3, X and m hâve the meanings defined in formula (I) and Y is agroup selected from NCO, NHCOCI, or NHCORa wherein Ra is an optionallysubstituted phenoxy or imidazole group with the amine R4NH2 (VIII) wherein R4has the meaning defined in formula (I). 10 (c) reacting a compound of formula (IX) wherein R3, R4, X and m hâve themeanings defined in formula (I).
15 with the compound R2R-,NCOCH2Z wherein Z is a leaving group; (d) Reacting an activated dérivative of the compound of formula (XV)wherein R3, R4, X and m hâve the meanings defined in formula (I) and Rbreprésente hydrogen 20

NHCONHR, (XV)

I 010236 37 with amine R-,R2NH; and thereafter if necessary or desired either before or after séparation into its stereochemical isomers converting one compound of formula (I) into another compound of formula (I).