AU608114B2 - Compounds - Google Patents

Description

011 k.

PCT

wc OPI DATE 19/07/89 APPLN. I] 29217 89 INTERNATIONAL APPLICATIO AOJP DATE 17/08/89 PCT NUMBER PCT/GB88/01140 (51) International Patent Classification 4 C08F 8/44, 8/32, A61K 31/745 SInt on Pub l l tion n be r: Int Itio TnaPPublic tion Date: WO 89/ 05828 29 June 1989 (29.06.89) (21) International Application Number: PCT/GB88/01140 (22) International Filing Date: 21 December 1988 (21.12.88) (31) Priority Application Number: 8730010 (74) Agent: GIDDINGS, Smith Kline French Laboratories Limited, Mundells, Welwyn Garden City, Hertfordshire AL7 IEY (GB).

(81) Designated States: AU, DK, FI, HU, JP, KR, NO.

Published With international search report.

Before the expiration of the time limit for amending the claims and to be republished in the event of the receipt of amendments.

(32) Priority Date: (33) Priority Country: 23 December 1987 (23.12.87) (71) Applicant: SMITH KLINE FRENCH LABORATO- RIES LIMITED [GB/GB]; Mundells, Welwyn Garden City, Hertfordshire AL7 IEY (GB).

(72) Inventors: JAXA-CHAMIEC, Albert, Andrzej 22 Beacon Way, Rickmansworth, Hertfordshire WD3 2PE HICKEY, Deirdre, Mary, Bernadette 19 Saxon Way, Saffron Walden, Essex CBI 4EQ (GB).

This document contains the amendments made under Section 49 and is correct for printing.

(54) Title: COMPOUNDS

-(CH

2 CH) a (CH 2 CH) b -(CH 2 CH) C R2 (CHCH 2 )b 1 ®N 3 b R R xG klil n (57) Abstract -t Polymers of structure in which, R 1 is a saturated or unsaturated C 6 to C 20 alkyl group; R 2 and R 3 are the same or different and are each C-4alkyl; X- is a physiologically acceptable counter ion; a, b, and c are numbers which indicate the relative molar percentages of the units present in a random distribution in said polymer, being from about 0.5 to about molar percent, and being from about 30 to about 99 molar percent; and n is a number indicating the number of repeating units in said polymer, processes for their preparation, intermediates useful in their preparation, pharmaceutical compositions containing them and their use in therapy for the lowering of serum cholesterol levels in humans.

K

I IX__ 89 1140 058281 I 4WO 89/05828 PCT/GB88/01140 1

COMPOUNDS

9.06.89) rch Lan City, ding the receipt The present invention relates to novel polystyrene anion exchange polymers, processes for their preparation, intermediates useful in their preparation, pharmaceutical compositions containing them and their use in the lowering of plasma cholesterol levels in humans.

Coronary Heart Disease (CliD) is one of the most serious health problems of contemporary society. Worldwide epidemiological studies have shown "hat the incidence of CHD is related to a number of independent risk factors, in particular, for example, high concentrations of serum cholesterol (hypercholesterolaemia). Such adverse factors lead to atherosclerosis, and ultimately, in severe cases, intermittent claudication, cerebrovascular insufficiency, thrombosis and cardiac arrest.

It is known that ion exchange polymers, in particular polystyrene polymers can be used as sequestering agents to bind non-absorbed bile acids and salts in the intestinal tract, forming complexes which are then excreted in the faeces. This sequestering leads to a decrease in the amount of bile acids returning to the liver via enterohepatic circulation. The synthesis of replacement bile acids from hepatic cholesterol depletes hepatic cholesterol, regulates hepatic LDL receptors and consequently reduces plasma cholesterol levels. Such sequestering polymers have been recognised as useful for the treatment of hypercholesterolaemia, and it is now proven that reducing serum cholesterol with bile acid sequestrants has a beneficial effect on protecting against the occurrence of coronary heart disease.

The such sequ a di- or trimethyl 5 discloses having of microp addition, polystyre 10 water abs polymer w polystyre groups, i 15 One which is in humans cholestyr linked an 20 ionisable backbone.

with a nu is unpala 36 g per 25 constipat its abili to the am It 30 compounds sequester sequester cholesteri WO 89/05828 ime or ite the about of re- -utical r

I

WO 89/05828 PCT/GB88/01140 WO 89/05828 15 Example Example 8 WO 89/05828 PCT/GB88/01140 2 The polystyrene polymers known in the art to have such sequestering activity are, in general, those bearing a di- or triloweralkyl ammonium group, such as a trimethylammonium group. For example, GB 1286949 discloses a series of macroporous polystyrene polymers having 5-20% cross-link, and GB 1579490 discloses a series of microporous polymers having 8-20% cross-link. In addition, GB 2026501 discloses a series of, inter alia, polystyrene polymers which are said to have particular water absorption capacities, i.e. 69-73% by weight of polymer weight. In each of the foregoing, the polystyrene polymers bear di- or triloweralkyl ammonium groups, in particular a trimethylammonium group.

One particular agent based..on a polystyrene polymer which is currently used to lower serum cholesterol levels in humans by binding bile acids in the intestinal tract is cholestyramine (GB 929391). Cholestyramine is a crosslinked anion exchange polystyrene polymer bearing an ionisable trimethylammonium group bound to the polymer backbone. However, the use of this agent is associated with a number of undesirable side-effects, for example, it is unpalatable and must be taken in large doses (up to 36 g per day) and causes, in some cases, bloating, constipation and other gut side-effects. In addition, its ability to bind bile acids is inefficient with respect to the amounts of polymer which it is necessary to use.

It is the object of the present invention to provide compounds which overcome the disadvantages of this known sequestering agent and provide improved bile acid sequestering agents which are useful for lowering serum cholesterol levels in humans.

Ir 1 'WO 89/05828 PCT/GB88/01140 3 The present'invention therefore provides in a first aspect, polystyrene polymers of structure (I)

-(CH

2

CH)

a

-(CHCH)

b

(CH

2 CH) c R 2 -(CHCH2)b- R 1 xO R in which, R is a saturated or unsaturated C 6 to C20 alkyl group; 2 3 R and R are the same or different and are each C alkyl; 1-4 X is a physiologically acceptable counter ion; a, b and c are numbers which indicate the relative molar percentages of theunits present in a random distribution in said polymer, being from about to about 10 molar percent, and being from about 30 to about 99 molar percent; and n is a number indicating the number of repeating units in said polymer.

Suitably, R is a saturated or unsaturated C 6 to

C

2 0 alkyl group. More suitably R is a saturated C 6 to C 20 alkyl group. Preferably R is a saturated C 8 to C14 alkyl group; most preferably a C12 alkyl group, in particular an unbranched C12 alkyl group.

.r WO 89/05828 PCT/GB88/01140 4 Suitably the groups R 2 and R 3 are the same or different and are each C 4alkyl; preferably they are 2 3 the same; most preferably R and R are both methyl.

Suitably is from about 0.5 to about 10 molar percent of said polymer, preferably is from about 1 to about 8 molar percent of said polymer; most preferably from about 1 to about 4 molar percent.

