WO2005009981A1

WO2005009981A1 - Multifunctional linking and cleavable solid-phase reagents and method for the production thereof

Info

Publication number: WO2005009981A1
Application number: PCT/DE2004/001684
Authority: WO
Inventors: Thomas RÜHL; Klaus Burger
Original assignee: Universität Leipzig
Priority date: 2003-07-23
Filing date: 2004-07-22
Publication date: 2005-02-03
Also published as: DE10333368A1; DE10333368B4

Abstract

The invention relates to surface-functionalized support materials, respectively comprising a polymer surface and at least one linker compound which is bonded to said surface in a covalent manner, as well as to the production and use thereof. In said carrier material, an a-amino, a-thiol or a-hydroxy group and a carboxy group are protected by hexafluoroacetone and, at the same time, linker compounds, activated by carboxy groups, are bonded to solid-phase reagents, having hydroxy and/or amine functions (for example, Wang resins), by ester, amide and/or urethane bridges. Such materials may be used for the covalent immobilization of biomolecules, for the creation of substance libraries in combinatorial chemistry, for the synthesis of amino acids, peptides, proteins or molecules with at least one peptide structure unit on solid phases in peptide chemistry and for the recovery of affinity-labeling derivatives.

Description

Multifunctional, releasable and cleavable base phase reagent and

Process for their production

The invention relates to surface-functionalized carrier materials, each having a polymer surface and at least one linker compound covalently bonded to it. Such materials can be used for the covalent immobilization of biomolecules, in particular amino acids, peptides or proteins or molecules with amino or carboxy groups, for creating substance libraries in combinatorial chemistry, for the synthesis of amino acids, peptides, proteins or molecules with at least one peptide structural unit Solid phases can be used in peptide chemistry and for the production of affinity labeling derivatives.

The invention further describes a method for producing the surface-functionalized carrier materials.

The rapidly growing field of combinatorial chemistry has reawakened interest in organic synthesis techniques for solid phase chemistry. In addition to the need to develop synthetic methods that are suitable for the construction of organic molecules on a solid phase, there is a need for improved and novel linkers for linking the molecules to the carrier phase.

It is known to carry out the synthesis of peptides or more complex molecules with peptide structural units in the form of so-called solid-phase syntheses. For this purpose, an amino acid, which is quasi a first molecular link of the peptide sequence to be produced, is covalently bound to a solid phase reagent, the surface of which carries suitable functional groups. A further chain extension takes place in that further amino acids are successively bound to the first amino acid or to the free end of the resulting peptide chain in accordance with the sequence to be built up. In addition to the pure chain extension, chemical modifications to the immobilized amino acid or the immobilized peptide are possible. Polystyrene is mainly used as the base material for the solid phase (the carrier material) (see F. Z. Dörwald, Organic Synthesis on Solid Phases, Wiley-Verlag Chemie, Weinheim 2000, p. 414 ff).

A distinction is made between two strategies with regard to the direction of synthesis of the peptide to be produced. In the Merrifield strategy, which is also referred to as the A-type extension, the surface functionalization of the polystyrene takes place by derivatization with Chloromethyl, hydroxymethyl or acrylamide groups (RB Merrifield, J. Am. Chem. Soc. 1963, 85, pp. 2149-2154; R. Arshady et al., J. Chem. Soc. Perkin Trans. I 1981, Pp. 529-537). The first amino acid is covalently linked to these groups via the carboxy group of the amino acid, ie C-terminal. The further chain structure includes a condensation of the next amino acid to the amino group (the TV terminus) of the already immobilized amino acid or - in the case of further synthesis - the immobilized peptide. According to the Merrifield strategy, the synthesis from the C to the IV Therminus of the peptide takes place. The Boc strategy derived from the Merrifield concept (R. Arshady et al., J. Chem. Soc. Pekin Trans. I 1981, 529-537) and the Fmoc strategy (LA Carpino, GY Hau, J Org. Chem. 1972, 37, 3404-3409), in which the amino group of the amino acids to be linked is protected by certain protective groups, the peptide is ultimately bound to the solid phase via the carboxy function of the first amino acid.

