ZA200104298B

ZA200104298B - Method of producing a polymer network.

Info

Publication number: ZA200104298B
Application number: ZA200104298A
Authority: ZA
Inventors: Klaus Gottschall
Original assignee: Gottschall Instruction Ges Fue
Priority date: 1998-11-30
Filing date: 2001-05-25
Publication date: 2002-05-23
Also published as: ATE349471T1; EP1149118B1; JP2002531599A; JP4737832B2; ES2280127T3; DE59914107D1; DE19860972A1; JP2010059433A; CA2352066C; AU1655900A; WO2000032649A1; EP1149118A1; CA2352066A1

Abstract

The invention relates to a method of producing a derivative of a polymer having at least one functional group. Said method is characterized in that it comprises step (i): reacting the polymer having at least one functional group with at least one activating reagent or at least one derivative of an activating reagent in the homogeneous phase.

Description

EEN

‘0 20014290 *

Process for the preparation of a polymeric network

The present invention relates to a process for the preparation of a polymeric network, the polymeric network per se which is obtainable by the process according to the invention, and the use of the polymeric network in various industrial application areas.

Polymeric structures which have pores in which a given substrate can be selectively bonded are of interest in a whole series of industrial applications. In this connection, reference may be made to substance separation processes, catalytic processes or the use of these polymeric structures as sensors.

WO 93/09075 describes a process for the preparation of a polymeric structure, in which a polymer is prepared by free-radical polymerization from the monomers in the presence of a crosslinking reagent and simultaneously in the presence of a substrate. Imprinting polymers are proposed for use in chromatography, in catalysis, as biosensors or as synthetic antibodies. G. Wulff gives an overview in

Angew. Chem., Int. Ed. Engl. 34 (1995) 1812-1832.

The imprinting technique, however, has a number of disadvantages. Thus the imprints show an unacceptable peak broadening and, as a rule, incomplete substance elution in the chromatographic test, the latter leading to the cross- contamination of further product fractions. Industrial application of the imprints in chromatography is thus essentially excluded. Likewise, the imprints as a rule have a low loading ability with substrate, essentially in the per thousand range, as a result of which, obviously, an only extremely small quantitative throughput can be achieved and an industrial separation process could only be carried out uneconomically.

In J. Org. Chem. 63 (1998) 7578-7579, Menger et al. describe a process in which, starting from a combinatorial mixture, polyacrylic anhydride is reacted with three or four amines selected more or less at random, 14 different amine combinations in 96 different concentration ratios leading to 1344 different polymers. The polymers were employed in the catalytic dehydrogenation of a beta-hydroxyketone, where, however, less than 1 per cent of all polymers were suitable for the catalytic process at all on account of the reaction rate. In connection with the experiments, it was i ,

Lo 20014298

J

-_ 2 _ + observed that the polymers gradually develop a better catalytic activity in the presence of the substrate. However, the polymeric structures with improved activity were not stabilized, but labile structures were obtained such that the catalytically more active structures were destroyed by changes in the pH or the temperature.

It was therefore an object of the present invention to make available a process which does not have the disadvantages of the abovementioned processes.

Accordingly, the present invention relates to a process for the preparation of a polymeric network, in which 6) one or more polymers are made available which can be crosslinked with one another intramolecularly or intermolecularly or intra- and intermolecularly by covalent or non-covalent bonding, (i) the conformation of at least one of the polymers is adapted to at least one template compound with obtainment of at least one preferred conformation of the at least one polymer and (iii) at least one of the preferred conformations obtained according to (ii) is fixed by crosslinkage. :

The term “template compound”, as is used in the context of the present application, includes all compounds on which at least one conformation of the at least one polymer employed is adaptable. Thus possible compounds are all those which lead to a preferred conformation of the at least one polymer employed by interaction with the polymer. The interaction must in this case not take place with the polymer per se, but can also take place with a polymeric structure which is derived from the polymer employed, as is described further below.

Accordingly, template compounds which are possible are both chemical compounds and also biological structures, such as microorganisms, with respect to which, inter alia, e.g. pathogenic organisms, preferably viruses, bacteria or parasites may be mentioned. Likewise, for example, cells, fragments or constituents of cells, epitopes, antigenic determinants or receptors may be mentioned.

In the process according to the invention, the concentration of the template compound employed in solution or suspension is basically freely selectable.

" . ‘® 20014298 ra] BN 3 B = Preferably, the concentration is in the range from 0.25 to 300 mmol/l, based on the solvent or solvent mixture employed. The solvents in which the template is employed are likewise essentially freely selectable. Organic and aqueous solutions are preferred, preferred solvents being, inter alia, chlorohydrocarbons having up to three C atoms, nitriles such as acetonitrile, esters such as ethyl acetate, ketones such as methyl ethyl ketone or acetone, open-chain or cyclic ethers such as THF or dioxane or aromatic compounds such as toluene or xylenes or mixtures of two or more of these compounds. The pH range of the solutions is essentially freely selectable and can be coordinated with the polymers and the template compound.

Preferably, the pH range during fixing is in the range from 3 to 12, preferably in the range from 4 to 9 and particularly preferably in the range from 6 to 8.

The term “preferred conformation”, as is used in the context of the present invention, designates a conformation of the polymer which results from one or more steps of the process according to the invention, the interaction enthalpy between the preferred conformation and the template compound being greater in amount than the interaction enthalpy between the template compound and that conformation which the polymer has before this one or before this number of steps.

