WO1996031476A1

WO1996031476A1 - Aromatic hydroxyethers

Info

Publication number: WO1996031476A1
Application number: PCT/GB1996/000854
Authority: WO
Inventors: Paul Joseph Cox; Christopher Gregory Newton; Malcolm Norman Palfreyman
Original assignee: Rhone-Poulenc Rorer Limited
Priority date: 1995-04-07
Filing date: 1996-04-04
Publication date: 1996-10-10
Also published as: IL117836A0; GB9507297D0; ZA962776B; AU5282796A

Abstract

This invention is directed to physiologically active compounds of formula (I), wherein R1 is optionally substituted alkyl, R2 is optionally substituted aryl or heteroaryl, A is cycloalkyloxy, cycloalkylalkoxy or alkoxy carrying one or more hydroxy moieties, the cycloalkyl moieties optionally further substituted by one or more methylene or lower alkyl groups, Q?1, Q2 and Q3¿ each represents CH or CX or nitrogen, Z1 is oxygen or sulphur, Z2 is an NH or methylene and X is halogen and N-oxides thereof, and their esters, and prodrugs, pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof. Such compounds inhibit the production or physiological effects of TNF and inhibit cyclic AMP phosphodiesterase. The invention is also directed to pharmaceutical compositions containing compounds of formula (I), their pharmaceutical use and methods for their preparation.

Description

AROMATIC HYDRQXYETHERS

This invention is directed to substituted aromatic hydroxyethers, their preparation, pharmaceutical compositions containing these compounds, and their pharmaceutical use in the treatment of disease states associated with proteins that mediate cellular activity. The invention is particularly directed to the compounds in a substantially pure form, to their pharmaceutical use, pharmaceutical compositions containing the compounds, and methods for their preparation.

Tumour necrosis factor (TNF) is an important pro-inflammatory cyto ine which causes hemorrhagic necrosis of tumors and possesses other important biological activities. TNF is released by- activated macrophages, activated T-lymphocytes, natural killer cells, mast cells and baεophils, fibroblasts, endothelial cells and brain astrocytes among other cells.

The principal in vivo actions of TNF can be broadly classified as inflammatory and catabolic. It has been implicated as a mediator of endotoxic shock, inflammation of joints and of the airways, immune deficiency states, allograft rejection, and in the cachexia associated with malignant disease and some parasitic infections. In view of the association of high serum levels of TNF with poor prognosis in sepsis, graft versus host disease and adult respiratory distress syndrome, and its role in many other im unologic processes, this factor is regarded as an important mediator of general inflammation.

TNF primes or activates neutrophils, eosinophils, fibroblasts and endothelial cells to release tissue damaging mediators. TNF also activates monocytes, macrophages and T-lymphocytes to cause the production of colony stimulating factors and other pro-inflamm-atory cytokines such ILχ, ILβ, I e and GM-CSF, which in some case mediate the end effects of TNF. The ability of TNF to activate T-lymphocytes, monocytes, macrophages and related cells has been implicated in the progression of Human Immunodeficiency Virus (HIV) infection. In order for these cells to become infected with HIV and for HIV replication to take place the cells must be maintained in an activated state. Cytokines such as TNF have been shown to activate HIV replication in monocytes and macrophages. Features of endotoxic shock such as fever, metabolic acidosis, hypotension and intravascular coagulation are thought to be mediated through the actions of TNF on the hypothalamus and in reducing the anti-coagulant activity of vascular endothelial cells. The cachexia associated with certain disease states is mediated through indirect effects on protein catabolism. TNF also promotes bone resorption and acute phase protein synthesis.

The discussion herein relates to disease states associated with TNF include those disease states related to the production of TNF itself, and disease states associated with other cytokines, such as but not limited to IL-1, or IL-6, that are modulated by associated with TNF. For example, a IL-1 associated disease state, where IL-1 production or action is exacerbated or secreted in response to TNF, would therefore be considered a disease state associated with TNF. TNF-alpha and TNF-beta are also herein referred to collectively as "TNF" unless specifically delineated otherwise, since there is a close structural homology between TNF-alpha (cachectin) and TNF-beta (lymphotoxin) and each of them has a capacity to induce similar biological responses and bind to the same cellular receptor. Cyclic AMP phosphodiesterase is an important enzyme which regulates cyclic AMP levels and in turn thereby regulates other important biological reactions. The ability to regulate cyclic AMP phosphodiesterase, including type IV cyclic AMP phosphodiesterase, therefore, has been implicated as being capable of treating assorted biological conditions. In particular, inhibitors of type IV cyclic AMP phosphodiesterase have been implicated as being bronchodilators agents, prophylactic agents useful against asthma and as agents for inhibiting eosinophil accumulation and of the function of eosinophils, and for treating other diseases and conditions characterized by, or having an etiology involving, morbid eosinophil accumulation. Inhibitors of cyclic AMP phosphodiesterase are also implicated in treating inflammatory diseases, proliferative skin diseases and conditions associated with cerebral metabolic inhibition. It has already been reported that certain substituted aromatic compounds have valuable pharmaceutical properties, in particular the ability to regulate proteins that mediate cellular activity, for example, type IV cyclic AMP phosphodiesterase and/or TNF, for example in the specifications of International Patent Application Publications Nos. WO 92/12961, WO 93/25517, WO 94/02465, WO 94/12461, WO 94/20455 and United States Patent Application No. 08/098178), but hitherto the compounds of the present invention have never been specifically described.

Furthermore, compounds within the present invention possess markedly superior properties in comparison with known compounds, for example, greater bioavailability, higher in-vivo activity, and/or greater freedom from toxic effects, e.g. tendency to cause emesis.

This invention is directed to compounds of the general formula: -

wherein

R! represents a straight- or branched-chain alkyl group of 1 to about 6 carbon atoms, optionally substituted by one or more halogen atoms;

R2 represents an optionally substituted aryl group or an optionally substituted heteroaryl group;

A represents a substituted cycloalkyloxy, cycloalkylalkoxy or alkoxy group wherein the cycloalkyloxy, cycloalkylalkoxy or alkoxy group is substituted by one or more hydroxy moieties, and wherein the cycloalkyloxy group or the cycloalkyl portion of the cycloalkylalkoxy group is optionally further substituted by one or more methylene (=CH2) or by one or more lower alkyl groups;

Q¹, Q² and Q³, which may be the same or different, each represents a CH or CX linkage or a nitrogen atom; Z¹ represents an oxygen or sulphur atom;

Z² represents an NH or methylene linkage; and

X represents a halogen atom; and N-oxides thereof, and their esters, and prodrugs, pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof.

In the present specification, the term "compounds of the invention", and equivalent expressions, are meant to embrace compounds of general formula (I) as hereinbefore described, including the N-oxides, the esters, the prodrugs, the pharmaceutically acceptable salts, and the solvates, e.g. hydrates, where the context so permits. Similarly, reference to intermediates is meant to embrace their N-oxides, esters, salts, and solvates, where the context so permits. For the sake of clarity, particular instances when the context so permits are sometimes indicated in the text, but these instances are purely illustrative and it is not intended to exclude other instances when the context so permits.

It will be understood by those skilled in the art that compounds wherein the group represented by the symbol A contains the moiety -0-C-O- are so unstable that they are excluded from the scope of the invention.

As used above, and throughout the description of the invention, the following terms, unless otherwise indicated, shall be understood to have the following meanings : -

"Patient" includes both human and other mammals.

"Alkyl" means straight- or branched-chain alkyl. Preferred alkyl groups and moieties have 1 to about

12 carbon atoms in the chain. "Lower alkyl" means straight- or branched-chain alkyl of about 1 to about 4 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl and t-butyl groups.

"Alkoxy" means straight- or branched-chain alkoxy. Preferred alkoxy groups have 1 to about 10, more especially 3 to 7, carbon atoms in the chain. In a cycloalkylalkoxy group the alkoxy moiety preferably has 1 to 3 carbon atoms, especially 1.

"Cycloalkyloxy" denotes a cycloalkyl group, attached by an oxygen atom to the rest of the molecule. "Cycloalkyl" denotes a non-aromatic mono- or multicyclic ring system of about 3 to about 10 carbon atoms. Preferred monocyclic cycloalkyl groups include cyclohexyl, cycloheptyl and, more especially, cyclopentyl. Examples of multicyclic cycloalkyl groups include perhydronaphthyl, adamantyl and norbornyl groups and spirobicyclic groups, e.g. spiro [4 , 4] non-2-yl .

"Aryl" denotes an aromatic carbocyclic moiety of about 6 to about 10 carbon atoms. Examples of suitable aryl groups include phenyl and naphthyl groups optionally substituted by one or more substituents selected from, for example, halogen atoms and alkyl, phenyl, phenylalkyl, hydroxy, hydroxyalkyl, alkoxy, phenoxy, phenylalkoxy, nitro and cyano groups.

"Heteroaryl" denotes an aromatic monocyclic or multicyclic organic moiety of about 5 to about 10 ring members in which one or more of the ring members is/are element(s) other than carbon, for example nitrogen, oxygen or sulphur. Examples of suitable heteroaryl groups include pyrazinyl, furyl, thienyl, pyridyl, pyrimidinyl, isoxazolyl, isothiazolyl, pyridazinyl, 1,2,4-triazinyl, quinolinyl, and isoquinolinyl groups, optionally substituted by one or more substituents selected from, for example, halogen atoms and alkyl, phenyl, phenylalkyl, hydroxy, hydroxyalkyl, alkoxy, phenoxy, phenylalkoxy, nitro and cyano groups. Preferred heteroaryl groups include optionally substituted pyridyl, pyrazinyl, thienyl, pyrimidinyl, isoxazolyl and isothiazolyl groups. Especially preferred heteroaryl groups are optionally substituted pyridyl groups, especially wherein the optional substituents are alkyl groups or, more particularly, halogen atoms .

"Halogen atoms" means fluorine, chlorine, bromine, or iodine atoms. Fluorine, chlorine or bromine atoms are preferred; fluorine or chlorine atoms are more preferred, and fluorine atoms are particularly preferred. "Prodrug" means a compound, for example an ester, which is convertible in vivo by metabolic means (e.g. by hydrolysis) to a compound of formula (I), including N-oxides thereof. Suitable esters are of many different types, for example acetates, citrates, lactates, tartrates, malonates, oxalates, salicylates, propionates, succinateβ, fumarates, maleates, methylene-bis-β- hydroxynaphthoates, gentisates, mesylatee, isethionates, di-p-toluoyltartrates, me hane- sulphonates, ethanesulphonates, benzenesulphonates, p-toluenesulphonates, cyclohexylsulphamates and quinates and, especially, 2,2-dimethylpropanoates. An especially useful class of esters may be formed from acid moieties selected from those described by Bundgaard et al., J. Med. Chern., ____, No. 12, (1989), 2503-2507, and include substituted (aminomethyl)benzoates, for example dialkylamino- methylbenzoates in which the two alkyl groups may be joined together and/or interrupted by an oxygen atom or by an optionally substituted nitrogen atom, e.g. an alkylated nitrogen atom, more especially (morph- olinomethyl)benzoates, e.g. 3- or 4-(morpholino- methyl)benzoates, and (4-alkylpiperazin-l-yl)benz- oates, e.g. 3- or 4- (4-alkylpiperazin-l-yl)benzoates. Some of the compounds of the present invention are basic, and such compounds are useful in the form of the free base or in the form of a pharmaceutically acceptable acid addition salt thereof. As will be self-evident to those skilled in the art, some of the compounds of this invention do not form stable salts. However, acid addition salts are most likely to be formed by compounds of this invention containing a nitrogen-containing heteroaryl group.

Acid addition salts are a more convenient form for use; and in practice, use of the salt form inherently amounts to use of the free base form. The acids which can be used to prepare the acid addition salts include preferably those which produce, when combined with the free base, pharmaceutically acceptable salts, that is, salts whose anions are non-toxic to the patient in pharmaceutical doses of the salts, so that the beneficial inhibitory effects inherent in the free base are not vitiated by side effects ascribable to the anions. Although pharmaceutically acceptable salts of said basic compounds are preferred, all acid addition salts are useful as sources of the free base form even if the particular salt, per se, is desired only as an intermediate product as, for example, when the salt is formed only for purposes of purification, and identification, or when it is used as intermediate in preparing a pharmaceutically acceptable salt by ion exchange procedures. Pharmaceutically acceptable salts within the scope of the invention include those derived from mineral acids and organic acids, and include hydrohalides, e.g. hydrochlorides and hydrobromides, sulphates, phosphates, nitrates, sulphamateβ, acetates, citrates, lactates, tartrates, malonates, oxalates, salicylateε, propionates, succinates, fumarates, aleates, methylene-bis-β-hydroxynaphthoates, gentisates, mesylates, isethionates, di-p-toluoyl- tartrates, methanesulphonates, ethanesulphonates, benzenesulphonates, p-toluenesulphonateβ, cyclohexyl- sulphamates and quinates .

As well as being useful in themselves as active compounds, salts of compounds of the invention are useful for the purposes of purification of the compounds, for example by exploitation of the solubility differences between the salts and the parent compounds, side products and/or starting materials by techniques well known to those skilled in the art.

It will be apparent to those skilled in the art that certain compounds of the invention can exhibit isomerism, for example optical isomerism and geometrical isomerism. All such iso ers of the compounds of the invention, and their mixtures, are within the scope of the invention. For example, centres of chirality can occur in the group A, especially at carbon atoms attached to oxygen atoms, and these chiral centres can of course be in the

R- or the S-configuration.

A particular embodiment of this invention are compounds of formula (I) wherein R^- represents a straight- or branched-chain alkyl group of 1 to about 6 carbon atoms, optionally substituted by one or more halogen atoms; R² represents an optionally substituted aryl group or an optionally substituted heteroaryl group; A represents a substituted cycloalkyloxy or alkoxy group wherein the cycloalkyloxy or alkoxy group is substituted by one or more hydroxy moieties, and wherein cycloalkyloxy denotes a non-aromatic mono- or multicyclic ring system of about 3 to about 10 carbon atoms, attached by an oxygen atom to the rest of the molecule; Q¹, Q² and Q³, which may be the same or different, each represents a CH or CX linkage or a nitrogen atom; Z¹ represents an oxygen or sulphur atom; Z² represents an NH or methylene linkage; and X represents a halogen atom; and N-oxides thereof, and their esters, and prodrugs, pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof. The compounds of formula (I) wherein R¹ contains 1 to 3 carbon atoms is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R¹ is unsubstituted is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R¹ is substituted by one or more fluorine atoms is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R¹ represents a methyl group is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R¹ represents a difluoromethyl group is a class of preferred compounds of the invention. The compounds of formula (I) wherein R² represents a phenyl group substituted on the 2-position or on both the 2- and 6-positions, and is optionally further substituted, is a class of preferred compounds of the invention. The compounds of formula (I) wherein R² represents a phenyl group which is halogenated on the 2-position or on both the 2- and 6-positions, and is optionally further substituted, is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a phenyl group substituted by one or two chlorine atoms, and is optionally further substituted, is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a heteroaryl group substituted on the position next to the position which is directly attached to Z² is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a heteroaryl group substituted on the position next to the position which is directly attached to Z², and is further substituted, is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a heteroaryl group substituted on both the positions next to the position which is directly attached to Z² is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a heteroaryl group substituted by one or two halogen atoms is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a heteroaryl group substituted by one or two chlorine atoms is a class of preferred compounds of the invention. The compounds of formula (I) wherein R² represents an optionally substituted pyridyl group is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² represents a pyridyl group substituted by one or two chlorine atoms is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² is a 3,5-dihalopyrid-4-yl moiety is a class of preferred compounds of the invention.

The compounds of formula (I) wherein R² is a 3,5-dichloropyrid-4-yl moiety is a class of preferred compounds of the invention.

The compounds of formula (I) wherein A is an alkoxy group of 3 to 7 carbon atoms carrying one or two hydroxy groups is a class of preferred compounds of the invention.

The compounds of formula (I) wherein A is a monocyclic cycloalkyloxy group of 4 to 8 carbon atoms carrying one or two hydroxy groups is a class of preferred compounds of the invention.

The compounds of formula (I) wherein A carries one hydroxy group is a class of preferred compounds of the invention. The compounds of formula (I) wherein A is a l-hydroxyprop-2-yloxy group is a class of preferred compounds of the invention.

The compounds of formula (I) wherein A is a 3-hydroxycyclopentyloxy group is a class of preferred compounds of the invention. The compounds of formula (I) wherein Q¹ and Q² and Q³ each represents a CH linkage is a class of preferred compounds of the invention.

The compounds of formula (I) wherein Q² represents a nitrogen atom and Q¹ and Q³ each represents a CH linkage is a class of preferred compounds of the invention.

The compounds of formula (I) wherein Q² represents a CX linkage and Q¹ and Q³ each represents a CH linkage is a class of preferred compounds of the invention.

The compounds of formula (I) wherein X represents a fluorine atom is a class of preferred compounds of the invention. The compounds of formula (I) wherein Z¹ represents an oxygen atom is a class of preferred compounds of the invention.

The compounds of formula (I) wherein Z² represents an NH linkage is a class of preferred compounds of the invention.

The compounds of the invention which are N-oxides is a class of preferred compounds of the invention.

The compounds of the invention which are N-oxides of compounds of formula (I) wherein Q² represents a nitrogen atom and Q¹ and Q³ each represents a CH linkage is a class of preferred compounds of the invention.

The compounds of the invention which are N-oxides of compounds of formula (I) wherein R² represents a heteroaryl group containing a nitrogen atom is a class of preferred compounds of the invention.

The compounds of the invention wherein R² represents a 3,5-dihalo-l-oxido-4-pyridinio group is a class of preferred compounds of the invention.