Suitably X- is a physiologically acceptable counter ion such as a sulphate, bicarbonate, carbonate, formate, acetate, sulphonate, propionate, malonate, succinate, malate, tartrate, citrate, maleate, fumarate, ascorbate, glucuronate, phosphate, or halide, or the anion of an amino acid such as aspartic or glutamic acid. More suitably X is a phosphate, sulphate or a halide ion; preferably a halide ion, in particular chloride.

n is a number indicating the number of repeating units in said polymer. Owing to the three dimensional cross-linkage precise figures cannot be given for n, but .in any case will be greater than 1,000.

The polystyrene polymers of the present invention are also characterised by their total exchange capacity i.e. the theoretical maximum capacity of the polymer if each counter ion were to be exchanged with. bile acid. In this specification the total exchange capacity is defined in terms of the number of milliequivalents of counter ion per gram of dry weight of polymer.

Suitable total exchange capacities are in the range of, for example where the counter ion X- is a halide ion such as chlorine, from 1.5 to 3.5 meq Cl per gram of polymer. Preferred within this range are polymers having a total exchange capacity of between 2 and 3 meq C1 /gram of polymer.

In addition, it is to be noted that the approximate molar percentages and are calculated from the monomer mixture or, in some instances from microanalytical data.

It is to be noted that the term 'bile acid' when used herein shall be taken to include bile acids, bile salts and conjugates thereof.

The polystyrene polymers of the present invention can be prepared by processes analogous to those known in the art. The present invention therefore provides, in a further aspect, a process for preparing the polystyrene polymers of structure which comprises reaction of a polymer of structure (II) -(CHCH)a-(CH 2 CH)b-(CH 2

CH)(

(II)

0 n

-(CHCH

2 )b

Y

in which a, b, c and n are as described for structure (I) and Y is a group displaceable by an amine, with an amine 1 2 3 1 3 of structure R R R N (III) in which R to R are as described for structure i .i WO 89/05828 PCT/GB88/01140 6 reaction of a compound of structure (IV)

-(CH

2 CH) a-(CHCH) -(CH 2H)

(IV)

-(CHCH

2 )b

Z

in which a, b, c and n are as described for structure (I) and Z is NR R or NR2R 3 in which R to R are as described for structure with a compound of structure R Y in which R 4 is a C 4alkyl group when Z is NR1R 2 or a saturated or unsaturated C6-20 alkyl group when Z is NRR 3 and Y is a group displaceable by an amine.

The reaction between a polymer of structure (II) and an amine of structure (III) can be carried out in a suitable solvent at elevated temperature. Suitable solvents include for example, a C14alkanol, N-methylpyrrolidone, sulpholane, dimethylformamide, nitromethane or tetrahydrofuran. Preferably the reaction is carried out in N-methylpyrrolidone at a temperature of between about 500 and 800 for up to 24 hours or until the reaction is complete.

The reaction between a polymer of structure (IV) and a compound of structure can be carried out in a suitable inert solvent such as a C 4alkanol, nitromethane, sulpholane, N-methylpyrrolidone, dimethylformamide or tetrahydrofuran at elevated temperature.

The intermediate polymers of structure (II) are available commercially or can be prepared from readily available materials by methods known to those skilled in the art. For example polymers of structure (II) in which Y is chlorine can be prepared by reaction of chloromethylstyrene, styrene and divinyl benzene in an aqueous suspension comprising polyvinyl alcohol in the presence of an initiator such as AIBN at elevated temperature.

Alternatively, the intermediate polymers of structure (II) can be prepared directly from polystyrene by methods analogous to those known in the art, for example where Y is chlorine by chloromethylation of polystyrene.

Certain intermediates of structure (IV) are novel and form a further aspect of the invention namely compounds of structure (IVA)

C

2 2 c

-(CH

2 CH) -(CH 2 CH)b-(CH CH)

(IVA)

-(CHCH

2 )b ZA in which a, b, c and n are as described for structure (I) and Z A is NRR 2 or NR1R 3 in which R 1 to R 3 are as described for structure The intermediate polymers of structure (IV) can be prepared from the polymers of structure (II) by reaction i a -a WO 89/05828 PCT/GB88/01140 8 1 2 with an amine of structure R2NH in which R 2 is R R or R2R 3 under the same or similar conditions as indicated for the reaction of a compound of structure (II) and a compound of structure (III).

Alternatively the intermediate polymers of structure (IV) can be prepared by polymerisation of styrene, divinylbenzene and a compound of structure (V)

(V)

A

in which Z a is as defined in structure (IVA), under standard polymerisation conditions. For example, polymerisation can be carried out in an aqueous suspension comprising, for example, polyvinyl alcohol in the presence of an initiator at elevated temperature. Suitable initiators include, for example, AIBN.

The intermediate compounds of structure can be prepared by reaction of an amine of structure R NH in which R2 is RR 2 or R R with a corresponding compound of structure in which Za is a group displaceable by an amine.

The polystyrene polymers of structure have been found to bind bile acids in in vitro experiments and in in vivo animal models they have been found to increase the amount of bile acids detectable in the faeces. In particular, when compared to the known sequestrants e.g.

I

'NYo 89/05828 PCT/GB88/01140 9 cholestyramine, the polymers of structure have surprisingly been found to have an unexpected profile of activity which is thought will provide advantages over the known compounds in the lowering of serum cholesterol levels in animals, in particular humans. More specifically, in in vitro experiments, when compared to cholestyramine the compounds of structure have been found to bind comparable amounts of bile acid per gram of polymer (at physiological concentrations of bile acids), and to bind the bile acid more strongly the bile acids have been found to dissociate more slowly from the compounds of the invention. It is expected that compounds having such qualities will be able to achieve significant lowering of plasma cholesterol levels at much lower dosages than has hitherto been possible with known sequestrants (currently given at up to 36 g/day).

As indicated earlier it is recognised that removal of bile acids from the intestinal tract in this way lowers serum cholesterol levels and also has a beneficial effect on protecting against atherosclerosis and its dependent clinical conditions. The present invention therefore provides in a further aspect, polystyrene polymers of structure for use in therapy, in particular for the lowering of serum cholesterol levels in. mammals, including humans. In addition the polymers of structure are expected to be of use in protecting.against atherosclerosis and its sequelae, and for example in the treatment of pruritus and diarrhoea.

In view of the foregoing the present invention also provides a method of lowering serum cholesterol levels in mammals which comprises administering to a mammal in need WO 89/05828 PCT/GB88/01140 10 thereof an effective serum cholesterol lowering amount of a polystyrene polymer of structure and a method of protecting against atherosclerosis.

When used in therapy in the methods of the invention, the polystyrene polymers of structure are in general administered in a pharmaceutical composition.

In a still further aspect of the present invention there is therefore provided a pharmaceutical composition comprising a polystyrene polymer of structure in association with a pharmaceutically acceptable carrier.

The compositions of the present invention can be prepared by techniques well known.to those skilled in the art of pharmacy and include all those known for the formulation of polystyrene polymers for human use.

The polymers are preferably administered as formulations in admixture with one or more conventional pharmaceutical excipients which are physically and chemically compatible with the polymer, which are nontoxic, are without deleterious side-effects but which confer appropriate properties on the dosage form.