Preferably, this difference in amount in the interaction enthalpy is greater than 0.1 kcal/mol, particularly preferably than 1 kcal/mol and very particularly preferably greater than 3 kcal/mol.

As already indicated and described in detail further below, the polymeric networks prepared according to the invention can be employed, inter alia, in substance separation processes. It is obviously possible here that substances which are to be separated under the reaction conditions which occur during the preparation of the polymeric network are not stable or only inadequately stable or, for example, are not available. In this case, for example, compounds which are homologous to or structurally related to the substances to be separated, preferably isosteric, can be employed as template compounds.

The adaptation of the conformation can be carried out here, inter alia, by interaction of the polymer with the template compound. It is also possible, for example, that two or more polymer strands of polymers which are identical to or different from one another are crosslinked and the resulting polymeric structure interacts with the at least one template compound and thus the conformation of the

¢ . he a -¢ polymeric structure and thus also the conformation of the polymer employed is adapted to the template compound.

It is furthermore possible that, by intramolecular crosslinkage of a polymer strand, a conformation is formed which interacts with the at least one template compound, the conformation of the polymer being adapted to the at least one template compound. Embodiments in which such an intramolecular crosslinking takes place are possible in the context of the present invention in the case of all suitable polymers. Polymers are particularly preferred which have a molar mass of more than 10 000 g/mol, further preferably of more than 30 000 g/mol and particularly preferably of more than 100 000 g/mol.

Furthermore, the present invention also includes embodiments in which the adaptation of the conformation is carried out such that, in the absence of the at least one template compound, polymeric structures are synthesized by inter- or intramolecular crosslinking whose conformations are adapted to the at least one template compound by specific selection of the at least one polymer and/or of the at least one crosslinking reagent employed. In this case it is possible, for example, that the steps (ii) and (iii), as mentioned above, can be carried out in one step. In the case of crosslinkage in the absence of the at least one template compound, however, it is also possible that polymeric structures are synthesized which are roughly adapted to the at least one template compound, where a more accurate adaptation can be carried out by further crosslinking steps in the presence or absence of the at least one template compound.

Accordingly, all polymers which can be crosslinked intra- and/or intermolecularly and which can interact per se or after crosslinking with the at least one template compound can be employed in the process according to the invention.

In the context of the present invention, the term “interaction” is understood as meaning all suitable covalent and non-covalent interactions.

Possible interactions of the at least one polymer or one polymeric structure employed, which is synthesized, for example, by intra- or intermolecular crosslinking, as described above, with the at least one template compound are, inter alia: - hydrogen bonds;

he Ss - - - dipole-dipole interactions; - Van der Waals interactions; - hydrophobic interactions; - charge-transfer interactions, eg. T-x interaction; - ionic interactions; - coordinative bonding, e.g. to transition metals; - combinations of these interactions.

Obviously, covalent bonds between polymer and/or polymeric structure and the at least one template compound are possible. If the polymer network prepared in the process according to the invention is used in, for example, substance separation processes, interactions between polymer and/or polymeric structure and the at least one template compound are particularly preferred, by means of which the at least one template compound is reversibly bonded.

If the at least one polymer employed interacts per se with the at least one template compound, according to the process according to the invention it has at least one functional group by means of which this interaction can be formed. If the conformation of the polymer which is formed by interaction with the at least one template compound is fixed by crosslinkage, it is possible, inter alia, that the crosslinkage takes place via the functional group via which the interaction with the at least one template compound was formed. Preferably, the polymer has at least one further functional group via which the crosslinkage takes place.

The term “functional group”, as is used in the context of the present invention, accordingly includes all chemical structures via which covalent and/or non- covalent interactions can take place. In particular, hydrocarbon chains and further structural units via which Van der Waals interactions can be formed also come under the term of functional group.

A particularly suitable structure of the polymers employed is accordingly present if the functional groups in the polymer which are capable of interaction are able, according to type and/or number and/or density and/or distribution, to bond a specific template compound. Very particularly suitable polymers are those which are able to bond a specific template compound bi-, tri-, oligo- and/or polyvalently in more than one molecular position. Accordingly, two or more functional groups

; @® CL

A which are optionally spatially separated by at least one group which is inert to an interaction can be responsible for the interaction.

The term “in the polymer”, as is used in the context of the present invention, relates, inter alia, to polymers in which the at least one functional group which is used for the formation of the interaction with the substrates and/or for the crosslinking is present in the polymer strand. The term likewise relates to polymers in which the at least one functional group is present in at least one side chain of the polymer strand, and also to polymers in which at least one type of a functional group is present both in the polymer strand and in at least one side chain of the polymer strand.

Accordingly, in the process according to the invention generally both derivatized and non-derivatized polymers can be employed.

The at least one functional group which is needed in the polymer for the formation of the interaction with the at least one template compound and/or for crosslinkage can accordingly already be present in the original polymer and does not necessarily have to be introduced into the polymer by subsequent derivatization. Inter alia, for example, the amino or formyl groups in polyvinylamine or, for example, the hydroxyl or acetyl groups in polyvinyl alcohol may be mentioned by way of example here.

In the process according to the invention it is possible, inter alia, to “designate” the receptor-template interaction using made-to-measure receptor groups by derivatization of at least one polymer which is employed in the process in derivatized form. In the context of the present invention, the degree of derivatization can be influenced here such that the best possible interaction with the template is achieved. It is likewise possible to designate the adaptation of the conformation of the at least one polymer in the absence of the at least one template compound by specifically introducing certain crosslinking possibilities or interaction possibilities into the polymer, for example by means of the functional groups introduced by derivatization.

