NZ334301A - Polycarboxylic based cross-linked copolymers and pharmaceutical compositions comprising the co-polymer as an inert support or excipient - Google Patents

Abstract

Cross-linked copolymers based on non cross-linked polycarboxylic polymers and a cross-linking agent comprising at least two amine functions. The copolymers comprise at least one polycarboxylic polysaccharide and at least one other non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide, and is preferably a polycarboxylic acrylic polymer. The cross-linking agent is chosen from diamines, natural or synthetic amino acids or polyamines. Also disclosed are pharmaceutical compositions containing the above copolymer as an inert support or excipient.

Description

% Intellectual Property Office of New Zealand Page: 1 of 1 IP Summary Report Date: 26 2000 Time: 12:04:41 (iprip02 2.00.21) Accepted Version number: 3 IP type: Patent PCT Inward (51) Classification: IPC Edition: IPC Status: 70 334301 A61K9/20, C08B37/06, CHent Ref.

C08B37/08, 10301 C08B37/10, C08K5/17, C08L33/06, C08L5/08 (86) International Application number: FR97/01534 Date actions completed: (87) WO Publication number: 98/08897 Application Accepted 26 May 2000 Elected: N Next renewal date: 29 August 200* ;(22) NZ Filing date: 29 August 1997 Date entered National phase: 22 February 1999 (30) Priority Data: (31)96 9610601 (32) 30 August 1996 (33) ;FR ;(71) Applicant: SOCIETE DE CONSEILS DE RECHERCHES ET ;□'APPLICATIONS SCIENTIFIQUES SCRAS, ;51/53 rue du Docteur Blanche, F-75016, Paris, ;France ;(72) Inventors: El Matni, Nada ;Labarre, Denis Fessim, Hatem Contact: P L BERRY & ASSOCIATES, A.E.Q Building, 61 Cambridge Terrace, Christchurch 1, New Zealand ;Prirr.ary Examiner: SARAH WONG ;Journal: 1452 I ;Office title: Polycarboxylic based cross-linked copolymers and pharmaceutical compositions comprising the co-polymer as an inert support or excipient (54) Applicant title: Polycarboxylic based cross-linked copolymers (57) Abstract: ;Patent 334301 ;Cross-linked copolymers based on non cross-linked polycarboxylic polymers and a cross-linking agent comprising at least two amine functions. The copolymers comprise at least one polycarboxylic polysaccharide and at least one other non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide, and is preferably a polycarboxylic acrylic polymer. The cross-linking agent is chosen 'rom diamines, natural or synthetic amino acids or polyamines. ;Also disclosed are pharmaceutical compositions containing the above copolymer as an inert support or excipient. _ ;" End of report* LLLECTUAL PROPERTY QFFIG OF N.Z. ' 2 r:3 1999 CROSS-LINKED COPOLYMERS ^ BASED ON POLY CARBOXYLIC POLYMERS The invent ion relates lo cross-linked copolymers based on non cross-linked polycarboxylic polymers, said copolymers containing at least one polycarboxylic polysaccharide. The invention also relates to a process for the preparation of these copolymers and their use in particular as a support in pharmaceutical compositions.

Certain compounds with a polymeric structure containing a polycarboxylic polysaccharide, optionally modified, have been described in the literature. For example. Patent Application W089/02445 describes a gel based on hyaluronic acid: but, in its structure, this gel only comprises hyaluronic acid and no other polycarboxylic polymer. Moreover, no cross-linking agent is used in the preparation of this gel. The 10 compound obtained in this way is mainly used in surgery. Patent Application W091/16881 describes, among others, the combination of an active ingredient with a matrix constituted by a modified polymer, i.e. to which saccharides are grafted. This modified polymer can be a natural polymer such as chondroitin sulphate. However, this matrix contains only one type of polymer.