In general, for liquid formulations, aqueous based pharmaceutically acceptable carriers such as water itself or aqueous dilute ethanol, propylene glycol, polyethylene glycol or glycerol or sorbitol solutions are preferred.

Such formulations can also include preservatives and flavouring and sweetening agents such as sucrose, fructose, invert sugar, cocoa, citric acid, ascorbic acid, fruit 1 1 _i i i x K) in which, R is a saturated or unsaturated C 6 to C 20 alkyl group; /2 WO 89/05828 PCT/GB88/01140 11 juices etc. In general, digestible oil or fat based carriers should be avoided or minimised as they contribute to the condition sought to be alleviated by use of the polymers. They are also subject to absorption by the polymers during prolonged contact, thus reducing the capacity of the polymer to absorb bile acids after administration.

The polymers can also be prepared as 'concentrates', for dilution prior to administration, and as formulations suitable for direct oral administration. They can be administered orally ad libitum, on a relatively continuous basis for example by dispersing the polymer in drinks or food.

Preferably, the polymers are administered in tablet form or in gelatin capsules containing solid particulate polymer or a non-aqueous suspension of solid polymer containing a suitable suspending agent. Suitable excipients for such formulations will be apparent to those skilled in the art and include, for example, for tablets and capsules lactose, microcrystalline cellulose, magnesium stearate, povidone, sodium starch glycollate and starches; and for suspensions in capsules, polyethylene glycol, propylene glycol and colloidal silicon dioxide.

Preferably the polymer is administered in unit dosage form, each dosage unit containing preferably from 0.3 g to 1 g of polymer.

The daily dosage regimen for an adult patient may be, for example, a total daily oral dose of between 1 and g, preferably 1-5 g, the compound being administered 1 -1 'ii ;L_-w WO 89/05828 PCT/GB88/01140 12 to 4 times a day depending on the size of individual dosage units. Suitably the compound is administered for a period of continuous therapy of one month or more sufficient to achieve the required reduction in serum cholesterol levels.

In addition the polymers of the present invention can be co-administered (together or sequentially) with further active ingredients such as HMGCoA reductase inhibitors and other hypocholesterolaemic agents, and other drugs for the treatment of cardiovascular diseases.

The following data and examples indicate the properties and preparation of the polymers of the present invention. Temperatures are recorded in degrees celsius.

The exchange capacity of the ammonium substituted polymers was determined by elemental analysis and/or potentiometric titration of chloride ion. Figures quoted are expressed as milliequivalents of exchangeable chloride ion per gram of dry polymer weight.

Chloromethylstyrene was used as a 60:40 m:p mixture and was washed free of inhibitor before use.

Divinylbenzene (DVB) was used as a 55% mixture with ethylstyrene and the weights given are that of the mixture.

The percent cross-link given is based on the percent of divinylbenzene (not the mixture) calculated to be in the monomer mixture. The molar percent of divinylbenzene (b) quoted is based on the molar percent of DVB in the monomer mixture.

,1 'WO 89/05828 PCT/GB88/01140 13 Example 1 Chloromethylstyrene (60:40 m.p. mixture) (26.7 g), styrene (22.4 divinylbenzene (0.91 and azobisisobutyronitrile (AIBN) (0.5 g) were mixed to give a homogenous solution and added to a solution of Foly(vinyl alcohol) 125,000) (1.0 g) in distilled water (500 ml). The mixture was then stirred at 800 under an atmosphere of nitrogen at such a rate as to maintain the monomers in suspension. After 7 hours the stirring was stopped and the mixture poured into distilled water. The resin formed was washed by decantation with cold and hot water, filtered, and washed with acetone, tetrahydrofuran, and acetone, and dried to give a 1% cross-linked chloromethyl-substituted polystyrene containing 3.3 m.eq.

Cl/g (36.6 This polymer was then sieved and the 53-106 VM fraction (15.8 g) used in further reactions.

This fraction of the chloromethylated polystyrene (3.3 g) was suspended in N,N-dimethylformamide (DMF) ml), N,N-dimethyloctylamine (4.96 g) added, and the mixture stirred at 650 for 24 hours. The polymer was then filtered off and washed with methanol and diethyl ether, and dried under vacuum to give N,N-dimethyl-Noctylammoniomethyl-substituted polystyrene, chloride salt, (4.35 (exchange capacity 2.17 m.eq. Cl1/g, 0.97 molar percent).

Examples Chloromethylated polystyrenes containing 1% divinyl benzene as crosslinking agent and containing 2.58, 3.95, 4.91 and 5.96 m.eq. Cl/g were prepared from chloromethyl i WO 89/05828 PCT/GB88/01140 -14 styrene, styrene and diviriylbenzene as in Example 1. The above polymers were then reacted with N,N-dimethyloctylamine in DMF and worked up, as in Example 1, to give N,N-dimethyl-N-octylammoniomethyl-substituted polystyrene, chloride salt, with the following exchange capacities:- 1.77, 2.34, 2.76, 3.15 m.eq. Cl /g and values of 0.93, 1.00, 1.08 and 1.17 molar percent respectively (Examples Example 6 Chloromethylated polystyrene cross-linked 4.91 m.eq. Cl/g), (1 g) as used in Example 4, was suspended in DMF (25 ml), N,N-dimethyldecylamine (1.8 g) added, and the mixture stirred at 700 for 24 hours. After work-up as in Example 1, the N,N-dimethyl-N-decylammoniomethylsubstituted polystyrene, chloride salt, was obtained as off-white beads (1.83 g) (exchange capacity 2.44 m.eq.

C1 1.08 molar percent).

Example 7 A 0.5% cross-linked chloromethylated polystyrene was prepared from chloromethylstyrene (31.29 styrene (18.25 and divinylbenzene (0.46 g) by the method described in Example 1. After sieving 7.52 g of material of particle size, 53-106 VM, containing 3.89 m.eq. Cl/g, was obtained. This polymer (2.0 g) was suspended in DMF ml), N,N-dimethyloctylamine (3.87 g) added, and the mixture stirred at 650 for 24 hours. After work-up as in Example 1, N,N-dimethyl-N-octylammoniomethyl-substituted polystyrene, chloride salt, (3.00 g) was obtained (exchange capacity 2.23 m.eq. Cl1/g, 0.51 molar percent).

_1 :L j WO 89/05828 PCT/GB88/01140 15 Example 8 A 2% cross-linked chloromethylated polystyrene was prepared from chloromethylstyrene (32.7 styrene (16.99 g) and divinylbenzene (1.82 g) by the method described in Example 1. After sieving 25.4 g of material of particle size, 53-106 pM, containing 3.86 m.eq. Cl/g, was obtained. This polymer (5 g) was reacted with N,N-dimethyloctylamine (5 DMF (80 ml) at 800 for 48 hours to give, after work-up as in Example 1, N,N-dimethyl-N-octylammoniomethylated-substituted polystyrene, chloride salt, (7.75 g) (exchange capacity 2.51 m.eq. Cl 0.98 molar percent).