In contrast, in the inverse strategy (B-type extension), chloroformic acid ester units immobilized on the surface of polystyrene are used as the starting point for peptide synthesis (RL Letsinger, MJ Komet, J. Am. Chem. Soc. 1963, 85, 2149-2144; R Matsueda et al., J. Am. Chem. Soc. 1975, 97, 2573-2575). The synthesis is carried out here by intravenous linkage of the amino acids protected as tert-butyl ester in the order given by the target sequence in the direction of the C-terminus of the peptide. A disadvantage of this prior art is that, depending on the desired synthesis strategy, differently functionalized support materials have to be used, since the chemical properties of the respective surface functions of the support material only permit an IV-terminal or a C-terminal linkage of the amino acid. Surface-functionalized carrier materials are known which allow the user to selectively bind a C-terninal or an IV-Tπninal binding of an amino acid or another molecule with corresponding functional groups (Sh. N. Khattab; A. El-Faha; AM El-Massry; EME Mansour; MM Abd. El-Rahman Letters in Peptide Science 2001, 7, 331-345; M. Lebl et al., US 5635598, Jun., 1997; P. Wessig et al., WO 02/051917 AI, July 2002).

The production of the known multifunctional, linkable and cleavable solid phase reagents has the disadvantage that additional steps for the protection and / or activation of the surface-functionalized carrier material have to be applied The surface functionalized carrier materials, which allow the user either a C-terminal or a H-terminal linkage of an amino acid or another molecule with corresponding functional groups, have the disadvantage that their production activates the eύien and protects the other C- The term and the protection of the IV terminus are required and additional steps have to be taken to split off the respective protecting group at the C and IV terminus and to activate the C terminus. Quantitative activation of the C-terminus can be difficult.

The cause of the disadvantages described can be seen above all in the fact that in the known multifunctional, linkable and cleavable solid-phase reagents, the coupling step of the linker to the solid phase is designed to be side-selective, so that the carboxy group available for subsequent derivatization is protected is. In addition, the IV terminal is protected in order not to react undesirably with the activated C-terminus during the coupling step to the solid phase.

The object of the invention is to provide easy-to-handle solid phase supports with at least one linker compound covalently bonded to them. The carrier should covalently immobilize molecules with amino or carboxy groups, especially amino acids, peptides or proteins, and thus z. B. simplify the production of substance libraries in combinatorial chemistry.

A method for producing the surface-functionalized carrier material is also to be provided.

According to the invention, the object is achieved by a surface-functionalized carrier material having a polymeric surface and at least one linker compound covalently bonded to it according to the general formulas (1) to (16) or (1 ') to (16'),

0 ') (2') (3 ") (4 ')

where in the general formulas (1) to (16) and (1 ') to (16') P is the polymeric surface, n is 1 to 12, R 1 and R ₂ independently of one another are H or an alkyl group and L is a Spacer means that links the linker compound to the polymeric surface.

The surface-functionalized carrier material according to the general formulas (1) to (16) according to the invention therefore consists of: 1. a polymeric surface (P) or a polymeric surface with a spacer (L) which links the linker compound to the polymeric surface 2. and one Hexafluoroacetone (F ₃ CC-CF ₃ ) protected linker compound. O In the corresponding surface-functionalized carrier material according to the general formulas (1 ') to (16'), the linker compound is protected by hexafluorothioacetone (F ₃ CC-CF ₃ ). S

The linker compounds consist of hexafluoroacetone or hexafluorothioacetone-protected and carboxyl-activated α-amino-dicarboxylic acids of the general formula (43),

in which n has the meaning 1 to 12 and R 1 and R ₂ independently of one another denote H or an alkyl group, α-hydroxy-dicarboxylic acids of the general formula (44),

in which n is 1 to 12 and Ri is H or an alkyl group,

α-mercapto-dicarboxylic acids of the general formula (45),

in which n is 1 to 12 and Ri is H or an alkyl group, iV-substituted Glycm derivatives of the general formula (46),

in which n has the meaning 1 to 12.

The hexafluoroacetone or hexafluorothioacetone simultaneously protects a carboxy group in the linker compound and either a -hydroxy-, an α-thiol (= - mercapto) or an α-amino group to form a five-membered heterocyclic lactone ring or thiolactone ring.

Due to the electron-withdrawing effect of the two trifluoromethyl groups on the lactone ring or thiolactone ring, the carboxy group is activated towards nucleophiles and is available in a reactive form.

The use of hexafluoroacetone or hexafluorothioacetone as a protective group advantageously makes it possible to combine protection and activation on the one hand and derivatization and deblocking on the other hand in one step, thereby saving synthesis steps.

The surface-functionalized support material according to the general formulas (1) to (16) or (1 ') to (16') can advantageously be water or molecules with hydroxyl, thiol or amino group at the C-terminal or molecules with carboxy-, sulfonyl - or either phosphoryl groups. O-, S-, or iV-terminal can be bound covalently.