If one or more polymers are first derivatized in the process according to the invention and then employed in the process according to the invention, the

® a derivatization can take place according to all suitable processes, for example processes known from the prior art.

In order to equip polymers which have functional groups with receptor groups and to derivatize them in this way, three routes can be mentioned, inter alia, which are listed in M. Antonietti, S. Heinz, Nachr. Chem. Tech. Lab. 40 (1992) No. 3, pp. 308-314. According to this publication, derivatized polymers are obtainable by means of random polymerization or copolymerization, by means of the preparation of block copolymers and by means of the preparation of surface-functionalized polymer particles. These preparation routes start from derivatized monomers from which the polymer is obtained.

A further possibility of derivatizing polymers is the polymer-analogous reaction of polymers having functional groups with derivatizing compounds.

Polymer derivatizations are carried out, for example, on solid surfaces by heterogeneous reaction. This group includes, inter alia, carrier activation and carrier immobilization, in which a nucleophilic substance is customarily heterogeneously bonded to a polymer, e.g. epoxy polyacrylic ester or BrCN- sepharose, as is described, for example, in P. Mohr, M. Holtzhauer, G. Kaiser,

Immunosorption Techniques, Fundamentals and Applications, Akademie Verlag,

Berlin (1992), pp. 34-40.

In a preferred embodiment, in the process according to the invention (i) a derivatized polymer can be made available which is prepared by reacting a polymer having at least one functional group with at least one activating reagent or a derivative of an activating reagent, where this reaction can take place homogeneously or heterogeneously, preferably homogeneously.

As a rule, the activating reagent is in this case selected such that the at least one functional group of the polymer reacts with the activating reagent during the reaction and is thus improved in its reactivity in a subsequent reaction with a derivatizing agent.

Accordingly, the present invention also describes a process in which the reaction product from the polymer having at least one functional group and the activating reagent is reacted with a derivatizing reagent.

@® .

Aa

In the context of this embodiment of the process according to the invention, the polymer having at least one functional group can be reacted simultaneously, i.e. in the sense of a “one-pot reaction” with at least one activated and/or at least one non- activated derivatizing reagent and/or an activating reagent.

By means of this reaction of the activated polymer having at least one functional group with a derivatizing reagent, a desired radical can be introduced into the polymer.

If a polymer was reacted here with different activating reagents, these activated functional groups can have different reactivity to one or more derivatizing reagents. Accordingly, it is possible in the context of the process according to the invention to derivatize functional groups selectively in this manner. The term “selective” in this connection means that a polymer which has, for example, two or more functional groups which are different from one another is reacted with, for example, two different activating reagents such that a subsequent reaction with a derivatizing reagent for derivatization takes place mainly to exclusively on the activated functional group(s) which is or are activated with one of these two activating reagents, as a rule on the functional group(s) more reactively activated with respect to the derivatizing reagent.

In the process according to the invention, it is furthermore possible to react the activating reagent before the reaction with the polymer having at least one functional group in order then to react this reaction product with the polymer having at least one functional group.

The present invention therefore also describes a process, as described above, in which the derivative of the activating reagent is obtained by prior reaction of the activating reagent with a derivatizing reagent.

A further embodiment of the present invention consists in reacting the polymer having at least one functional group with various produts from reactions of activating reagents and derivatizing reagents. Thus, for example, a mixture of compounds can be reacted with the polymer, the mixture of reaction products comprising an activating reagent and two or more different derivatizing reagents.

A mixture is likewise possible that comprises reaction products of a derivatizing r . 15) ) “081429; _ reagent and two or more different activating reagents. Of course, it is also possible, should this be necessary, to employ a mixture which comprises reaction products of two or more different activating reagents and two or more different derivatizing reagents. Obviously, it is also possible in the context of the present invention 10 react the different reaction products of activating reagent and derivatizing reagent not as a mixture, but individually and in the desired sequence with the polymer having at least one functional group.

Accordingly, the present invention also describes a process as described above, in which the polymer having at least one functional group is reacted with at least two different derivatives of an activating reagent and the reactions are carried out successively with one derivative in each case.

Activating reagents which can be employed in principle are all activating reagents known from the literature. The article by P. Mohr, M. Holtzhauer, G. Kaiser already cited above, which in this respect is included completely by way of reference in the context of the present patent application, gives, for example, an overview on a whole series of activating reagents which can be employed for the activation of various functional groups.

In particular, chloroformic acid esters and chloroformic acid esters having electron-withdrawing radicals may be mentioned here.

In particular, the present invention describes a process in which the activating reagent is derived from a compound of the following structure (I):

R, 0

Lo OH 0)

Re 0} Ig where R; and R, are identical or different and can be straight-chain, branched- chain or bridged to give a carbocycle or a heterocycle and are selected such that the activating reagent or the derivative of the activating reagent can be reacted in homogeneous phase with the polymer having at least one functional group.

@® - Rj and R; here can be, for example, cycloalkyl, cycloalkenyl, alkyl, aryl or aralkyl radicals having up to 30 C atoms.