The compounds of the invention wherein R² represents a 3, 5-dichloro-l-oxido-4-pyridinio group is a class of preferred compounds of the invention.

The compounds of the invention which are single optical isomers is a class of preferred compounds of the invention.

The compounds of the invention which are single geometric isomers is a class of preferred compounds of the invention. The compounds of the invention wherein A represents an optionally substituted hydroxycyclo- alkyloxy group wherein the bond connecting the hydroxy group to the cycloalkyl group is in the trans-configuration with respect to the bond connecting the cycloalkyl group to the rest of the molecule is a class of preferred compounds of the invention.

The compounds of the invention wherein A represents an optionally substituted hydroxycyclo- alkyloxy group wherein the bond connecting the hydroxy group to the cycloalkyl group is in the cis-configuration with respect to the bond connecting the cycloalkyl group to the rest of the molecule is a class of preferred compounds of the invention. The compounds of the invention which are in a substantially pure form is a class of preferred compounds of the invention. The compounds of the invention which have been prepared synthetically is a class of preferred compounds of the invention.

Certain of the compounds of the invention, namely those wherein R² represents an aryl or heteroaryl group optionally substituted by one, two or three substituents selected from halogen atoms and alkyl, hydroxy, alkoxy, nitro and cyano groups, Q¹, Q² and Q³ each represents a CH linkage, Z² represents an NH linkage and R¹, A and Z¹ are as hereinbefore defined, but which are not N-oxides nor esters nor pro-drugs, are within the broad scope of the specification of International Patent Application Publication No. WO 93/25517. However, they are not Exemplified in that specification and, as stated hereinbefore, hitherto they have never been specifically described.

A particularly preferred class of compounds of the invention includes the compounds of the general formula: -

wherein R¹ represents a methyl or difluoromethyl group; A represents a hydroxycycloalkyloxy group of 5 to 7 carbon atoms; represents a CH or CF linkage or a nitrogen atom; represents a nitrogen atom or a CH linkage;

X¹ and X² each represents a fluorine or, more particularly, chlorine atom; and

Z² represents an NH or methylene linkage; and N-oxides thereof, and pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof. A more particularly preferred class of compounds of the invention includes the N-oxides of the compounds of general formula (Ia) wherein

R¹ represents a methyl or difluoromethyl group;

A represents a hydroxycycloalkyloxy group of 5 to 7 carbon atoms;

Q² represents a CH linkage and Q⁴ represents a nitrogen atom; or Q² and Q⁴ both represent nitrogen atoms;

Z² represents an NH or methylene linkage; and pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof.

A more especially preferred class of compounds of the invention includes the compounds of formula

(Ia) wherein R¹ represents a methyl group.

A more especially preferred class of compounds of the invention includes the compounds of formula (Ia) wherein A represents a 3-hydroxycyclopentyloxy group, especially a trans-3-hydroxycyclopentyloxy group. A more especially preferred class of compounds of the invention includes the compounds of formula (Ia) wherein Q² represents a CH linkage.

A more especially preferred class of compounds of the invention includes the compounds of formula (Ia) wherein Q⁴ represents a nitrogen atom.

A more especially preferred class of compounds of the invention includes the compounds of formula (Ia) wherein X¹ and X² each represents a chlorine atom.

The invention includes compounds of the invention having any combination of the features referred to hereinbefore.

An especially preferred class of compounds of the invention, which possess markedly superior properties in comparison with known compounds, for example, greater bioavailability, higher in-vivo activity, and even greater freedom from toxic effects, includes the compounds of the general formula: -

wherein R¹ represents a methyl or difluoromethyl group; A represents a hydroxycycloalkyloxy group of 5 to 7 carbon atoms; and X¹ and X² each represents a fluorine or, more particularly, chlorine atom; and N-oxides thereof, and pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof .

A more particularly preferred class of compounds of the invention includes the N-oxides of the compounds of general formula (Ib) wherein

R¹ represents a methyl or difluoromethyl group; A represents a hydroxycycloalkyloxy group of 5 to 7 carbon atoms;

X¹ and X² each represents a fluorine or, more particularly, chlorine atom; and and pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof.

A more especially preferred class of compounds of the invention includes the compounds of formula (Ib) wherein R¹ represents a methyl group.

A more especially preferred class of compounds of the invention includes the compounds of formula (Ib) wherein A represents a 3-hydroxycyclopentyloxy group, especially a trans-3-hydroxycyclopentyloxy group.

(Ib) wherein X¹ and X² each represents a chlorine atom.

The invention includes compounds of the invention having any combination of the features referred to hereinbefore. Preferred compounds are selected from the following:

A (±) -N- (3 , 5-dichloro-4-pyridyl) -3- (cis-3-hydroxy¬ cyclopentyloxy) -4-methoxybenzamide; B (±) -N- (3, 5-dichloro-4-pyridyl) -3- (trans-3- hydroxycyclopentyloxy) -4-methoxybenzamide; C (±) -2- (3, 5-dichloro-4-pyridyl) -l-[3-(cis-3- hydroxycyclopentyl-oxy) -4-methoxyphenyl] ethanone; D (±) -2- (3, 5-dichloro-4-pyridyl) -1- [3- (trans-3- hydroxycyclopentyloxy) -4-methoxyphenyl] ethanone;

E (±) -2-(3,5-dichloro-l-oxido-4-pyridinio) -1- [3-

(cis-3-hydroxycyclopentyloxy) -4-methoxyphenyl] - ethanone;

F (±) -2- (3, 5-dichloro-l-oxido-4-pyridinio) -1- [3- (trans-3-hydroxycyclopentyloxy) -4-methoxyphenyl] - ethanone;

G (±) -2- (3, 5-dichloro-l-oxido-4-pyridinio) -1- [4- difluoromethoxy-3- (cis-3-hydroxycyclopentyloxy) - phenyl] ethanone; H (±) -2- (3, 5-dichloro-l-oxido-4-pyridinio) -1- [4- difluoromethoxy-3- (trans-3-hydroxycyclopentyloxy) - phenyl] ethanone;

I (±) -N- (3 , 5-dichloro-l-oxido-4-pyridinio) -3- (cis- 3-hydroxycyclopentyloxy) -4-methoxybenzamide; J (±) -N- (3, 5-dichloro-l-oxido-4-pyridinio) -3-

(trans-3-hydroxycyclopentyloxy) -4-methoxybenzamide; K (±) -2- (3 , 5-dichloro-4-pyridyl) -1- [3- ( 1-hydroxy- prop-2-yloxy) -4-methoxyphenyl] ethanone; L (±) -2- ( 3 , 5-dichloro-l-oxido-4-pyridinio) -1- [3- ( l-hydroxyprop-2-yloxy) -4-methoxyphenyl] ethanone; M ( - ) -N- ( 3 , 5-dichloro-4-pyridyl) -3- (trans-3- hydroxycyclopentyloxy) -4-methoxybenzamide; N (±)-N- (3, 5-dichloro-4-pyridyl) -4- (trans-3- hydroxycyclopentyloxy) -5-methoxypyridine-2- carboxamide;

0 (±) -N- (3, 5-dichloro-1-oxido-4-pyridinio) -4- (trans-3-hydroxycyclopentyloxy) -5-methoxy-l-oxido- pyridinium-2-carboxamide;

P (±) -N- (3, 5-dichloro-l-oxido-4-pyridinio) -3- [cis- 3- (2,2-dimethylpropanoyloxy)cyclopentyloxy] -4- methoxy-benzamide; Q (±)-2- (3,5-dichloro-4-pyridyl)-l- [4-difluoro- methoxy-3- (trans-3-hydroxycyclopentyloxy)phenyl] - ethanone;

R (±)-2-(3,5-dichloro-4-pyridyl)-l- [4-difluoro- methoxy-3- (cis-3-hydroxycyclopentyloxy)phenyl] - ethanone;

S (±)-N-(3,5-dichloro-4-pyridyl) -3- (1-hydroxyprop- 2-yloxy) -4-methoxybenzamide;

T (+) -N- (3, 5-dichloro-4-pyridyl) -3- (trans-3- hydroxycyclopentyloxy) -4-methoxybenzamide; U (±)-N- (3, 5-dichloro-4-pyridyl) -4- (tranβ-3- hydroxycyclopentyloxy) -5-methoxy-l-oxidopyridinium-2- carboxamide;

V (±) -N- (3, 5-dichloro-4-pyridyl) -3- [cis-3-(2,2- dimethylpropanoyloxy)cyclopentyloxy] -4-methoxy- benzamide;

W (±) -N- (3, 5-dichloro-4-pyridyl) -3-[cis-3-(3- morpholinomethylbenzoyloxy)cyclopentyloxy] -4-methoxy- benzamide;

X (±) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- [cis- 3- (3-morpholinomethylbenzoyloxy)cyclopentyloxy] -4- methoxybenzamide; Y (-) -N- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (trans-3-hydroxycyclopentyloxy) -4-methoxybenzamide; Z (+) -N-(3, 5-dichloro-1-oxido-4-pyridinio) -3- (trans-3-hydroxycyclopentyloxy) -4-methoxybenzamide; AA (±) -N- (3, 5-dichloro-4-pyridyl) -3- (trans-4- hydroxycyclohexyloxy) -4-methoxybenzamide; AB (±) -N- (3, 5-dichloro-4-pyridyl) -3- (cis-4- hydroxycyclohexyloxy) -4-methoxybenzamide; AC (±) -N- (3, 5-dichloro-l-oxido-4-pyridinio) -3- (trans-4-hydroxycyclohexyloxy) -4-methoxybenzamide;

AD (±) -N- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (cis- 4-hydroxycyclohexyloxy)-4-methoxybenzamide; AE (±) -N- (3, 5-dichloro-4-pyridyl) -3- (trans-3,3- dimethyl-5-hydroxycyclohexyloxy) -4-methoxybenzamide; AF (±)-N- (3, 5-dichloro-l-oxido-4-pyridinio) -3- (trans-3, 3-dimethyl-5-hydroxycyclohexyloxy) -4- methoxybenzamide;

AG N- (3, 5-dichloro-4-pyridyl) -3- (trans-2- hydroxycyclopentylmethoxy) -4-methoxybenzamide; AH N- (3, 5-dichloro-l-oxido-4-pyridinio) -3- (tranβ-2- hydroxycyclopentylmethoxy) -4-methoxybenzamide; AI (±) -2- (3, 5-dichloro-4-pyridyl) -1- [4-methoxy-3- (trans-3- (4-morpholinomethylbenzoyloxy)eyelopenty1- oxyphenyl]ethanone; AJ (±) -2- (3, 5-dichloro-4-pyridyl) -1- [4-methoxy-3-

(cis-3- (4-morpholinomethylbenzoyloxy)eyelopentyloxy- phenyl] ethanone;

AK (±) -N- (3, 5-dichloro-1-oxido-4-pyridinio) -4-

[trans-3- (4-morpholinomethylbenzoyloxy) cyclopentyl- oxy]hydroxycyclopentyloxy) -5-methoxy-l-oxido- pyridinio-2-carboxamide; AL (±) -2- (3, 5-dichloro-4-pyridyl) -1- [4-methoxy-3- (cis-3- (3-morpholinomethylbenzoyloxy)cyclopentyloxy- pheny1]ethanone; and

AM (±) -N- (3, 5-dichloro-4-pyridyl) -3- [cis-3- (4- morpholinomethylbenzoyloxy) cyclopentyloxy] -4-methoxy¬ benzamide.

The letters A to AM are allocated to compounds for easy reference in this specification.

A group of preferred compounds includes E, F, G, H, I, J and L.

A group of especially preferred compounds includes I and especially J, that is to say:- I (±) -els.- (3, 5-dichloro-l-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; and especially

J (±) -trans-(3,5-dichloro-1-oxido-4-pyridinio) -3- (3-hydroxycyclopentyloxy) -4-methoxybenzamide.

A class of compounds of importance consists of the single enantiomers of compounds I and J, namely (+) -cis- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; (-) -cis- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; (-) -trans- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; and

(+) -trans- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide.

The compounds of the invention exhibit useful pharmacological activity and accordingly are incorporated into pharmaceutical compositions and used in the treatment of patients suffering from certain medical disorders. The present invention thus provides, according to a further aspect, compounds of the invention and compositions containing compounds of the invention for use in therapy. Compounds within the scope of the present invention exhibit marked pharmacological activities according to tests described in the literature which tests results are believed to correlate to pharmacological activity in humans and other mammals. Detailed ___ vitro and in. vivo procedures are described hereinafter.

Compounds of the invention are inhibitors of tumor necrosis factor, especially TNF-alpha. Thus, in a further embodiment, the present invention provides compounds of the invention and compositions containing compounds of the invention for use in the treatment of a patient suffering from, or subject to, conditions which can be ameliorated by the administration of an inhibitor of TNF, especially of TNF-alpha. For example, compounds of the present invention are useful in joint inflammation, including arthritis, rheumatoid arthritis and other arthritic conditions such as rheumatoid spondylitis and osteoarthritis. Additionally, the compounds are useful in treatment of sepsis, septic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, asthma and other chronic pulmonary diseases, bone resorption diseases, reperfusion injury, graft vs. host reaction and allograft rejection. Furthermore, the compounds are useful in the treatment of infections such as viral infections and parasitic infections, for example malaria such as cerebral malaria, fever and myalgias due to infection, HIV, AIDS, cachexia such as cachexia secondary to AIDS or to cancer.

Compounds of the invention are also cyclic AMP phosphodiesterase inhibitors, in particular type IV cyclic AMP phosphodiesterase inhibitors. Thus, in another embodiment of the invention, we provide compounds of the invention and compositions containing compounds of the invention for use in the treatment of a patient suffering from, or subject to, conditions which can be ameliorated by the administration of an inhibitor of cyclic AMP phosphodiesterase, especially type IV cyclic AMP phosphodiesterase. For example, compounds within the present invention are useful as bronchodilators and asthma-prophylactic agents and agents for the inhibition of eosinophil accumulation and of the function of eosinophils, e.g. for the treatment of inflammatory airways disease, especially reversible airway obstruction or asthma, and for the treatment of other diseases and conditions characterized by, or having an etiology involving, morbid eosinophil accumulation. As further examples of conditions which can be ameliorated by the administration of inhibitors of cyclic AMP phosphodiesterase such as compounds of the invention there may be mentioned inflammatory diseases, such as atopic dermatitis, urticaria, allergic rhinitis, psoriasis, rheumatoid arthritis, inflammatory diseases (e.g. ulcerative colitis and Crohn's disease), adult respiratory distress syndrome and diabetes insipidus, other proliferative skin diseases such as keratosis and various types of dermatitis, conditions associated with cerebral metabolic inhibition, such as cerebral senility, multi-infarct dementia, senile dementia (Alzheimer's disease), and memory impairment associated with Parkinson's disease, and conditions ameliorated by neuroprotectant activity, such as cardiac arrest, stroke, and intermittent claudic tion.

Another group of conditions which may be treated with the compounds of the present invention includes diseases and disorders of the central nervous system such as brain trauma, ischaemia, Huntington's disease and tardive dyskinaesia

Other disease states which may be treated with the compounds of the present invention include pyresis, autoimmune diseases (e.g. systemic lupus erythematosus, allergic erythematosus, multiple sclerosis), type I diabetes mellitus, psoriasis, Bechet's disease, anaphylactoid purpura nephritis, chronic glomerulonephritis and leukemia.

A special embodiment of the therapeutic methods of the present invention is the treating of asthma.

Another special embodiment of the therapeutic methods of the present invention is the treating of joint inflammation.

According to a further feature of the invention there is provided a method for the treatment of a human or animal patient suffering from, or subject to, conditions which can be ameliorated by the administration of an inhibitor of cyclic AMP phosphodiesterase or of TNF, especially TNF-alpha, for example conditions as hereinbefore described, which comprises the administration to the patient of an effective amount of compound of the invention or a composition containing a compound of the invention. "Effective amount" is meant to describe an amount of compound of the present invention effective in inhibiting cyclic AMP phosphodiesterase and/or TNF and thus producing the desired therapeutic effect. According to another aspect of the invention, there is provided the use of a compound of the invention in the manufacture of a medicament for the treatment of a patient suffering from, or subject to, conditions which can be ameliorated by the administration of an inhibitor of cyclic AMP phosphodiesterase, especially type IV cyclic AMP phosphodiesterase.

According to a further aspect of the invention, there is provided the use of a compound of the invention in the manufacture of a medicament for the treatment of a patient suffering from, or subject to, conditions which can be ameliorated by the administration of an inhibitor of TNF, especially of TNF-alpha.

References herein to treatment should be understood to include prophylactic therapy as well as treatment of established conditions.

It is to be further understood that references herein to compounds of the invention and compounds of the invention are intended to include solvates (e.g. hydrates) of compounds of the invention. The present invention also includes within its scope pharmaceutical compositions comprising at least one of the compounds of the invention in association with a pharmaceutically acceptable carrier or excipient.

Compounds of the invention may be administered by any suitable means. In practice compounds of the present invention may generally be administered parenterally, rectally, orally or by inhalation, especially by the oral route.