The copolymers according to the invention based on polycarboxylic polymers contain at least one polycarboxylic polysaccharide and at least one other polycarboxyclic polymer which is not a polysaccharide. The combination of a polysaccharide with another type of polycarboxylic polymer allows the modulation of the properties of the polysaccharides such as the hydrophilicity. In this way, copolymers can be obtained 20 with appropriate degradation properties according to their uses. Moreover, the copolymers according to the invention are advantageously prepared in an aqueous medium. This is a real advantage as it is almost impossible to totally eliminate the 33 A 30 1 solvents in a polymer structure: the existence of traces of residual aqueous solvents is generally more easily acceptable and accepted than traces of residual organic solvents such as dimethylsulphoxide or dimethylformamide.

The present invention provides cross-linked copolymers based on non cross-linked polycarboxylic polymers and a cross-linking agent comprising at least two amine functions, said copolymers comprising at least one polycarboxylic polysaccharide and at least one other non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide.

The non cross-linked polycarboxylic polysaccharides can be chosen, for example, from glycosaminoglycans, pectinic acid, alginic acid, carboxylic derivatives of dextran such as carboxymethyldextrans, or the carboxylic derivatives of cellulose such as carboxymethylcelluloses. Among the glycosaminoglycans, there can be mentioned hyaluronic acid, chondroitin sulphate, heparin, dermatan sulphate, heparan sulphate, keratan sulphate or a mixture of the latter. Among the polycarboxylic polymers which are not polysaccharides, there can be mentioned poly(glutamic acid), poly(aspartic acid), poly(maleic acid), poly(malic acid) or poly(furmaric acid), the polycarboxylic acrylic polymers such as poly(acrylic acid), poly(methacrylic acid) or the copolymers of the latter such as the Eudragits© L and S. It will be appreciated that a partly or totally substituted derivative of the aforesaid polycarboxylic polymers may be selected, for example an ester, amide or salt thereof, to prepare co-polymers in accordance with the present invention.

One aspect of the present invention is cross-linked copolymers as defined above, characterized in that the polysaccharide is chosen from pectinic or alginic acid, glycosaminoglycans, and preferably hyaluronic acid, chondroitin sulphate, heparin, dermatan sulphate, heparan sulphate, keratan sulphate or a mixture of the latter. 4 00 ^1" o A second aspect of the present invention is cross-linked copolymers as defined above, characterized in that the non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide is chosen from polycarboxylic acrylic polymers, 5 poly(glutamic acid), poly(aspartic acid), poly(maleic acid), poly(malic acid) or poly(fumaric acid). The non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide is preferably a polycarboxylic acrylic polymer and more particularly poly(acrylic acid) or poly(methacrylic acid).

The polycarboxylic polymers according to the invention are linked together by a cross-10 linking agent. This cross-linking agent comprises at least two amine functions which are capable of reacting with the free carboxylic functions of said non cross-linked carboxylic polymers. It can be chosen, for example, from proteins, polyamines, triamines, diamines, natural or synthetic amino acids, or the derivatives of compounds as defined above such as, for example, their salts, esters or amides. Among the 15 amino acids there can be mentioned, for example, arginine, lysine, histidine and ornithine. Among the diamines there can be mentioned ethylenediamine, butanediamine, hexanediamine, heptanediamine, octanediamine or dodecanediamine. Among the polyamines there can be mentioned chitosan, poly(amino acids) such as polylysine or polyornithine, as well as the copolymers of these polyamines. The cross-20 linking agent can also be chosen from compounds such as spermine, spermidine, melamine, guanidine or diethylenetriamine. The cross-linking agent used is preferably an amino acid and advantageously lysine, ornithine or histidine.

A third aspect of the present invention is cross-linked copolymers as defined above, characterized in that the polycarboxylic polysaccharide is a polycarboxylic 25 polysaccharide which can be degraded by the microbial flora of the colon such as chonodroitin sulphate, hyaluronic acid, pectinic acid or heparin.

A fourth aspect of the present invention is cross-linked copolymers as defined above, characterized in that the polycarboxylic polysaccharide is chondroitin sulphate and the I I '! .j ,0 -;ji other said polcarbo.xylic polymer is chosen from polv(acrylic acidl and poly(mothacrylic acid), and the cross-linking agent is lysine or histidine.

