AU715774B2 - Intermediates for the production of pharmaceutically useful benzothiazolone derivatives - Google Patents

Description

AUSTRALIA

PATENTS ACT 1990 REGULATION 3.2 F 1594-DIV:1P AU Name of Applicant: Actual Inventor/s: ASTRA PHARMACEUTICALS LIMITED ROGER VICTOR BONNERT, ROGER CHARLES BROWN,

SEC

PETER ALAN CAGE,

O

FRAJJCIS INCE and 104 E.F. WELLINGTON CO., Patent and Trade Mark Attorneys, 312 St. Kilda Road, Melbourne, Southbank, Victoria, 3006.

Address for Service: Invention Title: "INTERMEDIATES FOR THE PRODUCTION OF PHARMACEUTICALLY USEFUL BENZOTHIAZOLONE DERIVATIVES" Details of Associated Provisional Applications Nos: The following statement is a full description of this invention including the best method of performing it known to us. IP Australia

CD

Documents received on: 0 09 APR 1999 Batch No: Batch No: 1 ]DNTRMIATES FOR THE PRODUCTTON OF PIUUMCEUrICALLY USEFUL BEZTHA LONE DERTVATIVE Field of the invention This invention relates to novel intermediates for the production of benzothiazolone derivatives, in particular the production of novel 7- (2aniinoethyl) -benzothiazolone derivatives, said novel comTpounds being dopamine IDA 2 receptor agonists and S -adrenoreceptor agonists, the subject of the parent Australian Patent Application No. 69371/96 herein and in respect of which the present application is a divisional application.

B acka-round Beazothiazolone derivatives are known. For example, international patent applications, publication numbers W092.'08708 and W093/23385 disclose biologically active amnines, amongz them bioioi callv active amninoethyl berizothiazolone derivatives which are02 adrenoreceptor agonists and dopamrine DA 2 -receptor agonists, and which are indicated in the treatment of obstructive airways diseases.

WO 93i24473 discloses 7-(2--ami'noethv!)-benzothiazolone compounds of formula (CH pX~CHqY (HrZ OH H 2R3 0weenXdYaeindependently hydrogre0-nnis0, or 2;k p,-6 qh andond ae5adeocctr a~st n dopamine DA 2 -recepIrtor agonists. and are indicated in the treatment of obstructive airways diseases.

1B The specification of the parent Australian Patent Application No. 69371/96 discloses that we have found a group of novel 7- (2-aminoethyl) -benzothiazolone derivatives which are useful as dopamine DA 2 -receptor agonists and I2-adrenoreceptor agonists, said derivatives being compounds of formula I, including optical isomers thereof:

R

HN(CH2)p-X-(CH2)q-O-(CH),-Y

S

HN

OH H wherein X represents -SO2NH- or -NHSO 2 p, q and r independently represent 2 or 3, Y represents thienyi optionally substituted by alkyl or halogen, or phenylthio- or phenyl optionally substituted by alkyi or halogen, and each R indeendently represents H or alkyl, and pharmaceutically acceptable salts, esters and amides thereof.

:The compounds are pharmacolozically active. They show both dopamine DA 2 -receptor agonism and (3-adrenoreceptor agonism. They exhibit little or no ac-adrenoreceptor agonism. The compounds have an advantageous duration of action and DA2/B2 ratio.

Preferably, q in formula I above is 2. r is preferably 2.

When Y is phenyl substituted by alkyl, the alkyl group is preferably a Ci6, for example a CI or C 2 group, most preferably methyl.

When Y is phenyl substituted by halogen, the halogen substituent is preferably a chloro- or fluoro-substituent.

Preferred compounds of the parent invention are compounds of formula I wherein X is S02NH, p is 3 and q and r are each 2. Other preferred compounds are compounds of formula I wherein X is NHS02, and p, q and r are all 2.

Outline of the present invention In accordance with the present invention there is provided novel intermediate compounds which can be utilized as hereinafter described for the production of the novel 7- (2-aminoethyl)-benzothiazolone derivatives of the parent invention, said novel intermediate compounds being of the formula VIII: R

A-(CH

2 pl-X-(CH 2 q-O-(CH)r-Y VIII wherein A is O=CH or is HOOC or the corresponding acid chloride, in which X represents -SO2NH- or NHSO 2 p, q and r independently represent 2 or 3, Y represents thienyl optionally substituted by alkyl or halogen, or phenylthio- or phenyl optionally substituted by alkyl or halogen, and each R independently represents H or alkyl.

Thus, the present invention provides intermediate compounds of formula III: R O =CH- (CH 2 p 1 (M2)q-0- (CH)r-Y III in which p, q, r, R, X and Y are as defined above, or intermediate "compounds of formula V:

R

I

HOOC- (CH 2 )p_ 1

(CH

2 q-0- (CH r -Y V or the corresponding acid chloride, in which p, q, r, R, X and Y are as defined above.

More particularly, the present invention provides intermediate compounds *of formula III or of formula V wherein q is 2; or wherein r is 2; or wherein Y is phenyl substituted by methyl; or wherein Y is phenyl substituted by a halogen substituent selected from a chloro- or fluorosubstituent; or wherein X is S02NH, p is 3 and q and r are each 2; or wherein X is NHSO 2 and p, q and r are all 2.

21 Suitable methods for the preparation of the intermediate compounds of formula VIII, namely, the intermediate compounds of formula III or the intermediate compounds of formula V, as disclosed in the description set out below, may comprise a process in which: when A is O=CH, reducing a corresponding ester compound, or (ii) when A is HOOC, hydrolysing a corresponding ester compound.

Conveniently, further reference to the intermediate compounds of the present invention, their preparation, and their use in producing the novel 7- (2-aminoethyl) -benzothiazolone derivatives of the parent invention, will be via the text and disclosures in the specification of the parent Australian Patent Application No. 69371/96 as reproduced herein. Thus, reverting to the parent specification, suitable pharmaceuticaily acceptable salts of the compounds of formula I include acid addition salts derived from inorganic and organic acids. The compounds may also form salts with suitable bases. Examples of suitable salts include the hydrochloride, citrate, D,Llactate, hemisulphate, hemitatrate, D-gluconate, methanesulphonate, p-toiuenesulphonate, hemifumarate, benzoate, xinafoate, hemisuccinate, 3-hydroxy-2-naphthoate, hemiembonate, hemimaleate, D-camphorsulphonate, 10-undecanoate, mandelate, naphthaiene-1-sulphonate, naphthalene-2-sulphonate, 4-methoxybenzoate, 4-chlorobenzoate, saccharinate, monomethvi suberate, hemisuberate and diphenyl acetate salts.

Suitable pharmaceutically acceptable esters of the compounds of formula I include phenylalkyi and alkvl esters.

Suitable amides include unsubstituted or mono- or di-substituted alkvi or phenvi amides.

The most preferred compounds of the parent invention are 3-[2-(4-Hydroxy-2-oxo-3 H- ,3-benzothiazol-7-yl)ethyamino]-N-[2-(2-phenylethoxy)ethyl] "propanesulphonamide; *9 N-[2-[2-(4-Hydroxy-2-oxo-3 H- I ,3-benzothiazol-7-yI)ethylaminolethyl]-2-(2phenvlethoxy)ethanesulphonamide; 3 -[2-(4-Hydroxy-2-oxo-3 H- 1,3 I-benzothiazol-7-yl)ethylamino]-N-[2-[2-(5-methyl-2thienyl)ethoxy] ethyl] propanesulphonamide; N-[2-[2-(4-Fluorophenyl)ethoxyj ethyl]-) -[2-(4-hydroxy-2-oxo-3 H-i, 3-benzothiazo-7r yl)ethylamino] prop anesulphonari de; i0 N- [2-(4-Chlorophenviethoxy] ethvlj-3 -[2--(4-hydroxy-2-oxo-3H- 1, 3-be.nzothiazol-7yl)ethyiamin o]propanesulphonanude;- 3 -[2-(4-Hydroxy-2-oxo-3 H- 1,3 -benzothiazol-7-yI)ethylan)ilN-[2-[2-(4methyiphenyl) ethoxv] ethyl] propanesulphonamnide; S)-3 -[2-(4-Hydroxv-2--oxo-3 1,3 -benzothi azol-7-yl)ethylainofl-N-[2-(2-phenyl- 1propoxy)ethvfllprop anesulp honamide, 3-[-(4-Hydrox'-2-oxo-3 H- 1.3-benzothiazo-7-y)ethylamil1N[2[-2 methylphenyl)ethoxy] ethyflpropanesulphonamide; and 3 -[2-(4-Hydroxv-2-oxo-3 H- 1,3 -benzotiazol-7-yl)ethyiamiflol-N-[2-(2phenylthioethoxy)ethyl] prop anesulphonamide; 2s preferablv in salt form and more preferably as the hydrochloride.

The parent invention also provides a method for the production of compounds of formula I, comprising selective reductive alkvlation of a compound of formula II,

NH

2

II

L 0

N

OH H with a compound of formula II,

R

O=CH-(CH

2

III

in which p, q, r, R, X and Y are as defined above, in the presence of a reducing agent.

The reducing agent may be, for example, hydrogen in the presence of a catalyst such as platinum, platinum oxide, palladium, palladium oxide, Raney nickel or rhodium, on a support e.g. charcoal, using an alcohol, e.g. ethanol, or an ester, e.g. ethyl acetate. or an ether, e.g. tetrahvdrofuran, or water, as reaction solvent, or a mixture of solvents. at normal or elevated temperature and pressure. The preferred temperature is room temoerarure. The preferred pressure is 1-3 atmospheres. Alternatively the reducing agent may be sodium ,i borohvdride or a metal hydride e.g. sodium cvanoborohydride. Suitable solvents for use with hydride reducing agents will depend on the particular hydride used and will be well known to the person skilled in the ar. Suitable solvents will include alcohols, for example ethanol or methanol.

20 The process may give rise to intermediate imine compounds, which may be reduced under the described conditions, to give compounds of formula I.

The compound of formula II may be prepared by known methods, for example by the method described in J. Med. Chem., 1987, 30, 1116.