Compositions according to the invention may be prepared according to the customary methods, using one or more pharmaceutically acceptable adjuvants or excipients. The adjuvants comprise, inter alia, diluents, sterile aqueous media and the various non- toxic organic solvents. The compositions may be presented in the form of tablets, pills, granules, powders, aqueous solutions or suspensions, injectable solutions, elixirs or syrups, and can contain one or more agents chosen from the group comprising sweeteners, flavorings, colorings, or stabilizers in order to obtain pharmaceutically acceptable preparations. The choice of vehicle and the content of active substance in the vehicle are generally determined in accordance with the solubility and chemical properties of the active compound, the particular mode of administration and the provisions to be observed in pharmaceutical practice. For example, excipients such as lactose, sodium citrate, calcium carbonate, dicalcium phosphate and disintegrating agents such as starch, alginic acids and certain complex silicates combined with lubricants such as magnesium stearate, sodium lauryl sulphate and talc may be used for preparing tablets. To prepare a capsule, it is advantageous to use lactose and high molecular weight polyethylene glycols. When aqueous suspensions are used they can contain emulsifying agents or agents which facilitate suspension. Diluents such as sucrose, ethanol, polyethylene glycol, propylene glycol, glycerol and chloroform or mixtures thereof may also be used.

For parenteral administration, emulsions, suspensions or solutions of the products according to the invention in vegetable oil, for example sesame oil, groundnut oil or olive oil, or aqueous-organic solutions such as water and propylene glycol, injectable organic esters such as ethyl oleate, as well as sterile aqueous solutions of the pharmaceut- icaily acceptable salts, are used. The solutions of the salts of the products according to the invention are especially useful for administration by intramuscular or subcutaneous injection. The aqueous solutions, also comprising solutions of the salts in pure distilled water, may be used for intravenous administration with the proviso that their pH is suitably adjusted, that they are judiciously buffered and rendered isotonic with a sufficient quantity of glucose or sodium chloride and that they are sterilized by heating, irradiation or microfiltration.

For administration by inhalation compounds of the invention may be dissolved or suspended in a suitable carrier for use in a nebulizer or a suspension or solution aerosol, or may be absorbed or adsorbed onto a suitable solid carrier for use in a dry powder inhaler. Solid compositions for rectal administration include suppositories formulated in accordance with known methods and containing at least one compound of the invention. The percentage of active ingredient in the compositions of the invention may be varied, it being necessary that it should constitute a proportion such that a suitable dosage shall be obtained. Obviously, several unit dosage forms may be administered at about the same time. The dose employed will be determined by the physician, and depends upon the desired therapeutic effect, the route of administration and the duration of the treatment, and the condition of the patient. In the adult, the doses are generally from about 0.001 to about 50, preferably about 0.001 to about 5, mg/kg body weight per day by inhalation, from about 0.01 to about 100, preferably 0.1 to 70, more especially 0.5 to 10, mg/kg body weight per day by oral administration, and from about 0.001 to about 10, preferably 0.01 to 1, mg/kg body weight per day by intravenous administration. In each particular case, the doses will be determined in accordance with the factors distinctive to the subject to be treated, such as age, weight, general state of health and other characteristics which can influence the efficacy of the medicinal product.

The compounds according to the invention may be administered as frequently as necessary in order to obtain the desired therapeutic effect. Some patients may respond rapidly to a higher or lower dose and may find much weaker maintenance doses adequate. For other patients, it may be necessary to have long-term treatments at the rate of 1 to 4 doses per day, in accordance with the physiological requirements of each particular patient. Generally, the active product may be administered orally 1 to 4 times per day. Of course, for some patients, it will be necessary to prescribe not more than one or two doses per day.

The compounds of the present invention may also be formulated for use in conjunction with other therapeutic agents such as agents which increase cyclic AMP production including β-agonists and PGE2 •

It is to be understood that the present invention includes combinations of compounds of the present invention with one or more of the aforementioned therapeutic agents.

Compounds of the invention may be prepared by the application or adaptation of known methods, which means methods used heretofore or described in the literature, for example as illustrated in the Examples and Reference Examples and chemical equivalents thereof.

Those skilled in the art will appreciate how to choose the reaction conditions in the following processes so as to avoid or minimise unwanted side- reactions .

For example, when a starting material contains ester moieties, any alcohol or ester used as reaction medium is preferably chosen so as to avoid transesterification. Those skilled in the art will also appreciate how to avoid or minimise unwanted side-reactions by the judicious and timely protection and deprotection of reactive groups, for example hydroxy groups, for example as illustrated in this specification. Thus, according to a feature of the invention, in a process (A), compounds of formula (I) in which the group represented by the symbol A contains one or more free hydroxy groups are prepared by the deprotection of the protected hydroxy groups in compounds of the general formula:-

.wherein R¹, R², Z¹, Z², Q¹, Q² and Q³ are as hereinbefore defined and A¹ represents a protected derivative of the group represented by the symbol A as just defined. General formula (II) may otherwise be written as : -

A.¹T ¹ (II) wherein T¹ represents a group of the formula:-

wherein R¹, R , Z¹, Z , Q¹, Q² and Q³ are as hereinbefore defined, and A¹ is as hereinbefore defined. The reaction may be carried out by the application or adaptation of known methods for the deprotection of such protected hydroxy groups. For example, suitable protecting groups include a silyl group substituted by three alkyl or aryl, e.g. phenyl, groups, for example a trialkylsilyl group, e.g. a tert-butyldimethylsilyl group, or an alkyl- diarylsilyl group, e.g. a tert-butyldiphenylsilyl group, and in those cases the deprotection can be carried out by reaction with an acid, for example by reaction with a Lewis acid, e.g. a quaternary alkyl ammonium halide, e.g. tetrabutylammonium fluoride, in an ethereal solvent such as tetrahydrofuran, at or near room temperature, or by reaction with an aqueous acid, e.g. toluene-p-sulphonic acid, formic acid, trifluoroacetic acid or acetic acid, neat or in the presence of a co-solvent, for example tetrahydrofuran or ethyl acetate, at a temperature from about room o temperature to about 100 C . According to a further feature of the present invention, in a process (B) , compounds of formula (I) wherein R¹, R² , R³ , Q¹, Q² ' Q³ and Z¹ are as hereinbefore defined, the group represented by the symbol A contains one or more free hydroxy groups and Z² represents an NH linkage, are prepared by the reaction of compounds of the general formula: -

wherein A¹, Q¹, Q², Q³ , R¹ and Z¹ are as hereinbefore described, and X³ represents a halogen, e.g. bromine or, preferably, chlorine, atom with compounds of the general formula: - R³NHR² (IV) wherein R² is as hereinbefore defined, and R³ represents a hydrogen atom or an acyl group, followed by removing any protecting groups present . R³ preferably represents a group R⁴C0, where R⁴ is an alkyl group containing up to 5 carbon atoms or a cycloalkyl group, preferably a methyl group. The reaction is preferably carried out in the presence of a base, for example an alkali metal hydride, such as sodium hydride, or an amine, preferably a tertiary amine, such as triethylamine, in an inert solvent, for example toluene, dimethylformamide, or an ether, such as tetrahydrofuran or diethyl ether, at a temperature from about 0 C to reflux, then with a second base, for example an amine, such as piperidine. It will be appreciated that if the starting material of formula (IV) is an N-oxide, then the product will also be an N-oxide.

According to a further feature of the present invention, in a process (C), compounds of formula (I) are prepared by interconversion of other compounds of the invention.

For example, the compounds of formula (I) wherein Q¹, Q² or Q³ represents a nitrogen atom and/or R² represents a heteroaryl group containing one or more nitrogen ring atoms, and Z¹ preferably represents an oxygen atom, can be oxidised to the corresponding N-oxides. The oxidation is carried out by means of reaction with a mixture of hydrogen peroxide and an organic acid, e.g. acetic acid, preferably at or above room temperature at 60-90°C. Alternatively, the oxidation is carried out by reacting a peracid, for example m-chloroperoxybenzoic acid, in an inert solvent such as dichloromethane, at a temperature from about room temperature to reflux, preferably at elevated temperature. Alternatively, the oxidation is carried out by reaction with hydrogen peroxide in the presence of sodium tungstate at temperatures between room temperature and about 60°C. This last method is preferred for compounds containing an acid-labile group.

As another example, an N-oxide group, in an N-oxide of a compound of formula (II) wherein Q¹, Q² or Q³ represents a nitrogen atom and/or R² represents a heteroaryl group containing one or more nitrogen ring atoms, Z¹ preferably representing an oxygen atom, can be reduced to a nitrogen atom. More particularly, one of the N-oxide groups in an N-oxide of a compound of formula (II) wherein Q¹, Q² or Q³ represents a nitrogen atom in its oxidised form and R² represents a heteroaryl group containing one or more nitrogen ring atoms in its oxidised form, can be reduced to a nitrogen atom. For example, the N-oxide group in the group R² in such a compound can be reduced to a nitrogen leaving the other N-oxide group unchanged. The reduction of an N-oxide group may be carried out by reaction with diphosphorus tetraiodide in an inert solvent, such as dichloromethane, preferably at or near room temp¬ erature, or by reaction with a chlorotrialkylsilane, preferably chlorotrimethylsilane, in the presence of zinc and an alkali metal iodide, e.g. potassium iodide, in an inert solvent, e.g. acetonitrile, at a temperature between about 0°C and about room temperature, preferably below room temperature.

According to a further feature of the invention, compounds of the invention containing hydroxy moieties are converted to their esters by the application or adaptation of known methods of esterification. For example, the appropriate acid may conveniently be converted to an acid halide, e.g. by reaction with thionyl chloride or oxalyl chloride, and then the acid halide is reacted with the appropriate alcohol of formula (I), preferably in the presence of a base, for example a tertiary amine, e.g. triethylamine. Alternatively, the appropriate alcohol of formula (I) may be reacted with the appropriate acid in the presence of compounds such as diisopropyl azodicarboxylate and triphenylphosphine, preferably in a dry ethereal solvent, e.g. diethyl ether or tetrahydrofuran, preferably at or near room temperature. These esterification routes are particularly applicable when Z² represents a methylene linkage. As another example, compounds of the invention containing hydroxy moieties are prepared by hydrolysis of esters of the invention. The hydrolysis may conveniently be by alkaline hydrolysis using a base, such as an alkali metal hydroxide or carbonate, in the presence of an aqueous/organic solvent mixture, using organic solvents such as dioxan, tetrahydrofuran or methanol, at a temperature from about ambient to about reflux. The hydrolysis of the esters may also be carried out by acid hydrolysis using an inorganic acid, such as hydro¬ chloric acid, in the presence of an aqueous/inert organic solvent mixture, using organic solvents such as dioxan or tetrahydrofuran, at a temperature from about 50°C to about 80°C. These routes are particularly applicable when Z² represents a methylene linkage.

According to a further feature of the invention, acid addition salts of the compounds of this invention are prepared by reaction of the free base with the appropriate acid, by the application or adaptation of known methods. For example, the acid addition salts of the compounds of this invention may conveniently be prepared either by dissolving the free base in aqueous or aqueous-alcohol solution or other suitable solvents containing the appropriate acid and isolating the salt by evaporating the solution, or by reacting the free base and acid in an organic solvent, in which case the salt separates directly or can be obtained by concentration of the solution.

The acid addition salts of the compounds of this invention can be regenerated from the salts by the application or adaptation of known methods. For example, parent compounds of the invention can be regenerated from their acid addition salts by treatment with an alkali, e.g. aqueous sodium bicarbonate solution or aqueous ammonia solution. Optical isomers and geometrical isomers of compounds of the invention can be separated from their mixtures, by the application or adaptation of known methods, for example chromatographic techniques and recrystallisation techniques, or they may be separately prepared from the appropriate isomers of their intermediates, for example by the application or adaptation of methods described herein.

The starting materials and intermediates may be prepared by the application or adaptation of known methods, for example methods as described in the Reference Examples or their obvious chemical equivalents .

For example, compounds of formula (II) wherein A¹, R¹, R² , Q¹, Q², Q³ and Z* are as hereinbefore defined and Z² represents a methylene group may be prepared by the reaction of compounds of the general formula : -

HO-T¹ (V) wherein T¹ is as hereinbefore defined and in which Z² represents a methylene group, with compounds of the general formula:- A^—H (VI) wherein A^ is as hereinbefore defined, preferably in the presence of compounds such as diisopropyl azodicarboxylate and triphenylphosphine, preferably in a dry ethereal solvent, e.g. diethyl ether or tetrahydrofuran, preferably at or near room temperature.

Compounds of formula (II) wherein A¹, R¹, R², Q¹, Q , Q³ and Z¹ are as hereinbefore defined and Z² represents an NH linkage may be prepared by the reaction of compounds of the general formula:-

T²-COX³ (VII) wherein X³ is as hereinbefore defined and T² represents a group of the formula:-

wherein A¹, R¹, Z¹, Q¹, Q² and Q³ are as hereinbefore defined, with compounds of the general formula:- R²NH2 (VIII) wherein R² is as hereinbefore defined, preferably in the presence of a base such as an alkali metal hydride, e.g. sodium hydride, or an amine, preferably a tertiary amine, e.g. triethylamine or pyridine, optionally in an inert solvent, for example dichloromethane, dimethylformamide, or an ether, e.g. diethyl ether or tetrahydrofuran, preferably at a temperature from 0°C to the reflux temperature or at the melting point of the reaction mixture. It will be appreciated that if the compound of formula (VIII) is an N-oxide, then the product of formula (II) will also be an N-oxide.

Compounds of formula (II), wherein A¹, R¹, R², Q¹, Q² and Q³ are as hereinbefore defined, Z² represents an NH linkage, and Z¹ represents an oxygen atom, may be prepared by the reaction of compounds of formula (VII) , wherein T² is as hereinbefore described, with compounds of formula (IV) wherein R² and R³ are as hereinbefore defined, preferably in the presence of a base, for example an alkali metal hydride, such as sodium hydride, or an amine, preferably a tertiary amine, such as triethylamine, in an inert solvent, for example toluene, dimethyl- formamide, or an ether, such as tetrahydrofuran or diethyl ether, at a temperature from about 0 C to reflux, then with a second base, for example an amine, such as piperidine. It will be appreciated that if the compound of formula (IV) is an N-oxide, then the product of formula (II) will also be an

N-oxide .

Compounds of formula (II), wherein A¹, R¹, R² , Z^, Q , Q² and Q³ are as hereinbefore defined and Z² represents a methylene linkage, may be prepared from compounds of the general formula:-

T²-COOR⁶ (IX) wherein T² is as hereinbefore defined and R"5 represents a lower alkyl, e.g. methyl, group by reaction with compounds of the general formula: - R²CH3 (X) wherein R² is as hereinbefore defined, in the presence of a strong base such as lithium diisopropylamide (usually prepared in situ from butyl lithium and diisopropylamine) , preferably at a low temperature.

Compounds of formula (II), wherein A¹, R¹, R², Z¹, Q¹, Q² and Q³ are as hereinbefore defined and Z² represents an NH linkage, may be prepared by the reaction of compounds of formula (IV), wherein R² is as hereinbefore defined and R³ represents a group R⁴CO in which R⁴ represents a lower alkyl, preferably methyl, group, with a base such as sodium hydride, preferably in a dry solvent such as dimethylform- amide, followed by reaction with compounds of the general formula:-

T²-CON3 (XI) wherein T² is as hereinbefore defined.

Compounds of formula (II) can also be prepared by interconversion of other compounds of formula (II) .

For example, the compounds of formula (II), wherein Q¹, Q² or Q³ represents a nitrogen atom and/or R² represents a heteroaryl group containing one or more nitrogen ring atoms, Z¹ preferably represents an oxygen atom, and R¹, R³ and Z² are as hereinbefore defined, can be oxidised to the corresponding N-oxides. The oxidation may be carried out by means of reaction with a mixture of hydrogen peroxide and an organic acid, e.g. acetic acid, preferably at or above room temperature at 60- 90°C. Alternatively, the oxidation is carried out by reacting a peracid, for example 3-chloroperoxy- benzoic acid, in an inert solvent such as dichloro- methane, at a temperature from about room temperature to reflux, preferably at elevated temperature. Alternatively, the oxidation is carried out by reaction with hydrogen peroxide in the presence of sodium tungstate at temperatures between room temperature and about 60°C. This last method is preferred for compounds containing an acid-labile group .

Compounds of formula (III) may be prepared, for example, from compounds of the general formula: -

wherein A^, Q , Q² , Q³, R^ and Z- are as hereinbefore defined, by the application or adaptation of known methods for the preparation of acid halides from carboxylic acids. For example, when X³ in the compound of formula (III) represents a chlorine atom, the reaction can be carried out by means of thionyl chloride in solution in a solvent such as toluene. Compounds of formula (XII) may be prepared, for example, from compounds of the general formula:-

wherein .⁶ and Z¹ are as hereinbefore defined, by the application or adaptation of known methods for the hydrolysis of esters, for example, methods as hereinbefore described, e.g. using potassium hydroxide in aqueous methanol, at or near room temperature.

Compounds of formula (XIII) wherein A^ is as hereinbefore defined may be prepared, for example, from compounds of the general formula:-

wherein A, Q¹, Q², Q³, R¹, R⁶ and Z¹ are as hereinbefore defined and A contains one or more hydroxy groups, by the application or adaptation of known methods for the protection of alcohols, for example, methods as hereinbefore described. Compounds of formula (XIV) may be prepared, for example, from compounds of formula (IX), wherein R⁶ and T² are as hereinbefore defined, by the application or adaptation of known methods for the deprotection of alcohols, for example, methods as hereinbefore described.

Compounds of formula (IV) may be prepared, for example, from compounds of formula (VIII), wherein ' is as hereinbefore defined, by the application or adaptation of known methods for the acylation of aromatic amines.