Examples 9-16, 18-21 and 25-32 A commercially available 1% cross-linked chloromethylated polystyrene (3.72 m.eq. Cl /g or 4.15 m.eq. Cl/g, 200-400 mesh) ("Bio Rad S-X1, chloromethylated") was used in these examples.

Examples 9-13 N,N-dimethylhexylamine, N,N-dimethyloctylamine, N,N-dimethyldecylamine, N.N-dimethyldodecylamine, and N,N-dimethyltetradecylamine were each reacted with the above chloromethylated polystyrene cross-linked, 3.72 m.eq. Cl in DMF at 700 to give, after work up as described in Example 1, the corresponding N,N-dimethyl- N-alkylammoniomethyl-substituted polystyrenes, chloride salts, with the following exchange capacities:- Example 9, hexyl, 2.62 m.eq. Cl Example 10, octyl, 2.40 m.eq.

Cl Example 11, decyl, 2.21 m.eq. Cl /g; WO 89/05828 PCT/GB88/01140 -16 Example 12, dodecyl, 2.11 m.eq. Cl Example 13, tetradecyl, 2.02 m.eq. Cl For each of these examples was 0.9 molar percent.

Example 13A Chloromethylpolystyrene (33.0 Kg) was washed twice with dichloromethane (2 x 340 L) and stirred with N-methylpyrrolidone (815 L) that had been purged under an atmosphere of nitrogen for 1 hour at 500. The mixture was warmed to 60-650 and N,N-dimethyldodecylamine (81.6 Kg) added. It was then further warmed to 65-700 for 5 hours.

The mixture was cooled to 55-600 and the product isolated by centrifugation. The wet product was reslurried with a mixture of N-methylpyrrolidone (250 L) and dichloromethane (250 then dichloromethane (500 followed by a mixture of dichoromethane (450 L) and ethyl acetate (450 The product was then further washed with ethyl acetate (2 x 910 L) and vacuum dried for 24 hours at room temperature followed by 500 for 72 hours, to afford N,N-dimethyl-N-dodecylammoniomethyl-susbtituted polystyrene chloride.

Example 14 Ethylchloroformate (18.3 ml) was added slowly to a solution of heptanoic acid (20.0 g) and triethylamine (26.7 ml) in tetrahydrofuran (250 ml), maintained at The resulting mixture was stirred at 100 for 0.75 hour, cooled to 50 and a 33% solution of dimethylamine in ethanol (41.6 ml) added dropwise. This mixture was then allowed to warm to room temperature and stirred for 4 hours. The mixture was filtered and the WO 89/05828 PCT/GB88/01140 17 filtrate evaporated to dryness. The residue was dissolved in chloroform (250 ml) and washed with 1% aqueous sodium carbonate solution (200 ml) and water, then dried and evaporated. The residual oil was chromatographed on silica gel to give N,N-dimethylheptanamide (14.1 g) as a colourless oil.

This amide (13.5 g) was treated with lithium aluminium hydride (4.88 g) in dry tetrahydrofuran (250 ml) to give, after work-up, N,N-dimethylheptylamine (7.10 g) as a colourless oil.

Chloromethylated polystyrene (3.0 g, 3.72 m.eq. Cl /g) was suspended in DMF, N,N-dimethylheptylamine (3.7 g) added and the mixture heated at 750 for 24 hours. After work-up as described in Example 1, N,N-dimethyl-N-heptylammoniomethyl-substituted polystyrene, chloride salt, was obtained as an off-white resin (4.49 (exchange capacity 2.50 m.eq. Cl 0.98 molar percent).

Example N,N-dimethylnonylamine was prepared from nonanoic acid by the method described in Example 14.

A suspension of chloromethylated polystyrene (3.0 g, 3.72 m.eq. C l/g) in DMF (50 ml) was treated with N,N-dimethylnonylamine (4.5 g) and the mixture stirred at 700 for 24 hours. Work-up as described in Example 1 gave N,N-dimethyl-N-nonylammoniomethyl-substituted polystyrene, chloride salt (4.72 g, exchange capacity 2.19 m.eq.

Cl 0.98 molar percent).

a ieit wuere one counter ion X is a halide ion WO 89/05828 PCT/GB88/01140 18 Example 16 N,N-dimethylundecylamine was prepared from undecanoic acid by the method described in Example 14.

A suspension of chloromethylated polystyrene (2.0 g, 3.72 m.eq. C 1/g) in DMF was treated with N,N-dimethylundecylamine (5.0 g) at 700 for 28 hours.

Work-up as described in Example 1 gave N,N-dimethyl-Nundecylammoniomethyl-substitited polystyrene, chloride salt (3.38 g, exchange capacity 2.05 m.eq. Cl -g, 0.98 molar percent).

Example 17 A 2% cross-linked, styrene-divinylbenzene copolymer, chloromethylated to give a Cl content of 4.45 m.eq.

Cl (5 g) was treated with N,N-dimethyloctylamine (9.15 g) at 700 for 27 hours. After work-up as described in Example 1, N,N-dimethyl-N-octylammoniomethylsubstituted polystyrene, chloride salt, was obtained as yellow beads (8.26 g, exchange capacity 2.5 m.eq.

Cl g, 1.61 molar percent).

Example 18 Sodium borohydride (6.62 g) was added in portions to propionic acid (52.5 ml), cooled to 50, maintaining the temperature below 200. N-methyloctylamine (5 g) was added dropwise, keeping the temperature at 200. The reaction was stirred at 800 for 3 hours, cooled and basified with 2N sodium hydroxide. The aqueous solution was extracted with dichloromethane (2 x 75 ml). The combined extracts 'WO 89/05828 PCT/GB88/01140 19 were dried with anhydrous magnesium sulphate and evaporated to dryness. The resulting oil was purified by column chromatography to give N-methyl-N-propyloctylamine as a colourless oil (4.09 g, 63%).

To a suspension of chloromethystyrene-styrenedivinylbenzene co-polymer w/w divinylbenzene, 4.15 m.eq. Cl-/g) (2 g) in DMF (50 ml) the above amine was added and the mixture stirred at 700 for 24 hours to give, after washing and drying as in Example 1, the corresponding quaternised polymer as off-white resin beads (3.11 g) (exchange capacity 2.26 m.eq. Cl 1 molar percent).

Example 19 N-methyl-di-n-octylamine (14.26 g) was reacted with 1% cross-linked chloromethyl polystyrene (5 g, 18.6 mmoles) in DMF (125 ml) at 70° for 24 hours, to give after washing as in Example 1 the corresponding quaternised polymer as off-white resin beads (9.05 g) (exchange capacity 1.87 m.eq. Cl 0.98 molar percent).

Example N,N-diethyloctylamine (13.7 g) was reacted with 1% cross-linked chloromethylstyrene-styrene-divinylbenzene co-polymer (10 g, 37.2 m.eq. Cl) in DMF (100 ml) at 800 for 12 hours. After work-up as in Example 1, the corresponding quaternised polymer was isolated as off-white resin beads (16.8 g) (exchange capacity 2.09 m.oq. Cl 0.98 molar percent).

i .L e L WO 89/05828 PCT/GB88/01140 20 ExamDls 21 N,N-dibutyloctylamine was prepared from dibutylamine and octanoyl chloride by a method similar to that in Example 14.

