CA2136196A1 - 8-substituted anthines as phosphodiesterase inhibitors - Google Patents

Abstract

A compound of formula (I) or if appropriate a pharmaceutically acceptable salt thereof, wherein R1 and R2 each independently represent a moiety of formula (a): -(CH2)m-A wherein m represents zero or an integer 1, 2 or 3 and A represents a substituted or unsubstituted cyclic hydrocarbon radical; R3 represents hydrogen, substituted or unsubstituted alkyl or an aralkyl group substituted or unsubstituted in the aryl moiety; and R4 represents hydrogen, alkyl or alkylcarbonyl; a process for preparing such a compound, a pharmaceutical composition containing such a compound and the use of such a compound in medicine.

Description

. WO 93/23401 P~/GB93/01014 , . .

~-SUBSTITUTED HANTHINES AS PHOSPHODIESTERAS~ INHIBITORS

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The present invention relates to certain novel compounds having phamlacological 5 activity, to a process for the preparation of such compounds, to pharmaceutical compositions containing such compounds and to Ihe use of such compounds and compositions in me~icine.

Molecular Pharmaco}ogy, Volume 6, No. 6, 1970, p.597-603 discloses 10 1,3-dimethyl-8-nitro-xanthine. This compound is disclosed as having lipolytic activity. Ann Chim, 47, 362-365 (1957) discloses 1,3-dimethyl-8-amino-xanthine and a process by which it may be prepared. Mo pharrnacological utility is disclosed for this compound. Drug Res. 27(1~ Nr 19, 1977, pages 4-14, ~an K.H. Klingler discloses certain 1,3-dimethyl- 8-substituted lS xanthines as interrnediates solely in the synthesis of phenylethyl aminoalkylxanthines. Drug Rcs. 31 (11), Nr. 12, ~981, R.G. Werner et a~, pages 2044-2048 discloses certain 1,3-dimethyl-8-substituted xanthines. No pharmacolo~ical activity is disclosed for these compounds.

European Patent Application, Publication Number 0369744 also discloses certain 1,3- or 1,3,7- 8-H cycloalkylalkylene xanthines, for use inter al~a as bronchodilators in the treatment of asthma.

It has now been discovered that a novel series of 8-substiluted xanthines show ~ -~
activity as phosphodiesterase inhibitors.

These compounds are indicated to be good inhibitors of induced blood eosinophilia and that they are therefore potentially useful in the treatment and/or prophylaxis of disorders associated with increased numbers of eosinophils, such as asthmaj and allergic disorders associated with atopy, such as urtic~lria, eczema and rhinitis. i~

These compounds are also indicated to have bronchodilator activity and thus eo be of potential use in the trea~ment of disorders of the respiratory tract, such as ~t re~ersibleairwaysobstruclionarldasthrna.

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WO g3/23401 ~ ~ ~ h 19 6 P~/GB93~01014 These compounds also have a protective effect against the consequences of cerebral metabolic inhibition. The said compounds improve data acquisition or retrieval following transient forebrain ischaemia and are therefore useful in the treatment of cerebral vascular and neuronal degenerative disorders associated ' -~
with learning. mernory and cognitive dysfunctions including cerebral senility, multi-infarct dementia, senile dementia of the Alzheimer type, age associated memory impairment and certain disorders associated with Parkinson's disease.
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These compounds are also indicated to have neuroprotectàrit activity. They are ~herefore useful in the prophylaxis of disorders associated with neuronal degeneration resulting from ischaemic events, including cerebral ischaemia due tO
cardiac arrest, stroke and also after cerebral ischaemic events such as those resulting from surgery and/or during childbirth. In addition ~ea~ment with the compound is indicated to be of benefit for the treatment of functional disordersresul~ing from disturbed brain function following ischaemia.

These cornpounds are also active in increasing the oxygen tension In ischaemic skeletal muscle. This property results in an increase in the nutritional blood flow through ischaemic skeletal muscle which in turn indicates that the compounds of the invention are of potential use as agents for the treatment of peripheral vascular disease such as intermit~ent claudication. I

The present compounds are also considered to be inhibltors of the in vivo !`;
production of Tumor Necrosis Factor (TNF) and hence they have potential for the ~,~
treatnent of diseases associated with excessive or unregulated TNF production ~`
including rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthIitis and other arthritic conditions; sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, :
cereb~.'al` malaria, chronic pulmonary inflammatory~ disease, silicosis, pulmonary sarcoldosis, bone reso~lption diseases, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as Inhuenza, 2 cachex~a secondary tO infection or malignancy, cachexia~ secondary to acquired immune deficiency syndrome ~AIDS), AIDS, ARC (AIDS related complex), Iceloid forma~ion, scar tissue fo~ma~ion, Crohn's disease, ulcerative colitis, or ~- :

WO 93/23401 ~ 619 ~ PCr/CB93/01014 pyresis. The present compounds are also useful in the treatment of vir~l infections ~hat proctuce TNF as a result of infection, or those which are sensitive to inhibition, such as by decreased replication, directly or indirectly, by the present compounds. Such viruses include for example HIV-l, HIV-2 and HIV-3, S Cytomegalovirus (CMV), Influenza, adenovirus and the Herpes group of viruses, such as but not limited to, Herpes Zoster and Herpes Simple~.

The above mentioned treatments of course include veterinary treatments and in particular they include the treatment of TNF mediated viral infections including, lO for example, feline immunode~lciency virus (FTV) or o~her retroviral infections such as equine infectious an~emia virus, caprine arthntis virus, visna virus, maedi virus and other lentiviruses.

Tl~ese compounds are also of potential use in the treatment of proliferative skin lS disease in human or non-human mammals.

Accordingly, the invention also provides a compound of formula (I):

F~NJ~--N q ,.
11 ~>~NH(CH2)2CH(CH20R )2 o~N - N
~2 (I) ~
;
or if appropriate a pharrnaceutically acceptable salt thereof, wherein R 1 and R2 each independently represent a moiety of forrnula (a):
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~(CH2)m~A (a) wherein m represents zero or an integer 1, 2 or 3 and A represents a substituted or unsubstituted cyclic hydrocarbon radical;
Ei~3 represents hydrogen, substituted or unsubstituted aLkyl or an aralkyl group30 substituted or unsubstituted in the aryl moiety; and R4 represer.[~ hydrogen, alkyl or alkylcarbonyl.

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WO93/23401 2136196 Pcr/GB93/0l0l4 ~ ~ ~;

Suitably, A is unsubstituted. Favourably, A represçnts a substituted or unsubstituted C3-8 cycloalkyl group, especi~lly a C3-6 cycloalkyl group. ~ -5 In particular, A represents a substi~uted or, preferably, unsubstituted cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl group. `

Favourably, A represents a cyclopropyl group or à"cyclobutyl group. ~`

10 Preferably, A represents a cyclopropyl group.

When R3 represents unsubstituted alkyl, suitable examples inciude methyl.