Compounds of formula (V) may be prepared, for example, by the reaction of compounds of the general formula: -

R⁷0-T¹ (XV) wherein T¹ is as hereinbefore defined and R⁷ represents an alkyl or cycloalkyl group, with hydrogen fluoride or a source thereof, e.g. "hydrogen fluoride-pyridine" [a material otherwise known as pyridinium poly(hydrogen fluoride), containing about 70% hydrogen fluoride and about 30^*% pyridine and considered to be a convenient source of anhydrous hydrogen fluoride, J. Fluorine Chem. , 3_7_, 343 (1987), commercially available from Aldrich Chemical Co.]. Compounds of formula (VI) may be prepared from compounds of the general formula:-

A-H (XVI) wherein A is as hereinbefore defined, by the application or adaptation of known methods for the protection of hydroxy groups. For example, when R³ represents a trialkylsilyl group, for example a tert-butyldimethylsilyl group, the compounds of formula (XVI) may be reacted with a trialkylsilyl halide, e.g. chloride, preferably in a solvent such as dichloromethane.

Compounds of formula (VII), wherein T² and X³ are as hereinbefore defined, may be prepared from compounds of the general formula:-

T²-COOH (XVII) wherein T² is as hereinbefore defined, by the application or adaptation of known methods for the preparation of acid halides from carboxylic acids. For example, when X³ in the compound of formula (VII) represents a chlorine atom, the reaction can be carried out by means of thionyl chloride or, preferably, oxalyl chloride in the presence of triethylamine. Compounds of formula (XVII), wherein T² is as hereinbefore defined, may be prepared by the oxidation of compounds of the general formula:-

T²-CHO (XVIII) wherein T² is as hereinbefore defined, e.g. by means of reaction with potassium permanganate, or with a mixture of sulphamic acid and sodium chlorite in acetic acid, or with sodium chlorite in the presence of sodium dihydrogen phosphate. Compounds of formula (XVIII), wherein T² is as hereinbefore defined, may be prepared from compounds of the general formula:-

wherein R¹, Ql, Q², Q³ and Z¹ are as hereinbefore defined, by reaction with compounds of the general formula: - ⁱ-X⁴ (XX) wherein A- is as hereinbefore defined and X⁴ is a halogen, preferably bromine, atom, preferably in the presence of a base, for example an alkali metal hydride, e.g. sodium hydride, an alkali metal hydroxide or carbonate, e.g. sodium hydroxide or carbonate, or an amine, preferably a tertiary amine, e.g. triethylamine or pyridine, optionally in an inert solvent, for example dichloromethane, dimethyl- formamide, or an ether, e.g. diethyl ether or tetra¬ hydrofuran, preferably at a temperature from 0°C to the reflux temperature, or alternatively by reaction with compounds of the general formula (VI), wherein T² is as hereinbefore defined, preferably in the presence of a compound such as diisopropyl azodicarb- oxylate.

Alternatively compounds of formula (XVII), wherein T² is as hereinbefore defined, may be prepared by the hydrolysis of compounds of formula (IX) wherein R⁶ and T² are as hereinbefore defined, e.g. by reaction with a base, such as an alkali metal carbonate or bicarbonate in the presence of water, followed by reaction with an aqueous acid such as dilute hydrochloric acid.

Compounds of formula (XI), wherein T² is as hereinbefore defined, may be prepared by the reaction of compounds of the general formula:-

T2-COOM¹ (XXI) wherein T² is as hereinbefore defined and M¹ represents an alkali metal, e.g. potassium, atom with compounds of the general formula:-

(R⁸0)2PON3 (XXII) wherein R⁸ represents an aryl, e.g. phenyl, group, preferably in a solvent such as dimethylformamide. Compounds of formula (XXI), wherein T² and M¹ are as hereinbefore defined, may be prepared from compounds of formula (IX) wherein T² and R⁶ are as hereinbefore defined, by the application or adaptation of known methods for the preparation of alkali metal salts from alkyl esters of carboxylic acids, for example, by reaction with an alkali metal trialkylsilanolate . Compounds of formula (IX), wherein R⁶ and T² are as hereinbefore defined, can be prepared from compounds of the general formula:-

(XXIII)

wherein R¹, R⁵, Q¹, Q², Q³ and Z¹ are as hereinbefore defined, by reaction with compounds of the formula (VI), wherein A¹ is as hereinbefore defined, preferably in the presence of diisopropyl azodicarb- oxylate and triphenylphosphine.

Compounds of formula (IX), wherein R⁵ and T⁴ are as hereinbefore defined, Z¹ represents an oxygen atom, Q¹ and Q³ represent CH and Q² represents N, may be prepared by the reaction of compounds of the general formula:-

wherein R¹ and R⁶ are as hereinbefore defined, with compounds of formula (VI), wherein A³- is as herein¬ before defined, in the presence of a dialkyl diazodi- carboxylate, for example diisopropyl diazodicarb- oxylate, and a phosphine, preferably a triaryl- phosphine, such as triphenylphosphine, in an inert solvent, for example toluene or an ether, such as tetrahydrofuran or diethyl ether, at a temperature from about -20°C to reflux.

Compounds of formula (XXIV) , wherein R¹ and R⁶ are as hereinbefore defined, may be prepared by the reaction of compounds of the general formula:-

wherein R^ is as hereinbefore defined, with compounds of the general formula:-

R⁶OH (XXVI) wherein R⁶ is as hereinbefore defined, in the presence of an acid, preferably a mineral acid, for example sulphuric acid, or preferably hydrogen chloride, at a temperature from about 0°C to reflux, preferably at an elevated temperature.

Compounds of formula (XXV) , wherein R¹ is as hereinbefore defined, may be prepared from compounds of the general formula:-

(XXVII)

wherein R⁸ represents R¹ or R⁹, R⁹ represents a protecting group, such as a benzyl group, and R¹ is as hereinbefore defined, by adaptation of the procedure described by Beyerman, Receueil, 7_7_/ 249- 57, (1958).

Compounds of formula (XXVII), wherein R^ is as hereinbefore defined, may be prepared from compounds of the general formula:-

(XXVIII)

wherein R⁹ is as hereinbefore defined, by adaptation of the procedures described by Beyerman, op . ci t . and European Patent Specification No. 204207, incorporated herein by reference.

Compounds of formulae (XV), (XIX) and (XXIII) may be prepared by the application or adaptation of known methods, for example as described in the specifications of International Patent Application Publications Nos. WO 94/02465 and WO 95/04045 and United States Patent Application No. 08/098178.

Intermediates of formulae (II), (III), (IX), (XI), (XII), (XIV), (XVIII), (XXI) are novel compounds and, as such, they and their processes described herein for their preparation constitute further features of the present invention. The present invention is further Exemplified but not limited by the following illustrative Examples and Reference Examples .

In the nuclear magnetic resonance spectra (NMR) the chemical shifts are expressed in ppm relative to tetramethylsilane. Abbreviations have the following significances: s = singlet; d = doublet; t = triplet; m = multiplet; dd = doublet of doublets; b = broad.

EXAMPLE 1

Compounds A. B. M and N

A stirred solution of 3, 5-dichloro-4-amino- pyridine (929mg) in dry dimethylformamide (50ml) was treated portionwise with sodium hydride (460mg; 60% dispersion in mineral oil; 11.5mmoles) and then it was stirred for 2 hours at room temperature. The suspension was treated dropwise with a solution of (±) -3- [cis-3- (tert-butyldimethylsilyloxy)cyclopentyl- oxy] -4-methoxybenzoyl chloride (1.8g; prepared as described in Reference Example 1) in dry dimethyl- formamide (5ml), and the solution was stirred for 18 hours. It was then treated with water (2ml), and concentrated in. vacuo to low bulk. Water (75ml) was added and the mixture extracted with ethyl acetate (100ml) . The organic phase was washed with hydrochloric acid (50ml;2N), dried over sodium sulphate and concentrated in. vacuo to give an oil. This oil (the crude tert-butyldimethylsilyl ether of the desired product) was dissolved in a solution of tetrabutylammonium fluoride in tetrahydrofuran (20ml; IM) and then stirred at room temperature for 18 hours. After concentration in vacuo the residue was dissolved in ethyl acetate (100ml) . The solution was washed with water (100ml), dried over sodium sulphate, and concentrated s. vacuo to give an oil, which was triturated with dichloromethane, to give (±) -N- (3, 5-dichloro-4- pyridyl) -3- (cis-3-hydroxycyclopentyloxy) -4-methoxy¬ benzamide (0.5g), m.p. 224-227 ^*C. [Elemental analysis:- C,54.2; H,4.7; N,6.8%; calculated:- C54.4; H,4.6; N,7.1%] .

By proceeding in a similar manner, but using the appropriate quantity of (±) -3- (trans-3- (tert-butyl- dimethylsilyloxy) cyclopentyloxy) -4-methoxybenzoyl chloride (prepared as described in Reference Example 1), there was prepared (±) -N- (3, 5-dichloro-4- pyridyl) -3- (trans-3-hydroxycyclopentyloxy) -4- methoxybenzamide, m.p. 154-157 ^*C. [Elemental analysis:- C54.6; H,4.6; N,6.8%; calculated: - C54.4; H,4.6; N,7.1%] . By proceeding in a similar manner but using the appropriate quantity of (- ) -3- (trans-3- (tert-butyldi- phenylsilyloxy) cyclopentyloxy) -4-methoxybenzoyl chloride (prepared as described in Reference Example 12), there was prepared (-) -N- (3, 5-dichloro-4- pyridyl) -3- (trans-3-hydroxycyclopentyloxy) -4-methoxy¬ benzamide, m.p. 158-159^*C. [Elemental analysis:- C54.2; H,4.5; N,6.9%; calculated: - C54.4; H,4.6; N,7.05%] .

By proceeding in a similar manner but using the appropriate quantity of (±) -4- (trans-3-tert-butyldi- phenylsilyloxycyclopentyloxy) -5-methoxypyridine-2- carbonyl chloride (prepared as in Reference Example 12) there was synthesised (±) -N- (3, 5-dichloro-4- pyridyl) -4- (trans-3-hydroxycyclopentyloxy) -5- methoxypyridine-2-carboxamide, m.p. 156-157^*C. [Elemental analysis:- C,51.1; H,4.3; N,10.4%; calculated:- C,51.3; H,4.3; N,10.55%].

EXAMPLE 2 Compounds C. D and K

A stirred solution of diisopropylamine (1.5ml) in dry tetrahydrofuran (100ml) at -78^*C under nitrogen was treated with a solution of n-butyl lithium in hexanes (6.71ml;1.6M) , and the solution was stirred for 0.25 hours. It was then treated with a solution of 3, 5-dichloro-4-methylpyridine (1.74g) in dry tetra-hydrofuran (10ml), dropwiβe, whilst maintaining the temperature at -78^*C, and the solution was stirred for a further period of 0.5 hours. It was then treated with a solution of methyl (±) -3- [cis-3- (tert-butyldimethylsilyloxy) - cyclopentyloxy] -4-methoxybenzoate (2.04g; prepared as described in Reference Example 3), in dry tetrahydro¬ furan (10ml), dropwise, and the mixture was allowed to warm to room temperature during 2 hours. It was then treated with saturated aqueous ammonium chloride solution (50ml). The aqueous phase and the organic phase were separated, and the aqueous phase was extracted twice with ethyl acetate (50ml). These extract was combined with the said organic phase, and the resulting mixture was dried over sodium sulphate and concentrated in vacuo, to give an oil.

This oil (the crude tert-butyldimethylsilyl ether of the desired product) was dissolved in a solution of tetrabutylammonium fluoride in tetra¬ hydrofuran (12ml; IM) , and then it was stirred at room temperature for 18 hours. It was then concentrated in. vacuo and the resulting residue was dissolved in ethyl acetate (100ml) . The solution was washed with water (100ml), dried over sodium sulphate, and concentrated a vacuo. to give an oil, which was subjected to flash chromatography on silica gel, eluting with a mixture of ethyl acetate and pentane (3:1, v/v), to give (±) -2- (3, 5-dichloro-4- pyridyl) -1- [3- (cis-3-hydroxycyclopentyloxy) -4- methoxyphenyl]ethanone (0.94g), m.p. 141-143 ^*C. [Elemental analysis:- C, 57.6; H,4.9; N,3.4%; calculated:- C,57.6; H,4.8; N,3.5%]. By proceeding in a similar manner, but using the appropriate quantity of methyl (±) -3- [trans-3- (tert- butyldimethylsilyloxy) cyclopentyloxy] -4-methoxy- benzoate (prepared as described in Reference Example 3), there was prepared (±) -2- (3, 5-dichloro-4- pyridyl) -1- [3- (trans-3-hydroxycyclopentyloxy) -4- methoxyphenyl] ethanone, m.p. 125-127 ^*C. [Elemental analysis:- C57.5; H,4.9; N,3.4%; calculated: - C57.6; H,4.8; N,3.5%] .

By proceeding in a similar manner, but using the appropriate quantity of methyl (±) -3- (1-tert-butyldi- phenylsilyloxyprop-2-yloxy) -4-methoxybenzoate (prepared as described in Reference Example 3), there was prepared (±) -2- <3, 5-dichloro-4-pyridyl) -1- [3- (1- hydroxyprop-2-yloxy) -4-methoxyphenyl] ethanone, m.p.127-128 'C. [Elemental analysis:- C55.2;

H,4.7; N,3.8%; calculated:- C,55.2; H,4.6; N, 3.8%] . EXAMPLE 3

Compound E. 0 and P

A mixture of (±) -2- (3, 5-dichloro-4-pyridyl) -1- [3- (cis-3-hydroxycyclopentyloxy) -4-methoxyphenyl] - ethanone (0.5g; prepared as described in Example 2) and 3-chloroperoxybenzoic acid (0.88g) in dichloro- methane (50ml) was heated at reflux for 6 hours. After cooling, the mixture was concentrated and the residue was subjected to medium pressure liquid chromatography on silica gel, eluting with a mixture of dichloromethane and methanol (9:1, v/v), to give (±) -2- (3, 5-dichloro-l-oxido-4-pyridinio) -1- [3- (cis-3- hydroxycyclopentyloxy) -4-methoxyphenyl] ethanone (O.llg), m.p. 185-187 ^*C. [Elemental analysis:- C,55.5; H,4.8; N,3.3%; calculated:- C55.4; H,4.7; N,3.4%] .

By proceeding in a similar manner but using the appropriate quantity of (±) -N- (3 , 5-dichloropyridin-4- yl) -4- (trans-3-hydroxycyclopentyloxy) -5-methoxy- pyridine-2-carboxamide (prepared as in Example 1) and a three fold excess of oxidant, there was obtained (±) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -4- (trans-3- hydroxyeyelo-pentyloxy) -5-methoxy-l-oxidopyridinium- 2-carboxamide, m.p. 243-4 ^*C. [Elemental analysis :- C,47.1; H,4.0; N,9.6%; calculated:- C,47.5; H,4.0;

N, 9.8%] .

By proceeding in a similar manner but using the appropriate quantity of (±) -N- ( 3 , 5-dichloropyridin-4 - yl)-3-(cis-3-(2,2, dime thy lpropanoy loxy ) eye lopenty 1- oxy) -4 -methoxybenzamide (prepared as described in

Example 13) there was synthesised (±)-N-(3,5- dichloro-1- oxido -4 -pyridinio) -3- [cis-3- ( 2 , 2 -dimethyl- propanoyloxy)cyclo-pentyloxy] -4-methoxybenzamide, m.p. 100-101'C. [Elemental analysis (as hydrate) :- C,54.1; H,5.0; N,5.8%; calculated:- C54.1; H,5.4; N, 5.5%] .

EXAMPLE 4 Compounds F & L

A stirred solution of (±)-2-(3,5-dichloro-4- pyridyl) -1- [3- (trans-3-hydroxycyclopentyloxy) -4- methoxyphenyl]ethanone (lg; prepared as described in Example 2) in glacial acetic acid (12ml) was treated dropwiβe with hydrogen peroxide (4.3ml; 27%), and then it was heated for 6 hours at 70-80^*C. The mixture was then neutralised to pH 7 by the addition of concentrated aqueous sodium hydroxide solution, and then extracted twice with ethyl acetate (50ml) . The combined extracts were dried over magnesium sulphate and concentrated in. vacuo and the resulting residue was subjected to flash chromatography on silica gel, eluting with a mixture of ethyl acetate and methanol (9:1, v/v), to give (±)-2-(3, 5-dichloro- l-oxido-4-pyridinio) -1- [3- (trans-3-hydroxycyclopent¬ yloxy) -4-methoxyphenyl] ethanone (0.9g), m.p. 164- 166'C. [Elemental analysis:- C55.3; H,4.5; N,3.4%; calculated:- C, 55.35; H,4.65; N,3.4%].

By proceeding in a similar manner, but using the appropriate quantity of (±) -1- [3- (l-hydroxyprop-2-yl- oxy) -4-methoxyphenyl] -2- (3, 5-dichloropyrid-4-yl) - ethanone (prepared as described in Example 2), there was prepared (±)-2- (3, 5-dichloro-l-oxido-4-pyrid- inio)-l-[3-(l-hydroxyprop-2-yloxy) -4-methoxy-phenyl] - ethanone, m.p. 188-189^*C. [Elemental analysis: C53.2; H,4.4; N,3.7%; calculated:- C52.9; H,4.4; N, 3.7%] .