In a preferred embodiment, the present invention describes a process in which the activating reagent is derived from a compound of the following structure (I')

Ry = I Ry 0 ] N=0_ oH ty) a Rs 0 )g

Rg where Rj to Rj can be identical or different and can be hydrogen, straight-chain or branched-chain alkyl, aryl, cycloalkyl, heterocyclic and aralkyl radicals having up to 30 C atoms, or else two or more of R3 to Ro can in turn be bridged to give a carbocycle or heterocycle and are selected such that the activating reagent or the derivative of the activating reagent can be reacted in homogeneous phase with the polymer having at least one functional group.

The present invention further describes a process in which the activating reagent has the following structure (II)

Rs i Ry ©

Ry 7] N=O_ a (mn

Ryo WE I

Rg where Rj to Rg are as defined above.

In a likewise preferred embodiment, the present invention describes a process in which the activating reagent is derived from a compound of the structure (II), as indicated above, where R3 to Rg is in each case hydrogen.

The compounds having the structures (I), (I) and (II) can be prepared by all customary processes known from the prior art. Such a process for ONB-CI is given, for example, in P. Henklein et al, Z. Chem. 9 (1986), p. 329 ft.

@ ” Using the activating reagents or the derivatives of activating reagents as described above, all polymers which have at least one functional group which is reactive with respect to the activating reagents can in principle be reacted.

Very generally, polymers which have as at least one functional group a group which has at least one nucleophilic unit are employed in the process according to the invention.

Preferred functional groups of the polymer having at least one functional group which may be mentioned are, inter alia, OH groups, optionally substituted amine groups, SH groups, OSO3H groups, SOsH groups, OPO3H, groups, OPOsHR\; groups, PO3;H, groups, PO;HR;; groups, COOH groups and mixtures of two or more thereof, where Rj; in each case is selected such that the activating reagent or the derivative of the activating reagent can be reacted with the polymer having at least one functional group in homogeneous and/or heterogeneous phase. Likewise, the polymers having at least one functional group can also contain further polar groups, for example —CN.

Both natural and synthetic polymers can be employed as the polymer having at least one functional group. Possible restrictions in the selection of the polymers only result in that the reaction of the polymer is performed in homogeneous phase in the context of the process according to the invention and from the later intended use of the derivatized polymer.

In the context of this invention, the term “polymer” here obviously likewise includes higher molecular weight compounds which are designated in polymer chemistry as “oligomers”.

Without wishing to be restricted to certain polymers, the following may be mentioned, inter alia, as possible polymers having at least one functional group: - polysaccharides, e.g. cellulose, amylose and dextrans; - oligosaccharides, e.g. cyclodextrins; - chitosan; - polyvinyl alcohol, poly-Thr, poly-Ser; - polyethyleneimine, polyallylamine, polyvinylamine, polyvinylimidazole, polyaniline, polypyrrole, poly-Lys; - poly(meth)acrylic acid (esters), polyitaconic acid, poly-Asp;

® - 12 - - - poly-Cys.

Likewise, in principle not only homopolymers, but also copolymers and in particular block copolymers and random copolymers, are suitable to be employed in the present process. Here, both copolymers having non-functionalized components such as, for example, co-styrene or co-ethylene or alternatively copolymers such as, for example, co-pyrrolidone may be mentioned.

If the polymers in the process according to the invention are derivatized in homogeneous liquid phase, then, in order to achieve optimum solubility, preferably mixed-functional or alternatively prederivatized polymers are employed. Examples of these which may be mentioned are, for example: - partially or completely alkylated or acylated cellulose; - polyvinyl acetate/polyvinyl alcohol; - polyvinyl ether/polyvinyl alcohol, - N-butylpolyvinylamine/polyvinylamine.

Likewise, polymer/copolymer mixtures can also be used. All suitable polymer/copolymer mixtures can be employed here, for example mixtures of the polymers and copolymers already mentioned above, where, inter alia, the following, for example, are to be mentioned here: - poly(acrylic acid)/co-vinyl acetate; - polyvinyl alcohol/co-ethylene; - polyoxymethylene/co-ethylene; - modified polystyrenes, e.g. copolymers of styrene with (meth)acrylic acid (esters); ‘ - polyvinylpyrrolidone and its copolymers with poly(meth)acrylates.

All of these abovementioned polymers, which are accessible to derivatization, can obviously also be employed in underivatized form in the process according to the invention.

If, as described above, the polymer having at least one functional group is reacted with an activating reagent such as a compound of the structure (II), then, as likewise described above, this reaction product can be reacted with a derivatizing reagent.

@® - Here, in principle, all reagents which can react with the activated polymer and lead directly or indirectly to the desired derivatized polymer can be used. Inter alia, compounds which have at least one nucleophilic group are employed in the process according to the invention as derivatizing reagents.

For example, derivatizing reagents are used which have the general composition

HY-Rj,. Here, Y is, for example, O, NH, NR; or S, where Ry, and Rj3 can generally be freely selected. For example, they are an alkyl or aryl radical which is optionally suitably substituted.

In addition, it is also possible to react the activated polymer with nucleophilic chiral compounds. Examples of such chiral nucleophiles which may be mentioned are, for example: borneol, (-)-menthol, (-)-ephedrine, a-phenylethylamine, adrenaline, dopamine.

A further possibility is to react the activated polymer with a mono- or polyhydric alcohol or thiol containing an amino group in the process according to the invention. If the polymer comprising at least one functional group is activated, for example, with ONB-CI, the mono- or polyhydric alcohol containing the amino group or the mono- or polyhydric thiol containing the amino group reacts selectively with the amino group. The OH or SH groups thus introduced into the polymer can then be activated again in a further step with, for example, one of the activating reagents described above, whereby chain extensions and branchings are facilitated, depending on the functionality of the alcohols or thiols originally employed.