The binding of water or molecules with hydroxyl, thiol or amino groups to the surface-functionalized support material according to the general formulas (1) to (16) or (1 ') to (16') is achieved by reaction with the hexafluoroacetone or Hexafluorothioacetone activated carboxy group enables.

The binding of molecules with carboxy, sulfonyl or phosphoryl groups in activated form or by a coupling reagent to the surface-functionalized support material according to the general formulas (1) to (16) or (! ') to (16 ⁵ ) is made possible by splitting off the hexafluoroacetone or hexafluorothioacetone group by reaction with the hydroxyl, thiol or amino group.

The surface-functionalized support material according to the general formulas (1) to (16) or (1 ') to (16') can thus advantageously be used for the covalent immobilization of photolabile victim groups, for the covalent immobilization of biomolecules, in particular amino acids, peptides or proteins or other molecules with amino or hydroxy, thiol (C-terminal immobilization) and / or carboxy groups (O-, S- or iV-terminal immobilization) can be used.

The surface-functionalized carrier material according to the general formulas (1) to (16) or (1 ') to (16') is therefore advantageously suitable for creating substance libraries in combinatorial chemistry, for the synthesis of amino acids, peptides, proteins or Molecules with at least one peptide structural unit on solid phases in peptide chemistry and for obtaining affinity labeling derivatives (J. Jenssen, K. Sewald, N. Sewald, Bioconjugate. Chem. 2004, 15, 594-600).

The surface-functionalized carrier material according to the general formulas (1) to (16) or (1 ') to (16') is advantageously suitable for use in photoaffinity labeling and its subsequent steps. The carrier material according to the invention serves as a carrier for a photosensitive group and a molecule with affine properties to form a partner molecule. The partner molecule is enriched, covalently bound by a photochemical reaction and broken down chemically or enzymatically. The polymer surface (P) is then removed via a predetermined breaking point by a cleavage reaction. The cleaning processes involved (filtration and washing) are simple here and the time required is considerably reduced compared to the methods known in solution.

The carboxy function of the surface-functionalized carrier material according to the general formulas (1) to (16) or (1 ') to (16') integrated in the lactone ring is capable of amide, thioester or ester formation. The amino, thiol or hydroxy function deprotected in the course of the amide, thioester or ester formation (by removing the hexafluoroacetone or hexafluorothioacetone group) is thus immediately available for derivatization. Complex cleaning steps such as flash Column chromatography (FSC), which are essential for synthesis in solution, is gone. In the synthesis on a solid phase, the quantitative removal of the hexafluoroacetone or hexafluorodioacetone hydrate is possible by simply washing with large amounts of water (advantageously dilute citric acid). The amine used and the acylation reagent used in the subsequent step are removed by simple washing with appropriate solvents, so the two reagents can be used in excess in the two reaction steps (3-5 equivalents), which considerably increases the yield.

For the cleavage of the hexafluoroacetone or hexafluorothioacetone protective group (ring opening), which takes place under mild conditions, two equivalents of nucleophile are required, water being used instead of the second equivalent of nucleophile to cleave the intermediate semi-amine, semi-acetal or semi-thioacetal intermediate , The corresponding amine, hydroxy or thiol function is deprotected. This means that, compared to the prior art, two steps, namely the derivatization of the carboxy group and the removal of protective groups on the corresponding amino, hydroxyl or thiol function, are combined to form one reaction step.

The surface-functionalized support material (33) according to the invention (Scheme 1) with high occupancy of linker compounds protected from hexafluoroacetone or hexafluorothioacetone can advantageously be used for a three-stage synthesis of (50) (Scheme 2).

After ring opening (e.g. by aminolysis), there is a reaction with electrophiles (e.g. acylation, sulfonation or phosphoryation) or, in the case of compounds (4), (8), (12) and (16), an oxidation (e.g. formation) of disulfides). The ring opening is preferably carried out with amines, even under aqueous conditions.

The spacer L is preferably an amino acid, a peptide or another compound which contains at least one amino or hydroxy function for the attachment of the linker compound and at least one carboxy function for the immobilization to the solid phase.

The carrier material preferably consists entirely or on its surface of an organic polymer. The polymeric surface (P) preferably contains hydroxyl groups (for example Wang resins) which enable the linker compound or the spacer L to be bound. The esters, amide or urethane bonds formed by the binding of the linker compound or the spacer L can advantageously be cleaved by simple methods known per se, after derivatization has taken place.

The organic polymer is preferably selected from polypropylene, polyethylene, polysulfone, polystyrene, polyvinyl chloride, polyacrylonitrile, cellulose, amylose, agarose, polyamide, polyimide, polytetrafluoroethylene, polyvinylidene difluoride, polyester, polycarbonate, polyacrylate, polyacrylamide or derivatives, copolymers or blends of these substances.