When R3 represents substituted alkyl, suitable examples include alkoxy methyl 15 such as methoxymethyl.

Suitably, R3 represents an aralkyl group, for example a benzyl group, substituted `
or unsubstitued in the aryl moiety.

20 When ~3 is a benzyl group, examples include unsubstitute~ benzyl or benzyl substituted in the phenyl moiety by methoxy groups, particular examples include 4-methoxy benzyl. `

Suitably R4 represents hydrogen.
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Suitably, R4 represents alkylcarbonyl.
'., Suitably alkycarbonyl groups are Cl 4 alkylcarbonyl groups for example acetyl. `:
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30 Suitable pharmaceutically,acceptable salts are pharmaeeutically acceptable base salts and pharmaceiutically acceptable acid addition salts. Suitable pharmaceutically acceptable base salts of the compounds of forrnula (I) include i -7-N base salts ineluding metal salts, such as alkali metal salts for example sodium salts, or organic amine salts such as that provided with ethylenediamine. ~ -: ' .

WO 93/23401 i~ i 3 fi l 9 6 PCI /CB93/01014 ~

Suitable acid addition salts of the comyounds of fonnu}a (I) are the acid addition salts includin~ pharmaceutically acceptdble inorganic salts such as the sulpha~e, nitrate, phosphate, borate, hydrochloride and hydrobromide and pharmaceutically acceptable organic acid addition salts such as acetate, tartrate, maleate, citrate, S succinate, benzoate, ascorbate, methanesulphonate, ~-lceto glutarate, a-glycerophGsphate and glucose- 1- phosphate. Preferably the acid additioll salt is a hydrochloride salt.

The phannaceutically acceptable salts of the compounds of fonnula (I) are prepared using conventional procedures.

When used herein the terrn 'cyclic hydrocarbon radical' includes single ring andfused ring, alicyclic hydrocarbons comprising up to 8 carbon atoms in each ring,suitably up to 6 carbon atoms, for example 3, 4, 5 or 6 carbon atoms.
Suitable optional substituents for any cyclic hydrocarbon radical includes a Cl-6 alkyl group or a halogen atom.

When used herein the term 'aryl' whether used alone or as part of another group (for example in an aralkyl group) includes phenyl and naphthyl optionally substituted with up to five, preferably up to three, groups selected from halogen, alkyl, phenyl, alkoxy, halo alkyl, hydroxy, amino, nitro, carboxy, alkoxycarbonyl, alkoxycarbonylalkyl, alkylcarbonyloxy, or alkylcarbonyl groups.
Optional substi~uents for any phenylene group include up to three of the substituents mentioned in relation to the aryl group.

Suitable optional substituents for the aryl moiety of any aralkyl group include those mentioned above in regard to the ` aryl' group and in particular include alkoxy groups, for example methoxy groups.
When us- d herein the ~errn 'alkyl' whether used alone or when used as part of 3 another group (for example as in an alkylcarbonyl group) includes straight and , - -branched chain alkyl groups, containing from 1 to 12 carbon atoms, suitably 1 to6 carbon atoms, for example methyl, ethyl, propyl or butyl. Suitable optional 7 W0 93/23401 2 i3 6 l pcr/GB93/olol4 f`

substituents for any alkyl group include up to five, preferably up to three of ~he substituents mentioned above in relation to the aryl group.

When used herein the expression 'proliferative skin diseases' means benign and ! ~ `
malignant proliferative skin diseases which are characte~ized by accelerated cell !, division in the epidermis, d~rmis or appendages thcre;o, associated with `-.
incomplete tissue differentiation. Such diseases include: psonasis, atopic :~
delmatitis, non-specific dermatitis, primary ~n-itant contact dermatitis, allergic contact dermatitis, basal and squamous cell carcinomas of the skin, lamellar ~ `
ichthyosis, epidelmolytic hyperkeratosis, premalignant sun induced keratosis, non-malignant keratosis~ acne, and seborrheic dermatitis in humans and atopic :~
dermatitis and mange in domesticated animals.

The compounds of formula (I) are preferably in pharrnaceutically acceptable :
form. By pha~naceutically acceptable forrn is meant, inter alia, of a phannaceutically acceptable level of purity excluding norrnal pharmaceutical additives such as diluents and carriers, and including no material considered toxic - ~:
at nor~nal dosage levels. A pharmaceutically acceptable level of purity will generally be at least 50% excluding norrnal pha~Traaceutical additives, preferably 75%, rIlore preferably 90% and still more preferably 95%. ` ~-~ , The invention ~rther provides a process for the preparation of a compound of -formula (I), which process comprises reacting a compound of formula (II): ;

R3a ~N~ ~

1 2a ~. .
R
i 1 ~ (II) ;, wherein Rla represents Rl as defined in relation to formula (I) or a group ~ -convertible to R l, R2a represents R2 as defined in relation to formula (I) or a . : .
group convertible thereto and R3a represen~s R3 as defined in relation to folmula ~-(T) or a group convertible to R3, '`~ WO 93/23401 ~ 1 ~ 619 6 P~/CB93/01014 wi~h a compound of formula (III):

LI-(cH~)7cHtcHoR5)2 (III) S wherein, E~5 represents a hydroxy protecting group and Ll represents a leavinggroup; and thereafter, if required carrying out one or more of the following optional steps:

(i) removing any protecting group;

(ii) conver~ing any group E~la to Rl andlor R2a tO R2 and/or R3a to R3;
.
(iii) converting a compound of ~ormula (I) into a further compound of formula (I);
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(iv) converting a compound of formula (I) into a pharrnaceutically accep$able salt thereof.

A suitable leaving group Ll is a halo atom, especially an iodine atom.
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The reaction ~etween compounds of formulae (II) and (III) may be camed out using conventional alkylation conditions, for example in an aprotic solYent suchas dimethoxyethane, dimethylforrnamide or tetrahyd~ofuran, at any temperature pro~iding a suitable rate of forma~ion of the required product, such as in the range 25 of ~ronn 0C to lû0C, conveniently in the range of from 40C to 80C~, ~or example 60C; and preferably in an inert atmosphere such as ni~ogen.

Suitably the 8-amino group of cornpound (II) is in an activated form, favourablyin an ionic fonn such as a salted form, for example an alkali metal salted fomn 30 provided by ~eating the compound of formula (Il) with an alkali metal base, ~or ` ~ example potassium t-butoxide.

A compound of forrnula (n~ may be prepared using methods described in Buropean Patent Application, Publication No.~ 0389282.