In another embodiment of the process according to the invention already described above, the polymer having at least one functional group is rcacted with an activated derivatizing reagent, the latter being obtained from the reaction of an activating reagent with the derivatizing reagent.

In the process according to the invention, activated derivatives of amines, alcohols, thiols, carboxylic acids, sulphonic acids, sulphates, phosphates or phosphonic acids are preferably reacted with the polymer having at least one functional group, where, in turn in a preferred embodiment, the compounds are activated with ONB-

CL i N a; “* «0014295 ) Inter alia, these activated derivatizing reagents which can be reacted with the polymer having at least one functional group thus have the following general structures (IIT) to (IX):

R.

Ry Ry 0

Rs 7 N=0_ _NRyH (Im

Rig Re bd

Rs

Re 0 0]

R

R; Ry 0

Ry

N=0_ oR; av)

Rig bd

Rs o 6

Re

R,

R, | Bs “7

N—O, SR \2

Rio RS bd

Ry

Re (0) 0]

EY: 7 H O

Ry

Bi N=0_ NN A (VD

YY

Ry

Re 0 O H

; Re

R, | Bs °

Ry

Bi N—O. 0. Rj (VID)

Ri XY

R | Re

O oO O

Rs

R, | Bs

Rg

B N=0_ Ry (vi

Rio RS bd

Re 0 ©

Re

Rs

R, | Rs QO

Rs

R 7] NTO (IX) 10 V/A

Rg Ry 0 0) Ryo

Rg where Rj to Ry are as defined above and Rj4 to Ryo are in general subject to no restrictions, for example can also have chirality, and in the process according to the invention are selected such that the reaction with the polymer having at least one functional group can be carried out in homogeneous phase. Here, the substituents

Ri4 to Ry as a rule are selected depending on the desired interaction with the subtrate. Here, R14 to Ryo can be identical or different and are radicals containing hydrogen, a straight-chain or branched-chain alkyl, aryl or aralkyl radical having upto 30 C atoms or corresponding heteroatoms.

Likewise, polyhydric amines, alcohols, thiols, carboxylic acids, sulphonic acids, sulphates, phosphates or phosphonic acids can be reacted with an activating reagent and this reaction product can be reacted with the polymer having at least one functional group, where here, in particular, polyols may be mentioned.

Obviously it is also possible to activate derivatizing reagents which have two or more different types of the abovementioned functional groups and to react them with the polymer having at least one functional group. Examples which may be mentioned here, inter alia, are, for example, aminoalcohols.

@ —_ 1 6 - 7 In the context of the present invention, such polyhydric derivatizing reagents can selectively be partially or completely activated using an activating reagent and reacted with the polymer having at least one functional group.

The reaction of the polymer having at least one functional group with an activated, polyhydric derivatizing reagent can also be used in the process according to the invention for polymer crosslinking and further for polymer stabilization and/or for polymer branching, in addition to the fact that a suitable polymer according to (i) is made available.

Both the reaction of the polymer having at least one functional group with an activated derivatizing reagent and the reaction of the polymer having at least one functional group with an activating reagent and subsequent reaction of the product with a derivatizing reagent by the process according to the invention make it possible to prepare polymer derivatives which have very different spatial arrangements and accordingly can be used for a large number of applications in which this spatial arrangement is of crucial importance.

Thus it is possible, for example, to realise arrangements which are constructed as hairy rods, comb polymers, nets, baskets, dishes, tubes, funnels or cages.

In a likewise preferred embodiment, the present invention describes a derivative of the type under discussion here, which has at least one receptor group which has a bonding unit decisive for the bonding of a biological or synthetic chemical substrate.

A made-to-measure derivative of this type for biological substrates then has corresponding receptor groups which have, for example, structures also occurring in nature or parts of structures of this type responsible for bonding, which can then interact with a biological substrate. Here in particular, for example, enzyme, amino acid, peptide, sugar, amino sugar, sugar acid and oligosaccharide groups or derivatives thereof may be mentioned. It is essential for the above receptor groups that the principle of bonding of a receptor with a substrate occurring in nature is exclusively retained here, such that, for example, synthetic enzymes, binding domains of antibodies or other physiological epitopes can be obtained by means of this embodiment. Inter alia, in the context of the present invention a derivative of a polymer having at least three functional groups is selected here, as described

@ - 17 = = above, in which at least one receptor group is an amino acid residue or an amino acid derivative residue. Possible amino acids are, for example: - amino acids having aliphatic residues such as glycine, alanine, valine, leucine, isoleucine; - amino acids having an aliphatic side chain which includes one or more hydroxyl groups, such as serine, threonine; - amino acids which have an aromatic side chain, such as phenylalanine, tyrosine, tryptophan; - amino acids which include basic side chains, such as lysine, arginine, histidine; - amino acids which have acidic side chains, such as aspartic acid, glutamic acid; - amino acids which have amide side chains, such as asparagine, glutamine; - amino acids which have sulphur-containing side chains, such as cysteine, methionine; - modified amino acids, such as hydroxyproline, y-carboxylglutamate, O- phosphoserine; - derivatives of the amino acids mentioned or optionally of further amino acids, for example amino acids esterified on the carboxyl group or optionally the carboxyl groups with, for example, alkyl or aryl radicals which can be optionally suitably substituted.