I The present invention further provides- a process lor the preparation of cross-linked copolymers as defined above, said process characterized in that said non cross-linked 5 polycarboxylic polymers constituting the cross-linked copolymer are reacted in the presence of an activator and a cross-linking agent comprising at least two amine functions, in an appropriate reaction medium. The preparation of cross-linked copolymers as defined above is preferably carried out in an aqueous medium. The expression aqueous medium means a medium only containing water or water mixed 10 with one or more solvents which are miscible with water such as. for example, acetone or lower alcohols such as ethanol. The aqueous medium preferably only comprises water. The implementation of the process according to the invention can be carried out in various manners. In fact, the process may consist in mixing non cross-linked polycarboxylic polymers and the cross-linking agent, then adding the activator. The 15 cross-linking process according to the invention can also consist in mixing together non cross-linked polycarboxylic polymers and the activator, then adding the cross-linking agent. The process may also consist in cross-linking ohe of the non cross-linked polycarboxylic polymers constituting the copolymer, mixing said polymer with the cross-linking agent then the activator, or with the activator then the cross-linking agent, 20 then adding at least one other non cross-linked polycarboxylic polymer to the reaction medium, in order to cross-link it with said polymer present in the reaction mixture. During the implementation of the process, the reagents introduced can previously be solubilized in the chosen reaction medium. The non cross-linked polycarboxylic polymers and the cross-linking agent are preferably mixed together in an aqueous 25 medium until solubilization then the activator is added. The process is implemented at a temperature comprised between -30 and 100°C, preferably between 0 and 40 °C and most preferably at ambient temperature. The implementation temperature for the cross- linking process is of course lower llian Ihe degradation or decomposition temperatures of the reagents introduced.

The relative proportions of the reagents constituted by the non cross-linkcd polycarboxylic polymers, the cross-linking agent and the activator can vary according 5 to the characteristics of the sought copolymers. The proportions of the non cross-linked polycarboxylic polymers are defined with respect lo the molar quantities of the carboxylic functions present per base unit. The non cross-linked polycarboxylic polymers can vary within a molar ratio comprised between 0.01 and 100. The molar ratio of the cross-linking agent with respect to the total carboxylic functions can vary 10 from 0 01 to 100. The molar ratio of the activator with respect to the total carboxylic functions can vary from 0.01 to 100.

The activator can be chosen from coupling agents in standard use in peptide synthesis. The activator can thus be chosen, for example, from carbodiimides, quinoline derivatives or mixed anhydrides. As examples of carbodiimides, there can be 15 mentioned hydrohalides such as N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC), N-cyclohexyl-N'-(2-morpholinoethyl) carbodiimide (CMC). As examples of quinoline derivatives, there can be mentioned 2-ethoxy-N-ethoxycarbonyl-1.2- dihydroquinoline (EEDQ), N-isobutoxycarbonyl-2-isobutoxy-l,2-dihydroquinoiine (IIDQ), N-isobutoxycarbony l-2-me!hoxy-1.2-dihydroquinoIine (IMDQ), N-isobutoxycarbonyl-2-ethoxy-l,2-dihydroquinoline (IEDQ). As examples of mixed anhydrides, there can be mentioned chloroformates and more particularly isobutylchloroformate (1BC). The activator used is preferably N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride.

The cross-linked copolymers according to the invention can be used, for example, in the 25 pharmaceutical, cosmetic, biomedical, veterinary, chemical, agro-chemical or agro-alimentary fields.

INTELLECTUAL PROPERTY OFFICE OF N.Z. 2 2 feb 1999 Intellectual mwtMi umui! OF N.Z. | 2 2 feb 1999 A more particular subject of the invention is a pharmaceutical comp( at least one active ingredient and. as an inert support or an excipient. at least one cross-linked copolymer according to the invention. The expression active ingredient designates any substance or mixture of substances having a therapeutic activity.