This amine (17 g) was reacted with 1% cross-linke chloromethylstyrene-styrene-divinylbenzene co-polymer g, 40.2 m.eq. Cl) in DMF (100 ml) at 800 for 1 hour.

Work-up as in Example 1, gave the corresponding quaternised polymer as off-white resin beads (15.0 g) (exchange capacity 1.90 m.eq. Cl 1 molar percent).

Example 22 Chloromethylstyrene (50 g) in ethanol (100 ml) was added dropwise to 33% dimethylamine in i.m.s (500 ml) over minutes and the temperature rose to 350. The reaction was stirred at room temperature for 20 hours. 33% Dimethylamine in i.m.s (50 ml) was added and the reaction was heated at reflux for 30 minutes. The reaction was cooled and evaporated to dryness. The resulting product was dissolved in dichloromethane and washed with 2N hydrochloric acid (2 x 500 ml). The combined extracts were basified with 2N sodium hydroxide and extracted with dichloromethane (2 x 200 ml). The combined organic extracts were washed with water, dried with anhydrous magnesium sulphate and evaporated to dryness. More inhibitor was added and the oil was purified by distillation to give 3(4)-(N,N-dimethylaminomethyl)styrene as a colourless oil, bp 660 at 0.1 Torr, (37 g, 78%).

'WO 89/05828 PCT/GB88/01140 -21 The above amine (31.17 styrene (17.93 g), divinylbenzene (0.87 g) and azobisisobutyronitrile (AIBN) g) were polymerised as in Example 1. After 7 hours the mixture was poured into distilled water. The resin formed was washed by decantation with cold and hot water, filtered and washed with acetone, ether and finally water.

Drying under reduced pressure gave a 1% w/w cross-linked N,N-dimethylaminomethyl-substituted polystyrene polymer containing 3.85 m.eq. N/g (11.01 g, 53-106 p, 22%).

The above cross-linked N,N-dimethylaminomethylsubstituted polystyrene (5 g) was suspended in DMF (100 ml), 1-iodododecane (15 g) was added and the reaction was stirred at 700 for 20 hours. The polymer was filtered off and washed with DMF and methanol. Anion exchange was accomplished by stirring the polymer in 2N HCI: MeOH (250 ml:250 ml) and standing overnight. The resin was then filtered off and washed with 2N HC1, water, methanol and ether. Finally dried under vacuum to give the corresponding quaternised polymer (4.26 g) (exchange capacity 1.82 m.eq. Cl 0.38 m.eq. I /g, 0.99 molar percent).

Example 23 To a mixture of N-methyldodecylamine (49.5 g) and anhydrous potassium carbonate (18.1 g) in ethanol (200 ml) was added chloromethylstyrene (20 g) in ethanol (75 ml) ove ?0 minutes. The reaction was stirred at room tem ture for 20 hours and heated at reflux for minutes. The reaction mixture was evaporated to dryness and the residue was dissolved in 2N hydrochloric acid and extracted with chloroform (200 ml). The chloroform WO 89/05828 PCT/GB88/01140 22 extract was washed with water, dried with anhydrous magnesium sulphate and evaporated to dryness.. The oil was purified by chromatography giving 3(4)-(N-dodecyl-Nmethylaminomethyl)styrene as an orange oil (27.5 g, 67%).

The above amine (11.46 styrene (3.22 g), divinylbenzene (0.33 g) and AIBN (0.2 g) were mixed to give a solution and then added to a solution of polyvinylalcohol 125,000) (0.5 g) in distilled water (200 ml). The mixture was stirred at 800 under an atmosphere of nitrogen at such a rate as to maintain the monomers in suspension. After 7 hours the mixture was poured into distilled water. The resin was filtered off and washed with water. Product was transferred to a beaker and washed by stirring and decantation with acetone (2 x 200 ml) and ether (2 x 200 ml). The resin was filtered off and drying under reduced pressure gave a 2% w/w cross-linked N-dodecyl-N- methylaminomethylsubstituted polystyrene resin (10 g, 66%).

c) The above 2% w/w cross-linked N-dodecyl-N-methylaminomethyl-substituted polystyrene (4 g) was suspended in DMF (50 ml) and stirred for 1 hour. Methyl iodide (1.75 ml) was added and the reaction was stirred at 600 for 22 hours. Additional methyl iodide (1.75 ml) was added at 19 hours. The polymer was filtered off and washed as in Example 22 to give, after drying under vacuum, the corresponding quaternised polymer (2.2 g, 106-212 (exchange capacity 2.11 m.eq. C1 /g, 0.065 m.eq. I 1.99 molar percent).

WO 89/05828 PCT/GB88/01140 23 Example 24 To a mixture of N-methyloctylamine (25 g) and anhydrous potassium carbonate (13.3 g) in ethanol (150 ml) was added chloromethylstyrene (13.3 g) in ethanol (10 ml) over 15 minutes. The reaction was stirred at room temperature for 24 hours, filtered and the filtrate was evaporated to dryness. The residue was dissolved in dichloromethane and washed with water. The organic layer was then dried with anhydrous magnesium sulphate and evaporated to dryness. The oil was purified by chromatography (with hexane:dichloromethane as eluent) to give 3(4)-(N-octyl-N-methylaminomethy)styrene as an oil (11.5 g, 51%).

The above amine (11.02 styrene (3.61 g), distyrene (0.37 g) and AIBN (0.2 were polymerised as in Example 23 to give, after drying, a 2% w/w crosslinked N-octyl-N-methylaminomethyl-substituted polystyrene resin (10.45 g, The above N-methyl-N-octylaminomethyl-substituted polystyrene (7 g) was suspended in DMF (100 ml), methyl iodide (5.3 ml) was added and the reaction was stirred at 600 for 9 hours. Additional methyl iodide (2 ml) was added at 5 hours. The polymer was filtered and washed as in Example 22 to give, after drying, the corresponding quaternised polymer (5.6 g) (exchange capacity 2.36 m.eq. Cl 0.10 m.eq. I 1.99 molar percent).

i WO 89/05828 PCT/GB88/01140 24 Example Diamylketone (12 g) was hydrogenated at 50 psi in the presence of 10% Pd/C (1.5 and benzylamine (8.31 g) at room temperature for 6 hours. Further catalyst (1.5 g) was added and the mixture was hydrogenated at 40° for hours. The reaction mixture was filtered and evaporated to dryness (combined with the product from a previous 12 g hydrog-nation). The oil was purified by column chromatography to give 6-undecylamine as an oil (9.1 g, 38%).

To 90% formic acid (11.9 ml), cooled to -50, was added the above amine (9.6 dropwise over 4f minutes keeping the temperature below 50. 37% Formaldehyde ml) was then added to the cooled reaction before heating at reflux for 6 hours. Concentrated hydrochloric acid (4 ml) was added and the reaction mixture was evaporated to dryness. The resulting oil was scratched under ethyl acetate to give a green solid. The solid was collected by filtration but this waxy solid rapidly became an oil on drying. This oil was dissolved in water, basified with sodium carbonate. The aqueous phase was extracted with ethyl acetate. The extracts were washed with water, dried with anhydrous magnesium sulphate and evaporated to dryness. The oil was distilled at 0.1 torr/250 0 in a Kugelrohr apparatus to give 6-(dimethylamino)undecane as a colourless oil (4.06 g, 36%).