6 Aldehydes of formula III may be prepared in a number of ways known per se. For example, isothiazolidine dioxides (as in Example Ic, for instance) may be reduced with DIBAL in toluene; acetals (as in Example 2b, for instance) may be hydrolysed with 70% acueous acetic acid; and esters (as in Example 3d, for instance) may be reduced with DIBAL in toluene. Specific syntheses of certain precursor compounds are described in the Examples and may be adapted to a variety of targets.

The aldehydes of formula 1I may also be prepared from the corresponding alcohols by partial oxidation using DMSO, DCC and anhydrous phosphoric acid; or using pyridinium to chlorochromate or pyridinium dichromate.

The parent invention also provides a further process for preparing compounds of formula I, comprising the selective reduction of a compound of formula IV, 0

R

HN (CH2)p-1-X-(CH2)-O-(C H),-Y

IV

S

S

O

N

OH H in which p, q, r, R, X and Y are as defined above.

Suitable reducing agents include electrophilic reducing agents, diborane and alane 20 (aluminium hydride), or nuciecphilic reducing agents, a complex metal hydride such as sodium bis(2-me:hoxvethoxy)aluminium hydride. The preferred reducing agent is diborane.

The solvent should be inert to the reaction conditions. Aprotic solvents are preferred, e.g.

tetrahydrofuran. diethyl ether, or 1,2-dimethoxyethane. The reaction may be carried out at a temperature of from about OC, to about 100 0 C, preferably at reflux temperature.

Compounds of formula IV may be prepared by coupiing of the amine of formula II and an appropriate acid, of formula V

R

I

SHOOC-(CH2) -X-CH 2 q v or corresponding acid chloride by conventional means. For example, the coupling may be performed in the presence of dicyclohexylcarbodiimide using the method of Sheehan and Hess, J Am. Chem. Soc., 1955, 77, 1067; or 1,1'-carbonvdiimidazoie as described by o0 Staab, Angew Chem. Int. Ed. Engl., 1962, 1, 351; or bromotripyrrolidinophosphonium hexafluorophosphate in a solvent such as DMF, following the procedure of Example le.

The acids required for the process may be obtained from the corresponding esters, by hydrolysis with lithium hydroxide in aqueous methanol, following the procedure of Example lb. Examples la, 2d, 3d, 4f, 5d, 6d, 7d, 8d and 9d describe specific processes for forming the esters, and these process may be adapted to give other esters, for forming further acids for coupling with formula II amines. The acid chlorides may be prepared from the acids for exampie bv reaction with oxalvi chloride or thionyl chloride in toluene at a temperature from ambient to reflux.

:0 The compounds of the arent invention may be prepared by several other methods as well.

0* Alkylation of the compound of formula II, or a salt, ester or amide thereof, with an alkviating agent of formula VI

R

L-(CH)p-X-(CH- 2 )q-O(CH)Y VI i* ft 8 in which p, q, r, R, X and Y are as defined above and L represents a good leaving group, for example a halide e.g. chloride, bromide or iodide, or an alkyl- or aryl-sulphonyloxy group, for example methanesulphonyloxy, is one such method.

The reaction may be carried out for example in the presence of a base, for example an inorganic base, sodium or potassium carbonate, or an organic base, triethylamine, N,N'-diisopropylethylamine or pyridine.

The reaction may be performed in a solvent, for example an ether, e.g. tetrahydrofuran or dioxan. a ketone. e.g. butanone or methyl isobutyl ketone, a substituted amide, e.g.

dimethylformamide, or a chlorinated hydrocarbon, e.g. chloroform, at a temperature of between ambient temperature and the reflux temperature of the solvent. Preferably the reaction is carried out at ambient temperature.

is The alkylating agent of formula VI may be prepared from the corresponding alcohol (i.e.

the compound in which L represents OH) by methods known to the person skilled in the art.

For example, the alcohol may be reacted with a halogenating agent to yield the compound of formula VI in which L represents a halogen atom. Suitable halogenating agents include.

for example, triphenylphosphine-tetrahalogenomethane adduct (conveniently formed in siru.

o r 20 e.g. by the reaction oftriphenylphosphine and carbontetrabromide). The reaction may take a place in the presence of a solvent such as acetonitrile, or a chlorinated hydrocarbon, e.g.

dichloromethane. for example at a temperature in the range of 0-30 0

C.

Another method is the selective reduction of a compound of formula VII, o o 9

R

I

HN(CH2)p-X-(CH2)q-O-(CH),-Y

HN

0 VII

S

I =o

N

OH H in which p, q, r, R. X and Y are as defined above.

Suitable reducing agents include e!ectrophiiic reducing agents, diborane and alane (aluminium hydride), or nucieophiiic, a complex metal hydride such as sodium bis(2methoxyethoxy)aluminium hydride. The preferred reducing agent is diborane. The solvent should be inert to the reaction conditions. Aprotic solvents are preferred, e.g.

tetrahydrofuran, diethyl ether, or 1,2-dimethoxvethane. The reaction may be carried out at a temperature of from about 0 0 C. to about 100°C, preferably at reflux temperature.

Compounds of formula VII may be prepared by coupling of an amine and an acid or acid chloride bv conventional means. For example, the coupling may be performed in the presence of dicyciohexvlcarbodiimide or 1,1'-carbonyldiimidazole or *0 o f ^i bromotripyrrolidinophosphonium hexafluorophosphate, as described above in relation to compounds of formula IV. The amines required for the coupling reaction may be prepared S: by reaction of compounds of formula VI, where L represents a good leaving group for example a halide such as chloride or bromide, with phthaiimide in the presence of a base.

The resulting imides may then be treated with hydrazine hydrate in ethanol to give o compounds of formula VI with the leaving group replaced by an amino group.

In the above processes it may be necessary for any functional groups. e.g. hydroxy or amino groups, present in the staring materials to be protected. Suitable protecting groups and methods for their removal are, for example, those described in "Protective Groups in Organic Synthesis" by T.W. Greene and P.G. M. Wuts, John Wiley and Sons Inc., 1991.

Another process for preparing compounds of formula I comprises removal of a protecting group from a corresponding protected compound of formula I in which one or more of the functional groups is protected, and where desired or necessary converting the resulting compound of formula I to a pharmaceutically acceptable salt, ester or amide thereof, or vice versa.

Pharmaceutically acceptable salts may be prepared for example by reacting the compound of formula I with an appropriate acid in the presence of a suitable solvent.

Pharmaceutically acceptable esters of the compounds of formula I may be made by conventional techniques, e.g. esterification or transesterification.

13 Pharmaceutically acceptable amides of the compounds of formula I may be made by conventional techniques, e.g. reaction of a compound of formula I with an acid or acid chloride.

20 The intermediates of formula IV are novel, thus according to a further aspect of the invention there are provided compounds of formula IV, 0 R HN (CH 2 )p--X-(CH 2

IV

S

ooo" lv>=o

N

OH H OH H 11 in which p, q, r, R, X and Y are as defined above.

The intermediates of formula VII as defined above are also novel and are thus also provided in accordance with the parent invention.

Further, the aldehydes of formula III as defined above are novel and are provided by the present invention.

Still further the acids of formula V and corresponding acid chlorides are novel and are 0o provided by the present invention.

The compounds of formula I and salts, esters and amides thereof are dopamine DA 2 receptor agonists. The binding affinities of the test compounds for the DA 2 receptor binding sites in bovine pituitary membranes may be determined from the displacement of [H]-N-n is -propylnorapomorphine and of['H]-spiperone in the absence or presence of nonhydrolysable GTP analogue respectively, D.R.Sibley, A. DeLean and I. Creese, Anterior Pituitary Dopamine Receptors, Demonstration of Interconvertible High and Low Affinity States of the D-2 Dopamine Receptor, J. Biol. Chem., 1982, 257(11), 6351-6361.

The DA2-receptor activity may also be demonstrated in a functional screen, the rabbit 20 isolated ear artery, as described by Brown and O'Connor, Br. J. Pharmacol., 1981, 73, 189P. The compounds are also P 2 -adrenoreceptor agonists. This activity may be demonstrated in the isolated trachea of the guinea pig, as described by I.G. Dougall, D.

Harper, D.M. Jackson, and P. Leff, Br. J. Pharmacol., 1991, 104, 1057. ac-Receptor activity may be analysed using the rabbit isolated ear artery screen described in Pharmacological Example herein.

The compounds of formula I and salts, esters and amides thereof are thus indicated for use in the treatment of the range of airways diseases, including conditions such as asthma, including bronchial asthma, allergic asthma, intrinsic asthma (for example late asthma and 12 airway hyper-responsiveness); and bronchitis and the like (see, for example, UK Patent No.

2022078 and Br. J. Pharmacol., 1987, 24, 4983).

The compounds of formula I and salts, esters and amides thereof are also indicated for use in the treatment of various other conditions, e.g. inflammatory and allergic skin disorders, cancer e.g. small cell lung cancer, congestive heart failure and glaucoma.

The term "treatment" as used herein includes prophylaxis as well as relief of the symptoms of disease.

AccordinQly, in a further aspect of the parent invention, there is provided the use of a compound of formula I, or a pharmaceutically acceptable salt, ester or amide thereof, in therapy.

Further, there is provided the use of a compound of formula I, or a pharmaceutically acceptable salt, ester or amide thereof, in the manufacture of a medicament for the treatment of obstructive airways disease, in particular for the treatment of asthma or chroric bronchitis.

o0 Still further, the parent invention provides a method of treatment of airways disease, which method comprises administering a therapeutically effective quantity of a compound of formula I, or a pharmaceutically acceptable salt, ester or amide thereof, to a patient suffering from or susceptible to such a condition.

:5 Typical daily unit doses may be for example I 1g 10mg for topical administration, preferably 10-500ug, for example divided two or three times, or 10ug 100mg for oral administration, preferably 100OOg-10mg, for example divided two or three times.

a The compounds of formula I and salts, esters and amides thereof may be used on their own or in the form of appropriate pharmaceutical compositions.

Administration may be by inhalation as well as by other routes, for example by oral or intraveneous administration.

Nasal or pulmonary administration may be achieved via a suitable inhalation device.