Such a composition can he produced from these different components by any standard technique known to a person skilled in the art. It can be presented, for example, in the form of matrix tablets, tablets coaled with the copolymers of the present invention, multi-layered tablets, matrix pellets, pellets or microparticles coated with the copolymers of the present invention. These microparticles and pellets may or may not 10 be contained in capsules. It can also be presented in the form of microparticles or nanoparticles at least one constituent of which is a copolymer of the present invention or else in any other form allowing oral administration. It can also be presented in any other form suited to the chosen or appropriate administration method such as suppositories or preparations for local application or injection. The quantity of active 15 ingredient allowing effective pharmacological action, in particular therapeutic action, can vary according to the type of active ingredient, the age and/or the illness of the patient to be treated.

A subject of the present invention is also the use of a pharmaceutical composition according to the invention for a sustained release of the active ingredient(s) it contains.

Such compositions can also possess other characteristics which optionally depend on the characteristics of the initial polycarboxylic polymers such as biointegration. Thus, a pharmaceutical composition according to the invention can also be used as a bioadhesive pharmaceutical system. A subject of the present invention is therefore also the use of a pharmaceutical composition according to the invention as a biointegration 25 system.

Compositions as defined above in which the polycarboxylic polysaccharide can be degraded by the flora of the colon can also be used as a specific release system at the INTELLECTUAL PROPERTY OFFIU OF N.Z. 2 2 feb 1999 specBiEiQ(iii.<lp4P ^ level of (he colon by I he aciion of l he microbial flora. The concept o (he level of ihc colon In the aciion of microbial flora is based on the properly of the colon to possess a very abundant microbial flora which, moreover, has the potential to metabolize substances which are slightly degraded or not degraded by the upper part of 5 the digestive tube. Such compositions are particularly suited lo conveying active ingredienis intended for the treatment of diseases of the colon, which allows their effectiveness to be increased and their side effects to be reduced. These activc ingredients include steroids such as dexamethasone and hydrocortisone, non-steroid anti-inflammatories such as 5-aminosalicylic acid, antineoplastics such as methotrexate. 10 tamoxifen, antispasmodics and chemotherapy agents. Such compositions are also particularly suited for conveying active ingredienis which are absorbed more efficiently at the level of the colon such as steroids or xanthine. Their direct administration at the level of the colon allows their effectiveness to be increased. Such compositions are also particularly suited to conveying active ingredients which are degraded in the upper parts 15 of (he digestive tube. Among these active ingredients, there can be mentioned peptides and proteins such as oral vaccines, insulin, contraceptive peptides, plasminogen activator peptides, growth peptides. LH/RH.

The following examples are presented in order to illustrate the above procedures and should in no event be considered as a restriction to the scope of the invention.

EXPERIMENTAL PART EXAMPLE 1 1.33 g of the sodium salt of chondroitin sulphate (A at 70%, C at 30 %) (CS), 0.29 g of the sodium salt of polymethacrylic acid (PMA) and 3.35 g of L-lysine monohydrochloride are mixed together in 9 ml of bidistilled water until a limpid 25 solution is obtained which is subsequently degassed. Then 4.59 g of N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) is added. The pH is maintained between 6 and 7 by successively adding 2.5 N hydrochloric acid. The • 8 - rcaciion is earned oui at amhieni temperature for 6 hours. Then the reaction medium is transferred to a dialysis apparatus (Speetra/por. cut-off threshold 12-14 KD) and dialysed 4 times against 4 litres of water each lime. The precipitate obtained in this way is washed with waier then dried. The sought chondroitin sulphate and 5 polymethacrylic acid copolymer is obtained with an average mass of 1.53 ± 0.12 g. The use of sulphur as a marker for chondroitin sulphate allows the definition by elementary analysis of the percentage by mass of chondroitin sulphate in the copolymer which is equal to 59 ± 2 %.