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wo 93~23401 2 1- 3 ~ l 9 6 -8- PCr/GB93/01014 i ~ ~ ~
The compounds of fo~mul;l (111) are known compounds or they may be prepared according to methods used tO prepare known compounds, for example those discussed in Tetrahedron (1990), 46, 6903.

S Conversions of one compound of formula (I) into another compound of fo~nula (I) includes converting one group R4 into another group R4, for example hydrolysing compounds wherein R4 represents an alkylcarbonyl group inlo a compound of fonnula ('i) wherein R4 represents a hydrogen atom 10 ln the said conversions the appropliate conventional procedures are suitably employed? hence in the abovementioned hydrolysis conventional hydrolysis `
conditions are used, for example the hydrolysis of an a~kylcarbonyl group is effected by using mild basic hydrolysis preferably by means of potassium ;-`
carbonate in an ethanolic solution such as methanol, conveniently at ambient 1 5 temperature.

Suitable values for Rla and R2a include Rl and R2 respectively or nitrogen protecting groups such as benzyl, nitrobenzyl or trimethoxybenzyl groups.
Suitable values for R3a include R3.
;~
Suitably, when R is substituted or unsubstituted aralkyl, R1a and R2a represent nitrogen protecting groups which can be inserted and removed without affecting :~
R3, for example trimethylsilyl groups~ ;~

25 Preferably, when R3 is substituted or unsubstituted aralkyl then Rla is Rl and R2a is R2.

When ~ 1 a, R2a or R3a represents other Ihan R 1, R2 or R3 repectively7 the abovementioned conversions of R1a into Rl, R2a to R2 and R3a into R3 may be 30 calTied out using the appropriate conventional procedure. For example when R1a (or R2a) represents a nitrogen protecting group, such as a benzyl group, the protecting group may be removed using the appropriate con~/entional procedure, such as catalytic hydrogenation, and the resul~ing product reacted wilh a ~ -compound of fo~ ula ~IV):

WO 93/2340l pcr~GB93/olol4 ,;
. .

x-(cH2)m-A (IV) , .
wherein A and m are as defined in relation to formula (IA) and X represents a leaving group, such as halide, for example bromide or iodide.
The protection of any reactive group or atom, such as the xanthine nitrogen at~?m may be calned out at any appropriate stage in the aforementioned process Suitable protecting groups include those used conventionally in the art for the particular group or atom being protected, for example suitable protecting groups10 for the xanthine nitrogen atoms are alkylsilyl groups, especially trimethylsilyl or ~-butyldimethylsilyl groups.

Protecting groups may be prepared and removed using the appropnate conventional procedure: ~?or example, alkylsilyl protecting groups may be 15 prepared by treating the compound of formula (II) with an appropriate alkylsilyl halide, for example trimethylsilyl chloride for trimethylsilyl groups and 1-butyldimethylsilyl chloride for t-butyldimethylsilyl groups. The silyl protec~ing groups may be removed by ~eatment with t-butylammonium fluoride in a suitable solvent, such as tetrahydrofuran conveniently at an ambient temperature.
As rnentioned above the compounds of the invention are indicated as having useful therapeutic properties: the present invention accordingly provides a compound of forrnula tl) or where appropriate a pharrnaceutically acceptable salt thereof ancVor a pharmaceutically acceptable solvate thereof, for use as an active 2S therapeutic substance.
.,.

Thus the present invention provides a compound o~ forrnula (I) or where appropriate a pharmaceutically acceptable salt ~hereof and/or a ph~naceutically acceptable solvate thereof, for use in the treatment of and/or prophylalcis of 30 disordersl associated with increased numbers of eosinophils, such as asthma, and allergic disorders associated with atopy, such as urticana, ec~ema and rhinitis.
In a further aspect the present invention also provides a compound of fo~mula (I) or where appropriate a pharrnaceutically acceptable salt thereof andlor a .~ .

WO93/23401 2~3~ 196 PC~/GB93/01014 ~

1 ( ) Opharmaceutically accept~ble solvate thereof, for use ~s a phosphodiesterase inhibitor. ;
" ' In a p~rticular aspect, as indicated hereinbefore, the present invention provides a 5 compound of forrnula (I) or where appropriate a phannaceutically acceptable salt thereof and/or a pharmaceutically acceptable solvatc thereGf, for use in the treatmen~ of disorders of the respiratory tract, such as reversible airways :.
obstruction and asthma.
:
lO ln a further particular aspect, the present invention provides a compound of forrnula (I) or where appropriate a pharmaceutically acceptable salt thereof and/or a pharmaceutically acceptable solvate thereof, for use in the treatments mentioned hereinbefore, such as cerebral vascular and neuronal denerative .
disorders associated with learning, memory and cognitive dysfunctions, ::
15 peripheral vascular disease or proliferate skin disease or for the prophylaxis of disorders associated with neuronal degeneration resulting from ischaemic events. ;~

In a further aspect there is provided a compound of formula (I) or if appropriate a -pharmaceutically acceptable salt thereof and/or a pharmaceutically acceptable 20 solvate thereof, for use as an inhibitor of the in vivo production of Tumor Necrosis Factor (TNF).

Particularly, there cornpound of formula (I) or if appropriate a pharrnaceutically acceptable salt thereof and~or a pharmaceutically acceptable solva~e thereof, for ~5 use ill the treatment of and/or prophylaxis of diseases associated ~ith excessive or unregulated TNF production.

Diseases associated with excessive or unregulated TNF production include rheurnatoid arthritis, rheuma~oid spondylitis, osteoar~hritis, gouty arthritis and 30 other arthritic conditions; sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, cerebral malaria, chronic pulmona~ inflammatory disease, silicosis, pulmonary ;:
sarcoidosis, bone resorption diseases, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as influenza, 35 cachexia secondary ~o infection or malignancy, cachexia, secondary to acquired ~`;.
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, ` WO 93/23401 21~ fi 1~ 6 pcrJcB93/o1ol4 `

immune deficiency syndrollle (AIDS), AIDS, ARC (AIDS related complex), keloid formation, scar tissue formation, Crohn's disease, ulcerative colitis, orpyresis.
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In a further aspect there is provided a compound of formula (I) or if appropriate a i `
pharrnaceutically acceptable salt ~hereof and/or a phamlaceulically a~ceptable solvate thereof, for use in the treatment and/or prophylaxis of viral infections that produce TNE~- as a result of infection, or those which are sensitive to inhibition, such as by decreased replication, directly or indirectly, by Ihe present compounds.
Such viruses include for example HIV-I, HIV-2 and HIV-3, Cytomegalovirus (CMV), Influenza, adenovirus and the Herpes group of viruses, such as but not limited to, Herpes Zoster and Her~es Simplex.