For example metered dose inhaler devices may be used to administer the compound, dispersed in a suitable propellant and with or without additional excipients such as ethanol, surfactants, lubricants and stabilising agents.

Suitable propellants include hydrocarbon, chlorofluorocarbon and hydrofluoroalkane propellants, or mixtures of any such propellants. Especially preferred propellants are P134a and P227 each of which may be used alone or in combination with other propellants and/or surfactants and/or other excipients, for example in combination with each other.

Nebulised aqueous suspensions or, preferably, solutions may also be employed. with or without a suitable pH and/or tonicity adjustment, either as a unit-dose or multi-dose device.

Dry powder inhalers may be used to administer the compound, alone or in combination with 20 a pharmaceutically acceptabie carrier, in the latter case either as a finely divided powder or as an ordered mixture. The dry powder inhaler may be single dose or multi-dose and may utilise a dry powder or a powder-containing capsule.

Metered dose inhaler, nebuiiser and dry powder inhaler devices are well known and a 25 variety of such devices are available.

The invention is illustrated, but in no way limited, by the following Examples, in which temperatures are in degrees Celsius. Where necessary, the reactions were performed under an inert atmosphere of either nitrogen or argon. Where necessary, preparative HPLC separations were generally performed using a Novapak Bondapak or Hypersil column packed with BDSC-18 reverse phase silica. Flash chromatography was carried out using Fisher Matrix 60 silica, 3 5-70 micron.

Example I 3 -[r2-(4-Hvdro-xv-2 -oxo- 3H- 1, .3-enzothizol- 7-vl)ethv arino -N22 -p henv e:oxv ethvIl prop anesulp honamide hydrochloride a) Methyl 3-[2-(2-p~henviethoxv,,ethvlaminosuIahonvflarooanoate a 2-(2-Phenylethoxy)ethanane 1(1.95 g) was stirred in dichioromethane at room temperature, Triethylarnine (3.36 mil) followed by methyl 3-(chlorosulphonyl)propanoate 2were added and the mixture stirred overnight at room temp~erature. The mixture was diluted with a ifirther amount of dichi oro methane, washed with dilute hydrochloric acid then water, then dried (MgSO 4 The solvent was removed in vacua to yield a pale yellow oil which was further purified by flash chromatcc_-aphy (01% ethanol: dichloromethane as eiuant) to give the subtitle compound as pale ye;iow o*1 (1.68 g), Mass spectrum: F.AB 3 16 (Mf-H'- IrH (3 60 N'fl-z, CDCI3,) 5: 2.75 (2K 2.85 (2H, 3.18-3.35 (4H. mn), 3.46-3.57 in), 3.60-3.77 (5K, in), 4.44 (1K, brt), 7.12-7.35 (5K in).

1 Chem. Ber., 1964, 97, 510-519.

2 J. Am. Chem. Soc., 1950, 72., 128-132.

b) 3-f 242 -Phenyl eth")ethv amino sul hoflvl ro paoi c acid The product of step a) (1.68 a) was dissolved in methanol (30 mil). Lithium hydroxide (0.45 g) in water (30 ml) was added and the mixture stirred overnight at room temperature. Water was added and the mixture washed with ether. The aqueous layer was acidified with dilute hydrochloric acid and extracted with ether. The ether extract was washed with water then brine, then dried (MgSO 4 The solvent was removed in vacuo to give the subtitle compound as a white solid (0.97 g) which was used without further purification.

mp 80-82°; Mass spectrum: ESI 300 H nmr (360 MHz, CDC13 5: 2.77-2.95 (4H, 3.20-3.47 (4H, 3.51-3.58 (2H, 3.69 (2H. 4.66 (1H, 7.18-7.38 (5H, m).

c) 2-f2-(2-Phenvlethoxv)ethvil-3-isothiazolidinone-1 1 '-dioxide The product of step b) (34 g) was dissolved in dimethylformamide (200 ml). To this is stirred solution was added 1,1'-carbonyldiimidazole (20.12 g) and the mixture stirred for 2 hours. Triethylamine (15.7 ml) was added and the mixture was stirred at room temperature for 60 hours. The mixture was poured into dilute hydrochloric acid and extracted with ethyl acetate The combined organic extracts were washed with saturated sodium bicarbonate solution then brine, dried (MgSO 4 and the solvent 0 removed in vacuo to yieid a pale yellow oil which was further purified by flash chromatography (50% ethyl acetate:petrol as eluant) to give the subtitle compound as an oil (27.4 g).

9 Mass spectrum: ESI 301 (M-NHQ); S 25 H nmr (360 MNE, CDhI) 5: 2.87 (2H, 2.99 (2H, t) 3.51 (2H, 3.64-3.68 (4H, m), 3.76 (2H, 7.18-7.43 (5H, m).

d) 3-[2-(-Hvdroxv-2-oxo-3H-1,3-benzothiazol-7-viethvlaminol-N-f2-(2phenvlethoxv'ethvl] orooanesutlhonamide hvdrochloride 16 The product of step c) (0.62 g) was stirred in toluene (20 ml) then cooled to Diisobutylaluminium hydride (1.6 ml of a 1.5 M solution in toluene) was added over min keeping the temperature below The mixture was stirred for 10 min after which time tic showed that no starting material remained. Ethyl acetate (9 ml) was cautiously added keeping the temperature below -600. The mixture was allowed to warm to room temperature and a 10% aqueous solution of sodium potassium tartrate added.,.After stirring for 1 hour the mixture was extracted with ethyl acetate The combined organic extracts were dried (MgSO 4 and then about 70% of the solvent was removed in vacuo. Methanol (20 ml) was added to the mixture and again about 70% of the solvent was removed in vacuo, this was repeated twice more. This solution was diluted with methanol (20 ml) and 7-(2-aminoethyl)-4-hydroxy-1,3-benzothiazol- 2(3H)-one hydrobromide (0.64 g) added. The pH was adjusted to pH 4 with glacial acetic acid. Sodium cyanoborohydride (0.14 g) was added followed by sodium sulphate mg) and the mixture stirred for 72 hours. The mixture was made basic by the i addition of concentrated aqueous ammonium hydroxide solution. The volatiles were remove in vacuo and the residue purified by flash chromatography (10-25% methanol in chloroform as eluant). The material was further purified by reverse phase HPLC (25% methanol in 0.1% aqueous trifluoroacetic acid as eiuant) to ive. after conversion to the hvdrochloride salt, the title compound as a white solid (0.417 g).

S* mp 205-206°; Mass spectrum: FAB 480 :1H nmr (360 MHz, d 6 DMSO) 5:2.00 (2H, 2.73-2.92 (4H, 2.96-3.19 (8H, m), 3.44 (2H, 3.60 (2H, 6.75 (1H, 6.85 (1H, 7.13-7.35 (6H, 8.92 (2H, s), 25 10.41 (1H, 11.77 (lH, s); Analysis Found: C,51.48; H,6.25; N,8.41; S,12.50% Required for C22H2 9 N305S2.HC: C,51.20; H,5.86; N,8.14; S,12.43% In an alterative method, steps a) and b) above were repeated, followed by steps e) and f) below: 17 e) N-f2-(4-Hvdroxv-2-oxo-3H-1,3-benzothiazol-7-vl)ethvyl-3-f2-(2phenviethoxv'ethviaminosulphonvl]propanamide A solution of the product from step b) (3.89 7 -(2-aminoethyl)-4-hydroxy-1,3benzothiazol-2(3H)-one hydrochloride (3.22 bromotripyrrolidinophosphonium hexafluorophosphate, PyBroP, (6.32 g) in dimethylformamide (50 ml) was cooled to C and diisopropylethylamine (9.0 ml) was added dropwise over 5 minutes. The solution was stirred at -15° C for 5 min then allowed to warm to 130 C over 4 hours.

This mixture was then added dropwise over 40 min to dilute hydrochloric acid (2N, 500 ml) and after stirring over the weekend the solid was collected bv filtration. This solid was dried in vacuo to give the subtitle compound (4.5 g).

Mass spectrum FAB 492 1 f) 3-12-(4-Hvdroxv-2-oxo-3H- 1.3-benzothiazol-7-vl)ethviaminol-N-12-(2phenvlethoxv'ethvilpropanesulphonamide hvdrochloride S* To a solution of the product from step e) (0.28 g) in tetrahydrofuran (2 mi) was added borane tetrahydrofuran (2.44 ml of 1 M solution) over 5 min. The mixture was heated 20 at reflux for 3 hours and, after cooling, methanol (1 mi) was added cautiously. The volatiles were removed in vacuo, the residue was redissolved in methanol (5 ml) and concentrated hydrochloric acid (1 ml) was added. The volatiles were again removed in vacuo. The residue was partitioned between water and ethyl acetate, the acueous laver collected and extracted again with ethyl acetate. The aqueous layer was then made basic with sodium hydrogen carbonate and extracted four times with chloroform. The combined chloroform extracts were dried and the volatiles removed in vacuo to provide after conversion to the hydrochloride salt, the title compound (0.070 g).

Example 2 Example 2 18 N-f 2-[2-(4-Hvdroxv-2-oxo-3 H-I.3 -benzothiazol-7-v Jethvlarninolethvll-242phenviethoxv~ethanesulohonarnide hydrochloride a) 2-(2-Phenviethoxv~ethanesulphonvichloride A stirred suspension of 2-(2-phenylethoxy)ethanethiol (1.0 g) in water (40 rnl) was saturated at 5-10O1 with chlorine over 20 min. The mixture was flushed under a stream of nitrogzen to remove any excess chlorine. The mixture was then extracted with dichlorornethane the combined organic extracts were washed with water then dried (CaCl 2 The soivent was removed in vacuo to give an oil which was azeotroped with toluene to aive the subtitle compound as a yellow oil (1.36 g) which was used without further purification.