EXAMPLE 2 The operation is carried out in the same manner as in Example 1. but using 1.77 g of L-lysine monohydrochloride and 2.76 g of EDC. The mass of the copolymer obtained is 1.06 ± 0.15 g: the percentage by mass of CS in the precipitate is 55 ± 2.

EXAMPLE 3 The operation is carried out in the same manner as in Example 1. but using 7.06 g of L-15 lysine monohydrochloride and 8.21 g of EDC. The mass of the copolymer obtained is 1.61 ± 0.12 g; the percentage by mass of CS in the precipitate is 61 ± 1.

EXAMPLE 4 The operation is carried out in the same manner as in Example 1 but using 3 g of histidine instead of L-Lysine. The mass of the copolymer obtained is 1.94 ± 0.01 g; the 20 percentage by mass of CS in the precipitate is 48 ± 3.

EXAMPLE 5 The operation is carried out in the same manner as in Example 1. but using 0.43 g of PMA, 4.5 g of L-lysine monohydrochloride and 5.82 g of EDC. The mass of the copolymer obtained is 1.86 ± 0.05 g; the percentage by mass of CS in the precipitate is 25 58 ± 2.

INTELLECTUAL PROPERTY OFFICE OF N1. 2 2 feb 1999 Dtrrc ivrn . <■) - EXAMPLE 6 The operation is carried wui in I he same manner as in Example 1. but using 0.58 g of PMA. 5.45 g of L-lysine monohydrochloride and 7.05 g of EDC. The mass of the copolymer obtained is 2.07 ± 0.01 g: the percentage by mass of CS in the precipitate is 5 54 ± 2.

EXAMPLE 7 Tests on the .solubilization of the copolymer of Example 1 are carried out in the following solvents and mixtures of solvents: water at pH 3 and 7. acetonitrile. eihanol, tetrahydrofuran. dichloromethane, dimethylsulphoxide. dimethylacetamide. acetone. 10 dioxane. triethylamine. chloroform, petroleum ether, hexane. dimethylformamide. benzyl alochol. heptane, isopropyl alcohol. 1.2-propanediol. water / acetone mixture (50%/50%), water / ethanol mixture (50%/50%).

The copolymer is insoluble in all these solvents, which demonstrates its cross-linked character.

Study of the enzymatic degradation of copolymers 1 - spectroscopic study We are here studying the degradation of the copolymers of the invention based on chondroitin sulphate by chondroitinases, enzymes of the microbial flora of the colon.

Suspensions of the copolymers of Examples 1 to 6, in a buffer (acetate/tris/albumin) at pH 7,3. are prepared and agitated for a few hours in order to stablize them. The suspensions contain 67 mg of CS/ml of buffer. A solution of chondroitinases is added at a rate of 3.10"^ EU (Enzymatic Unit) for each mg of CS contained in the suspension.

The mixture is incubated at 37°C. At determined times, a suspension is centrifuged at 4°C then filtered. A study of the UV absorbance of the supernatant is carried out. The disaccharides originating from the degradation of the CS have a maximum absorption at 230-240 nm (Yamagata, T.et a\.,J.Biol.Chem., 243(7): 1523-1535(1968): Salyers, A. et INTFI \ Fr.TUAl PROPERTY OFFtUh OF N.Z. 2 2 feb 1999 INTELLECTUAL PROPERTY Ui I OF N.Z. 2 2 feb 1999 received ;il.. J.Back'rio/.. 143(2): 772 780)). The control is a solution of non cross-linked CS prepared under the same operating conditions as above.

The kinetics of the appearance in solution of the disaccharides originating from the degradation of the non cross linked CS and the copolymer obtained in Example 1 are 5 shown in Figure 1 below.

These results show thai (he copolymer of Example 1 is degraded by (he enzymes. Comparison of the degradation of the copolymer of Example 1 with that of the control shows that the copolymer, although cross-linked, is rapidly degraded by the enzymes.

The same tests are carried out on the copolymers of Examples 2 to 6; the results show 10 that these copolymers containing the CS are degraded by the chondroitinases. 2- rheological study The enzymatic degradation of the copolymers leads to the appearance of molecular chains of smaller sizes and should therefore lead to a reduction in the viscosity of the medium in which they are suspended.