Instead of the amino acid, the use of one or more di- or oligopeptides is also possible, where in particular homopeptides, which are only synthesized from identical amino acids, may be mentioned. An example of a dipeptide which may be mentioned is, for example, hippuric acid. Furthermore, beta-, gamma- or other structurally isomeric amino acids and peptides derived therefrom such as depsipeptides can also be used.

Very generally, the activating reagents employed in the process according to the invention can be compounds of the general structure (X) 0

R;

N—O_ Ry (X)

Sage oO 0)

which are characterized in that Ry is a halogen atom or a radical (X')

Rr" 0—N Xx)

Ry" 0] and R;', Ry," R;" and R," are identical or different and are hydrogen, straight-chain or branched-chain alkyl, aryl, cycloalkyl, heterocyclic or aralkyl radicals having up to 30 C atoms or either Ry' and Ry’ or R;" and R," or both R;' and R;' and R;" and :

R," are linked to at least one carbocycle or to at least one heterocycle or to at least one carbocycle and to give at least one heterocycle. In particular, compounds may be mentioned by way of example here which have the following structures (X;) to (X39): 0

OSV : xy

Fo) lo} 0 hE : 0

Q- Cl (Xs)

LY

; ® - 19 -

[0] [0] of 0) (0, 4)]

SRE

[0] [0] 0 I 0 0 0]

T

[0] | [0]

Y [0] [0] Y 0

Bes Cl (Xs)

C1 0]

BL - Cl ) (Xo)

0

BL © (X10) 0] 0

Q

N—0— Xu)

HO Cl

O

0]

HO o

N— , 0]

HO x X12) lo)

OH

(0)

OH

0) 8) (Dod

Cl X13) {y° [@) . 9 ole

AN

(7) o) N (Xia)

@ 0 lo) n—0—~< (X15) —{CH cl 17 0 0 lo) n—o— (Xue) ~{cH, Cl le) 0 0) n—0—{ CX

Cre] | Cl

Vi fo) n—o0— (X15) on ci 13 te) 0 lo) n—0—{ . ere] y bs (is) . (, ,

HO3S P oO

N—o— (X20)

Cl to)

} - 22 = o .

HO3S [®)] n—0— 0 0 Xa) o) N

A HO38 :

Oo ; oT —0O 0 }

So

X22)

Cl 0

O

N—0— 0

Fo) “N (X23) 0 (8) —. N . 0

F 0) n—o0—< (X24)

F Ct (0) 0}

F P

N—0—< 0

F O_

O N (Xs) © F

F

- - 2 3 = 0 @) vo \ 0 (X20) 0 N ’

A

0] 0] n—o0—¢ (X27)

Cl 0] 0 lf

N —_—

Cl (X23) y 0]

Oo © 3

OQ

0

N— 4 (X29)

Ci 0 le} .

Ha5C12”

o) lo) n—0—< 0 a ;

HzsC12” ° (X30) 0) ?

Ciy2Hzs o) n—0— (X31) —0 lo] fo) 0) vo _ 0) fo) 0 0 X32) 0) lo}

AN lo) ~N 0

N lo)

Cre Xx) lo] te)

® 0p 4240 fo) 0) vo \ oO (X34) fo) > ° fo) § 0)

N—0— 29) 7 ci 0) fo) fo) aN

Cpe fo) (0) bol (X36) =

Va

N

HaC—O P aC— fo) al x

N Xs

H3C—0 Cl lo)

0

N—0—4 ° 0 N . (X38) o “CH

Oo “CHa 0

Oo

Nn—o0—< [o] I (X39)

O Oo

Oo 3 ‘where R" is hydrogen or a straight-chain or branched-chain, optionally substituted alkyl, aryl or aralkyl radical having up to 30 C atoms.

In the context of the present invention, the crosslinkage according to (iii) can be achieved, for example, in that two or more strands of derivatized or underivatized polymer are reacted directly with one another. This can be achieved, for example, in that the groups introduced by derivatization are constituted such that covalent and/or non-covalent bonds can be connected between these groups. Very generally, it is possible that these covalent and/or non-covalent bonds are formed between groups which are attached to one polymer strand, and/or are formed between groups which are attached to two or more polymer strands, such that two or more polymer strands can be connected to one another via one or more sites by crosslinkage.

Obviously, the bonding of the at least one polymer to the carrier material can also take place via functional groups which are present in the polymer itself or have been introduced into the polymer by suitable derivatization, as described above.

Likewise, it is also possible to employ for crosslinkage one or more suitable crosslinking reagents with which, as described above, groups within a polymer strand and/or groups which are attached to a number of strands of optionally

@ - different, optionally derivatized polymers can be crosslinked in a covalent and/or non-covalent manner.

Inter alia, it is possible here, even in the selection of the at least one polymer, to design its composition for later crosslinkage. Furthermore, it is in particular possible in the context of the present invention to design the derivatizing reagent with respect to its chemical composition, inter alia, with regard to later crosslinkage even during derivatization. In particular, the derivatizing reagent can contain groups which are selective for covalent and/or non-covalent crosslinkage.