A compound of forrnula (I) or where appropriate a phannaceutically acceptable salt thereof and/or a pharrnaceutically acceptable solvate thereof, may be administered ~ se or? preferably, as a pharmaceu~ical composition also comprising a pharrnaceutically acceptable carrier.

Accordingly, the present invention provides a phalmaceutical composition `
20 comprising a compound of forrnula ~I) or where appropnate a pha~maceutically acceptable salt thereof and/or a pharrnaceutically acceptable solvate thereof, and a phalmaceutically acceptable carrier.
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The active compound may be formulated for administration by any suitable route, 25 the preferred route depending upon the disorder for which treatment is required9 and is preferably in unit dosage form or in a form that a hurnan patient may administer to himself in a single dosage. Advantageously, the composition is suitable for oral, rec~al, topical, parenteral, intravenous or intramusc~
administration or through the respiratory tract. Preparations may be designed to i 30 give slo~ release of the active ingredient. ` ~ l ' .
The compositions of the invention may be in the forn of tablets, capsules, }~
sachets, vials, powders, granules, lozenges, suppositories, reconstitu~able ~ ;
powdiers, or liquid preparations such as oral or sterile parenteral solutions or ' 3S suspensions. Topical forrrulations are also envisaged where appropriate.

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W0 93/23401 2 1 3 6 1 9 ~ - 12- PCr/G B93/0 1014 In order to obtain consistency of administration it is prefe~red that a composition of the invention is in the form of a unit dose.
.~ , .
S Unit dose presentation forTns for oral administration may be tablets and capsules and may contain conven~ional excipients such as blnding ag.,n;s, for exarnple syrup, acacia, gelatin, sorbitol, tragacanth, or po!yvinylpyrrolidone; illlers, for example lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine;
tabletting lubricants, for ex~nple magnesium stearate; disintegrants, for example 10 starch, polyvinylpyrrolidone, sodium starch glycollate or microcrystalline cellulose; or phannaceutically acceptable wetting agents such as sodium lauryl sulphate.

The solid oral compositions may be prepared by conventional methods of -15 blending, filling, tabletting or the like. Repeated blending operations may be used eo distribute the active agent throughout those compositions employing large quantities of fillers :

Such operations are of course conventional in the art. The tablets may be coated20 according tO methods well known in normal phalmaceutical practice, in particular with an enteric coating.

Oral }iquid preparations may be in the form of, for cxample, emulsions, syrups, or elixirs, or may be presented as a dry product for reconslitution with water or other 25 suitable vehicle before use. Such liquid preparations may contain conven~ional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminium :
stearate gel, hydrogenated edible fats; emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible 30 oils), for ~xample almond oil, fractionated coconut oil, oily esters such as esters of glycerine, propylene glycol, or ethyl alcohol; preselvatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid; and if desire~ conventional ~;
flavouring or colouring agents~ ~

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~361~6 i~ Wo ~3~23401 pcr/GB93/o1ol4 Compositions may also ~uitably be presented for administration to the respiratory tract as a snuff or an aerosol or solution for a nebulizer, or as a microfine powder for insumation, alone or in cornbination with an inert carner such as lactose. In such a case Ihe par~icles of active compound suitably have diameters of less than 50 microns~ such as from 0.1 to 50 microns, preferably less than 10 microns, forexaMple from 1 to lO microns~ l to S microns or frorn 2 tO 5 microns. Where appropriate, small amounts of other anti-asthmatics and bronchodilators, for example sympathomimetic amines such as isoprenaline, isoetharine, salbutamol, phenylephrine and ephednne; corticosteroids su~h as prednisolone and adrenal }O stimulants such as AC~H may be included.

For parenteral adminis~ration, fluid unit dosage fonns are prepared utilizing the compound and a sterile vehicle, and, depending on the concentration used, can be .-either suspended or dissolved in the vehicle. In preparing solutions the compound can be dissolved in water for injection and filter sterilized before filling into a suitable vial or ampoule and sealing.
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Advantageously, adjuvants such as a local anaesthetic, a preservative and buffering agents can be dissolved in the vehicle. To enhance ~he stability, the composition can be frozen after filling into the vial and the water removed under vacuum. Parenteral suspensions are prepared in substantia!ly the same manner, except that the cornpound is suspended in the vehicle instead of being dissolved, and sterilization cannot be accomplished by filtration. The compound can be sterilized by exposure to ethylene oxide before suspending in the stenle vehicle.
Advantageously, a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the compound.

The compositions may contain from 0.1% to 99% by weight, preferably from 10-60% by weight, of thè active material, depending on the method of administration.

Compounds of formula (1), or if appropnate a pharrnaceutically acceptable salt thereof and/or a pharrnaceutically acceptable solvate thereof, may also be adrninistered as a topical formulation in combination with conv~ntional topical ; ~ 35 excipients.

: ' 2-~36~9~
WO 93/2340 1 ^ pcr/G B~3/0 1 0 1 4: - -Topical formulations may be presented as, for instance, ointments, c.reams or lotions, impregnated dressings, gels, gel sticks, spray and aerosols, and may contain appropriate conventional additives such as preservatives, solvents to assist S drug penetration and emollients in ointments and creams. The formulations may contain compatible conventional carriers, such as cream or ointment bases and ethanol or oleyl alcohol for lotions.
~ : .
Suitable cream, lotion, gel, sticlc, ointment, spray or aerosol formulations that 10 may be used for compounds of formula (I) or if appropriate a pharmaceuticallyacceptable salt thereof, are conventional folmulations well known in the art, for example, as described in standard text books of pharrnaceutics and cosmetics, .
such as Harry's Cosmeticology published by Leonard Hill Books, Remington's Pharmaceutical Sciences, and the British and US Pharmacopoeias Suitably, the compound of formula (I), or if appropriate a pharmaceutically acceptable salt thereof, will comprise from about 0.5 to 20% by weight of the formulation, favourably from about 1 to 10%, for example 2 to 5%.

20 The dose of the compound used in the treatment of the in~ention will vary in the usual way with the seriousness of the disorders, the weight of the suf~erer, and the relative ef~lcacy of the compound. However, as a general guide suitable unit doses may be 0.1 to 1000mg, such as 0.5 to 200, 0.5 to 100 or 0.5 tO 1O mg, for example 0.5, 1, 2, 3, 4 or 5 mg; and such uni~ doses may be administered more ~5 than once a day, for example 2, 3, 4, 5 or 6 times a day, but preferably 1 or 2 times per day, so that the total daily dosage for a 70kg adult is in the range of about O.l to 1000 mg, that is in the range of about 0.001 to 20 mglkg/day, such as .
0.007 t~ 3, Q.007 to 1.4, 0.007 to 0.14 or 0.01 to 0.5 mg/kg/day, for example 0.01, 0.02, 0.04, 0.05, 0.06, 0.08, O.l or 0.2 mg/kg/day; and such therapy may 30 extend for a number of weeks~or months~

When used herein the tenn 'pharrnaceutically acceptable' encompasses materials suitable for both human and veterinary use. No toxicological effects have been established for the compounds of formula (I) in the abovementioned dosage 35 ranges.