Mass speczrum: EI 248/2-50 13 1 nxnr (360 MI-z, CDCI 3 2.90 (2K, 3.74 (21-L 3.88 (2HK 3.99 (21-L, t), 7. 1 3-7.36 (5HK m).

b) iN-(2,2-Dirretho\xve,-hv l)'-(2-phenvlethoxv)ethanesuiphonami de '0 A stirred solution of the product of step a) (1.0 g) in dichloromethane (20 ml) and pyridine (0.3 58 ml) was treated dropwise over 5 nmin with a solution of aaminoacetaldehyde dim ethyl ac etal (0.438 ml) in dichloromethane (5 mlJ). The mixture was stirred at room tem;Derature for 2 days. The mixture was washed with water then dried (CaCl27). The volatiles were removed in vacuo to yield an orange oil which was 25 further purified by flash chromatography (ether as eluant) to give the subtitle compound as a yellow oil (0.42 2).

Mass spectrum: 335 (MN-H.

4 IHnrnr (360 vll-z, CDCI 3 5: 2.90 (2K 3.13 (2H, t) 3.26 (2K 3.38 (6K s), 3.73'(2H, 3.85 (2K1, 4.38 (lH, 4.46 (1K, 7.14-7.35 (5K in).

c) N-2-(2-(4-Hvdroxv-2-oxo-3H-1.3-benzothizol-7-vl)ethvlamino)ethvl-2-f2phenvlethoxv')ethanesulhonamide hydrochloride s A solution of the product of step b) (0.26 g) in 70% aqueous acetic acid (5 ml) was heated to 1000 for 2 hours after which time tic indicated that no starting material remained. The solvent was removed in vacuo. The residue was taken up into methanol ml) and to this stirred solutiorrwas added 7-(2-aminoethyl)-4-hydroxy-l,3benzothiazol-2(3H)-one hydrobromide (0.238 sodium cyanoborohydride (0.038 g) followed by acetic acid (1 drop). The mixture was stirred at room temperature for 24 hours. The mixture was made basic by the addition of concentrated aqueous ammonium hydroxide solution. The volatiles were removed in vacuo and the residual material purified by flash chromatography (17% ethanol in dichloromethane as eluant) to give a light yellow gum. This was further purified by reverse phase HPLC (30-45% is acetonitrile in 0. 1% aqueous trifluoroacetic acid as eluant) to give, after conversion to the hydrochloride salt, the title compound as a white powder (0.080 g).

Mass spectrum: 466 H nmr (360 MHiz, d 6 DMSO) 5: 2.81-2.90 (4H, 3.04-3.09 (4H, brd), 3.26-3.28 20 (2H, 3.24 (2H H20), 3.63 (2H, 3.74 6.78 6.87 7.19- 7.30 (5H, 7.43 (1H, 9.10 10.16 (1H, 11.78 (1H, s); Analysis Found: C,49.35, K5.69; N,8.30; S, 11.94% Required for C2 H2 7

N

3 05S2.HC1.0.5 H 2 0: C,49.31; H,5.67; N,8.22; S,12.52% s. In an alternative method, step a) was repeated, followed by steps d) to g) below: d) Methyl f2-(2-henvethoxvethvlsulphonviamino]acetate A suspension of glycine methylester hydrochloride (2.52 g) in dichloromethane (30 ml) was stirred at -18' C and diisopropylethyl amine (8 mi) was added over 10 min. To this was added the product from part a) (2.64 g) in dichloromethane (10 ml) dropwise over minutes keeping the temperature below C. The cooling bath was removed and the stirred mixture allowed to warm to room temperature. After a further 50 minutes the mixture was washed with 5% aqueous potassium hydrogen sulphate the dried (Na 2 S0 4 and the volatiles removed in vacuo to give the subtitle compound as a brown oil (58.5 g).

Mass spectrum FAB 302 (M+H) 1H nmr (360 MHz, CDCL 3 2.90 (2H, 3.36 (2H, 3.72-3.85 (7H, 3.96 (2H, 4.70 (1H, brs), 7.20-7.31 (5H, m).

e) 2-f(2-Phenvlethoxv)ethvlsulphonvlaminoacetic acid

S..

C 20

CC..

25

C

C

C.

C

.CC.

A solution of the product from part d) (3.2 g) in methanol (30 mi) was cooled in an ice bath and treated with a solution of lithium hydroxide.hydrate (1.06 g) in water (7 ml) over 5 minutes. The cooling bath was removed and the mixture stirred for 16 hours.

The mixture was acidified with concentrated hydrochloric acid (3 ml) and then concentrated in vacuo to approximately 15 ml. The residue was mixed with sodium bicarbonate and extracted with ether. The aqueous phase was then reacidified with satuated aqueous potassium hydrogen sulphate solution and extracted with ether. This extract was dried (MgSO 4 and volatiles removed in vacuo to give a solid. The product was further purified by recrystallisation from dichloroethane-toluene mixture to give the subtitle compound as a crystalline solid (1.57 g).

Mass spectrum FAB 238 IH nmr (360 MHz. CDCL3): 2.90 (2H, 3.32 (2H, 3.7--3.78 (4H, 3.89 (21, 4.66 (1H, 7.20-7.33 (5H, 8.07 (1H, brs).

f) N-r2-(4-Hvdroxv-2-oxo-3H-1,3-benzothiazol-7-vl)ethvll-2-r2-(2phenvlethoxv)ethvlsulhonviaminolacetamide A solution of the product from part e) (70 g) in dimethylformamide (605 mi) was cooled to -10° C and 1,1'-carbonyldiimidazole (39.5 g) was added. The mixture was stirred for 119 minutes then 7-(2-aminoethyl)-4-hydroxy-1,3-benzothiazol-2(3H)-one hydrochloride (68.87 g) was added followed by triethylamine (34 ml) drop wise over 3 minutes keeping the temperature at -10° C. After a further 10 minutes at -10° C the mixture was allowed to warm to room temperature and stirred for 24 hours. This mixture was then added dropwiseTo dilute hydrochloric acid (2N, 1870 ml) and aqueous mixture was extracted with ethyl acetate. The organics were then washed with dilute hydrochloric acid, aqueous sodium bicarbonate and then dried (MgSO 4 The volatiles were removed in vacuo to provide the subtitle compound as a foam (92.21 g).

Mass spectrum FAB 480 H nmr (360 NMHz, d6DMSO): 2.61 (2H, 2.81 (2H, 3.29-3.32 (4H, m 1i overlapping water), 3.56 (2H, 3.61 (2H, 3.74 (2H, 6.70 (IH, 6.80 (1H, d), 7.16-7.53 (5H. 8.03 (IH, brt).

g) N-[f-(2-f2-14-Hvdroxv-2-oxo-3H-1,3-benzothiazol-7-vl)ethvlamino)ethvll-2-(2phenviethoxviethanesuichonamide hvdrochloride 0 A solution of the product from part f) (38.7 g) in tetrahydrofuran (200 ml) was cooled in an aceton'eice bath and lithium borohydride (727 ml, 2.0M in tetrahydrofuran) was added over 30 minutes. After 15 minutes the cooling bath was removed and trimethylsilvi chloride (205 mi) was added and the stirring continued for 162 hours at 2 room temperature. The mixture was cooled to -20° C and methanol carefuliv added.

After this addition a fir-uher 50 ml of methanol saturated with hydrogen chloride was added and the mixture allowed to warm to room temperature. After 1 hour the mixture was heated at reflux for 30 minutes then cooled and the volatiles were removed in vacuo. Water (400 ml) was added and the flask thoroughly shaken and, after standing in an acetone/ice bath for 30 minutes, the water was decanted off. A further amount of water was added (100 ml) and the process repeated. The semi solid remaining was then dissolved in hot ethanol (40 ml), treated with activated charcoal (3 g) and stirred for 1 hour. The mixture was filtered and the solvent removed in vacuo. The residue was redissolved in hot ethanol (80 ml) and allowed to stand at room temperature for 16 hours. The crystals that had precipitated out were then loosened and broken up with a spatula, swirled and left to stand for a further 16 hours. The solid was then collected by filtration and washed with ethanol, ether then dried under vacuum to provide the title compound (23 g).

Example 3 3-r2-(4-Hvdroxv-2-oxo-3H- 1.3-benzothiazol-7-v)ethvlamino1-N-r2-r2-(5-methvi-2thienvl)ethoxvlethvllpropanesulphonamide hvdrochloride Is a) 2 -12-(5-Methvl-2-thienvl)ethoxv]acetic acid Sodium hydride (0.622 g) was suspended in dry dimethylformamide (5 mi) and treated dropwise with a solution of 2-(5-methyl-2-thienyl)ethanol (1.0 g) in dry dimethylformamide (5 ml). The mixture was stirred at room temperature under nitrogen 20 for 2 hours then a solution of choroacetic acid (0.664 g) in dry dimethylformamide ml) added. The mixture was stirred at room temperature overnight. The volatiles were removed in vacuo and the residue quenched with water and extracted with ethyl acetate. The aqueous layer was adjusted to pH 2 using dilute hydrochloric acid and then extracted with ethyl acetate. This extract was washed with water, brine then dried S* 25; (MgSO 4 The solvent was removed in vacuo to give a brown oil (1.51 g) which was used without further purification.

Mass spectrum: 200 H nmr (360 MHz, CDCI 3 5: 2.45 (3H, 3.05-3.09 (2H, 3.75-3.83 (2H, 4.17 (2H, 6.56 (1H, 6.63 (1H, d).

b) 2-r2-(5-Methvl-2-thienvl)ethoxv]acetamide The product of step a) (6.48 g) was dissolved in toluene (55 ml) and oxalyl chloride s (2.877 ml) added dropwise at room temperature under nitrogen. A drop of dimethylformamide was added and the mixture stirred for 3 hours after which time tic showed no starting material. The volatiles were removed in vacuo to provide a brown oil (6.7 g) which was added dropwtse to a stirred solution of concentrated ammonium hydroxide (50 ml) at The mixture was allowed to warm to room temperature and stirred for 4 hours. A brown solid precipitated out. This solid was collected by filtration and washed with water to give the subtitle compound (2.02 g).

Mass spectrum: 200 H nmr (360 MHz, CDC13) 5: 2.45 (3H, 3.04 (2H, 3.73 (2H, 3.97 (2H, s), 5.61 (2H, brs), 6.57 (1H, 6.63 (1H, d).

c) 2-[2-(5-Methyl-2-thien.vi)ethoxvlethanamine

S.