Possible crosslinking reagents in principle are all suitable compounds known from the prior art. Accordingly, the crosslinkage can be carried out, for example, in a covalently reversible manner, in a covalently irreversible manner or in a non- covalent manner, where in the case of crosslinkage in a non-covalent manner, for example, crosslinkages via ionic interaction or via charge-transfer interaction may be mentioned. Crosslinking processes or reagents of this type are described, inter alia, in Han, K.K,, et al., Int. J. Biochem., 16, 129 (1984), Ji, T.H., et al., Meth.

Enzymol., 91, 580 (1983) and Means, G. and Feeney, R.E. Bioconj. Chem., 1,2 (1990).

With respect to non-covalent crosslinkage, an example which may be mentioned is, for example, crosslinkage by shifting the pH, when at least one basic and at least one acidic group are crosslinked with one another. Likewise, for example, non- covalent crosslinkage can take place when, in the case where two basic groups of, for example, polyallylamine are crosslinked with one another, a dibasic acid such as glutaric acid is added, or in the case where two acidic groups of, for example, polyacrylic acid are to be crosslinked with one another, a bifunctional such as cthylencdiamine is added. Likewise, a non-covalent crosslinkage can be formed by way of example by complex-forming metal ions or by metal complexes with free coordination sites. Non-covalent crosslinkage is preferably reversible and can therefore be employed in a preferred use of the polymeric networks prepared according to the invention for rapid systematic interaction studies. Very generally, with respect to non-covalent crosslinkage, reference can be made to all possible interactions which have already been presented above with respect to the interaction between template and polymeric structure.

@ - 28 =- ) With respect to covalently reversible attachment, inter alia, bonding via disulphide bridges or via labile esters or imines such as Schiff’s bases or enamines may be mentioned by way of example.

The chain length of the crosslinking reagents is in general arbitrary and can be adapted to the requirements of the particular process. Preferably, the chain length in the case of crosslinking reagents which have a carbon chain is in the range from 2 to 24 C atoms, preferably in the range from 2 to 12 C atoms and particularly preferably in the range from 2 to 8 C atoms.

Crosslinking reagents which may be mentioned which can lead to covalently irreversible crosslinkage are, inter alia, bi- or polyfunctional compounds such as diols, diamines or dicarboxylic acids. Here, for example, bifunctional crosslinkers are reacted with the activated polymer derivative or the at least bifunctional activated crosslinking reagent is reacted with the non-activated polymer derivative.

A covalently reversible crosslinkage can be realized, for example, by connecting a sulphur-sulphur bond to a disulphide bridge between two groups attached to one or two polymer strands or by formation of a Schiff’s base. Crosslinking via ionic interaction can take place, for example, via two radicals, of which one, as a structural unit, has a quaternary ammonium ion and the other has, as a structural unit, for example -COO" or -SO5°

A crosslinkage via hydrogen bridges can be formed, for example, between two complementary base pairs, for example via the following structure:

H

\

N—H-------0

Coad

N-------H—N

N—H-------0 /

H

@ - Very generally, polymers to be crosslinked non-covalently can be synthesized in a complementary manner with respect to the crosslinking sites, structural units complementary to one another being, for example, acid/ triamine or uracil/melamine. Likewise, in the case of non-covalent crosslinkage the crosslinking reagent can be complementary to the crosslinking sites on the polymer strand. An example of this which may be mentioned would be, for example, an amine group on the polymer strand and a dicarboxylic acid as a crosslinking reagent. Itis necessary in the context of the process according to the invention, with respect to a crosslinking step, to activate at least one of the functional groups which are involved in the crosslinkage, thus this is essentially possible according to all processes which are known from the prior art. In particular, the activation of a functional group can be carried out according to a process as is described in detail above in the activation and derivatization of polymers.

If, in the process according to the invention, the crosslinkage takes place via the use of at least one crosslinking reagent, this crosslinking reagent can in particular be a condensation compound which is prepared by reaction of at least one functional group of a first low molecular weight compound having at least two functional groups with at least one functional group of at least one further second low molecular weight compound having at least two functional groups, which can be identical to the first or different from the first low molecular weight compound, with obtainment of a condensation compound, the process being characterized in that at least one of the functional groups involved in this reaction has been activated before the reaction by reaction with a compound of the structure (X) 0] 2 lo)

Crp (1) Kn) © 0 oO as defined above. (Activated) crosslinking reagents which may be mentioned are, by way of example, compounds of the following structures (XI;) to (XI;7) mentioned below;

Q

1% o)

Beas ey ou 0 fo 0 ®)

OO 0 —

N—O o 4 (XIy) lo)

Q

1% 0) i GOSS) d : (X14) 0 0 (pe 0 o

Oo (Xs)

0 0] 0 o Y

Lore 8

N—0" “ch,-c—4{ lL, "o—N (0) 0 fo) 2 c 0 0 (el ox

Ng _

RW lo} &F 0—N bg) (XI)

C co” 0 0 N 0 oT ~"o_ ~c

C0 FOTN

NY 0 w (Xs)

Ca 0 &° 0 ‘N—o0—& le) X, % So NNO 6

A & (XI)

F fo) RN \ ty 0] H oO & < X10)

SNOT d (0)

fo) RY \Y sq)

C—O0—N \

NT Y% fo) RY e—o—~ el) \,

NEN ¢ pa J H (XL)

Sd / 0

OL

°N js) 0 sss od (XL13) oC or

CN 5 0 Q H

N O—N

N—O — h— (X14) 0 o 0 : 0

N O—N (pe Y os o fo! lo} 0 ~~ TN XL)