.~WO 93/23401 2 ~ 3 61 9 6 pcr/GB93/olo14 ~ :
., ' , The followlng phaimacolog;cal data and examples illustrate the inven~ion. The following preparations illustrate the prep3ration of intermediates to the novel ! ~:
compounds of ~ormula (I).
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;., ;-W093~23401 z~36~9 PCT/GB93/010l4 ~

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R-t4-AcetQXy-~=~9L5~o 11~L)~ut ylaminn~-1,3-di( ~ ~enzyl)xanthine S

~\N ~ 2 ~1 ;' Potassium t~butoxide ~0.35 g, 3.13 mmol) was added to a solution of 8-amino-1,3 di~cyclopropylmethyl)-7-~4-methoxybenxyl)xanthine (0.99 g, 2.5 mmol) in dimethoxyethane (DME 10 ml) at 60 C under nitrogen. After 3h a solution of 4-acetoxy-3-(acetoxymethyl)butyl iodide ~1.6~ g, 5.4 mmol) in DME (3 ml) was slowly added over 5 min~ After stirring for 18 h the mixture was cooled, poured : into eth~.~ acetate and the organic solution ~ashed with lS water, dried and e~aporated. Chromatography (acetone/hexane 1:7) on silica gave 8-~4-acetoxy-(3-acetoxymethyl)butyl- . .
amino~- 1,3~di(cyclopropylmeth~ 7-(4-methoxyben2yl)-xanthine (0.63 g, 43%) mp 129-129.5 C;
:
~ (CDCl3) O.q3-O.S0 (8H, m), 1.26-1.38 (2H,m), 1.58 (2H,m), 1.91 ~}H,m), ~.OS (6Hts), 3~47 (2H,m), 3.79 (3~,s), 3~90 (2H,d,J=3.9Hz~ 3.93 (2H,d,J=3.9Hz), 4.03 ~4H,m), 4.30 ~lH~tt~=6~0Hz) ~ So29 (2H,s)~ 6.88 (2H,d,J=8.5Hz) and 7.22 ~2H,d,J=8.5~z); l .
i.. -vmax (KBr) 3272 (m), 1742 (s), 1693 (s), 1651 ~s), 1617 ,~
(5~ t 1566 (s~, 1250 (s), 1240 (s), 1221 (s) and 1040 's) cm~l; '' , m/e (FA~) 121 ~100~), 145 (34), 10~ (27), 5~2 ~MH~,25) and ~ 604 (MNa~, 153;

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WO 93~23401 213 61 9 ~ PCl/~B93/01014 - 17 - !
Found C, 61~7~; H, 6.89; N, 11.82; C30H3~NsO7 requires C, 61.94; H, 6 76; N, 12.04 followed by starting material (0.41g, 42%) identical t~o an . :~
5 authentic sample. ~ -.
1 . .

~XA~ ` ' ) i ( cyc 1 op rn~y l met hy 11 ~ -hy c1 ro ~ y - ~ -hydroxymethyl)~llt.yla ~ o~1-7-(4-methQxyhton7yl)xanthin~

~2~)2 O ~ , ''' : .

:~` ''' .
~; 8-(4-Acetoxy-3-~acetoxymethyl)butylamino)-1,3-di(cyclopropylmethyl)-7-(4-methoxybenzyl~xanthine ~0.22 g, ` 0~3~ mmol) and potassium carbonate tO~05 g, 0.037 mmol) were stirred in me~hanol (8 ml) at room temperature for 5 ~.
~; h. The mixture was ne~tralised with cHCl and the solvent removed under reduced pressure. Chromatography of the ~: 20 residue (250 mg~ on silica ~exane/acetone gradient) ~` ylelded 1,3-di(cyclopropylmethyl)-8-~4~hydroxy~3 (hydroxymethyl)butylamino~-7-(4-methoxy ~enzyl)xanthine ~0.11 g, 59%), mp. 141-2C; ~.

~ (CDC13) 0.3~-0.49 ~8H,m), 1.,28~1.38 (2H,m~, 1.61-1.65 (3E~,m), 2.29 ~2El,t,J=4~5Hz), 3.4:6 ~2H,m), 3~68 (4H,hr s), 3.80 (3H,s~, 3.92 (4H,t (overlapping d) ,J=6.5Hz), 4.56 ' (lH,t,3=5.3~Iz~; 5~28 (2H,s~, 6.88 (2H,d,J=8.5Hz) and 7.23 .
(2H,d,J=8.SHz);
max (KBr~ 3314 (m~ t 3250 (s), 1695 Is), 1685 ~s), 1634 ;

WO~3/23401 ~36~9 PCT/GB93/01014 , .

(s), 1611 (s), 1607 (s), 1578 (m), 1030 ~m) and 756 (m) cm~
l; ; .

m/e ~CI) 498 (MH~, 10~%), 35 (40), 448 (12), 396 ~lO)~and 121 (7); ~:

Found C, 62.92; H, 7 10; N, 14.22; C26H3sNss requires C~
62.76; H, 7.09; N, 14.08%

10 I~hF, d;~yclopropylmethyl~-7-me~hylx~,nthine '.r,~ 2 8-~4-Acetoxy-3-~acetoxymethyI)butylaminoJ-1~3 di(cyclopropylmethyl)-7-methylxanthine was prepared from 8-Amino-1,3-di~cyclopropylmethyl)-7-methylxanthine .in 32~, 20 yiel d in an identical manner to Example 1 , mp 163-4C;

S(CDC13) 0~41-0.49 ~8H,m), 1.24~1.37 ~2H,m3, 1.72 ~ (2H,~,J=6.9Hz), 2.08 (6H,s), 2.13 (lH,t,J=~.3Hz), 3.59 : (2H,dt,J=6.9,6.0Hz), 3.6B (3H,s), 3.90 (4H,~ (overlapping d)1 J=7-0 Hz3, 4106-4~20 (4H,mt, 4.50 ~lH,t,J=6.0Hz);;

Found C, 58.05; H, 6~97; N, 14.53; C23H33N5~6 requires C~
58.09; H, 6.99; N, 14.73%.

; ~ , ~''` !, WO ~3~23401 2 ~ ~ 6 I ~ 6 PCl /GB93/01014 1 9 -- - `
. , "
, .

ni (c:ycLQprQpy] methYl2 -B- ~ 4-hydrc?xy~

~ , ,'.
. ~ .