Borane-tetrahydrofuran solution (1.0 M in THF, 21.7 ml) was added dropwise to a 20 stirred solution of the product from step b) (1.25 g) in dry tetrahydrofuran (100 ml).

The reaction was heated at reflux under an inert atmosphere for 5 hours. The reaction *was cooled and methanol (10 ml) added cautiously. The solvents were removed in *5 vacuo and the residue dissolved in methanol (100 ml) to which was added concentrated hydrochloric acid (sg. 1.18. 0.45 ml). This solution was heated at reflux for 15 min then 25 the solvent removed in vacuo. The residue was purified by flash chromatoeraphy (dichloromehane: 5% methanol as eluant) to give the subtitle compound as a white solid (0.916 g).

Mass spectrum: 186 IHnmr (360 MI~z, CDCI 3 5: 2.45 3.04 (2H, 3.73 (2K 3.97 (2K s), 5.61 (2K brs), 6.57 (1H, 6.63 (1H, d).

d) Methyl 3 -r2-r2-(5 -methvl-2-thienvflethoxvlethvaminosulphonvlphroTanoate The product of step c) as the hydrochloride salt (1.50 g) was stirred under nitrogen in dichloromethane (25 ml). Triethylarnine (2.21 m.1) was added followed by methyl 3- (chlorosulphonyl)propanoate 1 8g). The mixture was stirred at room temp~erature overnight. The solution was diluted with a further amount of dichiloromethane and the organics washed with dilute hydrochloric acid then water, then dried (MgSO 4 The mixture was filtered and the *'olatiles removed in vacuo to give the subtitle compound as an oil (1.4 g).

Mass spectrum: 336 13 H nrnr (360 MHz, CDCL 3 6: 2.38 (3F1, 2.80 (21-L 3.00 (2K 3.26,3.36 (4K in), 3.56 (2K 3.64 (2K 3.72 (3K 4.71 (1K 6.57 (1K, 6.60 (1KL d).

e) 3-{2-(4-Hvdro-xv-2-oxo-3H- 1 .3benzothiazcl-7-vl~ethviaminol-N-r2-42-(5-rnethvl-2'thienvlethc-xvlethv]Ilprooanesulphonamiude hydrochloride :The product of ste:pi) (0.60 g) was dissolved in toluene (30 ml) under nitrogen and cooled to Diisobutylalumriniumhydride (1.5 M in toluene, 1.78 ml) was added dropwise and the mixture kept at -78' for 10 mlm. The reaction was quenched with ethyl acetate followed by a 10% aqueous sodium potassium tartrate solution. The mixture was warmed to room temperature and after stirring for 1 hour e.xtracted with toluene The combined organic extracts were washed with water, dried (MgSO 4 ~:.then about 70% of the solvent removed in vacuo. Methanol (20 ml) was added to the *mixture and azain about 70% of the solvent removed in vacuo, this was repeated twice more. This solution was diluted with methanol (20 ml) and 7-(2 -amino ethyl)- 4 hydroxv-1,3-benzothiazol-2-(3H)-one hydrochloride (0.5 16 iz) added. The pH1 was adjusted to pH 4 with glacial acetic acid. Sodium cyanoborohydride 114 g) was added and the mixture stirred under nitrogzen for 2 hours. The mixtre was made basic by the addition of concentrated aqueous ammonium hydroxide solution. The volatiles were removed in vacuo and the residue purified by flash chromatography (5-20% s methanol: dichloromethane as eluant). The material was further purified by reverse phase H-PLC (25-85% methanol in 0. 1 aqueous trifluoroacetic acid as eluant) to give, after conversion to the hydrochloride salt, the title compound 12 g) as a white solid.

mp210-2 12'; Mass specrn'm- FAB 500 1Hnmr (360 MI-z, d 6 DMSO) 5: 2.00 (2K q, 2.37 (3K, 2.85 (2H, 2.93 (2Kt), 1-3. 17 (8-L in), 3.46 (2K 3.58 (2H, 6.60 (1KL 6.65 (IH, 6.75-6.88 7.31 (lIi 8.93 (2K 10. 15 (1K, 11.7 (1K brs).

Example 4 -r 2 -(4IFlu oroc henvi) et hoxv] ethyllJ- 3- r2-(4-hvdroXV-2-Oxo-3 H- 3 -b enzot hiazo 1-7v)ethvlarninolpropanesulchonainide hydrochloride a) t-Butvl 3-r2-(.-fluoro 'nenvflet-hoxvltvropanoate 2-(4-Fluorophenvl)ethanol (20.5 a) and Triton-B (2.4 ml, 40% inmethanol) were mixed together and the methanol removed in vacuc. t-Butyl acrvlate (21.38 ml) was ~added and the solution heated at 500 for 2 hours then stirred room tern erature over'aht. The mixture was diluted With water and extracted with diethvi ether. The S.....combined orzanic ex tracts were washed with brine, dried (Mg6O4) and filtered. The volatiles were removed in vacuo to give the subtitle compound (37.91 g) which was used without further purification.

1H nmr (360 MHz, CDCl 3 6: 1.46 (9H, 2.47 (2H, 2.84 (2H, 3.55-3.69 (4H, 6.93-6.98-(2H, 7.15-7.18 (2H, m).

b) 3-[2-(4-FluoroDhenvlnethoxv]propanoic acid The product of step a) (37.91 g) was dissolved in dichloromethane (50 ml) and trifluoroacetic acid (50 ml) added. The solution was stirred at room temperature for 1 hour then diluted with water and extracted with ethyl acetate The combined organic extracts were washed with water (x4) then brine, dried (MgS04) and filtered.

t0 The volatiles were removed in vacuo to give an oil, this oil was taken into diethyl ether and extracted with aqueous sodium bicarbonate solution The combined aqueous layers were acidified with concentrated hydrochloric acid then extracted with diethyl ether The combined organic extracts were washed with brine, dried (MgSO4) and filtered. The volatiles were removed in vacuo to give the subtitle compound (19.95 g) i3 which was used without further purification.

Mass spectrum: 212 H nmr (360 NMIz, CDCI 3 5: 2.62 (2H, 2.85 (2H, t) 3.66 (2H, 3.73 (2H, t), 6.94-6.99 (2H. 7.14-7.18 (2H, 7.37 (1H, brs).

c) 2-[2-(4-Fluorophenviethoxv1lethvlisocvanate The product of step b) (15.49 triethylamine (11.2 mi) and diphenyiphosphoryl aide (15.7 ml) were heated in toluene (150 ml) at 80' for 5 hours under nitrogen. The mixture was allowed to cool, then left at room temperature overnight. The mixture was diluted with water and extracted with ethyl acetate The combined organic extracts were washed with aaueous sodium bicarbonate, dilute hydrochloric acid, brine then dried (MgSO 4 and filtered. The volatiles were removed in vacuo and the residue further purified by flash chromatography (50% diethyl ether:iso-hexane as eluant) to give the subtitle compound (5.54 g).

Mass spectrum: 209 H nmr (360 MHz, CDC13) 5: 2.89 (2H, 3.38 (2H, 3.56 (2H, 3.68 (2H, t), 6.95-7.01 (2H, 7.17-7.21 (2H, m).

d) Methyl N-[2-(2-(4-fluorophenvi)ethoxv)ethvl carbamate The product of step c) (9.0 g) was dissolved in methanol (300 ml) and to this stirred solution was added sodium methoxide (4.65 The mixture was stirred at room temperature for 3 hours. The volatiles were removed in vacuo and the residue was extracted with ethyl acetate. The ethyl acetate was washed with brine, dried (MgSO 4 and filtered. The volatiles were removed in vacuo and a portion of the residue was further purified by flash chromatography (60% diethyl ether: iso-hexane as eluant) to give the subtitle compound (0.375 g).

Mass spectrum; FAB 242 H nmr (360 MHz, CDC 3 5: 2.84 (2H, 3.34 (2H, 3.49 (2H, 3.63 (2H, t), 3.67 (3H, 6.95-7.00 (2H, 7.14-7.18 (2H, m); Analysis Found C,59. 1 H,6.S3, N,5.83% :0 Required for C 1 zH 6 FNO3: C,59.74; H,6.68; N,5.81%.

e) 2-2-(4-Fluorophenvhethoxvlethanamine The product of step d) (8.0 g) was dissolved in ethylene glycol and to this was added 25 potassium hydroxide (43 g) and hydrazine hydrate (8.3 ml). The stirred mixture was heated to 140° for 4 hours then allowed to cool to room temperature overnight. The mixture was then diluted with water and extracted with diethyl ether The combined organic extracts were washed with brine, dried (MgOS4) and filtered. The volatiles were removed in vacuo to give the subtitle compound (5.36 g).

28 Mass spectrum: 184 (M-fH), IH rmr (360 Nfv-iz, CDC1 3 5: 2.82-2.88 (4H, in), 3.47 (21L 3.64 (2K-1 6.95-6.99 (2H, in), 7.16-7.20 (2H, in).

s f Methyl 3- r2-[r2-4-fluorop henvl)ethoxv] ethvl aminosulphonvilpropano ate The subtitle compound (8.32 g) was prepared according to the method of Example 3d) using 2-[2-(4-fluorophenvl)ethoxylethanainne (5.36 triethylamine (4.6 ml) and methyl 3-(chlorosulphonvl)propanoate (5.6 g) in diethyl ether (150 ml).

mp 490;- Mass spectrum: 334 IHnmr (360 MI-Iz, CDCI 3 2.78-2.87 (4K in), 3.24-3.28 (2H, 3.34 (2K t), 3.51-3.56 (121- mn), 3.65 (2H, 3.73 (3K, 6.97-7.01 (2K in), 7.15-7.19 (2KL m); Analysis Found: C,50.52, K6.24; N,4.16; S,9.40% Required for C 1 4 HNPFN0 5 S. C,50.44; K,6.05; N,4.20; S,9.62%.