The above amine (3 g) was reacted with 1% crosslinked chloromethylated polystyrene (2 g, 7.44 mmoles) in DMF (50 ml) at 700 for 20 hours to give, after washing and )Y 89/05828 PCT/GB88/01140 25 drying as in Example 1, the corresponding quaternised polymer as off-white resin beads (2.96 g) (exchange capacity 2.00 m.eq. Cl 0.98 molar percent).

Example 26 To a solution of potassium t-butoxide (34.68 g) in anhydrous diethyl ether (750 ml), under nitrogen, 7-carboxyheptyltriphenylphophohonium bromide (75 g) was added and the mixture was heated at reflux for 2 hours.

The diethyl ether was removed by distillation and 6-undecanone (40 g) was added over 10 minutes. After minutes, anhydrous diethyl ether (200 ml) was added to aid stirring. The mixture was heated at reflux for 2 hours, cooled and acidified with 2N HC1. The aqueous solution was extracted with dichloromethane (300 ml). The organic extracts were combined and washed with 2N NaOH (2 x 200 ml). The aqueous extracts were acidified with 2N HC1 and extracted with dichloromethane (2 x 200 ml).

The organic extracts were combined, dried with anhydrous magnesium sulphate and evaporated to dryness. The resulting oil was triturated with light petroleum. The light brown precipitate was removed by filtration and the filtrate was evaporated to dryness. The oil was purified by column chromatography to give 9-pentyl-8-tetradecenoic acid as an oil (13.94 g, 30.4%).

To a solution of the above acid (13.8 g) and triethylamine (7.8 ml) in dry tetrahydrofuran (200 ml), cooled to 50, was added ethyl chloroformate (5.4 ml) dropwise over 15 minutes. The reaction mixture was stirred at 100 for 2 hours. t Dimethylamine in industrial methylated spirits (125 ml) was added keeping

I

WO 89/05828 PCT/GB88/O0 26 the temperature at 10°. The reaction mixture was then stirred at room temperature for 4 hours and evaporated to dryness. The resulting oil was dissolved in diethyl ether and washed with IN NaOH, water and then dried with anhydrous magnesium sulphate and evaporated to dryness.

(This was then combined with the product from a previous reaction of the above acid (10.4 The oil was purified by column chromatography (with dichloromethane as eluent) to give 9-pentyl-8-tetradecenamide'as a coloured oil (14.91 g, 57%).

The above amide (14.6 g) in dry tetrahydrofuran was added dropwise to a suspension of lithium aluminium hydride (2.6 g) in dry tetrahydrofuran (150 ml), under an atmosphere of nitrogen. The temperature rose to 380 during the addition. The reaction was stirred at room temperature for 1.5 hours and heated at reflux for 1 hour.

The reaction was cooled and water (2.6 ml), 15% aq. sodium hydroxide (2.6 ml), water (7.8 ml) were added. The reaction was stirred for 1 hour and then filtered. The filtrate was evaporated to dryness. The oil was purified by distillation to give l-dimethylamino-9-pentyl-8tetradecene as an oil (bp 0.01 torr, 161-1680) (10.79 g, 77%).

The above amine (3 g) was reacted with 1% crosslinked chloromethylated polystyrene (1.5 g, 6.23 mmoles) in DMF (60 ml) at 600 for 24 hours to give, after washing and drying as in Example 1. the corresponding quaternised polymer as off-white resin beads (3.18 g) (exchange capacity 1.88 m.eq. Cl 1 molar percent).

WO 89/05828 PCT/GB88/01140 27 Example 27 l-Dimethylamino-9-pentyl-8-tetradecene (Example 26c) g) was hydrogenated in ethanol (75 ml) at 45 psi in the presence of 10% Pd/C (0.7 g) at room temperature for hours. The reaction mixture was filtered and evaporated to dryness. The resulting oil was purified by distillation at 0.01 torr (in a Kugelrohr apparatus at 2100) to give l-dimethylamino-9-pentyltetradecane as a colourless oil (5.7 g, 76%).

The above amine (5.5 g) was reacted with 1% crosslinked chloromethylated polystyrene (3 g, 12.45 mmoles) in DMF (100 ml) and methanol (20 ml) at 600 for 28 hours to give, after washing and drying as in Example 1, the corresponding quaternised polymer as off-white resin beads (6.6 g) (exchange capacity 1.81 m.eq. Cl 1 molar percent).

Example 28 Geranylbromide (20 g) in ethanol (50 ml) was added to 33% dimethylamine in industrial methylated spirits (240 ml) and the mixture was heated at reflux for 8 hours. The reaction mixture was evaporated to dryness, mixed with 2N sodium hydroxide (50 ml) and then extracted with diethyl ether (2 x 100ml). The ether extracts were combined and washed with water (50 ml), then dried with anhydrous magnesium sulphate and evaporated to dryness.

The resulting orange oil was purified by chromatography (with a gradient CH Cl :MeOH system as eluent) to give l-dimethylamino-3,7-dimethyl-2,6-octadiene as an oil (12.1 g, 72%).

ii -4 WO 89/05828 PCT/GB88/0114Q 28 The above amine (3.76 g) was reacted with 1% crosslinked chloromethylated polystyrene (2 g, 8.3 mmoles) in DMF at 700 for 20 hours to give, after washing and drying as in Example 1, the corresponding quaternised polymer as off-white resin beads (3.1 g) (exchange capacity 2.44 m.eq. Cl 1 molar percent).

Example 29 1-Dimethylamino-3,7-dimethyl-2,6-octadiene (Example 28a) (12 g) was hydrogenated in ethanol (100 ml) at 40 psi in the presence of 10% Pd/C (1.2 g) at room temperature for 5 hours. The reaction mixture was filtered and evaporated to dryness. The residue was dissolved in 2N sodium hydroxide and extracted with diethyl ether (2 x 100 ml). The ether extracts were combined and washed with water, dried with anhydrous magnesium sulphate and evaporated to dryness. The oil was purified by chromatography (10:1 dich-loromethane:methanol as eluent) to give 1-dimethylamino-3,7-dimethyloctane as a wax (7.14 g, b) The above amine (3 g) was reacted with 1% crosslinked chloromethylated polystyrene (2 g, 8.3 mmoles) in DMF (60 ml) at 70° for 24 hours to give, after washing and drying as in Example 1, the corresponding quaternised polymer as off-white resin beads (3.24 g) (exchange capacity 2.31 m.eq. C1 1 molar percent).

NOW

,NO 89/05828 PCT/GB88/01140 29 Example To 90% formic acid (24.2 ml), cooled to 100, was added isoamylamine (10 g) in portions keeping the temperature below 200. 37% Formaldehyde (20 ml) was added to the cooled reaction mixture. The reaction was then heated at 700 for 6 hours. The reaction mixture was cooled and concentrated sulphuric acid (2 mole equivalents) was added and the reaction mixture evaporated to dryness. Isopropanol (25 ml) was added to the oil with cooling and this was diluted with anhydrous diethyl ether.