1,3 Di(cyclopropylmethyl)-8-[4-hydroxy-3- .:;
(hydroxymethyl)butylamino~-7-methylxanthine was prepared from 8-~4~acetoxy-3-(acetoxymethyl)butylarnino~-1,3-di(cyclopropylmethyl~-7-methyl xanthine in 59~ii yield in an -identical manner to Example 2, mp 173C;
:''.`' ~ (CDCl3) 0.39-0.48 (8H,m), ~.25-1.39 (2H,m~, 1.69 :~ lS (~H~q,J-6.6Hz), 1.83 (lH,m), 3.49 t2HIm~, 3.64 (3H,s), 3.67 (4H,t,J=5.5Hz), 3.87 (2H,d,J=7.2Hz), 3.91 (2H,d,J=7.2Hz), 'I;

4~04 (2H~t,J=5.5Hz~, 6.17 ~lH,t,J=5~5Hz)~
....
1~ Found C, 58.11; H, 7.56; N, 17.98; C1gH29NsO4 requi~es C, ~1 20 58~29; H, 7~47; N, 17~89%.

i,~ .

,~

. ~r, ` ` ' . :'.

:

: ~ ~
' W093/~340~ 36~,9~ - 20 - PCI/CB93/01014 ~~ ~

. , ,:

.. . ~ ..

~/\N~H (CH2~ Z~ (Q~zOAd z ~1 .
The title compound (mp 98-99C) was prepared using an analogous procedure to that described in Example 1.
' `

, ' '.

~ w093/23401 ~131;~96 PC~/CB93/OiO14 ~ ~

PHAR~IACOLOGICAL DATA

Inhibition of Rhosphodiesterase Isolation of phosphodiesterases ~;

The Ca2~/calmodulin-stimulated PDE~ (PDE I, see Table I and Beavo and Reifsynder ~1990) for nomenclature) was prepared from bovine cardiac ventricle.
Following chromatography on a Mono Q column, the fractions showing 10 stimulation of PI)E activity by Ca2~ and calmodulin were pooled and further puri~ed on a calmodulin-affinity column. cGMP-stirnulated PDE (PDE II), `
cGMP-inhibited PDE (PI:~E III) and cAMP-specific PDE (PDE IV) were all isolated from guinea-pig cardiac ventricle. Initial chromatography on a 20 rr~
Mono Q column resolved PDE III from a peal; that contained both PDE II and 15 PDE IV. The la~ter were separately rechromatographed on a 1 ml Mono Q
column. cGMP-selective PDE tPDE V) was obtained from porcine lung using chromatography on DEAE-cellulose and ~ono Q columns; a calmodulin-affinity column was used to remove residual PDE I activisy.

20 Character;sticsof phosphodiesterase isoenzymes With the exception of PDE II, which displayed positive cooperativity, all the preparations showed simple Michaelis-Menton kinetics (see Table 1).

PDE I The activity of this isoenzyme was stimulated by the Ca2+- calm~ulin 25 complex. The isoenzyme could hydrolyse both cAMP and cGMP, the latter was the prefe~ed substrate. ;

PDE II The activity of this isoenzyme with cAMP as a substrate was stimulated by c&MP. The isoen~yme could hydrolyse both cAMP and cGMP, the latter was 30 the prefe~ed substrate unde~ basal conditions. The activity of this isoenzyme was unaffected by the Ca2+-calmodulin complex.
t~ -~
PDEIlIThe activity of this isoenzyme with cAMP as a subs~rate was inhibited by cGMP. The isoen:~yme could hydrolyse bo~h cAMP and cGMP, the former was ,...

'.

W093/23401 `~36~9 pcr/GB93/olol4 '~

the preferred substrate. The activity of this isoenzyme was unaffected by the Ca2+calmodulin complex.

~ ' ' PDE IV This isoenzyme had high affinity for cAMP, the hydrolysis of S which was not inhibited by cGMP. The activity of this isoenzyme was unaffected by the Ca2-calmodulin complex.
` ."
PDE VThis isoenzyme had high affinity for cGMFi. The activity of this isoenzyme was unaffected by the Ca2~-calmodulin complex.
Assay of phosphodiesterase activity PDE activity was assayed by the boronate colurnn method as previously described (Reeves et. al., 1~87). The enzymes were assayed by incubation at 37C for 4-30 min. in 50 mM Tris, S mM MgC12, pEI 7.5 with 3H-labelled cyclic nucleotide (4 x 105 disintegrations min -l) and 14C-labelled nucleotide 5'-monophosphate (3 x 103 disintegrations min~l). The assay was stopped by boiling and the 3H-la~elled 5'-monophosphate product separated from substrate on boronate columns. The reaction mixture was diluted with 0.5 rnL lO0 mM HEPES
[N-(2-hydroxyethyl)piperazine-Nl-2-ethanesulfonic acid], 100 mM NaCl, pH
8.5, and applied to the column. The colurnn was extensively washed ~vith the same buffer, and the 5'-nucleotide elu~ed with 6 rnL of 0.25 M ace~ic acid. The recovery of product as judged by 14C-recovery was approximately 8()%. All assays were linear with time of incubation and concentration of enzyme over the range used in these experiments.
ICso values (the concentration of inhibitor required for S0% inhibition of activity) were obtaine~ by incubation of the isoen~yme using 1 ~M cGMP as a substrate for PDE I (in ~he absence of Ca2+ and calmodulin), PDE II and PDE V
and with 1 ,uM cAMP as a substrate ~or PDE III and PDE~ IV.
, 30 A range of inhibitor concentrations from O. l x ICso to l O0 x ICso was used.

~, '.

`: W0 93/23qO1 ~ ~ 3 61 9 6 PCr/CB93/01014 ~ ~-23- ! ~`
~e~e~ enc~s BEAVO, J.A. and D.H. REIFSNYDER, Primary sequence of cyclic nucleotide , .
phosphodiesterase isozymes and the design of selective inhibitors. Trends.
Pharrnacol. Sci. l 1, 150 155 ( l 990).

REEVES M.L., B.K. LEIGH and P.J. ENGLAND, The identification of a new `~
cyclic nucleotide phosphodiesterase activity in human and guinea-pig cardiac ventricle. Biochem. J. 241, 535-541 (1987).
.

~.

WO 93123401 2 ~ t9 6 PCl /GB93/01014 T~ble 1: Kinetic properlies of phosphodiesterase isoenzymes Isoenzyme Km ~IlM) Vmax cAMP
cAMP cGMP V-nax cGMP
I. Ca~/calmodulin- 36 5 5 stimulated Il. cGMP-stimulated 45 14 III. cGMP-inhibi~ed 0.5 0. l 5 IV. cAMP-specific 2 > n d.
V. cGMP-specific > l N.d.

a enzyme displayed positive cooperativity lS > Km>l00~M
n.d. not deterrnined, due to inability of PDE ~o hydrolyse one of the substlates.