Q) iN-f2-r2-(-FlTuoroohen,\'I)etho-<vethvil-3-r2-(4-hvdroxv-2--oxo-3H- I 3 -benzothiazol-7- *viethvlam-Lhorooanesuiohonamj de hydrochloride The title compound (0.092 g) was prepared according to the method of Example 3e) using methyl 3-[2 7-[2-(-fluorophenvl)ethoxy]ethylarrinosulrhonvl]propanoate (1.0 g), diisobutvlaiunulniumnhydride (1.5 M in toluene, 4 ml), 7-(2-aininoethvi)-4-hydroxy-1,3benzothiazoi-2-(3H)-one hydrochloride (0.875 g) and sodium cyanoborohydride (O.354 g).

mrp 195-7; Mass spectrum: FAB 498 Inmr (360 MfHz, d 6 DMSO) 56:1.99 (2K 2.78-2.87 (41-L m) 3.04-3.16 (8HKr), 3.45 (2K, 3.59 (2H, 6.86 (1K 6.85 (1H, 7.01 (2K 7.26-7.3 1(3,n, 8.90 (2H, 10. 15 (1 K 11. 77 brs); Analysis Found: C,49.01 I K,-5.74; N, 7.96; S, 11. for C2-,2FN 3 0 5 S2.HCL.0.5 Hi0: C,49.48; K5.47; N,7.87; S;12.01%.

Examole N-r2-f2-(4-Chlorochenviehoxvlethvll-3-r2-(4-hvdroxv-2-oxo-3)H-1, 3-benzothiazol-7vl)ethvlaminoloropanesulphonamide hydrochloride.

a) 2- r2 -(4-Chlororhenv1)ethoxv1 acetic acid 2-(4-Chlorochenvl)ethanol (10 g) was stir-red in 50 aqueous sodium hydroxide ml), tetrabunrlammnoniumbromiude (1.4 g) was added and the mixture stirred for 1 hour.

t-Butvibromoacetate (2S.6 ml) in toluene (140 m.1) was added and stirring continued for 18 hours. Water- (50 mli) was added and after 2 hours the mixture was coojed in ice and 0: acidified to pH 1 with concentrated hydrochloric acid. The organic layer was separated and the aq ueous extracted with ethyl acetate. The combined organic extracts were washed with brine and dried (MSO 4 The solvent was removed in vacuo to ive a pale yellow oil This was dissolved in dichoromethane (100 ml) and trifluoroacetic acid (100 ml) added, the mixture was heated at reflux for I hour. The volatiles were removed in vacuo and the residue taken up in sodium hydroxi'de and washed With ethyl acetate. The aqueous layer was then acidified with concentrated hydrochloric acid and extracted wih ethvl acetate, this ethyl acetate was washed with water, dried 9S 04) and the volatiles removed in vacuo to provide the subtitle comound as a buff solid (16.4 g) which was used without fuirther purification.

Mass spectrum El 214/6 b) 2-r2-(4-Chlorophenvl)ethoxvlacetamide The product from part a) (16.4 g) was dissolved in toluene (300 ml) and oxalyl chloride (13 ml) was added dropwise at room temperature and under nitrogen. The mixture was stirred for 1 hour and then dimethylformamide (0.3 ml) was added. After a further 2 hours the volatiles were removed in vacuo to provide a brown oil which was added dropwise to a stirred solution of concentrated ammonium hydroxide (60 ml). The solid which precipitated out was collected by filtration and washed with water and isohexane to yield the subtitle compound (6.6 g).

Mp. 106-9° C, Mass spectrum FAB 214/6 is H nmr (360 MHz, CDCl 3 2.39 (2H, 3.73 (2H, 3.93 (2Hs), 5.87 (1H, brs), 6.22 (1H, brs), 7.16 (2H 7.28 (2H,d).

c) 2-r2-(4-ChlorophenvlYethoxy'Iethanamine The product from step b (6 g) was added portion wise to a stirred solution of boranetetrahvdrofuran (1.0 M in THF, 85 ml). The reaction was then heated at reflux under an inert atmosrhere for 3 hours. The reaction was cooled and methanol (10 mis) was cautiousiv added. The solvents were removed in vacuo and the residue redissolved in methanol (100 ml) to which was added concentrated hydrochloric acid (sg. 1.18, 4 ml). This solution was heated at reflux for 30 minutes and then the solvent removed in vacuo. The residue was taken up into water and washed with ether. Sodium bicarbonate was added and the aqueous extracted with ethyl acetate. The ethyl acetate was washed with water, brine and dried (MgS04). The volatiles were removed in vacuo to provide the subtitle compound as a oil (5.5 g) which was used without further purification.

31 Mass spectrum El 200/2 d) Methyl 3-[2-r2-(4-Chlorophenvl)ethoxvlethvlaminosulphonvl]propanoate The product from part c (1.1 g) was stirred under nitrogen in dichloromethane (30 mi), triethylamine (1.52 mi) was added followed by methyl 3-(chlorosulphonyi)propanoate (2.04 The mixture was stirred a room temperature for 3 hours. The volatiles were removed in vacuo and the residue purified by flash chromatography over silica ether/petrol) to provide the subtitle compound as an oil (1.1 g).

Mass spectrum FAB 350/2 H nmr (360 ,MHz, CDCI 3 2.80 (2H, 2.82 (2H, 3.26 (2H, 3.33 (2H, 3.54 (2H, 3.67 (2H, 3.72 (3H, 4.63 (1H, 7.14 (2H, 7.27 (211 d).

e) N-r2-r2-(4-Chlorophenvi)ethoxv1ethvll-3-2-(4-hvdroxv-2-oxo- 3 H-1I3-benzothiazol-7vl)ethvlaminoplropanesulphonamide hvdrochloride.

The product from part d (1.1 g) was dissolved in toluene (50 ml) under nitrogen and 20 cooled to -78°C. Diisobutylaluminiumhydride (1.5 M in toluene, 2.6 ml) was added dropwise and the mixture kept at -78 0 C for 10 minutes. The reaction was quenched with 10% hydrochloric acid in methanol and the mixture allowed to warm to room temperature. The mixture was poured into 10% hydrochloric acid and extracted with ether. The combined organic extracts were washed with water then dried over magnesium sulphate and then about 70% of the solvent was removed in vacuo.

Methanol (20 ml) was added to the mixture and again the about 70% of the solvent was removed in vacuo, this was repeated twice more. The solution was then diluted with methanol (20 mi) and 7-(2-aminoethyl)-4-hydroxy-1,3-benzothiazol-2(3H)-one hydrochloride (0.92 g) added. The pH was adjusted to pH 4 with glacial acetic acid.

Sodiumcyanoborohydride (0.247 g) was added and the mixture stirred under nitrogen for 2 hours. The mixture was made basic by the addition of concentrated aqueous ammoniumn hydroxide solution. The volatiles were removed in vacuo and the residue purified by flash chromatography (2.5-10% methanolldichloromethane). The material was further purified by reverse phase HPLC using 50% to 100% methanol in 0. 1% aqueous trifluoroaceric acid as eluant to yield, after conversion to the. hydrochloride salt, the title compound (0.089 g) as a white solid.

Mp. 190-3'C; Mass spectrum: FAB 5 14/6 1Hnmr (360 MIHz., d 6 DMSO) 1. 99 (2K in), 2.82 (4K, in), 3 12 (8K rn), 345 (2K t), 3.60 (2K, 6.76 (1KL 6.87 (11-1 7.30 (5K rn), 8.87 (21-L brs), 10.14 (1K s), 11.77 b rs); Analysis Found: C, 46.68; K 5.55; N, 7.47; S, 10.86%. Required for C7 H 28

N-O,

5 S-Y.HCI. H20O; C,46-47; H, 5.50; N, 7.39; S, 11.28%.

Examole 6 3-f 2-(4-Hvdroxv-2-oxo-3H- 1.3-benzothiazol-7-vl)ethvlarninol-N-[2-r2-(4methvichenvflethoxvlethvl larap anesul phonam-ide hydrochloride.

a) 2-f 2-(4-M lvethvlp~henvlehoxvacetic acid 23 2 ethyl ph enyi) ethanol1 (8.16 g) was stirred in 50% aqueous sodium hydroxide a. ml).t-Butyibromnoacetate (9.05 ml) in toluene (30 mnl) was added together with tetrabutvlamnmoniuibrornide (2.2 g) and the mixture stirred for 18 hours. Iced water (100 ml) was added followed by ether, the organic layer was separated and then the aqueous layer acidified wi*th hydrochloric acid. The acidified aqueous was extracted with ether and this extract was washed with brine, dried (M.-SO 4 The solvent was removed in vacua to give the subtitle compound as a white solid 1 g) which was used without further purification.

nmH r (360 Nvfl1z, CDCI 3 2.3 2 (3 H, 2.91 (2K 3.77 (2K 4 10 (21-L S), 7.11 (4K, in).

b) 2-f 2-(4-MethviTnhenvflethoxvlacetamide The subtitle compound (0.77 a) was prepared according to the procedure in example 1 part b using 2-[2-(4-merhylphenvl) ethoxy] acetic acid (1.1I oxalyl chloride (1.44g) concentrated ammonium hvdroxide (20 ml), and toluene (40 nil).

mp 106-7 0

C;-

Mass spectrum FAB 194 100).

c) 2 -f2-( 4 -MehvlpherwflIetho\-vlethanarnjne hydrochoride subtitle compound (6.25 z) was prepared according to the proce-dure in example part c using 2-[2-(4-inehvlp henivl)ethoxyjaceaxnid (5.79 gY), borane-tetrhvdrofuran solution (1.0 M in THF, 75 ml2), and tetrahydrofliran (40 ml).

Mass spectrum FAB 1S0 (MI-H, 100); H (i (360 Ii7- CDCl1 3 )1 2.3 0(3 K 2.8 7(2K 317 (2KL 3. 6 -3 73 (4K in), 7.11 (4K in), 8.30 (2KL brs).

Methyl 3 -r,--r2-(4-methvlothe-i,,ietho-<vlethviarinosulohonvilur-onoiat-, The subtitle compound (2.49 g) was prepared according to the procedure in example part d using 2--[2-(4-me,.hylphenyl)ethoxy] ethanamine hydrochloride (3.5 g) triethylamnine 4.74 ml) methyl 3 -(chlo rosulphonyl) pro panoate (3.08 and dichloromethane (80 ml).