EXAMPLE 5 Compounds G. H. 0 & R

A solution of triphenylphosphine (l.βlg) in dry tetrahydrofuran (45ml) was treated dropwise with a solution of diisopropyl azodicarboxylate (1.39g) in dry tetrahydrofuran (10ml) . The resulting creamy precipitate was stirred at room temperature for 0.5 hours, and then it was treated dropwise with a solution of (±) -trans-3-tert-butyldimethylsilyloxy- cyclopentanol (1.49g; prepared as described in Reference Example 4) in dry tetrahydrofuran (10ml) . It was then treated with 2- (3, 5-dichloro-l-oxido-4- pyridinio) -1- (4-difluoromethoxy-3-hydroxyphenyl) - ethanone (1.2g; prepared as described in Reference Example 5) in one batch. The resulting solution was then heated at 80-85^*C for 24 hours, and then it was cooled to room temperature and the solvent was removed under reduced pressure. The resulting residue was subjected to flash chromatography on silica gel, eluting with diethyl ether, to give (±) -2- (3 , 5-dichloro-l-oxido-4-pyridinio) -1- [4- difluoromethoxy-3- (cis-3-tert-butyldimethylsilyloxy- cyclopentyloxy)phenyl] ethanone (contaminated with triphenylphosphine oxide) in the form of a waxy solid. This waxy solid was dissolved in a solution of tetrahydrofuran and water (8ml; 20:1, v/v), and treated with p-toluenesulphonic acid monohydrate

(0.76g). The resulting mixture was stirred at room temperature for 6 hours, after which time it was treated with a saturated aqueous solution of sodium hydrogen carbonate (5ml). The mixture was stirred for 0.25 hours, and then it was extracted with ethyl acetate (100ml). The extract was dried over magnesium sulphate and the solvent was removed under reduced pressure. The resulting residue was subjected to flash chromatography on silica gel, eluting with a mixture of ethyl acetate and pentane (75:25, v/v), to give (±)-2-(3, 5-dichloro-l-oxido-4- pyridinio) -1- [4-difluoromethoxy-3- (cis-3-hydroxy- cyclopentyloxy)phenyl]ethanone (0.75g), in the form of a white solid, m.p 176^*C. [Elemental analysis:- C,50.43; H,3.85; N,3.05%; calculated:- C50.91; H,3.82; N,3.12%]. By proceeding in a similar manner, but using the appropriate quantity of (±)-cis-3-tert-butvldimethyl- silyloxycyclopentanol (prepared as described in Reference Example 4), there was prepared (±)-2-(3,5- dichloro-l-oxido-4-pyridinio) -1- [4-difluoromethoxy-3- (trans-3-hydroxycyclopentyloxy)phenyl]ethanone, m.p

172"C. [Elemental analysis:- C50.55; H,3.85; N,2.96%; calculated:- C, 50.91; H,3.82; N,3.12%] .

By proceeding in a similar manner but using the appropriate quantities of (±) -cis-3-tert-butyldi- methylsilyloxycyclopentanol (prepared as described in

Reference Example 4) and 2- (3, 5-dichloro-4-pyrid¬ inio) -1- (4-difluoromethoxy-3-hydroxyphenyl) -ethanone (prepared as described in Reference Example 5) there was prepared (±) -2- (3, 5-dichloro-4-pyridyl) -1- [4- difluoromethoxy-3- (trans-3-hydroxycyclopentyloxy) - phenyl]ethanone, m.p. 124-125^*C. [Elemental analysis:- C52.8; H,4.0; N,3.3%; calculated: - C52.8; H,4.0; N, 3.2%] .

By proceeding in a similar manner but using the appropriate quantities of (±) -trans-3-tert-butyldi- methylsilyloxycyclopentanol (prepared as described in Reference Example 4) and 2- (3, 5-dichloro-4-pyrid¬ inio) -1- (4-difluoromethoxy-3-hydroxyphenyl) ethanone (prepared as described in Reference Example 5) there was prepared (±) -2- (3 , 5-dichloro-4-pyridyl) -1- [4- difluoromethoxy-3- (cis-3-hydroxycyclopentyloxy) - phenyl]ethanone, m.p. 117-119^*C. [Elemental analysis:- C,52.9; H,3.9; N,3.2%; calculated:- C.52.8; H,4.0; N,3.2%] .

EXAMPLE 6

Compound I

A stirred suspension of (±) -N- (3, 5-dichloro-4- pyridyl) -3- (cis-3-hydroxycyclopentyloxy) -4-methoxy¬ benzamide (0.28g; prepared as described in Example 1) in dichloromethane (13ml) at room temperature was treated with metachloroperbenzoic acid (0.9g; 50%), and the mixture was heated at reflux for 6 hours . It was then concentrated in. vacuo. and the resulting residue was subjected to flash chromatography, eluting with a mixture of ethyl acetate and methanol (9:1, v/v), to give (±) -N- (3, 5-dichloro-l-oxido-4- pyridinio) -3- (cis-3-hydroxycyclopentyloxy) -4-methoxy- benzamide (20mg), m.p. 104-106^*C. [NMR (CDCI3) :- 8.26(s,2H), 7.62(bs,lH), 7.57(d,lH), 7.52(dd,lH), 6.96(d,lH), 5.0(m,lH), 4.38(m,lH), 3.93(s, 3H) , 2.2- i.88 (m, 4H) , 1.6(bs,2H) . [Elemental analysis:- C51.8; H,4.9; N,6.1%; Calculated:- C52.3; H,4.4; N, 6.8%] .

EXAMPLE 7 Compound J

A solution of (±) -N- (3, 5-dichloro-l-oxido-4- pyridinio) -3- (trans-3-tert-butyldimethylsilyloxy- cyclopentyloxy) -4-methoxybenzamide (0.4g; prepared as described in Reference Example 8) in a mixture of tetrahydrofuran and water (10ml; 20:1, v/v) was treated with p-toluenesulphonic acid (150mg), and the resulting mixture was stirred for 3 hours. The solution was then diluted with water (30ml) and concentrated in. vacuo to low bulk. The aqueous residue was extracted three times with ethyl acetate (50ml) and the combined extracts were dried over magnesium sulphate and concentrated in. vacuo. The resulting residue was subjected to flash chromat¬ ography, eluting with a mixture of ethyl acetate and methanol (9:1, v/v), to give (±) -N- (3, 5-dichloro-l- oxido-4-pyridinio) -3- (trans-3-hydroxycyclopentyloxy) - 4-methoxybenzamide (0.06g), m.p. 149-151^*C. [NMR(CDCl3) :- 8.24(s,2H), 7.78(bs,lH), 7.48- 7.52(m,2H), 6.93 (d, J=8HZ, IH) , 5.0-5.7 (m, IH) , 4.52- 4.6(m,lH), 3.92(s,3H), 2.27-2.37 (m, IH) , 2.1- 2.2(m,3H), 1.90-1.97 (m,lH) , 1.65-1.75 (m, IH) . [Elemental analysis:- C52.1; H,4.5; N, 6.1%; Calculated:- C,52.3; H,4.4; N,6.8%] . EXAMPLE 8 Compound S

(±) -N- (3, 5-Dichloro-4-pyridyl) -3- ( 1-tert-butyl- diphenylsilyloxyprop-2-yloxy) -4-methoxybenzamide (0.6g) (prepared as described in Reference Example 8) was treated with a solution of tetrabutylammonium fluoride (2.5ml) in tetrahydrofuran (30ml). The resulting solution was stirred at room temperature for 18 hours then evaporated. The residue was dissolved in ethyl acetate (100ml) and the solution washed with water (100ml) . The organic phase was dried over magnesium sulphate and evaporated. The residual oil was subjected to flash chromatography on silica gel eluting with ethyl acetate to give (±) -N- (3, 5-dichloro-4-pyridyl) -3- (l-hydroxyprop-2- yloxy) -4-methoxybenzamide (80mg), m.p. 146-7 'C. [Elemental analysis:- C.51.8; H,4.3; N,7.55%; calculated:- C51.5; H,4.45; N,7.5%].

EXAMPLE 9

Compound T

A solution of 3, 5-dichloro-4-aminopyridine (0.73g) in dry dimethylformamide (10ml) at room temperature under nitrogen was treated portionwise a 60% oil dispersion of sodium hydride (0.2g) . After stirring at ambient temperature for 2 hours the mixture was cooled to 0^*C and then treated with (+) -3- (trans-3-hydroxycyclopentyloxy) -4-methoxy- benzoyl azide [prepared by treating a solution of (+) -3- (trans-3-hydroxycyclopentyloxy) -4-methoxy- benzoic acid (0.5g) (prepared as in Reference Example 10) and diphenylphosphoryl azide (0.5ml) in dry dimethylformamide (10ml), under nitrogen and at ambient temperature, with triethylamine (0.35ml) dropwise and stirring for 2 hours] . The reaction mixture was then allowed to warm to room temperature, stirred for 5 hours and then allowed to stand overnight at room temperature. The solution was concentrated in vacuo. water (50ml) was added and the pH of the mixture adjusted to 5 by the addition of 2N hydrochloric acid. The aqueous solution was extracted twice with ethyl acetate (200ml). The combined organic layers were washed with hydrochloric acid (200ml;2N), then with brine (100ml), dried over magnesium sulphate and evaporated. The residual oil was subjected to flash chromatography on silica gel eluting with ethyl acetate to give (+) -N- (3, 5-di- chloro-4-pyridyl) -3- (trans-3-hydroxycyclopentyloxy) - 4-methoxybenzamide (0.3g). m.p. 158-159^*C. t^D] at 23^*C + 16.13 (c=0.0012g/ml) . [Elemental analysis:- C54.4; H,4.6 ;N,6.9%; calculated:- C54.4; H,4.6; N,7.05%].

EXAMPLE 10 Compound U

To a stirred solution of N- (3, 5-dichloro-4- pyridyl) -4- (trans-3-hydroxycyclopentyloxy) -5-methoxy- pyridine-2-carboxamide (1.35g; prepared as in Example 1) in dichloromethane (50ml) at room temperature was added m-chloroperbenzoic acid (1.03g). The resulting mixture was stirred for 24 hours and then a quantity of m-chloroperbenzoic acid (500mg) was added and stirring was continued for a further 6 hours. The reaction mixture was allowed to stand at ambient temperature for 5 days then treated with aqueous sodium bicarbonate (100ml). After stirring for 1 hour the layers were separated. The organic phase was dried over magnesium sulphate and evaporated. The resulting oil was subjected to flash chromatography on silica gel eluting with a mixture of ethyl acetate and methanol (95:5, v/v) to give (±) -N- (3, 5-dichloro- 4-pyridyl) -4- (trans-3-hydroxycyclopentyloxy) -5-meth- oxy-l-oxido-pyridinium-2-carboxamide (0.34g), m.p. 174-175"C. [Elemental analysis:- C49.12; H,4.1; N,9.9%; calculated:- C,49.3; H,4.1; N,10.1%].

EXAMPLE 11 Compound V A solution of (±) -N- (3, 5-dichloro-4-pyridyl) -3- (cis-3-hydroxycyclopentyloxy) -4-methoxybenzamide (prepared as in Example 1) (0.5g) in tetrahydrofuran (12ml) , under nitrogen, was treated with trimethyl- acetic acid (0.13g), diisopropylazodicarboxylate (0.25ml) and triphenylphosphine (0.34g). The mixture was stirred at ambient temperature for 4 hours. The solution was evaporated and the residue subjected to flash chromatography on silica gel eluting with a mixture of ethyl acetate and pentane (1:1, v/v) to give (±) -N- (3, 5-dichloro-4-pyridyl) -3- [cis-3- (2, 2- dimethylpropanoyloxy)cyclopentyl-oxy] -4-methoxy- benzamide (0.46g), m.p. 104-106"C. [Elemental analysis:- C,57.5; H,5.4; N,5.8%; calculated:- C57.4; H,5.4; N.5.9%] . EXAMPLE 12 Compounds W and X

A stirred solution of 3, 5-dichloro-4-amino- pyridine (0.18g) in dry dimethylformamide (5ml) was treated with sodium hydride (O.lg, 60 % dispersion in mineral oil) . After stirring at ambient temperature for 3 hours the mixture was cooled to 0^*C then treated with (±) -3- [cis-3- (3-morpholinomethyl)benz- oyloxycyclopentyloxy] -4-methoxybenzoyl azide [prepared by treating a solution of (±) -3- [cis-3- {3- (morpholino methyl)benzoyloxy)cyclopentyloxy] -4- methoxybenzoic acid (0.5g) (prepared as in Reference Example 10) and diphenylphosphoryl azide (0.33g) in dry dimethylformamide (5ml), under nitrogen and at room temperature, dropwise with triethylamine (1.3ml) and stirring the resulting solution at ambient temperature for 3 hours] portionwise over 1 hour. When the addition was complete the solution was stirred at room temperature overnight and then evaporated. The residue was partitioned between ethyl acetate (100ml) and water (100ml) . The aqueous phase was extracted with ethyl acetate (25ml) . The combined organic phases were washed with brine (25ml), dried over magnesium sulphate then evaporated. The residue was subjected to flash chromatography, eluting with a mixture of dichloro- methane and methanol (19:1, v/v), to give (±)-N-(3,5- dichloro-4-pyridyl) -3- [cis-3- ( 3-morpholinomethylbenz- oyloxy) cyclopentyloxy] -4-methoxybenzamide (0.28g) , m.p. 71^*C. [Elemental analysis as 1.2ml water: -

C58.0; H,5.2; N,6.5%; calculated:- C,57.9; H,5.4; N, 6.75%] . By proceeding in a similar manner but using the appropriate quantity of N-(3, 5-dichloro-l-oxido-4- pyridinio)acetamide there was synthesised (±)-N-(3,5- dichloro-l-oxido-4-pyridinio) -3- [cis-3- (3-morpholino- methylbenzoyloxy)cyclopentyloxy] -4-methoxybenzamide, m.p. 96-98^*C. [Elemental analysis:- C,58.25; H,5.3; N,6.4%; calculated:- C,58.4; H,5.1; N,6.8%].

EXAMPLE 13 Compounds Y and Z

A suspension of (-) -N-(3, 5-dichloro-4-pyridyl) - 3-(trans-3-hydroxycyclopentyloxy) -4-methoxybenzamide (0.15g, prepared as described in Example M) in dichloromethane (10ml) was treated with peracetic acid (2ml, 37%). After stirring at ambient temperature for 3 days the reaction mixture was evaporated and the residue treated with water (10ml) . The insoluble material was recrystallised from ethyl acetate to give (-)-N- (3,5-dichloro-l-oxido-4- pyridinio) -3- (trans-3-hydroxycyclopentyloxy) -4- methoxybenzamide (0.04g) as a white solid, m.p. 90- 95^*C with decomposition.

A suspension of (+) -N- (3, 5-dichloro-4-pyridyl) - 3- (trans-3-hydroxycyclopentyloxy) -4-methoxybenzamide (O.lg, prepared as described in Example T) in dichloromethane (5ml) was treated with peracetic acid (lml, 37%). After stirring at ambient temperature overnight the mixture was treated with another aliquot of peracetic acid (lml) and stirring was then continued for 2 days. The reaction mixture was evaporated and the residue treated with water (5ml). The insoluble material was recrystallised from ethyl acetate to give (+) -N- (3, 5-dichloro-l-oxido-4- pyridinio) -3- (trans-3-hydroxycyclopentyloxy) -4- methoxybenzamide (O.Olg) as a white solid, m.p. 91- 98^*C with decomposition.

REFERENCE EXAMPLE 1 A stirred solution of crude (±)-3- [cis-3-(tert- butyldimethylsilyloxy)cyclopentyloxy] -4-methoxy- benzoic acid (2g; prepared as described in Reference Example 2) and tert-butyldimethylsilyl chloride

(0.95g) in dichloromethane (30ml), under nitrogen and at room temperature, was treated with 1,8-diazabi- cyclo[5.4.0]undec-7-ene (0.93ml), dropwise, and the resulting solution was stirred for 18 hours. It was then was concentrated _____ vacuo and the resulting residue was partitioned between water (75ml) and diethyl ether (100ml) . The aqueous layer was washed with diethyl ether (100ml), and the combined organic layers were dried over sodium sulphate and concentrated.

The resulting residue was dissolved in dichloromethane (15ml) containing dimethylformamide (4 drops), at 0^*C under an atmosphere of nitrogen, and treated, dropwise, with oxalyl chloride (0.75ml). The resulting solution was allowed to warm to room temperature during 2 hours, and then it was concentrated in. vacuo. The residue was then treated with toluene (100ml), and the solution was concentrated i vacuo again, to give (±)-3- [cis-3- (tert-butyldimethylsilyloxy) cyclopentyloxy] -4-meth- oxybenzoyl chloride (1.8g).

This product was used directly in Example 1. By proceeding in a similar manner, but using the appropriate quantity of (±) -3- [trans-3- (tert-butyldi- methylsilyloxy)cyclopentyloxy] -4-methoxybenzoic acid, prepared as described in Reference Example 2, there was prepared (±) -3- [trans-3- (tert-butyldimethylsilyl- oxy)cyclopentyloxy] -4-methoxybenzoyl chloride.

This product was used directly in Example 1.

REFERENCE EXAMPLE 2 A solution of methyl (±) -3- [cis-3-(tert-butyl- dimethylsilyloxy)cyclopentyloxy] -4-methoxybenzoate (4g; prepared as described in Reference Example 3) in methanol (70ml) and water (7ml) was treated with potassium carbonate (3g), and the resulting solution was heated at reflux for 24 hours. The mixture was cooled and filtered, the filtrate was concentrated in vacuo. and the resulting residue was treated with ethyl acetate (150ml). The resulting organic solution was washed with hydrochloric acid (100ml;2N) and then with saturated aqueous sodium bicarbonate solution (100ml), dried over sodium sulphate, and concentrated in vacuo, to give (±) -3- [cis-3-(tert- butyldimethylsilyloxy)cyclopentyloxy] -4-methoxy- benzoic acid (3g) . [NMR(CDCl3) :- 7.69(dd,lH), 7.49(d,lH), 6.85(d,lH), 4.71(m,lH), 4.21(m,lH), 3.88(s,3H), 2.4(m,lH), 2.0(m,2H), 1.8(m,3H), 0.85(s,9H), 0.02(s,3H), 0.01(s,3H)].