RESULTS
EXAMPLE NO. INHIBITION OF: ;
PDE VA PDE IV
(IcsollM) 0.2 3 2 0.2 9 ":'.

.

t ` '' ~

WO~3/23~101 ~13 li1 9 6 PClr/GB93/01014 ~ ~

Inhibitory Eff~ct (>f compouncls of Formula (I) on I ;`
itro TNF production by Hum~n Monocytes Sççti~n l: A~SRY set-~lp ~ ;
The effects of compounds of Folmula (l) on the in vitro production of TNF !
by human rnonocytes ~as ex~mined using the following protccol.
Human peripheral blood monocytes were isolated and purified from either :
blood bank buffy coats or plateletpheresis residues, according to the procedure of Colotta, R. et al., ~. lmmunol., 132(2):936 (1984). The monocytes were plated ata density of I X 106 cells/ml medium/well in 24-well multi-dishes. The cells were ~;~
allowed to adhere for l hour after which time the supernatant was aspirated and 1 Tnl fresh medium (RPMI- l 640 (Whitaker Biomedical Products, Whitaker, CA) containing 1% fetal calf serum and penicillin and streptomycin at 10 units/ml was added. The cells were incubated for 45 minutes in the presence or absence of test compounds at lnM-lOuM dose ranges (compounds were solubilized in Dimethyl- ;
sulfoxide/E~hanol such ~hat the final solvent concentration in the culture medium was 0.5% Dimethyl sulfoxidelO.5% Ethanol). Bacterial lipopolysaccharide (E.
coli O55:B5 [LPS] from Sigma Chemicals Co.) was then added at 10~) ng/ml in 10 ~:
ml Phosphate Buffered Saline (PBS) and cultures incubated for 16-18 hours at ;
37C in a 5% C02 incubator. At the end of the incubation period, culture supernatants were removed from the cells, centrifuged at 3000 revolutions per minute (rpm) to remove cell debris and .05 ml of the supernatant assayed for TNFactivity using the radioimmunoassay described below.

2~ Radioimmunoas~aY procedure fQr TNF activitv The assay buffer consisted of O.OlM NaP04, O.l5M NaCl, 0.025M EDT~
and 0.1% sodium azide at pH 7.4. Human recombinant T~F (rhTNF) ob~ained ;, using the procedure of Chen et al., Nature, 330:581-583 (1987) was iodinated by a modified Chloramine-T method describ~d in Section III below. To samples (50 ~l i culture supematants) or rhTNF standards, a 119000 dilution of polyclonal rabbit anti-rhTNF (Genzyme1 Boston, MA) and 8000 cpm of 125I-TNF was added in a : :
~Inal volume of 400 yl buffer and incubated overnight (18 hours~ at 4C:. Normal ~}~-rabbit serum and goat anti-rabbit IgG (Calbiochem) were titrated against each other for maximum precipitation of the anti-rhTNF. The appropriate dilutions sf camer normal rabbit serum (l/200), goat anti-rabbit IgG (1/4) and 25 Units WO 93/23401 ~ " PC~/GB93/01û14 ~
~, ~ 3 6 --2~-- !
hep.~r~n tCal~iochem) were allowed tO precipitate and 200 ~1 of this complex wasadded per assay tub~ and incubated overnight at 4C. Tubes were centrifuged for 30 minutes at 2000 rpm, supernatants were carefully aspirated, and radioactivityassociated with the pellets measured in a Beckman Gamma 5500 counter. The S logit-log linear transformation curve was used for the calculations. The concentrations of TNF in the samples was read frorn a standard curve of rhT~F
that was linear in the 157 to 20,000 pg/ml range.
;
~s~n Tll: R~dioiodination of rhTNF
Iodination of rhTNF was performed using a modified chloramine-T
rnethod of Frolik et al., J. Biol. Chem., 259:10995-11000 (1984). Bnefly, 5 mg of rhTNF in 5 ml of 20MM Tns ph 7.5, was diluted with 15 ml of 0.5M KPO4 and 10 ml of carrier free 1251tlOOmCi/ml;ICN). To initiate the reaction, a 5ml aliquot of a 100mg/ml (aqueous) chloramine-T solution was added. After 2 minutes at room temperature, an additional 5 ml aliquot was added followed l.S minutes later by a final 5 ml addition of chloramine-T. The reaction was stopped 1 minute later by sequential addition of 20 ml of 50mM Sodium Metabisul~lte, 100 ml of 120rnM Potassium Iodide and 200 ml of 1.2 mg/ml Urea. The contents were mixed and the reaction mixture was passed over a pre-packed Sephadex G-25 column tPD 10 Pharmacia), equilibrated and eluted with Phosphate Buffered Saline pH 7~4 containing 0.25% gelatin. The peak radioactivity containing ~ractions were pooled and stored at -20C. Specific activity of 125I^TNF was 80 100 mCilmg protein. Biological activity of iodinated Tl~- was measured by the L929 cytotoxicity assay of Neale, M.L. et al., Eur. J. Can. Clin Oncol, 25tl):133-137 tl989) and was found to be 80% that of unlabeled TNF.
Section rv: Mea~urement of TNF- EL,ISA-l~vels of TNF were also measured using a modification of the basic sandwich E;LISA assay method described in Winston et al., (~rr~t P~~
Molec~llar Biolog~ Page l 1.2.1, Ausubel et al., Ed. (1987) John Wiley and Sons,New Yorkl USA The ELISA employed a murine monoclonal anti-human TNF
antibody, described below, as the capture antibody and a polyclonal rabbit anti-human TNF, described below, as the second antibody. For detection, a peroxidase-conjugated goat anti-rabbit antibody (Boehringer Mannheim, Indianopolis, Indiana, USA, ~atalog # 605222~ was added followed by a subs~ate f~r peroxidase (lmg/rnl orthophenylenedlamine with 0.1% urea peroxide). TNF -' Wo 93/23401 21 3 61 9 ~ P~/GB93/01 01 4 levels in samples we~e calculated from a standard curve generated with recombin~nt human TNF produced in E. Coli (obtained from SmithKline Beecham Pharmaceuticals, King of Prussia, PA, USA).
Seçtion V: Productic)n Qf ~nti-h~man TNF anti~odi~:
S Monoclonal antibodies to human TNF were prepared from spleens of BALB/c mice immunized with recombinallt human TNF usin~ a modi~lcation of the method of Kohler and Millstein, Nature 256:495 (1975), the entire disclosureof which is hereby incorporated by reference. Polyclonal rabbit anti-human TNF
antibodies were prepared by repeated immunization of New Zeland Whi~e (NZW) rabbits with recombinant human TNF emulsified in complete Freund's adjuvant (DIFCO, ~L., USA).
.'~
It has been deterrnined that 8-(4~Acetoxy-3-acetoxymethyl)-l-butylamino-l,3-di(cyclopropylmethyl)-7-p-methoxybenzyl xanthine demonstrated an ICso of about 0.30 !~M in the in-vitro TNF production assay system.