Mass spectrum FAB 330 (M4-H 119 (100).

e) 3 -[2-(4-hvdroxv-2-oxo-3 H- .3 -benzothiazol-7-vl)ethvl aminol-N-[2-r2-(4methvl phenvi)ethox-v1 ethvll pro panesul phonarmide hydrochloride.

The title compound (0.2 g) was prepared according to the procedure in example 5 part e using methyl 3-[2 [2 -(4-methylphenyl)ethoxy] ethylamninosulpho nyi]prop anoate (2.49 diisobutvlaluminiumhvdride (1.5 M in toluene, 7.5 ml), 7-(2-aminoethyl)-4-hydroxy- 1,3-benzothiazol-2(3H)-one hydrochloride (2.2 g) and sodium cyanoborohydride (0.48 g).

m.p. 213-5'C, Mass spectrum FAR 494 (M-t-H; nmr (360 MIHiz., d 6 DMSO) 1.99 (2K in), 2.25 (3K 2.74-2.84 (4H-L 3.06- 3.15 (SH. mn), 3.45 (211, 3.57 (2K, 6.75 (lII, 6.87 (Ili, 7.10 (4HL 7.30 ~:.Example 7 3 -r 2 -(4-Hvd ro xv-2 -oxo 3H- 1, .3-b enzo thi azo I- 7-vi) et hviamino 22 -0h envl Ipro pov) ethvl p rot)anesuir'nonaid e hydrochloride.

a) 2-(2-Phenvi-1I-propoxv~acetic acid The subtitle compound (14.95 a) was prepared according to the procedure in example 6 part a using 2-phenyi-1-propanol (13.6 t-butylbroinoacetate (19.5 g), tetrabutvlammrroniurnbromride (3.2 toluene (70 nil), 50% aqueous sodium hydroxide ml).

I H nmr (360 M'11z, CDCI 3 1.32 (3HK 3.08 (1H, in), 3.67 (21-L t) 4.07 (2K S), 7.23-7.34 (5K in).

b) 2-(2-Phenvi- I-propoxv~acetamide The subtitle compound (4.39 g) was prepared according to the procedur-e in example part b using henvl-lI-propoxy)acetic acid 0 oxaMy chloride 4. 5 ml), concentrated ammonium hydroxide (20 ml), and toluene (30 ml).

Mass spectrum GC 134 (M-59); 1Hnmr (360 MVI-z, CDCI 3 1.3 1 (3K, 3.05 (111 in), 3.61 (2K in), 3.90 (2HK q), ~7.22 (Hin,7.33 (2H, in).

c) (2--Phenvl- -rop)oxv' ,ethanan'ne hydrochloride .0 The subtitle compound (3.58 g) was prepared according to the procedure in example *part c using henvl-1I-pro poxy)acetamide (3.86 borane-tetrahvdrofuran solution (1.0 M in 50 ml), and tetrahydrofuiran (40 ml).

Mass spectrum GC 1S0 Methyl 3 f-(2 -phenvi I proo cxv) ethyl arninosu I phonyll pro panoate The subtitle compound (2.0 gY) was prepared according to the procedure in example part d using >-7(2-7phenyilI-propoxy)ethanam~ne hydrochloride (3.58 triethylamine (5.65 mi) methyl (3-chlorosulphonyl)propanoate (3.73 and dichloromethane ml).

Mass spectruIm GC 224 (M-105); 1 H nmr (360 rvlIz, CDCI 3 1.27 2.75 (2K 3.02 (1K in), 3.20-3.29 (4K mn), 3.45-3.56 (41-L mn), 3.72 4.42 (IIHLt), 7.21-7.34 (5K, in).

e) 3-f 2-(4-Hvdroxv-2-oxo-3 H-i .3 beiizothiazol-7-vP~ethviaminol-,N-[2-(2-ohenvl- propoxv) ethyll prop anesulp honainide hydrochloride.

The title compound 164 g) was prepared according to the procedure in example part e using methyl 3-[2 -phenl- 1 -prop oxv)ethylamino sulphonyl] propanoate (1.01 diisobutvlaluminiumnhvdride (1.5 M in toluene, 3 7-(2-anoethvl)-4-hydroxy- 1,3-benzothiazol-2(3)-one hydrochloride (0.89 g) and sodium cyanoborohydride 1s 19 g).

m.p. 1833-A 0

C,

Mass spectrum FAB 494 (M-vi-H); Analysis Found:- C, 52.42; 6.3 1; N, 8.28; S, 11.74%. Required for

C-)

3

H

3 jN-,O5S-.HCl; C, 52.11;1 H, 6.08; N, 7.93; S, 12.10%.

Examnle 8 :s -r2-(4-Hvdroc,,--2-oxo-3H- I .3-belzothiazol-7-vflethviamninol--N-r2-r2-(2- .99 methvlnhenvl~etho-xvlethvll~rcoanesulphonarnide hdoho~e a) 2-r2-(2-Methvlphenvl)etho-xvIacetic acid The subtitle compound (8.91 g) was prepared according to the procedure in example parn a using 2-(2-methylphenyl)ethanol (5.0 t-butylbromo acetate (5.47 mi), tetrabutylammoniumbromide (0.78 toluene (80 ml), 50% aqueous sodium hydroxide ml), trifluoroacetic acid (20 ml) and dichloromethane (20 ml).

1 H nrnr (360 MU-z, CDC 3 2.34 (3K 2.97 (2K 3.75 (2K 4. 1' (2K 7. 12- 7.17 (4K m) 8. 14 (1iiK brs).

b) 2-f 2-4'2-Methviphenvi)ethoxvlacetamide The subtitle compound (5.02 g) was prepared according to the procedure in part b using 2-[2 -(2-mer.hvlphenvl)ethoxy] acetic acid (6.726 oxalyl chloride (6.22 ml), concentrated armonium hydroxide (60 ml1), and toluene (60 ml).

Mass spectrum FAB 194 (GA-H, IHnmr (360 MN-z, CDCl 3 Z2 2 2.86 (2K 3.60 (2K 3.30 (2K S), 7.06-7.19 (4K-1 in), 7.25 (2K brs).

2o c) 2-f 2-(2-Methviphenvethox<v~ehanamine hydrochloride.

The subtitle compound (5.4 g) was prepared according to the procedure in example part c using 2-[2-(2-methyiphenvl)ethoxyjacetamiide 13 borane-tetavdrofturan solution (1.0 M in TI-F, 53.5 mi), and tetrahvdroftiran (100 ml).

Mass spectrum F.AB 180 (M Methyl 3-f 2-[2-(2-ineth-vlIhenvF!)ethoxvlethvlam-i n osulphonvil prop anoat g The subtitle compound (5.4 g) was prepared according to the procedure in example pan c using 2-[2-(2-methylphenyl)ethoxy]ethanamine hydrochloride (5.4 g) triethylamine (8.73 ml) methyl 3-(chlorosulphonyl)propanoate (4.66 and dichloromethane (100 ml).

Mass spectrum FAB 330 H nmr (360 MHz, CDC1 3 2.32 3H, 2.71 (2H, 2.91 (2H, 3.24 (2H, 3.34 (2H, 3.53 (2H, 3.62 (2H, 3.70 (3H, 4.64 (1H, 7.14 (4H, m).

e) 3-f2-(4-Hvdroxv-2-oxo-3H-1,3-benzothiazol-7-vl)ethvlaminol-N-r2-r2-(2methvlphenvl)ethoxvl ethvtloropanesulphonamide hvdrochloride.

The product from part d (2.0 g) was dissolved in toluene (100 ml) under nitrogen and cooled to -78 0 C. Diisobutylaluminiumhydride (1.5 M in toluene, 6.22 ml) was added 13 dropwise and the mixture kept at -78 0 C for 10 minutes. The reaction was quenched with ethyl acetate followed by a 10% aqueous sodium potassium tartrate solution. The mixture was warmed to room temperature and after stirring for one hour the mixture was extracted with toluene. The combined organic extracts were washed with water then dried over maenesium sulohate and then about 70% of the solvent was removed in 20 vacuo Methanol (50 ml) was added to the mixture and again about 70% of the solvent was removed in vacuo, this was repeated twice more. This solution was then diluted with methanol (50 ml) and 7-(2-aminoethyl)-4-hydroxy-l,3-benzothiazol-2(3H)-one hydrochloride (1.8 g) added. The pH was adjusted to pH 4 with glacial acetic acid.

Sodium cyanoborohydride (0.398 g) was added and the mixture was stirred under :5 nitrogen for 18 hours. The mixture was made basic by the addition of concentrated aqueous ammonium hydroxide solution and the volatiles were then removed in vacuo :i The residue was purified by flash chromatography over silica (1% methanol/dichloromethane). The material was further purified by reverse phase HPLC using 35%-85% methanol in 0.1% aqueous trifluoroacetic acid as eluant to yield, after conversion to the hydrochloride salt, the title compound (0.38 g) as a white solid.

m.p. 184-7 0

C,

Mass spectruim FAB 494 nm= (360 MI-z., d 6 DMSO) 1.74-1.77 (2K in), 1.99 (2K 2.27 (3'K 2.79-2.85 (4K 3.07-3. 16 (6H, in), 3.45-3.48 (2K in), 3.55-3.61 (2K, mn), 6.74 (1K 6.88 (1K 7.08-7.17 (4K 7.32 (1K 8.81 (1K 10.17 (1K, 11.77 (1K, brs).

Exainvle 9 3 -[2-(4-Hv-drox-v-2--oxo-3 H-I .3-benzothiazol-7-vl)-ethvlaminol-N'-[2-(2phenvlthioethoxv)ethvlThroT~anesulThonarnide hydrochloride.

a) t-.Butvl- 2-(2 -p henlthicet hoxv) acetate 2-Phenvithic ethanol 1 t-butvlbromoacetate (7.9 mld), tetrabutvlammioniumbromide (1.3 toluene (80 ml), and 75% aqueous sodium hydroxide (40 ml1) were stirred together for 72 hours. The organic layer was separated .0 and the aqueous laver extr-acted with dichio ro methane. The combined organics were **washed w'ith brine, dried (IMgS04) and the solvent was removed in vacuo to provide the subtitle compound (12.07 g).