By proceeding in a similar manner, but using the appropriate quantity of methyl (±) -3- [trans-3- (tert- butyldimethylsilyloxy)cyclopentyloxy] -4-methoxy- benzoate, prepared as described in Reference Example 3, there was prepared (±) -3- [trans-3- (tert-butyldi- methylsilyloxy)cyclopentyloxy] -4-methoxybenzoic acid. [NMR (CDCI3):- 7.69(dd,lH), 7.52(d,lH), 6.84(d,lH), 4.89(m,lH), 4.41(d,lH), 3.85(s,3H), 2.22(m,lH), 2.05- 1.93(m,3H), 1.8(m,lH), 1.57(m,lH), 0.82(s,9H), 0.0 (s,6H)].

By proceeding in a similar manner, but using the appropriate quantity of methyl (-)-3- [ (trans-3-tert- butyldiphenylsilyloxy)cyclopentyloxy) -4-methoxy- benzoate (prepared as in Reference Example 3), there was prepared (-) -3- [trans-3- (tert-butyldiphenyl- silyloxy)cyclopentyloxy] -4-methoxybenzoic acid. [NMR(CDCl3) :- 7.73-7.28 (m,13H), 4.85(m,lH), 4.45(m,lH), 3.68(s,3H), 2.23-1.54 (m, 6H) , 1.0(s,9H)]. By proceeding in a similar manner, but using the appropriate quantity of methyl (±) -4- (trans-3-tert- butyldiphenylsilyloxycyclopentyloxy) -5-methoxy- pyridine-2-carboxylate (prepared as in Reference Example 3) there was prepared (±) -4- (trans-3-tert- butyldiphenylsilyl-oxycyclopentyloxy) -5-methoxypyr- idine-2-carboxylic acid, m.p. 106-7^*C. [NMR (CDCI3) :- 8.07(s,lH), 7.72-7.34 (m,HH), 5.1(m,lH), 4.48(m,lH), 3.94(s,3H), 2.42-1.72 (m, 6H) , 1.07(s,9H)].

REFERENCE EXAMPLE 3 A stirred solution of methyl 3-hydroxy-4- methoxybenzoate (2.53g), (±) -trans-3-tert-butyldi- methylsilyloxycyclopentanol (3g; prepared as described in Reference Example 4) and triphenyl¬ phosphine (3.64g) in dry tetrahydrofuran (90ml) was treated, dropwise, with diisopropyl azodicarboxylate

(2.76ml), and the mixture was heated at reflux overnight. The solution was then concentrated in vacuo, triturated with a mixture of diethyl ether and pentane, and filtered. The filtrate was diluted with ethyl acetate (200ml), washed with aqueous sodium hydroxide solution (2x200ml;2N) , dried over sodium sulphate and concentrated. The resulting residue was subjected to medium pressure liquid chromatography on silica gel, eluting with a mixture of pentane and ethyl acetate (9:lv/v), to give methyl (±) -3- [cis-3- (tert-butyldimethylsilyloxy) cyclopentyl- oxy]-4-methoxybenzoate (4g) . [NMR(CDCl3 ) :- 7.65 (dd,lH), 7.49(d,lH), 6.87(d,lH), 4.73(m,lH), 4.24 (m,lH), 3.89(s,3H), 3.88(s,3H), 2.42(m,lH), 2.1-1.93 (m,2H), 1.83(m,3H), 0.88(s,9H), 0.06(s,3H), 0.05 (s,3H) ] . By proceeding in a similar manner, but using the appropriate quantity of (±) -cis-3-tert-butyldimeth- ylsilyloxycyclopentanol, prepared as described in Reference Example 4, there was prepared methyl (±) -3- [trans-3- (tert-butyldimethylsilyloxy) cyclopent- yloxy] -4-methoxybenzoate. [NMR(CDCl3) : -

7.6(dd,lH), 7.47(d,lH), 6.81(d,lH), 4.89(m,lH), 4.4 (quintet, IH) , 3.84(s,3H), 3.83(s,3H), 2.22(m,lH), 2.03-1.93(m,3H) , 1.8(m,lH), 1.57 (m, IH) ₍ 0.82(s,9H), 0.0 (s, 6H) ] . By proceeding in a similar manner, but using the appropriate quantity of (±) -1-tert-butyldiphenyl- silyloxypropan-2-ol, prepared as described in Reference Example 9, there was prepared methyl (±) -3- ( 1-tert-butyldiphenylsilyloxyprop-2-yloxy) -4- methoxybenzoate. [NMR(CDC13) : - 7.69-7.6 (m, 6H) ,

7.45-7.33(m, 6H) , 6.87 (d,lH), 4.58(m,lH), 3.92- 3.68(m,2H), 3.88(s,3H), 3.87 (s,3H), 1.37(d,3H), 1.03 (s, 9H) ] .

By proceeding in a similar fashion but using (±) -l-tert-butyldiphenylsilyloxypropan-2-ol (prepared as described in Reference Example 9) there was prepared methyl (±) -3- (1-tert-butyldiphenylsilyloxy- prop-2-yloxy) -4-methoxybenzoate. [NMR(CDCl3) :- 7.68-7.63(m,6H) , 7.43-7.33 ( , 6H) , 6.88(d,lH), 4.58(m,lH), 3.88(s,3H), 3.87(2,3H), 3.72(q,2H), 1.38(d,3H), 1.02(s,9H)] .

By proceeding in a similar manner but using the appropriate quantities of 3-hydroxy-4-methoxybenz- aldehyde and (+) -cis-3-acetoxycyclopentanol (prepared as described in Reference Example 11), there was prepared (+) -3- (trans-3-acetoxycyclopentyloxy) -4- methoxy-benzaldehyde. [<«D] at 24^*C + 7.8^*

(c=0.016g/ml; dichloromethane) . [NMR(CDCI3 ) : - 9.84(s,lH), 7.45(dd,lH), 7.48(d,lH), 6.97(d,lH), 5.34(m,lH), 4.98(m,lH), 3.94(s,3H), 2.34-2.16 (m, 4H) , 2.07(s,3H), 1.97(m,lH), 1.82(m,lH)] .

By proceeding in a similar manner, but using the appropriate quantity of (+) -cis-3- (tert-butyl- diphenylsilyloxy)cyclopentanol (prepared as described in Reference Example 13), there was prepared methyl ( - ) -3- [ (trans-3-tert-butyldiphenyl- silyloxy) cyclopentyloxy] -4-methoxybenzoate. [NMR (CDCI3) :- 7.73-7.32(m,12H) , 6.84(d,lH), 4.98(m,lH), 4.5(m,lH), 3.9(s,3H), 3.83(s,3H), 2.36-1.66 (m, 6H) , 1.06(s, 9H) ] . By proceeding in a similar manner but using the appropriate quantities of (±) -cis-1-tert-butyldi- methylsilyloxy-3-hydrocyclopentane (prepared as described in Reference Example 15) and 3-hydroxy-4- ethoxybenzaldehyde, there was prepared (±) -3- (trans- 3-tert-butyldimethylsilyloxycyclopentyloxy) -4- methoxybenzaldehyde. [NMR(CDCl3) :- 7.38(dd,lH), 7.32(d,lH), 6.9(d,lH), 4.91(m,lH), 4.4(m,lH),

3.88(s,3H), 2.3-1.53 (m, 6H) , 0.83(s,9H), 0.0(s,6H)]. By proceeding in a similar manner to Reference Example 3 but using (±) -3- (trans-3-hydroxycyclo- pentyloxy) -4-methoxybenzaldehyde and 3-(4-morpholino- methyl)benzoic acid there was prepared (±) -3- [cis-3-

(3-morpholinomethylbenzoyloxy)cyclopentyloxy] -4-meth- oxybenzaldehyde. [NMR(CDCl3) :- 9.73 (s,lH), 7.98 (m,2H), 7.56(d,lH), 7.46(dd,lH), 7.4(m,2H), 6.989 (d,lH), 5.44(m,lH), 4.93(m,lH), 3.9(s,3H), 3.7(t,4H), 3.53 (s,2H), 2.5θ( , IH) , 2.44(m,5H), 2.2(m,4H)].

By proceeding in a similar manner but using the appropriate quantities of methyl (±) -4-hydroxy-5- methoxypyridine-2-carboxylate and 3-tert-butyldi- phenylsilyloxycyclopentanol (prepared as in Reference Example 15), there was prepared methyl (±) -4- (trans-

3-tert-butyldiphenylsilyloxycyclopentyloxy) -5- methoxypyridine-2-carboxylate. [NMR(CDCl3) :- 8.17(s,lH), 7.7-7.35 (m,11H) , 5.05(m,lH), 4.48(m,lH), 3.99(s,3H), 3.92(s,3H), 2.42-1.7 (m, 6H) ] .

REFERENCE EXAMPLE 4 A mixture of cis/ (±) -trans-cvclopentane-1.3-diol (5g) and tert-butyldimethylsilyl chloride (6.6g) in dichloromethane (30ml) at room temperature was treated, dropwise, with 1, 8-diazabicyclo[5.4.0]undec-

7-ene (6.6ml). The mixture was stirred at room temperature for 3 hours, then it was diluted with dichloromethane (30ml), washed with hydrochloric acid (50ml;2N)and then with saturated aqueous sodium bicarbonate solution (50ml), dried over sodium sulphate, and concentrated in. vacuo. The resulting oil was subjected to medium pressure liquid chromatography on silica gel, eluting with a mixture of dichloromethane and methanol (9:lv/v), to give, firstly, (±)-cis-3-tert-butvldi- methylsilyloxycyclopentanol (lg) [NMR(CDCl3) : - 4.4(m,lH), 4.27(m,lH), 3.05(d,lH), 1.96-1.62(m,6H) , 0.88(s,9H), 0.08(8,6H)]; and then (±) -trans-3-ter - butyldimethylsilyloxycyclopentanol (4g) [NMR (CDCI3):- 4.42(m,2H), 2.14-1.96(m,2H) , 1.83(m,2H), 1.52(m,2H), 0.87(8, 9H), 0.04(s,6H)]. By proceeding in a similar manner, but using the appropriate quantities of hydroxyacetone and tert- butyldiphenylsilyl chloride, there was prepared l-tert-butyldiphenylsilyloxypropan-2-one. [NMR(CDC1₃) :- 7.75-7.35(m,10H) , 2.2(s,2H), 1.65 (bs,lH) , 1.12(B,9H)] .

REFERENCE EXAMPLE 5 Hydrogen fluoride-pyridine [a material otherwise known as pyridinium poly(hydrogen fluoride), containing about 70% hydrogen fluoride and about 30% pyridine and considered to be a convenient source of anhydrous hydrogen fluoride, J. Fluorine Chem. , 37, 343 (1987), commercially available from Aldrich Chemical Co.; (18ml)] at 0^*C was stirred and treated with 1- (3-cyclopentyloxy-4-difluoromethoxyphenyl) -2-

(3, 5-dichloro-1-oxido-4-pyridinio)ethanone (3.89g; prepared as described in the specifications of International Patent Application Publication No. WO 94/02465 and United States Patent Application No. 08/098178) . The resulting solution was allowed to warm to room temperature and stirred for 5 hours. The mixture was basified to pH9 by treatment with aqueous sodium hydroxide solution (IM), and it was then acidified to pH4 by treatment with concentrated hydrochloric acid. The aqueous solution was then extracted twice with ethyl acetate (100ml), the combined organic extracts were dried over magnesium sulphate and the solvent was removed under reduced pressure. The resulting residue was triturated with diethyl ether, to give 2-(3,5-dichloro-l-oxido- 4-pyridinio) -1- (4-difluoromethoxy-3-hydroxyphenyl) - ethanone (2.77g), m.p 195-197^*C. [NMR(CDCl3) : - 9.80(bs.lH), 8.23(s,2H), 7.67(d,lH), 7.54(dd,lH), 7.24(d,lH), 6.67(t,lH), 4.56(s,2H)].

By proceeding in a similar manner but using the appropriate quantity of 1- (3-cyclopentyloxy-4- difluoromethoxyphenyl) -2- (3, 5-dichloro-4-pyridyl) - ethanone (prepared as described in the specifications of International Patent Application Publication No. WO9402465), there was prepared 2- (3, 5-dichloro-4-pyr- idyl) -1- (4-difluoromethoxy-3-hydroxyphenyl)ethanone, m.p. 124-125^*C. [Elemental analysis:- C52.8;

H,4.0; N,3.3%; calculated:- C,52.8%; H,4.0; N,3.2%].

REFERENCE EXAMPLE 6 A stirred solution of methyl (±)-3- [trans-3- (tert-butyldimethylsilyloxy) cyclopentyloxy] -4- methoxybenzoate (lg: prepared as described in Reference Example 3) in dry toluene (50ml) under nitrogen was treated with potassium trimethylsilan- olate (0.44g), and the mixture was heated on a steam bath for 5 hours. It was then cooled, and concen¬ trated in. vacuo, to give potassium (±) -3- [trans-3- (tert-butyl-dimethylsilyloxy) cyclopentyloxy] -4- methoxybenzoate, in the form of a white gum.

The whole of this product was used directly in Reference Example 7.

By proceeding in a similar fashion but using methyl (±) -3- (l-tert-butyldiphenylsilyloxyprop-2- yloxy)-4-methoxybenzoate [prepared as described in Reference Example 3] there was prepared potassium (±)-3- (l-tert-butyldiphenylsilyloxyprop-2-yloxy) -4- methoxybenzoate, which was used directly in Reference Example 7.

REFERENCE EXAMPLE 7 A stirred solution of potassium (±)-3- [trans-3- (tert-butyldimethylsilyloxy)cyclopentyloxy] -4-meth- oxybenzoate (the whole of the product prepared as described in Reference Example 6) in dry dimethyl- formamide (25ml) was treated, dropwise, with a solution of diphenylphosphoryl azide (0.81g) in dry dimethylformamide (5ml) and the mixture was stirred for 3 hours. A clear solution of (±)-3- [trans-3- (tert-butyldimethylsilyloxy)cyclopentyloxy] -4-meth- oxybenzoyl azide was formed, the whole of which was used directly in Reference Example 8.

By proceeding in a similar manner but using potassium (±) -3- (l-tert-butyldiphenylsilyloxyprop-2- yloxy) -4-methoxybenzoate [prepared as described in Reference Example 6] there was prepared (±)-3-(l- tert-butyldiphenylsilyloxyprop-2-yloxy) -4-methoxy- benzoyl azide, which was used directly in Reference Example 8.

REFERENCE EXAMPLE 8

A stirred solution of N-(3,5-dichloro-l-oxido-4- pyridinio)acetamide (0.66g) in dry dimethyl- ormamide (24ml) was treated, portionwise, with sodium hydride (430mg; 60% dispersion in mineral oil; 10.75mmoles) and the mixture was stirred for a further 3 hours after the addition was complete. It was then treated, portionwise, with the whole of the solution of (±) -3- [trans-3- (tert-butyldimethylsilyloxy)cyclo¬ pentyloxy] -4-methoxybenzoyl azide prepared in Reference Example 7, and then it was stirred for a further 5 hours and allowed to stand at room temp¬ erature overnight. The solution was concentrated in vacuo. and the resulting residue was dissolved in ethyl acetate (150ml) and washed with saturated aqueous ammonium chloride solution (75ml). The aqueous phase was extracted with ethyl acetate

(2x30ml) and the combined organic phases were dried over magnesium sulphate and concentrated in vacuo. to give a brown oil, which was subjected to medium pressure liquid chromatography on silica gel, eluting with ethyl acetate, to give the (±)-N-(3, 5-dichloro- l-oxido-4-pyridinio)-3- (trans-3-hydroxycyclopentyl¬ oxy) -4-methoxybenzamide (0.44g) [NMR(CDCl3) :- 8.3 (s,2H), 7.6(bs,lH), 7.4-7.5 (m, 2H) , 6.9(d,J=8HZ, IH) , 4.92-5.0 (m,lH), 4.4-4.5(m, IH) , 3.91(s,3h), 2.2-2.35 (m,lH), 1.8-2.Km,4H) , 1.58-1.7 (m, IH) , 0.87(s,9H), 0.05 (s, 6H) ] . By proceeding in a similar fashion but using 4-amino-3,5-dichloropyridine and (±) -3- (1-tert-butyl- diphenylsilyloxyprop-2-yloxy) -4-methoxybenzoyl azide [prepared as described in Reference Example 7], there was prepared (±) -3- (1-tert-butyldiphenylsilyloxyprop- 2-yloxy) -N- (3, 5-dichloropyrid-4-yl) -4-methoxybenz- amide. [NMR(CDCl3) :- 8.57(s,2H), 8.43 (s,lH)_. 7.68-7.2(m,12H) , 6.94(d,lH), 4.63(m,lH), 3.91(s,3H), 3.74(q,2H), 1.49(d,2H), 1.0(s,9H)].

REFERENCE EXAMPLE 9 A stirred solution of 1-tert-butyldiphenylsilyl- oxypropan-2-one (5.3g; prepared as described in Reference Example 4) in methanol (100ml) was treated portionwise with sodium borohydride (1.25g), and the resulting solution was stirred for 30 minutes. The solution was concentrated in. vacuo. and then it was treated with water (100ml) and the resulting residue was extracted with ethyl acetate (100ml) . The organic layer was washed with aqueous sodium hydroxide solution (100ml;2N) , dried over magnesium sulphate and concentrated ±__ vacuo. to give (±) -l-tert-butyldiphenylsilyloxypropan-2-ol (4.9g) , in the form of a yellow oil. [NMR(CDCl3) :- 7.75- 7.65(m,5H), 7.47- 7.35(m,5H), 3.92 (m.lH), 3.65-3.43 (m,2H), 2.58(bs,lH), 1.01(d,3H), 1.05(s,9H)].