The resulting thick, white precipitate was collected by filtration and washed with cold anhydrous diethyl ether.

This was quickly dried under reduced pressure but gave an oil. Sucked dry at 0.01 torr for 24 hours and this gave l-dimethylamino-3-methylbutane as the sulphate salt (11.1 g, The above amine sulphate salt (4.5 g) was reacted with 1% cross-linked chloromethylated polystyrene (2 g, 7.44 mmoles) and anhydrous potassium carbonate (4.1 g) in DMF (50 ml) at room temperature for 24 hours and at 600 for 20 hours. The product was filtered off and washed with DMF, methanol, water, 2N hydrochloric acid, water, methanol and ether to give, after drying under reduced pressure, the corresponding quaternised polymer as offwhite resin beads (2.28 g, exchange capacity 2.76 m.eq.

Cl 0/98 molar percent).

Example 31 3-Pentylamine (20 g) w;.s added to 90% formic acid (48.4 ml), cooled to 00, keeping the temperature below

I'

WO 89/05828 PCT/GB88/01149 30 100, followed by 37% formaldehyde (40 ml). The reaction mixture was heated at reflux for 7 hours, cooled and concentrated hydrochloric acid (2 mole equivalents) was added. The reaction mixture was evaporated to dryness.

Isopropanol (25 ml) was added and the solution was cooled and diluted with anhydrous diethyl ether. The resulting cream coloured solid was collected by filtration and dried under reduced pressure giving 3-dimethylaminopentaneethylpropane hydrochloride as a cream powder (m.p.

144-1450) (26.1 g, The above amine (9 g) was reacted with 1% crosslinked chloromethylated polystyrene (4 g, 16.6 mmoles) and anhydrous potassium carbonate (8.2 g) as in Example to give the corresponding quaternised polymer (5.12 g) (exchange capacity 2.30 m.eq. C1 1 molar percent).

Example 32 N,N-dimethylheptadecylamine 1170 0.15 mmHg) was prepared from dimethylamine and heptadecanoic acid by the method described in Example 14.

This amine (11.7 g) was reacted with 1% cross-linked chloromethylstyrene-styrene-divinylbenzene co-polymer g, 21 m.eq. Cl) in DMF (100 ml) at 800 for 12 hours.

Work-up as in Example 1 gave the corresponding quaternised polymer as off-white resin beads (9.78 g) (exchange capacity 1.93 m.eq. C1l/g, 1 molar percent).

r !l i :i

,P

'NYO 89/05828 PCT/GB88/01140 31 Example A A liquid formulation for oral administration prepared from the following: Compound of Example 12 Avicel RC591 Antifoam emulsion Flavours Sodium saccharide Preservatives Methyl Parabenz Propyl Parabenz Sorbitol syrup (70%) Glycerin Water (w:v) 1.25% 0.05% 0.02% 0.01% 0.12% 0.04% to 100% Example B A capsule formulation for oral administration is prepared by incorporating the following into a soft gelatin capsule Compound of Example 12 (500 mg), Aerosil 200 (5 mg), Magnesium Stearate (5 mg), Avicel PH101 (40 mg), and, optionally, sodium starch glycollate (10 mg).

Example C A food additive formulation for example a sachet for reconstitution or mixing with food, is prepared by incorporating into a powder formulation, compound of Example 12 (2500 mg), sodium carboxymethylcellulose mg), sucrose (2400 mg) and flavours (50 mg).

WO 89/05828 PCT/GB88/01140 -32 Biological Data In the binding assays the polymers used contained 5-15% H20 w/w. and all weights used in the assays refer to these undried materials.

Eouilibrium binding of Bile Acids (as sodium salts) to resins The compounds of the examples (10 mg/tube) were incubated with a range of C radiolabelled sodium taurochenodeoxycholate (TCDC) or sodium glycocholate (GC) solutions (0.05 mM-24 mM) of known specific activity, in Krebs Henseleit buffer (2.5 ml final volume). The resin was allowed to equilibrate with the bile salt solution for a period of three hours at 370 in a shaking water bath.

The resin was removed from the incubation medium by centrifugation (15 minutes at 3,000 The radioactivity of the supernatant was determined, and from this the free and bound quantities of bile salt were calculated. These were expressed as moles of bile salt bound per gram of resin.

Results The compounds of examples 1-32 at a concentration of 5.7mM GC were found to bind in the range of from 0.63 to 1.24 mmoles GC per gram weight of resin; and at a concentration of 5.7mM TCDC were found to bind in the range of from 0.53 to 1.34mmoles TCDC per gram weight of resin.

i i, .L ~YYLI-LI~ SWO 89/05828 PCT/GB88/01140 33 In vitro Dissociation assays.

In the method of Table I of test compound (20 mg) was shaken in Krebs' buffer for 3 hours in the presence of 14 sodium C] glycocholate (5 ml). In the method of Table 2, test compound (150 mg) was equilibrated with 5 mM sodium glycocholate (30 ml) in the same buffer. The compound was separated by centrifugation and the total bound determined by subtraction of the amount in the supernatant from the total bile acid used. Dissociation was measured either by continuous perfusion of the preloaded compound with Krebs' buffer and determination of the radioactivity in the perfusate (Table 1) or by resuspending the compound in Krebs' buffer, shaking and sampling the mixture through a filter at several time points up to 20 minutes.

Radioactivity and hence bile acid dissociated was determined in the filtrate (Table 2).

Table 1 In vitro dissociation continuous.

Example GC Bound (mmoles/g) t=0 t=2h Cholestyramine 0.86 0.11 1 0.94 0.31 2 0.75 0.34 3 0.92 0.28 4 0.94 0.26 0.92 0.22 6 0.95 0.38 7 1.02 0.38 8 0.92 0.25 9 0.95 0.24 0.98 0.34 11 0.83 0.47 12 0.78 0.53 13 0.94 0.49 14 0.95 0.21 0.95 0.37 WO 89/05828 -34 Table 2 In vitro dissociation equilibrium.

PCT/GB88/0 1140 Example GC Bound (mmoles/g) t=z0 t=2min Cholestyramine 9 210 112 12 1.6 2.8 22.

22 23 24 26 2 7 31 0.76 0.76 0.80 0.83 0.82 0.82.

0.62.

0.63 0.77 0.74 0.74 0.78 0.77 0.72 0.73 0.42 0.71 0.74 0.76 0.76 0. 0.74 0.55 0.75 0.69 0.71 0.71 0.73 0.68 0.58

Claims (12)

1-4 X is a physiologically acceptable counter ion; a, b and c are numbers which indicate the relative molar percentages of the units present in a random distribution in said polymer, being from about 0.5 to about 10 molar percent, and being from about 30 to about 99 molar percent; and n is a number indicating the number of repeating units in said polymer.