A suspension of the copolymer of Example 1 in the buffer mixture (tris/aceiate/albumin) is prepared under the same operating conditions as those used in the spectroscopic study as presented above. Then, 4 ml of suspension is incubated in the cylinder of the viscometer (Haake RS100) maintained at 37°C. Measurement of the initial viscosity (h) is carried out. Then. 0.8 EU of chondroitinases dissolved in 160 ml 20 of water are added to the suspension. The control is a suspension of the copolymer of Example 1 prepared under the operating conditions previously described, without adding any enzyme and diluted in 160 ml of waier. The evolution of the viscosity is monitored over time. The experiment is carried out twice for each test.

Figure 2 is a semi-logarithmic illustration of the evolution of the viscosity of the 25 suspension of the copolymer of Example 1 in the presence of enzymes (continuous line) or in the absence of en/vmes (dotted control line). The viscosity of the control, which is of the order of 17± 3 niPn.s. does not vary over lime. On the other hand, in the presence of en/vmes. the viscosity progressively drops from 17 mPa.s to 3 mPa.s over 55 minutes then becomes quasi-stable. This significant drop in viscosity is explained by the degradation of the copolymer by the enzymes.

Moreover, following the above spectroscopic and rheological studies carried out under the same operating conditions, there can be observed, alter incubation in the presence of enzymes for 55 minutes, a virtually total drop in viscosity although only part of the disaccharides originating from the the degradation of the CS which is detected in solution. The degradation of a few sites of the copolymer by the enzymes is sufficient to entail a collapse of the three-dimensional network of the copolymer.

Study of sustained release tablets The cross-linked copolymers of Examples 1 to 6 are sieved then mixed with aminosalicylic acid (5ASA) and magnesium stearate (mass ratio 79.5/20/0.5). Then 250 mg tablets of hardness > 100 N are prepared by direct compression.

Dissolution tests are carried out on the tablets prepared in this way. in a device with a rotating vane (DISSOLUTEST) at 37°C under agitation at 50 revolutions/minute. The dissolution media used are a buffer mixture of pH 1.2 and 7.5 respectively corresponding to the artificial gastric and intestinal media (without enzymes). For each formula and in each medium, the test is carried out three times. At determined times, a sample of the dissolution medium is taken and filtered. Dosage of the 5ASA is carried out by UV spectroscopy.

Table 1 below summarizes the time (in hours) taken to release 50% of the initial dose of 5ASA (t50%). obtained in artificial gastric and intestinal media.

INTELLECTUAL PROPERTY UFNOt OF N.Z. 2 2 feb 1999 recf'ved_ Table 1 Excmplc 150% 150% (gastric medium) (intestinal medium) 1 2.88 7.66 2 1.42 1.61 3 6.48 8.29 4 1.22 1.59 7.94 8.65 6 7.96 11.05 In a gastric medium, the t50%'s vary from 1.2 to 8 hours thus allowing the release of the active ingredient to be modulated according to the type of copolymer. Among these copolymers, the copolymers of Examples 3, 5 and 6, respectively having 150%'s of 6.5, 7.9 and 8 hours, significantly moderate the release of the active ingredient.

In an intestinal medium, the t50%'s vary from 1.6 to 11 hours, also allowing modulation of the release of the active ingredient according to the type of copolymers. Moreover, a significant moderation of the release of the active ingredient is obtained with the copolymers of Examples 1. 3. 5 and 6. In fact, the t50%'s obtained with these copolymers are 7.7, 8.3, 8.7 and 11 hours respectively.

The synthesized copolymers therefore allow the creation of sustained release pharmaceutical systems according to the characteristics of the cross-linked copolymers. More particularly, those which possess the property of significantly moderating the release of the active ingredient and being degradable by chondroitinases appear to be 15 useful candidates for creating sustained release systems at the level of the colon by action of the microbial flora.