REFERENCE EXAMPLE 10 A stirred solution of (+) -3- (trans-3-acetoxy- cyclopentyloxy) -4-methoxybenzaldehyde (7g) (prepared as described in Reference Example 3) was treated with sulphamic acid (4.3g) and the solution was cooled to 0^*C. A solution of sodium chlorite (4.4g) in water (40ml) was added dropwise keeping the temperature between 13-15^*C. The solution was allowed to reach room temperature and stirred for a further 2 hours. Water (250ml) was added and the solution extracted twice with ethyl acetate (200ml) and then with 2N sodium hydroxide (300ml) . The aqueous layer was acidified to pH5 with hydrochloric acid and extracted twice with ethyl acetate (200ml) . The combined organic layers were dried over magnesium sulphate and evaporated to give (+) -3- (trans-3-hydroxycyclopent¬ yloxy) -4-methoxybenzoic acid as a white solid (7.3g), m.p. 159^*C. [°-=]D at 20^* + 12.2^* (c=0.0098g/ml; methanol) . [Elemental analysis:- C, 61.5; H,6.4%; calculated:- C,61.2; H, 6.2%].

By proceeding in a similar manner but using the appropriate quantity of (±) -3- [cis-3- (3- (morpholino- methyl) benzoyloxycyclopentyloxy] -4-methoxybenzald¬ ehyde (prepared as in Reference Example 3) there was synthesised (±) -3- [cis-3- (3- (morpholinomethyl) - benzoyloxycyclopentyloxy] -4-methoxybenzoic acid. [NMR(CDCl3) :- 8.14(m,2H), 7.77(dd,2H), 7.68(d,lH), 7.5 (t,lH), 6.93(d,lH), 5.4.4 On, IH) _ 4.95(m,lH), 4.08- 3.92 (m,4H), 3.91(s,3H), 2.9(m,4H), 2.37-2.16 (m, 6H) ] .

REFERENCE EXAMPLE 11

A suspension of (+) -cis-3, 5-diacetoxycyclo- pentane (5.4g) (prepared as described by Laumen and Schneider, Tet . Lett. 2_5_ 5875-8 (1984)) and potassium azodicarboxylate (74g) in methanol (400ml) at 0'C was treated with glacial acetic acid (38ml) in methanol (400ml) . After stirring at ambient temperature for 7 hours and allowing the suspension to stand at room temperature overnight the mixture was treated with a further quantity of potassium azodicarboxylate (37g) . The reaction mixture was stirred for a further 4 hours then treated with water (200ml) . The mixture was concentrated to low bulk and the residue extracted three times with ethyl acetate (100ml). The organic layers were combined, dried over magnesium sulphate then evaporated to give (+) -cis-3- acetoxycyclopentanol (5g) . [°^C]D ^{t 23}' + 5.8^* (c=0.012g/ml; chloroform). [NMR(CDCl3) : - 5.18 (m,lH), 4.34(m,lH), 3.75(t,lH), 2.16(m,lH), 2.04 (s,3H), 2.03-1.6(m,6H)] .

REFERENCE EXAMPLE 12

A solution of (-) -3- [trans-3- (tert-butyldi- phenylsilyloxy) cyclopentyloxy] -4-methoxybenzoic acid (4.5g) (prepared as described in Reference Example 2) in a mixture of dichloromethane (30ml) and dimethyl- formamide (3 drops), under nitrogen and at room temperature, was treated dropwise with oxalyl chloride (1.6ml) . After stirring for 3 hours, the solution was concentrated to give (-) -3- [trans-3- (tert-butyldiphenyIsilyloxy)cyclopentyloxy] -4- methoxybenzoyl chloride which was used directly in

Example 1.

By proceeding in a similar manner, but using the appropriate quantity of (±) -4- (trans-3-tert-butyldi- phenylsilyloxycyclopentyloxy) -5-methoxypyridine-2- carboxylic acid (prepared as in Reference Example 2) there was prepared (±) -4- (trans-3-tert-butyldiphenyl- silyloxycyclopentyloxy) -5-methoxypyridine-2-carbonyl chloride, which was used directly in Example 1.

REFERENCE EXAMPLE 13 A solution of (+) -cis-3-tert-butyldiphenyl- silyloxycyclopentylacetate (7.8g) (prepared as described in Reference Example 14) in ethanol (100ml) was treated with a solution of potassium hydroxide (2.0g) in water (25ml) . The mixture was stirred at room temperature overnight then evaporated. The residue was treated with water (100ml) and the aqueous solution extracted twice with ethyl acetate (200ml) . The organic extracts were combined, dried over magnesium sulphate and evaporated to give (+) -cis-3-tert-butyldiphenylsilyloxycyclopentanol (6.7g) as a colourless oil. [NMR(CDCl3 ) : - 7.75- 7.33 (m,10H), 4.8(m,lH), 4.26(m,lH), 2.95<m,lH), 1.97-1.83(m,3H) , 1.68-1.57 (m, IH) , 1.06(s,9H)] .

REFERENCE EXAMPLE 14

A solution of (+) -cis-3-acetoxycyclopentanol (4.5g) (prepared as in Reference Example 11) and tert-butyldiphenylsilyl chloride (9.5g) in dichloro¬ methane (50ml) was treated dropwise with 1,8-diazabi- cyclo[5.4.0]undec-7-ene (4.68ml). The resulting solution was stirred at room temperature for 1 hour then allowed to stand at room temperature overnight. The mixture was concentrated and the residue was dissolved in ethyl acetate (50ml). The organic solution was washed with water (50ml), then with 2N hydrochloric acid (50ml), then with aqueous sodium bicarbonate (50ml) . The organic layer was dried over magnesium sulphate and evaporated, to give (+) -cis-3-tert-butyldiphenyIsilyloxycyclopenty1 acetate (8.2g) . [NMR(CDCl3 ) : - 7.74-7.33 (m, 10H) , 5.0(m,lH), 4.23(m,lH), 2.12-1.6 (m, 6H) ] .

By proceeding in a similar manner but using the appropriate quantity of (±) -ciβ-l-acetoxy-4-hydroxy- cyclopent-2-ene [prepared as described by Deardorff and Myles, Organic Synthesis, 6_9_ 13-16 (1989)], there was prepared (±) -cis-l-acetoxy-4-tert-butyldiphenyl- silyloxycyclopent-2-ene. [NMR(CDCl3) : - 7.75-7.35 (m,10H). 5.92(m,lH), 5.85(m,lH), 5.36(m,lH), 4.68 (m,lH), 2.65(m,lH), 2.07(s,3H), 1.75(m,lH). 1.06 (s,9H)] .

REFERENCE EXAMPLE 15 A stirred solution of (±) -cis-l-acetoxy-4-tert- butyldiphenylsilyloxycyclopent-2-ene (66.8g) (prepared as in Reference Example 14) in ethanol (660ml) was treated with a solution of potassium hydroxide (11.8g) in water (50ml) at room temperature. After stirring for 3 hours, the mixture was treated with 5% palladium on charcoal (4g) and ammonium formate (12.6g) and was heated at 50 ^*C with vigorous stirring under a nitrogen atmosphere for 3 hours. The mixture was cooled to room temperature, filtered and the filter cake washed twice with ethanol (200ml) . The combined filtrate and washings were concentrated to give a yellow paste which was partitioned between ethyl acetate (1000ml) and water (500ml) . The organic phase was washed with water (500ml), dried over magnesium sulphate then evaporated. The resultant oil was subjected to flash chromatography on silica gel eluting with dichloromethane to give (±) -cis-1-tert-butyldiphenyl- silyloxy-3-hydroxycyclopentane (26g) . [NMR(CDCl3) : - 7.7-7.36(m,10H) , 4.38 (m,lH), 4.26(m,lH), 2.93(d,lH), 1.98-1.56(m, 6H) , 1.07 (s,lH)].

REFERENCE EXAMPLE 16 A stirred solution of (±) -3- (trans-3-tert- butyldimethylsilyloxycyclopentyloxy) -4-methoxy- benzaldehyde (13.48g) (synthesised as described in Reference Example 3) in tetrahydrofuran (30ml) was treated with tetrabutylammonium fluoride (65ml; IM solution in tetrahydrofuran) dropwise and the solution was stirred for 7 hours at room temperature. The solution was concentrated and the residue dissolved in ethyl acetate (100ml) and washed with water (100ml). The organic phase was dried and evaporated. The residue was subjected to flash chromatography on silica gel eluting with ethyl acetate to give (±) -3- (trans-3-hydroxycyclopentyl¬ oxy) -4-methoxybenzaldehyde (8.46g). [NMR(CDCl3) : - 7.46(dd,lH), 7.38(d,lH), 6.97(d,lH), 5.03(m,lH), 4.57 (m,lH), 3.94(s,3H), 2.38-1.38(m, 6H) ] .

IN VITRO AND IN VIVO TEST PROCEDURES

1. Inhibitory effects of compounds on PDE IV activity 1.1 Preparation of PDE from guinea pig macrophages

The method is described in Turner et al. (Br. J. Pharmacol, 108 876-883, 1993) . Briefly, cells are harvested from the peritoneal cavity of horse-serum treated (0.5ml i.p.) Dunkin Hartley guinea pigs (250- 400g) and the macrophages purified by discontinuous

(55%, 65%, 70% v/v) gradient (Percoll) centrifug- ation. Washed macrophages are plated out in cell culture flasks and allowed to adhere. The cells are washed with Hank's balanced salt solution, scraped from the flasks and centrifuged (1000 g) . The supernatant is removed and the pellets stored at -80°C until use. The pellet is homogenised in 20mM tris (hydroxymethyl) aminomethane HCl, pH7.5, 2mM magnesium chloride, lmM dithiothreitol, 5mM ethylenediaminetetraacetic acid, 0.25mM sucrose, 20μM p-tosyl-L-lycine chloromethyl ketone, lOμg/ml leupeptin and 2000U/ml aprotinin. 1.2 Measurement of PDE activity.

PDE activity is determined in macrophage homogenates by the two-step radioisotopic method of

Thompson et al., (Adv. Cyclic Nucl. Res., __Q_, 69-92, 1979) . The reaction mixture contains 20mM tris (hydroxymethyl)aminomethane HCl (pH8.0), lOmM magnesium chloride, 4mM 2-mercaptoethanol, 0.2mM ethylenebis (oxyethylenenitrilo) tetraacetic acid and

0.05 mg of bovine serum albumin/mL. The concentration of substrate is lμM. The IC₅₀ values (i.e. concentrations which produce 50% inhibition of substrate hydrolysis) for the compounds examined are determined from concentration-response curves in which concentrations range from 0.InM to 40μM. 1.3 Results.

Compounds within the scope of the invention produce up to about 50% inhibition of guinea pig macrophage cyclic AMP-specific phosphodiesterase (PDE IV) at concentrations from about 10"⁹M up to about 10^"5M, preferably from about 10"⁹M up to about 10^-7M. The compounds of the invention are from about 10,000-fold to about 50-fold more selective for cyclic AMP phosphodiesterase IV than cyclic nucleotide phosphodiesterase types I, III or V.

2. In vivo bronchodilator actions of compounds 2.1 Measurement of bronchodilatation.

Bronchorelaxant activity is measured in in vivo tests in the anaesthetized guinea-pig or rat according to the method described in Underwood et al., Pulm. Pharmacol. 5., 203-212, (1992) in which the effects on bronchospaβm induced by histamine (or other spasmogens such as methacholine or leukotriene D4) is determined. Compounds are administered orally 1 hour prior to administration of spasmogen.

3. In vivo actions of compounds on antiσen (ovalbamin. -induced eosinophilia in σuinea-ϋiσs 3.1 Treatment of animals and measurement of eosinophil numbers. Male Dunkin-Hartley guinea-pigs weighing 200- 250g are sensitized using lOμg ovalbumin in ImL of a lOOmg/mL suspension of aluminium hydroxide, i.p.

28 days after sensitization guinea-pigs are dosed orally. 23 Hours later this procedure is repeated and 60 minutes later the guinea-pigs are challenged with nebulised saline or ovalbumin (1% in saline) for 15 seconds. 24 Hours after challenge the guinea-pigs are killed and the lungs are lavaged with warm saline. Total and differential cell counts are made.

4. Inhibitory effects of compounds against antiσen- induced eosinophilia in the rat in vivo 4.1. Treatment of rats and measurement of eosinophil numbers.

Male Brown Norway rats weighing 150-250g are sensitized on days 0, 12 and 21 with ovalbumin (lOOμg, i.p.) . Rats are challenged on any one day between days 27-32.

24 hours and 1 hour before antigen challenge rats are dosed orally. Rats are challenged by exposure for 30 minutes to nebulized saline or ovalbumin (1% in saline) . 24 hours after challenge, rats are killed and the airways are lavaged with physiological salt solution. Total and differential cell counts are made .

5. In Vitro Inhibitory Effects on TNF-alpha Release bv Human Monocytes The effects of compounds on TNF-alpha production by human peripheral blood monocytes (PBMs) are examined as follows .

5.1. Preparation of blood leukocytes. Blood is drawn from normal donors, mixed with dextran, and the erythrocytes allowed to sediment for 35 minutes at 37°C. Leukocytes are fractionated by centrifugation through a discontinuous (18, 20 and 22%) metrizamide gradient. The mononuclear cell fraction comprising 30-40% PBMs is suspended in

Hank's balanced salt solution and stored at 4°C until use .

5.2. Measurement of TNF-alpha.

Cells from the PBM-rich metrizamide fraction are spun down (200 g for 10 minutes at 20°C) , resuspended at lO^PBMs/mL of medium; RPMI 1640 containing l%v/v FCS, 50U/mL penicillin and 50mgmL streptomycin (Gibco, U.K.), then plated out in 96 well plates at 2xl0⁵ cells/well. The medium (200μL) is changed to remove any non-adherent cells and the remaining, adherent PBMs left in the incubator overnight (18 hours) . One hour prior to challenge, the medium is changed to that containing compound for test or drug vehicle. Control treatments and compounds for test are assayed in quadruplicate wells. Compounds are tested within the concentration range of 3xlO"¹⁰M to 3x10~⁶M. Medium (50μL) with or without lOng/ml LPS (E. Coli, 055 B5 from Sigma, U.K.) is then added. The incubation is then continued for a further 4 hours. Cell supernatants are removed for storage at -20°C. TNF-alpha levels in cell supernatants are quantified using a standard sandwich ELISA technique. ELISA plates (Costar, U.K.) are coated overnight at 4°C with 3 mg/mL polyclonal goat anti-human TNF-alpha antibody (British Biotechnology, U.K.) in pH9.9 bicarbonate buffer. Rabbit polyclonal anti-human TNF-alpha antiserum (Janssen Biochimicha, Belgium) at 1/500 dilution is used as the second antibody and polyclonal goat anti-rabbit IgG horseradish peroxidase (Calbiochem, U.S.A.) at 1/8000 dilution is used as the detection antibody. Color development is measured by absorbance at 450nm using a Titek plate reader.

TNF-alpha levels are calculated by interpolation from a standard curve using recombinant human

TNF-alpha (British Biotechnology U.K. ) (0.125-8ng/mL) . Data (log-cone. vs. log-resp) are fitted by linear regression (p > 0.99) using a Multicalc (Wallac Pharmacia, U.K.) software program. Basal TNF-alpha levels are less than lOOpg/mL whilst LPS (lipopoly- saccharide) stimulation of the PBMs increases TNF-alpha levels to 3-10ng/mL. 5.3 Results.

Compounds within the scope of the invention produce 50% inhibition of LPS-induced TNF-α release from human PBMs at concentrations within the range of about 10^-9M to about 10~⁶M. , preferably about 10"⁹M to about 10"⁷M. 6. Inhibitory effects of compounds on antiσen-induced bronchoconstriction in the conscious cruinea-piσ

6.1 Sensitisation of guinea-pigs and measurement of antigen-induced bronchoconstriction.

Male Dunkin-Hartley guinea-pigs (550-700g) are sensitized as above. Specific airways resistance (SRaw) is measured in conscious animals by whole body plethysmography using a variation of the method of Pennock et al. , (J. APPI. Phvsiol.. 46 ,399, 1979). Test compounds or vehicle are administered orally 24 hours and 1 hour before antigen challenge. 30 Minutes before challenge the animals are injected with mepyramine (30mg/kg i.p.) to prevent anaphyl- actic collapse and placed into the plethysmography chambers where SRaw is determined at 1 minute intervals. Resting SRaw is then determined. Animals are challenged with an aerosol of ovalbumin and SRaw is determined every 5 minutes for 15 minutes.

7. Inhibitory effects of compounds aσainst antiσen- induced bronchoconstriction in the anaesthetized rat in vivo 7.1. Treatment of rats and measurement of antigen-induced bronchoconstriction.

Male Brown Norway rats weighing 150-250g are sensitized on days 0, 12 and 21 with ovalbumin (lOOμg, i.p.). Rats are challenged on any one day between days 27-32.

24 hours and 1 hour before antigen challenge rats are dosed orally. Rats are anaesthetized to allow recording of lung function (airway resistance and lung compliance) using respiratory mechanics software. Rats are challenged with ovalbumin i.v. and the peak changes in airway resistance and lung compliance are determined.

The ED50 of the compounds is typically found to be between 1 and lOOmg/kg animal body weight.