2. A polystyrene polymer as claimed in claim in 12 which R1 is a saturated C 8 to C 14 alkyl group, and R and R are each methyl. 20 polymer compris (a)r I WO 89/05828 PCT/GB88/01140 36

3. A polystyrene polymer as claimed in claim 2 in which is from about 1 to about 4 molar percent of said polymer.

4. A polystyrene polymer as claimed in claim 1 which is N,N,dimethyl-N-nonylammoniomethyl-substituted polystyrene, chloride salt N,N,dimethyl-N-octylammoniomethyl-substituted polystyrene, chloride salt N,N,dimethyl-N-decylammoniomethyl-substituted polystyrene, chloride salt NN,Ndimethyl-N-undecylammoniomethyl-substituted polystyrene, chloride salt, or N,N,dimethyl-N-dodecylammoniomethyl-substituted polystyrene, chloride salt. A process for the preparation of a polystyrene polymer as claimed in any one of claims 1 to 4 which comprises reaction of a polymer of structure (II) -(CH 2 CH) -(CH 2 CH)b-(CH CH) c 6 0J n -(CHCH2 )b Y WO 89/05828 PCT/GB88/01140 37 in which a, b, c and n are as described in claim and Y is a group displaceable by an amine, with an amine of structure R R R N (III) in which R to R are as described for structure or reaction of a compound of structure (IV) -(CH 2 CH) -(CH 2 CH)b-(CH 2 CH)- I (IV) n -(CHCH 2 )b Z in which a, b, c and n are as described for structure (I) and Z is NR R or NR R 3 in which R to R are as described for structure with a compound of structure R 4 Y in which R 4 is a C alkyl or a saturated or unsaturated 1-4 C 6 20 alkyl group and Y is a group displaceable by an amine.

6. A pharmaceutical composition comprising a polystyrene polymer of structure as claimed in claim 1 in association with a pharmaceutically acceptable carrier.

7. A polystyrene polymer as claimed in claim 1 for use as a therapeutic agent.

8. A polystyrene polymer as claimed in claim 1 for use in the lowering of serum cholesterol levels. -tr-0 k.40 Ul) was added keeping WO 89/05828 PCT/GB88/01 14Q

9. A polystyrene polymer Of structure (Iv -(CH 2 CH) (CH 2 CH)b-(CH 2 CH) c I (IVA) in wh a, b c ad n are as described forsrctr I an Z a NR or NR R3 whichRl and R in~~e i n 2 m1 V Q J Z)A41 0 ON i i i i i;_il 38 9. A polystyrene polymer of structure (IVA) H)CH 2 CH) a-(CH 2 CH) b (CHC H) c n -(CHCH 2 A in which a, b, c and n are as described for structure (I) and Z A is NR R 2 or NR2R 3 in which R 1 and R 2 are as described for structure in claim 1. 0*

10. Polystyrene polymers of structure processes for preparing them and pharmaceutical compositions containing them substantially as hereinbefore described with reference to any one of the Examples. Dated this 7th day of December, 1990 SMITH KLINE FRENCH .**LABORATORIES LIMITED By Its Patent Attorneys DAVIES COLLISON *0 0 INTERNATIONAL SEARCH REPORT International Application No PCT/GB 88/01140 I. CLASSIFICATION OF SUBJECT MATTER (II several classification symbols apply, indicate all) According to International Patent Classification (IPC).or to both National Classification and IPC IPC 4 C 08 F 8/44; C 08 F 8/32; A 61 K 31/745 II, FIELDS SEARCHED Minimum Documentation Searched 7 Classification System Classification Symools IPC C 08 F; A 61 K Documentation Searched other than Minimum Documentation to the Extent that such Documents are Included in the Fields Searched III. DOCUMENTS CONSIDERED TO BE RELEVANT' Category' I Citation of Document, with indication, where appropriate, of the relevant passages I2 Relevant to Claim No X FR, A, 2352547 (ETABLISSEMENT VIRIDIS) 1-9 23 December 1977 see claims 1-5; page 2, line 24 page 6, line X WO, A, 83/01002 (DYNAPOL SHAREHOLDERS' 1-9 LIQUIDATIONS TRUST) 31 March 1983 see claims 1-9; page 9, line 5 page 11, lin.e X GB, A, 2134529 (CIBA-GEIGY) 1-9 August 1984 see claims 1-11 A GB, A, 857193 (CLINICAL PRODUCTS) 1 29 December 1960 see claims 1-12 A FR, A, 988486 (RESINOUS PRODUCTS 1 CHEMICAL CO.) 28 August 1951 see summary Special categories of cited documents: 1o later document published after the international filing date d men defining the ener ateo the art which is not or priority date and not n conflict with the application but document defining the general ate o the art which i not cited to understand the principle or theory underlying the considered to be of particular relevance invention earlier document but published on or after the International document of particular relevance; the claimed invention filing date cannot be considered novel or cannot be consiaered to document which may throw doubts on priority claim(s) or involve an inventive step which is cited to establish the publication date of another document of particular relevance; the claimed invention citation or other special reason (as specified) cannot be considered to involve an inventive step when the document referring to an oral disclosure, use, exhibition or document is combined with one or more other sucn docu- other means ments, such combination being obvious to a person skilled document published prior to the international filing date but in the art. later than the priority date claimed document member of the same patent family IV. CERTIFICATION Date of the Actual Completion of the Intarnational Search Date of Mailing of this International Search Report 2 0. 04, 83 3rd April International Searching Authority Si tulan 6oTAuthorized l ler EUROPEAN PATENT OFFICE DER PUTEN Form PCT/ISA/210 (second sheet) (January 1985) ANNEX TO THE INTERNATIONAL SEARCH REPORT ON INTERNATIONAL PATENT APPLICATION NO. GB 8801140 SA 26235 This annex lists the patent family members relating to the patent documents cited in the above-mentioned international search report. The members are as contained in the European Patent Office EDP file on 17/04/89 The European Patent Office is in no way liable for these particulars which are merely given for the purpose of information. Patent document Publication Patent family Publication cited in search report date member(s) date FR-A- 2352547 23-12-77 BE-A- NL-A- DE-A,C JP-A- US-A- GB-A- CA-A- 855044 7705746 2723638 53010386 4198395 1579490 1104775

16-09-77

29-11-77 08-12-77

30-01-78 15-04-80 19-11-80 14-07-81 WO-A- 8301002 31-03-83 EP-A- 0087473 07-09-83 US-A- 4482680 13-11-84 US-A- 4532128 30-07-85 GB-A- 2134529 15-08-84 BE-A- AU-A- SE-A- DE-A,C JP-A- NL-A- EP-A,B FR-A,B LU-A- US-A- CH-B- OA-A- CA-A- AU-B- DE-A- SE-B- 898649 2323084 8400098 3400491 59134759 8400098 0122219 2542735 85171 4510128 655507 7628 1218077 570230 3474152 457959 11-07-84 19-07-84 13-07-84 12-07-84 02-08-84 01-08-84 17-10-84 21-09-84 24-10-84 09-04-85 30-04-86 23-05-85 17-02-87 10-03-88 27-10-88 13-02-89 GB-A- 857193 None FR-A- 988486 None For more details about this annex see Official Journal of the European Patent Office, No. 1282 For more details about this annex see Official Journal of the European Patent Office, No. 12/82 L, I-