Mass spectrm FAB 2-69 b) 2-(2-Phenvlthjoethoxvlacetainide The product from part a (12.07 g) was dissolved in dichoroinethane (50 ml) and trifluoroacetic acid (50 ml) was added, the mixture was stirred for 2 hours. The volatiles were removed in vacuo and the residue taken up in aqueous sodium bicarbonate and washed with ether. The aqueous layer was then acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The ethyl acetate was washed with brine, dried (MgS04) and the volatiles removed in vacuo. The residue was dissolved in toluene (50 ml) and oxalyl chloride (50 ml) was added dropwise at room temperature and under nitrogen. Dimethylformamide (0.3 ml) was added dropwise and the mixture was stirred for 45 minutes. Volatiles were removed and the crude acid chloride was added dropwise to a stirred solution of concentrated ammonium hydroxide (50 ml) at -10 C. The solid which precipitated out was collected by filtration and washed with water and ether to yield the subtitle compound (3.53 g).

Mass spectrum FAB 212 H nmr (360 MHz, CDC13); 3.13 (2H, 3.72 (2H, 3.95 (2H, 5.58 (1H, brs), 6.56 (1H, brs), 7.20-7.39 (5H, m).

1, c) 2-(2-Phenvithioethoxv)ethanamine hvdrochloride.

The subtitle compound as its hvdrochloride salt (6.0 g) was prepared according to the procedure in example 5 part c using 2-(2-phenylthioethoxy)acetamide (5.5 g), 20 boranetetrahydrofuran solution (1.0 M in THF, 60 ml), and tetrahydrofuran (60 ml).

d) Methyl 3-f2-(2-phenvlthioethoxv)ethviaminosulohonvi]procanoate The subtitle compound (2.85 g) was prepared according to the procedure in example 25 part d using 2-(2-phenyithioethoxy)ethanamine hvdrochloride (6 triethyiamine (4 ml) methyl 3-(chlorosulphonyl)propanoate (5.8 and dichloromethane (50 mi).

Mass spectrum FAB 348 (M-105).

41 e) 3-f 2-(4-Hvdroxv-2-oxo-3 H-I.3 -benzothiazoI-7-v~ethvlamino1-N-r2-(2phenvlthioethoxv~ethvloropanesuIphonamide hydrochloride.

The title compound 106 g) was prepared according to the procedure in example 8 s part e using methyl 3 -phenylthi oetho xy) ethylamino sul phonyl] prop ano ate (1.4 g), diisobutylaluminiumnhydride (1.5 M in toluene, 3 mnl), 7-(2-arminoethyl)-4-hydroxv-1,benzothiazol-2(3'H)-ore hydrochloride (1.2 g) and sodium cyanoborohydride (0.3 a).

rn.p. 193-4'C1- Mass spectrum FAB 512 nrHn= 360 MfHz., d 6 DMSO) 2.06 (2,L in), 2.85 (2K in), 3.08 (6-L in), 3.15 (4K in), 3.46 (2K 3.59 (2K 6.76 (1K 7.19 (1K 7.3 1-7.36 (4K in), 9.06 (2K s), 17 11. 78 (1KH, br).

Pharmacolomical Example The bindina affinities of comp.Iounds of the above Examples for the DA 2 receptor bindig sites in bovine pituitary membranes were determnined from the displacement of VKil-N-n -propyinorapomorphine and of[ 3 K1-spiperone in the absence or presence of nonhydrolysable GTP anaiczue respectively, D.R.Sibley, A. DeLean and I. Creese, Anterior Pituitary Dopamine Receptors, Demonstration of Interconvertible High and Low Affinity States of the D-2 Doparimine Receptor, J. Biol. Chein., 1982, 257(11), 6351-6361.

The DAi-receptor activity ,va also demonstrated in a fu~nctional screen, the rabbit isolated ear artery, as describ.ed by Brown and O'Connor, Br. JI Pharmacol., 1981, 73, 189P.

f3 2 -adreno receptor activity was demonstrated in the isolated trachea of the guinea pig, as described by I.G. Dougall, D. Harper, D.M. Jackson, and P. Leff, Br. J. Pharmacol., 1991, 104, 1057.

ca-Receptor activity was determined in the rabbit isolated ear artery using the following method: Rabbit isolated ear artery Male NZW rabbits (2.5-3.0 kg) were killed by intravenous injection of pentobarbitone sodium (60 mg/kg). The ears were removed and the proximal portion of the middle ear artery exposed and cannulated using a polypropylene cannula (0.75mm external diameter).

After removal, the artery was cleared of adherent connective tissue and 6 rings, 5mm wide, were prepared preserving the plane of the circular smooth muscle. Tissues were mounted on fine tungsten wire hooks (0.25mm diameter) in 20 ml organ baths containing Krebs solution of the following composition (mM) NaCI 117.56; NaHCO 3 25.00; KCI 5.36; NaH 2

PO

4 0.89; MgSO 4 1.18; glucose 11.10 and CaCI 2 2.55. Cocaine (30uM) and propanolol (1LM) were included in the Krebs solution to block neuronal uptake and Preceptors respectively. Ascorbate (100uM) was also added to prevent catecholamine oxidation. This solution was maintained at 37°C and continuously gassed with 95% Oz0 20 5% CO 2 The upper wire hook was attached to an Ormed force displacement transducer, the lower hook being attached to a stationary support in the bath. Changes in isometric force were recorded on Advance Bryans AB500 flat-bed recorders.

Experimental General At the beginning of each experiment, a force of 1.0g was applied to each tissue. This force was reinstated two or three times during a stabilisation period of approximately 60 min.

until it remained constant. At the same time as the force was reinstated the baths were washed out. Agonist concentration-effect, curves were constructed by cumulative additions ofagonist at 0.5 logio increments. Responses (contractions) were recorded as a percentage of the maximum response of the standard agonist.

Quantification of agonism Phenylephrine has been adopted as the standard agonist. An curve to phenylephrine was constructed first. The phenylephrine was then washed out and an curve to the test compound was constructed. Responses of compounds that produced agonism were expressed as a percentage of the maximum response to phenylephrine. The value of the asymptote of the test compound curve relative to phenylephrine indicated the intrinsic 1o activity of the compounds. (Phenylephrine was assumed to have an intrinsic activity of 1).

The p[Aso] value is a measure of agonist potency. It is the negative logarithm of the agonist concentration which produces a response that is half the maximum response. For compounds with intrinsic activities significantly less than 1, i.e. it is possible to is calculate efficacy values and affinity (pKA) values using the comparative method of analysis. This analysis assumes that phenylephrine is acting as a full agonist in this system and thus uses it to define the operational model parameters E, and n (ref. Leff, et al., "Estimation ofagonist affinity and efficacy by direct and operational model fitting.," J.

Pharmacol. Methods., 1989, 23, 225-237). These parameters can then be utilised to perform 20 a comparative analysis on the test compound to be made. Affinity is expressed as a pKA (the negative logarithm of the agonist concentration that occupies half of the receptors).

99* Quantification of antagonism Compounds that did not demonstrate agonism were investigated as antagonists by 25 incubatine tissues with as high a concentration of the compound as possible and subsequently constructing phenylephrine curves curves. The degree of rightward shift of these phenylephrine curves compared to the control phenylephrine curve allowed an estimation of the affinity of a test compound to be made. Such affinity estimates are shown as pA 2 values (negative logarithm of the concentration of antagonist that produces a 2-fold rightward displacement of the control curve).

Confirmation of a,-mediated agonism Prazosin has been adopted as the standard a, antagonist. If a test compound showed agonism then, upon reaching the asymptote of the test compound E/[AJ curve, prazosin (1pM) was added to see if' the response was reversed. If an antagonist reverses the response of the test compound this suggests that the agonism is a, mediated.

Claims (9)

1. Compounds of formula VIII: R I A-(CH2)p_-X-(CH 2 q-O-(CH)r-Y VIII wherein A is O=CH or is HOOC or the corresponding acid chloride, in which X represents -SO 2 NH- or NHSO 2 p, q and r independently represent 2 or 3, Y represents thienyl optionally substituted by alkyl or halogen, or phenylthio- or phenyl optionally substituted by alkyl or halogen, and each R independently represents H or alkyl.

2. Compounds of formula VIII according to claim 1 which are compounds of formula III: R O=CH-(CH 2 p- 1 (CH2) qO III in which p, q, r, R, X and Y are as defined in claim 1.

3. Compounds of formula VIII according to claim 1 which are compounds 15 of formula V: R I HOOC- (CH 2 pi-X-(CH 2 V or the corresponding acid chloride, in which p, q, r, R, X and Y are as defined in claim 1. S *e *5 S

4. A compound as claimed in any one of claims 1 to 3, wherein q is 2. 20 5. A compound as claimed in any one of claims 1 to 3, wherein r is 2.

6. A conmpound as claimed in any one of claims 1 to 3, wherein Y is phenyl substituted by methyl.

7. A compound as claimed in any one of claims 1 to 3, wherein y is phenyl substituted by a halogen substituent selected from a chloro- or fluoro-substituent.

8. A compound as claimed in any one of claims 1 to 3, wherein X is SO 2 NH, p is 3 and q and r are each 2.

9. A compound as claimed in any one of claims 1 to 3, wherein X is NHSO2, and p, q and r are all 2. A process for the preparation of a compound of formula VIII as defined in claim 1, which comprises: when A is O=CH, reducing a corresponding ester compound, or (ii) when A is HCOC, hydrolysing a corresponding ester compound.

11. Compounds of formula VIII as defined in claim 1, when obtained by the process of claim DATED this 9th day of April 1999 ASTRA PHARMACETICALS LIMITED, By its Patent Attorneys, E. F. WELLINGION CO., 4 *4 4 4e S. 4 (Bruce Wellington) A/KA~4694 .4 .4.