INTELLECTUAL PROPERTY OFFICl OF N.Z. 2 2 feb 1s39 ____r f ^1 - I.? -

Claims (3)

1. CLAIMS 1- Cross-linked copolymers based on non cross-linked polycarboxylic polymers and a cross-linking ageni comprising ai leasi two amine functions, said copolymers comprising at least one pol\carboxylic polysaccharide and at least one other non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide. 5 2- Copolymers according to claim I. characterized in that the polycarboxylic polysaccharide is chosen from glycosaminoglycans. pectinic or alginic acid. 3- Copolymers according to one of claims 1 to 2, characterized in that the polycarboxylic polysaccharide is a glycosaminoglycan chosen from hyaluronic acid, chondroitin sulphate, heparin, dermatan sulphate, heparan sulphate, keratan sulphate. 10 4- Copolymers according to one of claims 1 to 3. characterized in that the non cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide is chosen from polycarboxylic acrylic polymers, poly(glutamic acid), poly(aspartic acid), poly(maleic acid), poly(malic acid) or poly(fumaric acid). 5- Copolymers according to any one of claims 1 to 4, characterized in that the non 15 cross-linked polycarboxylic polymer which is not a polycarboxylic polysaccharide is a polycarboxylic acrylic polymer. 6- Copolymers according to claim 5, characterized in that the polycarboxylic acrylic polymer is poly (aery I ic acid) or poly(methacrylic acid). 7- Copolymers according to any one of claims 1 to 6, in which the cross-linking agent is 20 chosen from diamines, natural or synthetic amino acids or polyamines. INTELLECTUAL PROPERTY OFFICE OF N.Z. 2 2 feb 1599 u 8- Copolymers according lo claim 7 in which ihe amino acid is chosen from lysine, hislidine or ornithine. 9- Copolymers according io claim 7 in which the diamine is chosen from ethylenediamine, bulanediamine. hexanediamine. heptanediamine. ociancdiaminc and 5 dodecanediamine. 10- Copolymers according lo claim 7 in which the polyamine is chosen from chitosan. polyornithine or poly lysine. 11- Copolymers according to one of claims 1 to 10, characterized in that the polycarboxylic polysaccharide can be degraded by the flora of the colon. 10 12- Copolymers according to claim 11. characterized in that the polycarboxylic polysaccharide is chosen from chondroitin sulphate, hyaluronic acid, pectinic acid or heparin. 13- Copolymers according to any one of claims 1 to 8. 11 and 1

2. characterized in that the polycarboxylic polysaccharide is chondroitin sulphate, the other said polycarboxylic 15 polymer is poly (acrylic acid) or poly (methacry lie acid) and the cross-linking agent is lysine or histidine. 14- Process for the preparation of copolymers according to any one of claims 1 to 13, characterized in that said non cross-linked polycarboxylic polymers are reacted in an aqueous medium, in the presence of an activator and of said cross-linking agent. 20 15- Process according to claim 14, in which the activator is chosen from carbodiimides, quinoline derivatives and mixed anhydrides. 16- Pharmaceutical composition containing at least one active ingredient and, as an inert support or excipient, at least one copolymer according to one of claims 1 to 10. INTELLECTUAL PROPERTY OFFICE OF N.Z. 2 2 feb 1939 -15- ; «» " ^ v 17 - Pharmaceutical composition containing at least one active ingredient and as an inert support or excipient, at least one copolymer according to one of claims 11 to 1

3. 13 - A cross-linked copolymer based on non cross-linked polycarboxylic polymers and 5 a cross-linking agent comprising at least two amine functions substantially as hereinbefore described with reference to any one of Examples 1-7. 19 - A process for the preparation of copolymers in accordance with claim 1 substantially as hereinbefore described with reference to any one of Examples 1-7. 20 - A pharmaceutical composition containing at least one active ingredient and, as an 10 inert support or excipient, at least one copolymer according to claim 1, substantially as hereinbefore described with reference to any one of Examples 1-7. p.- n y f > ••• • L-. U v' *****