8. Inhibitory effects of compounds on serum TNF-alpha levels in LPS-challenσed mice

8.1. Treatment of animals and measurement of murine TNF-alpha. Female Balb/c mice (age 6-8 weeks, weight 20-22g from Charles River, U.K.) in groups of five or more animals are dosed p.o. with compounds suspended in 1.5% (w/v) carboxymethyl cellulose then challenged after a minimum period of 30 minutes with 30mg of LPS i.p. After 90 minutes the animals are killed by carbon dioxide asphyxiation and bled by cardiac puncture. Blood is allowed to clot at 4°C, centrifuged (12,000 g for 5 minutes) and serum taken for TNF-alpha analysis. TNF-alpha levels are measured using a commercially available murine TNF-alpha ELISA kit, purchased from Genzyme (Cat. no. 1509.00), as recommended by the manufacturer. Values for TNF-alpha are calculated from a recombinant murine TNF-alpha standard curve.

The ED50 of the compounds is typically found to be between 1 and lOOmg/kg animal body weight. 9. Systemic bioavailabilitv in female Balb/c mouse Intravenous administration :

Following surgery to expose the jugular vein for dosing, a solution of test compound in dimethylsulph- oxide is added at a dose of lmg/kg body weight. Oral administration :

A suspension of test compound in 1.5% aqueous carboxymethylcellulose is introduced into the stomach by gavage at a dose of lmg/kg body weight. Following either i.v. or oral dosing, blood is obtained by cardiac puncture following carbon dioxide asphyx¬ iation and is obtained at a single time post-dose for each animal. Three animals are sacrificed at each time point. Blood samples are obtained at the following times after dosing by both the i.v. and oral routes; 5 minutes (i.v. only), 0.25, 0.5, 1, 2, 3, 4, 5.5, 7 and 24 hours. Corresponding plasma is obtained by centrifugation of each blood sample. The drug content in the plasma samples is then determined using conventional methods. 9.1 Metabolism (i)Preparation of mouse liver homogenate

Fresh mouse liver is homogenised in sucrose- phosphate buffer. Following centrifugation the resulting supernatant (liver homogenate) is used fresh or frozen in liquid nitrogen for one minute and stored at -30°C to -40°C prior to use.

(ii) Incubation of compounds with mouse liver homogenate To 0.5ml of mouse liver homogenate is added

0.5ml taken from a vortexed mixture of 8mg NADPH added to a mixture of aqueous magnesium chloride (lml, 0.15M) nicotinamide (lml, 0.5M) and pH 7.4 tris buffer (8.5ml, 0.1M). The compound is added at a concentration of lμg/ml in lOμl of solvent. Incubates are maintained at 37°C. Samples are taken at 0 minutes, 5 minutes, 10 minutes, 20 minutes and 30 minutes and the incubation stopped by the addition of lOOμl acetonitrile. The drug content in the incubation samples is determined using conventional methods . 9.2 Results

Compounds within the scope of the invention have _tl/2 values from about 60 minutes to greater than 600 minutes. The preferred compounds in this invention have a t^1/f2 minimum of about 100 minutes.

10. Streptococcal Cell Wall-Induced Arthritis in Rats 10.1 Preparation of S. pyoσenes purified cell wall

Purified S. pyoσenes cell wall is prepared from the cell pellet of a log-phase culture of S. pyoσenes. group A, strain D-58. The whole bacteria are homogenized by grinding with glass beads and the crude cell wall collected by centrifugation and subsequently washed with 2% sodium dodecyl sulphate in phosphate buffered saline followed by phosphate buffered saline to remove contaminating proteins and nucleic acids. The cell wall is further purified by sonication and differential centrifugation to obtain a purified preparation which pelleted at 100,000 g. This material is suspended in sterile phosphate buffered saline and the quantity of cell wall determined by measuring the rhamnose content of the preparation (purified cell wall contains 28% rhamnose by weight) . The material is filtered through a 0.22μM filter and stored at 4°C until used for arthritis induction

10.2 Arthritis Induction and measurement of joint diameters

Female Lewis rats weighing 140-160g are injected intra-articularly into the left or right tibio-tarsal joint on day 0 with purified S. pyoσenes cell wall extract (10 μg in lOμl sterile saline) . On day 20, rats received an intravenous injection of purified cell wall (100 μg in lOOμl sterile saline) via the lateral vein of the tail. Joint diameters are measured with calipers across the lateral and medial malleoli of the previously intra-articularly injected joint immediately prior to the i.v. injection and then daily through day 24. The net joint diameter is determined by subtracting the value for the contralateral joint. Body weights are also measured daily. Compounds or vehicle are administered by oral gavage on days 20-23. Typically, 8-10 animals are used per group. For each dose, the total daily dose is divided into two equal aliquots which are given at approximately 9 a.m. and 3 p.m.

The value of the compounds of the invention is enhanced by their very low mammalian toxicity levels.

The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof.

Claims

A compound of formula (I)

wherein

R^- represents a straight- or branched-chain alkyl group of 1 to 6 carbon atoms, optionally substituted by one or more halogen atoms;

R² represents an optionally substituted aryl group or an optionally substituted heteroaryl group;

Q^, Q² and Q³, which may be the same or different, each represents a CH or CX linkage or a nitrogen atom;

Z¹ represents an oxygen or sulphur atom;

Z² represents an NH or methylene linkage; and

2 . A compound of f ormula ( Ia )

where in

R! represents a methyl or difluoromethyl group; A represents a hydroxycycloalkyloxy group of 5 to 7 carbon atoms;

Q² represents a CH or CF linkage or a nitrogen atom;

Q⁴ represents a nitrogen atom or a CH linkage; X^ and X² each represents a fluorine or chlorine atom; and

Z² represents an NH or methylene linkage; and N-oxides thereof, and pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof.

3 . A compound of formula ( Ib)

( lb) wherein

R! represents a methyl or difluoromethyl group;

A represents a hydroxycycloalkyloxy group of 5 to 7 carbon atoms; and

X^- and X² each represents a fluorine or chlorine atom; and N-oxides thereof, and pharmaceutically acceptable salts, and solvates (e.g. hydrates), thereof.

4. A compound of formula (I) as claimed in Claim 1, 2 or 3 which is in a substantially pure form.

5. A compound of formula (I) as claimed in Claim 1, 2, 3 or 4 which has been prepared synthetically.

6. A compound of formula (I) as claimed in Claim 1, 4 or 5 in which R¹ represents a methyl or difluoromethyl group.

7. A compound of formula (I) as claimed in Claim 1, 4, 5 or 6 in which R² represents a heteroaryl group substituted on both the positions next to the position which is directly attached to Z².

8. A compound of formula (I) as claimed in Claim 7 in which R² represents a heteroaryl group substituted by one or two halogen atoms.

9. A compound of formula (I) as claimed in Claim 8 in which R² represents a heteroaryl group substituted by one or two chlorine atoms.

10. A compound of formula (I) as claimed in Claim 1, 4, 5, 6, 7, 8 or 9 in which R² represents an optionally substituted pyridyl group.

11. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 10 in which A represents an alkoxy group of 3 to 7 carbon atoms carrying one or two hydroxy groups.

12. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 10 in which A represents a monocyclic cycloalkyloxy group of 4 to 8 carbon atoms carrying one or two hydroxy groups.

13. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 12 in which A represents a l-hydroxyprop-2-yloxy group.

14. A compound of formula (I) as claimed in any of Claims 1 to 12 in which A represents a 3-hydroxy¬ cyclopentyloxy group.

15. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 14 in which Q¹ and Q² and Q³ each represents a CH linkage.

16. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 14 in which Q² represents a nitrogen atom and Q¹ and Q³ each represents a CH linkage .

17. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 16 in which Z¹ represents an NH linkage.

18. A compound of formula (I) as claimed in any preceding claim which is an N-oxide.

19. A compound of formula (I) as claimed in Claim 1 or any of Claims 4 to 18 in which R² represents a 3, 5-dichloro-l-oxido-4-pyridinio group.

20. A compound of formula (I) as claimed in any preceding claim which is an ester.

21. A compound of formula (I) as claimed in Claim 20 which is a 3- or 4-(4-alkylpiperazin-l-yl)benzoate.

22. A compound of formula (I) as claimed in Claim 20 which is a 2,2-dimethylpropanoate.

23. A compound of formula (I) as claimed in any preceding claim which is a single geometric isomer.

24. A compound of formula (I) as claimed in any preceding claim which is a single optical isomer.

25. A compound of formula (I) as claimed in any preceding claim which is a prodrug.

26. A compound of formula (I) as claimed in Claim 1 selected from: (±) -N- ( 3 , 5-dichloro-4-pyridyl) -3- (cis-3-hydroxycyclo¬ pentyloxy) -4-methoxybenzamide;

(±) -N- (3 , 5-dichloro-4-pyridyl) -3- (trans-3-hydroxy¬ cyclopentyloxy) -4-methoxybenzamide; (±) -2- (3 , 5-dichloro-4-pyridyl) -1- [3- (cis-3-hydroxy¬ cyclopentyloxy) -4-methoxyphenyl] ethanone;

(±) -2- (3 , 5-dichloro-4-pyridyl) -1- [3- (trans-3-hydroxy¬ cyclopentyloxy) -4-methoxyphenyl] ethanone;

(±) -2- (3, 5-dichloro-1-oxido-4-pyridinio) -l-[3-(cis-3- hydroxycyclopentyloxy) -4-methoxyphenyl] ethanone;

(±) -2- (3, 5-dichloro-1-oxido-4-pyridinio) -1- [3- (trans- 3-hydroxycyclopentyloxy) -4-methoxyphenyl] ethanone; (±) -2- (3, 5-dichloro-l-oxido-4-pyridinio) -1- [4- difluoromethoxy-3- (cis-3-hydroxycyclopentyloxy) - phenyl] ethanone;

(±) -2- (3, 5-dichloro-l-oxido-4-pyridinio) -1- [4- difluoromethoxy-3- (trans-3-hydroxycyclopentyloxy) - phenyl] ethanone; (±) -N- ( 3 , 5-dichloro-1-oxido-4-pyridinio) -3- (cis-3- hydroxycyclopentyloxy) -4-methoxybenzamide;

(±) -N- ( 3 , 5-dichloro-1-oxido-4-pyridinio) -3- (trans-3- hydroxycyclopentyloxy) -4-methoxybenzamide;

(±) -2- (3, 5-dichloro-4-pyridyl) -1- [3- ( l-hydroxyprop-2^■ yloxy) -4-methoxyphenyl] ethanone; (±) -2- (3, 5-dichloro-1-oxido-4-pyridinio) -1- [3-(l- hydroxyprop-2-yloxy) -4-methoxyphenyl] ethanone; ( - ) -N- (3 , 5-dichloro-4-pyridyl) -3- (trans-3-hydroxy¬ cyclopentyloxy) -4-methoxybenzamide; (±) -N- (3 , 5-dichloro-4-pyridyl) -4- (trans-3-hydroxy- cyclopentyloxy) -5-methoxypyridine-2-carboxamide; (±) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -4- (trans-3- hydroxycyclopentyloxy) -5-methoxy-l-oxidopyridinium-2- carboxamide;

(±) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- [cis-3- (2, 2-dimethylpropanoyloxy) cyclopentyloxy] -4-methoxy¬ benzamide;

(±) -2- (3 , 5-dichloro-4-pyridyl) -1- [4-difluoromethoxy- 3- (trans-3-hydroxycyclopentyloxy)phenyl] ethanone; (±) -2- (3 , 5-dichloro-4-pyridyl) -1- [4-difluoromethoxy- 3- (cis-3-hydroxycyclopentyloxy)phenyl] ethanone;

(±) -N- (3 , 5-dichloro-4-pyridyl) -3- (l-hydroxyprop-2- yloxy) -4-methoxybenzamide;

(+) -N- (3, 5-dichloro-4-pyridyl) -3- (trans-3-hydroxy¬ cyclopentyloxy) -4-methoxybenzamide; (±) -N- (3, 5-dichloro-4-pyridyl) -4- (trans-3-hydroxy¬ cyclopentyloxy) -5-methoxy-l-oxidopyridinium-2- carboxamide;

(±)-N- (3, 5-dichloro-4-pyridyl)-3- [cis-3-(2,2- dimethyl-propanoyloxy) cyclopentyloxy] -4- methoxybenzamide;

(±)-N- (3, 5-dichloro-4-pyridyl) -3-[ciε-3-(3- morpholino-methylbenzoyloxy) cyclopentyloxy] -4- methoxybenzamide; (±) -N- (3, 5-dichloro-1-oxido-4-pyridinio) -3- [cis-3- (3- morpholinomethylbenzoyloxy) cyclopentyloxy] -4-methoxy- benzamide;

( - ) -N- ( 3 , 5-dichloro-1-oxido-4-pyridinio) -3- (trans-3- hydroxycyclopentyloxy) -4-methoxybenzamide; and

( +) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- (trans-3 - hydroxycyclopentyloxy) -4-methoxybenzamide and N-oxides thereof, and their esters, and prodrugs, pharmaceutically acceptable salts, and solvates thereof .

27. A compound of formula (I) as claimed in Claim 1 selected from:

(±) -N- (3 , 5-dichloro-4-pyridyl) -3- (trans-4-hydroxy¬ cyclohexyloxy) -4-methoxybenzamide; (±) -N- (3 , 5-dichloro-4-pyridyl) -3- (cis-4-hydroxy- cyclohexyloxy) -4-methoxybenzamide;

(±) -N- (3, 5-dichloro-1-oxido-4-pyridinio) -3- (trans-4- hydroxycyclohexyloxy) -4-methoxybenzamide; (±) -N- (3, 5-dichloro-l-oxido-4-pyridinio) -3- (cis-4- hydroxycyclohexyloxy) -4-methoxybenzamide; (±) -N- (3, 5-dichloro-4-pyridyl) -3- (trans-3, 3-dimethyl- 5-hydroxycyclohexyloxy) -4-methoxybenzamide; (±) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- (trans- 3 , 3-dimethyl-5-hydroxycyclohexyloxy) -4-methoxy- benzamide; N- (3 , 5-dichloro-4-pyridyl) -3- (trans-2-hydroxycyclo- pentylmethoxy) -4-methoxybenzamide; N- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- (trans-2- hydroxycyclopentylmethoxy) -4-methoxybenzamide; (±) -2- (3 , 5-dichloro-4-pyridyl) -1- [4-methoxy-3- (trans- 3- (4-morpholinomethylbenzoyloxy) cyclopentyloxy- pheny1] ethanone; (±) -2- (3, 5-dichloro-4-pyridyl) -1- [4-methoxy-3- (cis-3-

(4-morpholinomethylbenzoyloxy) cyclopentyloxyphenyl] - ethanone; (±) -N- (3 , 5-dichloro-1-oxido-4-pyridinio) -4- [trans-3-

(4-morpholinomethylbenzoyloxy) cyclopentyloxy] - hydroxycyclopentyloxy) -5-methoxy-l-oxido-pyridinio-2- carboxamide;

(±) -2- (3 , 5-dichloro-4-pyridyl) -1- [4-methoxy-3- (cis-3- (3-morpholinomethylbenzoyloxy) cyclopentyloxyphenyl] - ethanone; and

(±) -N- (3, 5-dichloro-4-pyridyl) -3- [cis-3- (4-morphol¬ inomethylbenzoyloxy) cyclopentyloxy] -4-methoxy- benzamide and N-oxides thereof, and their esters, and prodrugs, pharmaceutically acceptable salts, and solvates thereof .

28. A compound of formula (I) as claimed in Claim 1 selected from: (+) -cis- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; (-) -cis- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; ( -) -trans- { 3 , 5-dichloro-1-oxido-4-pyridinio) -3- (3- hydroxycyclopentyloxy) -4-methoxybenzamide; and

( +) -trans- (3 , 5-dichloro-1-oxido-4-pyridinio) -3- ( 3- hydroxyeyelopentyloxy) -4-methoxybenzamide and N-oxides thereof, and their esters, and prodrugs, pharmaceutically acceptable salts, and solvates thereof.

29. A compound of formula (I) as claimed in Claim 1, 2 or 3 which is (±) -trans- (3.5-dichloro-1-oxido-4- pyridinio) -3- ( 3-hydroxycyclopentyloxy) -4- methoxybenzamide, or an ester, prodrug, pharmaceutically acceptable salt, or solvate thereof.

30. A compound of formula (I) as claimed in Claim 26 which is (±) -trans- (3, 5-dichloro-l-oxido-4- pyridinio) -3- (3-hydroxycyclopentyloxy) -4- methoxybenzamide.

31. A pharmaceutical composition containing a compound of formula (I) as claimed in any preceding claim in association with a pharmaceutical carrier or excipient.

32. A compound of formula (I) as claimed in any preceding claim for use in therapy.

33. A compound of formula (I) as claimed in any preceding claim for use in the treatment of a patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of TNF.

34. A composition as claimed in Claim 31 for use in the treatment of a patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of TNF.

35. A compound of formula (I) as claimed in any preceding claim for use in the treatment of a patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of type IV cyclic AMP phosphodiesterase.

36. A composition as claimed in Claim 31 for use in the treatment of a patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of type IV cyclic AMP phosphodiesterase .

37. Use of a compound of formula (I) as claimed in any preceding claim in the manufacture of a medicament for the treatment of a patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of TNF.

38. Use of a compound of formula (I) as claimed in any preceding claim in the manufacture of a medicament for the treatment of a patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of type IV cyclic AMP phosphodiesterase.

39. A method for the treatment of a human or animal patient suffering from, or subject to, a condition which can be ameliorated by the administration of an inhibitor of TNF or of type IV cyclic AMP phosphodiesterase comprising administering to said patient an effective amount of a compound of formula (I) as claimed in any preceding claim.

40. A process for the preparation of a compound of formula (I) as claimed in any preceding claim substantially as hereinbefore described.

41. A compound substantially as hereinbefore described with reference to the Examples .