Od” =, « lo}

0

O H

2 _N__o-—N —_0—S8~ XI

N hi (XLi7) oO 0] o) 0

An example of a crosslinking reagent to be used according to the invention which may be mentioned below is a dimeric crosslinker which is prepared from phenylalanine and leucine by the process described above:

NH: ©

NJ AN

N° V4 ee M N NC . /

HO % . H HY

H N 5" H ONB-CI H ~S

I 1 ~

Phe-Leu-crosslinker

Examples for the synthesis of a condensation compound to be used as a crosslinking reagent by the process according to the invention which may be

) mentioned are the following reaction routes (A) and (B), in which the radical BNO represents the following structural unit (XII): fo}

Crp (XI) lo}

Reaction route (A):

Q

°o QA on I) H

NZ \ ( + SS NH, (wo od 2 0 ! NH

H ~~ 0 0 N

I VW \ & =

Petatal

S$ o

H O

/N 0 ‘1 H ~~ ON -— Q N— le} {

SEA, ay eno” N=" Oo

HO

Va n \ +2 H / ROSNY

N a NH, 0} N— 0] 3 7 eno” \—r/ bo) etc. 5

Reaction route (B):

jo] pe

Ne 2 N—0 or 1 o o H 0] ? NPP / lo}

OH

+1 H o

MM, 0 H

MN A { 0” ™N

BNO ISN o]

RY etc.

By means of this process, in which activated or non-activated crosslinking reagents can be prepared, it is of course also possible specifically to prepare polymers which can be employed in the process according to the invention and whose conformation can be adapted to at least one template compound. It is possible here that by means of this process, in which a condensation compound is synthesized, a polymer is prepared which is derivatized by the process already described above.

Likewise, it is also possible to prepare an already derivatized polymer.

Very generally, in the case of covalent crosslinkage, inter alia, ester, amide, carbonate, hydrazide, urethane or urea compounds or thio-analogous or nitrogen- homologous bonds can be formed.

In a preferred embodiment of the process according to the invention, the conformation of at least one of the polymers is adapted in the presence of at least one of the template compounds according to (ii).

. ® - 36 -

Accordingly, the present invention relates to a process such as described above, characterized in that the adaptation according to (ii) is carried out in the presence of at least one of the template compounds.

Inter alia, it is possible here to dissolve or to suspend the at least one polymer in one or more suitable solvents and to mix it together with the at least one template compound.

Inter alia, at the same time it is possible to add the at least one polymer to a solution in which the at least one template compound is present dissolved in at least one solvent, where the template compound can also be present in suspended form. Microparticles, for example, may be mentioned as a template compound which can be present in suspended form.

Inter alia, it is further possible to add at least one template compound to a solution in which the at least one polymer is present in dissolved or suspended form in at least one solvent.

Obviously, two or more solutions can also be mixed together, where in at least one solution the at least one polymer is present in dissolved or suspended form and in at least one further solution the at least one template compound is present in dissolved or suspended form.

If two or more polymers which are different from one another and/or two or more different template compounds are employed, each polymer and/or each template compound can be dissolved or suspended separately in one or more suitable solvents and the individual solutions and/or suspensions can be mixed together.

In this connection, embodiments are also possible in which a solution is employed which contains two or more solvents, in which the at least one polymer and/or the at least one template compound are both present in dissolved or in suspended form.

Obviously, it is also possible to start from a template compound which is already present bonded to at least one polymer, for example in the form of a complex or covalently, preferably covalently reversibly bonded. At the same time, the polymer to which the template compound is bonded can be a polymer whose conformation

Claims

a - Patent Claims

1. Process for the preparation of a polymeric network, in which (i) one or more polymers are made available which can be crosslinked with one another intramolecularly or intermolecularly or intra- and intermolecularly by covalent or non-covalent bonding, (ii) the conformation of at least one of the polymers is adapted to at least one template compound with obtainment of at least one preferred conformation of the at least one polymer, (iii)at least one of the preferred conformations obtained according to (ii) is fixed by crosslinkage.

2. Process according to Claim 1, characterized in that the adaptation according to (ii) is carried out in the presence of at least one of the template compounds.

3. Process according to Claim 1 or 2, characterized in that the adaptation of the conformation of the at least one polymer takes place in at least two steps.

4. Process according to Claim 3, characterized in that after each step the preferred conformation obtained from this step is fixed by crosslinkage.

5. Process according to Claim 3 or 4, characterized in that at least one step is carried out in the absence of the at least one template compound.

6. Process according to one of Claims 1 to 5, characterized in that the polymeric network is prepared on at least one support material.

7. Process according to Claim 6, characterized in that the at least one polymer is applied to the at least one support material in layers in at least two successive steps.

8. Process according to Claim 7, characterized in that the application in layers leads to a primarily crosslinked polymer network which has a conformation which is adapted to the at least one template compound in at least one

Le . oe ~ 68 - ) further step in the presence of the at least one template compound and is fixed by crosslinkage.

9. Polymeric network, preparable according to one of Claims 1 to 8.

10. Use of a polymeric network, preparable according to one of Claims 1 to §, in substance separation processes, substance conversion processes, substance preparation processes, substance recognition processes or for the detection of signals.

. vi \ -68/A

11. Process according to Claim 1, substantially as herein described and . exemplified.

12. Polymeric network according to Claim 9, substantially as herein described and exemplified.

13. Use according to Claim 10, substantially as herein described and exemplified. AMENDED SHEET