Endotoxin Shock in D~ Sensiti~.ed Mi~e The protocol is essentia}ly as described in Galanos et al., Proç Nat'l Aca~
Sci USA, ~ 5939-43 (l979) whose disclosure is herein incorporated by reference.
Briefly, D-gal (D(+) Galactosidase) sensitizes various strains of mice tO the lethal effec~s of endotoxin. The adrninistration of D-gal (300-SOOmg/kg~ intra-venously(i.v.) sensitizes the mice to doses of lipopolysaccharide (LPS) as low as O.l,ug.
Briefly, male C57BL/6 mice, obtained from Charles River Laborator~es (Stone Ridge1 New York, USA) of 6-l2 weeks of age are injected i.v. with O.l ~g o~LPS
from Salmonella ~hosa (Difco Laboratories, Detroit, Michigan, USA) admixed with 1:)(+) gal (Sigma; 500 mglkg) in 0.20-0.25 ml pyrogen-free saline.
Compounds to be tested are administered at various times prior to or following the i.v. injection of LPSlD-gal. In this model, the control animals usually die 5-6 hr.
following the injection of LPS, although on occasion deaths are seen between 24 and 48 hr.
~leasurement of T~F Activitv Plasma levels of TNF are measured using a modification of the basic sandwich E~ISA methocl descnbed in Wins~on et al., Current Prot~cols in ~ -Molecular B;QIO~Y~ Pg. l E~. l, Ausubel et al., Ed. (1987) John Wiley and Sonst ', 35 New York t USA. The Elisa employed a hampster monoclonal anti-mouse TNF

WO 93/23401 ~ 6 ~9 PCl/GB93tO1014 (Genzyme, Boston, ~vIA7 IJSA ) as the caplure anlibQdy and a polyclonal rabbit anti-murine TNF (Genzyme, Boston, MA, USA ) as the detecting antibody. TNF
levels in mouse samples are calculated from a s~andard curve generated with recombinant murine TNF~ (Genzyme, Boston, MA, USA). TNF levels determined 5 by ELISA correlated with levels detected by the L929 bioassay of Rut-f et. al., L
ImmunQL 125:1671-1677 (1980), with 1 Unitof activity in the bioassay co~Tesponding to 70 picograms (pg) of TNF in the ELISA. The ELISA detected levels of TNF down to 25 pg/ml. :
Active compounds provide a positive in-vivo response in the above noted 10 model and for example demonstrate an EDso for reduction of serum TNF of about 2 50 mg/lcg orally. .

:.

, .

.' , ~, `~ ' ';

Claims (16)

1. A compound of formula (I):
(I) or if appropriate a pharmaceutically acceptable salt thereof, wherein R1 and R2 each independently represent a moiety of formula (a):

-(CH2)m-A (a) wherein m represents zero or an integer 1, 2 or 3 and A represents a substituted or unsubstituted cyclic hydrocarbon radical;
R3 represents hydrogen, substituted or unsubstituted alkyl or an aralkyl group substituted or unsubstituted in the aryl moiety; and R4 represents hydrogen, alkyl or alkylcarbonyl.

2. A compound according to claim 1, wherein A represents a cyclopropyl group.

3. A compound according to claim 1 or claim 2, wherein R3 represents an aralkyl group.

4. A compound according to any one of claims 1 to 3, wherein R3 represents a substituted or unsubstituted benzyl group.

5. A compound according to any one of claims 1 to 4, wherein R3 represents a 4-methoxy benzyl group.

6. A compound according to any one of claims 1 to 5, wherein R4 represents hydrogen.

7. A compound according to any one of claim 1 to 5, wherein R4 represents alkylcarbonyl.

8. A compound according to claim 1 selected from any one of Examples 1 to 28 herein.

9. A process for preparing a compound of formula (1), which process comprises reacting a compound of formula (II):

(II) wherein R1a represents R1 as defined in relation to formula (I) or a group convertible to R1, R2a represents R2 as defined in relation to formula (I) or a group convertible thereto and R3a represents R3 as defined in relation to formula (I) or a group convertible to R3, with a compound of formula (III):
L1-(CH2)2CH(CHOR5)2 (III) wherein, R5 represents a hydroxy protecting group and L1 represents a leaving group; and thereafter, if required carrying out one or more of the following optional steps:

(i) removing any protecting group;
(ii) converting any group R1a to R1 and/or R2a to R2 and/or R3a to R3;
(iii) converting a compound of formula (I) into a further compound of formula (I);
(iv) converting a compound of formula (I) into a pharmaceutically acceptable salt thereof.

10. A process for preparing a compound of formula (I), wherein L1 is a halo atom, especially an iodine atom.

11. A pharmaceutical composition comprising a compound of formula (I) or if appropriate a pharmaceutically acceptable salt thereof and/or a pharmaceutically acceptable solvate thereof, and a pharmaceutically acceptable carrier.

12. A compound of formula (I) or if appropriate a pharmaceutically acceptable salt thereof and/or a pharmaceutically acceptable solvate thereof, for use as an active therapeutic substance.

13. A compound of formula (I) or if appropriate a pharmaceutically acceptable salt thereof and/or a pharmaceutically acceptable solvate thereof, for use as an inhibitor of the in vivo production of Tumor Necrosis Factor (TNF).

14. A compound of formula (I) or if appropriate a pharmaceutically acceptable salt thereof and/or a pharmaceutically acceptable solvate thereof, for use in the treatment of and/or prophylaxis of diseases associated with excessive or unregulated TNF production.

15. A use according to claim 14, wherein the treatment is the treatment and/or prophylaxis of rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions; sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, cerebral malaria, chronic pulmonary inflammatory diseaser silicosis, pulmonary sarcoidosis, bone resorption diseases, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as influenza, cachexia secondary to infection or malignancy, cachexia, secondary to acquired immune deficiency syndrome (AIDS), AIDS, ARC (AIDS related complex), keloid formation, scar tissue formation, Crohn's disease, ulcerative colitis, orpyresis.

16. A use according to claim 14, wherein the treatment is the treatment and/or prophylaxis of viral infections that produce TNF as a result of infection, or those which are sensitive to inhibition, such as by decreased replication, directly orindirectly, by the present compounds. Such viruses include for example HIV-1, HIV-2 and HIV-3, Cytomegalovirus (CMV), Influenza, adenovirus and the Herpes group of viruses, such as but not limited to, Herpes Zoster and Herpes Simplex.