The organic polymer is particularly preferably a Wang resin, a PEGA resin (2-acrylamidoprop-l-yl- (2-aminoprop-l-yl) polyethylene glycolsoo and dimethylaciylamide cross-linked with bis 2-acrylamidoprop-l-yl polyethylene glycolsoo) or a Rink Amide resin.

In an alternative embodiment, the carrier material is an inorganic and / or mineral material, preferably a glass, a silicate, a ceramic material or a metal or a composite of at least one inorganic and / or mineral material and at least one organic polymer.

The carrier material is preferably in the form of a membrane, a film, a plate, a microtiter plate, a reaction vessel, a slide, a fiber, a hollow fiber, a nonwoven, a tissue, a powder, a granulate or particles and is porous or non-porous , It is preferably a powder. In the form of a membrane, it has a symmetrical or asymmetrical pore structure. It preferably has pores with a diameter of 1 nm to 10 μm. The invention further relates to a process for producing the surface-functionalized carrier material according to the invention with a polymeric surface and at least one linker compound covalently bonded to it according to the general formulas (1) to (16) or (1 ') to (16').

For this purpose, the pipe connections according to the general formulas (17) to (32),

(17) (18) (19) (20)

(22) (21) (23) (24)

(25) (26) (27) (28)

in which n, R 1 and R _{2 have} the above meaning and L is a spacer which is intended to link the linker compounds to the polymer surface, or the linker compounds according to the general formulas (17 ') to (32')

(21 ') (22') (23 ') (24')

(25 ') (26') (27 ') (28')

in which n, Ri and R and L have the above meaning,

with a polymeric surface (P) of a support material (solid phase support), which contains s linkable functional groups.

In solution systems, hexafluoroacetone has long been known as a protective group for α-amino, α-hydroxy, α-mercapto-dicarboxylic acids and iV-substituted glycine derivatives (E. Windeisen, dissertation TU Munich 1993; K. Burger, H. Neuhauser, A. Worku, Z. Naturforsch. 1993, 48b, 107-120 K. Burger, C. Böttcher, G. Radics, L. Hennig, TetrahediOn Lett. 2001, 42, 3061-3063; C. Böttcher, K Burger , TetrahediOn Lett. 2002, 43, 9711-9714; C. Böttcher, K. Burger, Tetrahedron Lett. 2003, 44, 4223-4226). The lactone ring is activated by the two trifluoromethyl substituents. So far, the ring opening in solution systems with amines, hydroxy compounds or water. When the ring opening of hexafluoroacetone-protected α-amino-dicarboxylic acids with excess strong nucleophiles at room temperature in solution, an elimination rank reaction disadvantageously dominates. Due to the often stable half-aminal or half-thioacetal intermediate stage, a large amount of water is required for quantitative elimination, which makes additional washing and extraction steps necessary, which are expensive to carry out in solution systems, before working up by flash column chromatography.

Since the thiol which has been deprotected after removal of the hexafluoroacetone is frequently decomposable, a process which enables rapid processing and further processing is desirable. In particular, if macromolecular nucleophiles (e.g. those with enzyme-substrate interactions) are used for ring opening, the prior art requires extensive work-up, e.g. B. by the streptavidin / biotin technique or SDS-PAGE (sodium dodeca-sulfate polyacrylamide electrophoresis).

The carrier material according to the general formulas (1) to (16) and (1 ') to (16') according to the invention enables rapid workup after simple removal of the hexafluoroacetone or hexafluorothioacetone by simply washing the carrier material.

The transfer of the hexafluoroacetone protective grapple technology according to the invention to solid-phase systems has so far failed because hexafluoroacetone is a very toxic and aggressive gas, the hydrates of which are also regularly used as solvents for H-containing polymers.

In the alternative synthesis for obtaining the surface-functionalized support material according to the general formulas (1) to (16) via a hexafluoroacetone reaction with an α-amino, α-thiol, α-hydroxy-dicarboxylic acid or a glycine derivative, the polymer is in the hexafluoroacetone and hexafluoroacetone hydrate present in the solvent. The dissolved polymer makes it impossible to isolate the product by filtration. According to the invention, the surface-functionalized support material according to the general formulas (1) to (16) or (1 ') to (16') is therefore, in order to avoid dissolving the polymeric surface, not by direct reaction of the linker compound bound to the support material with hexafluoroacetone or hexafluorothioacetone.

To this end, the linker compounds are reacted in excess with linkable functional groups of solid-phase reagents (e.g. Wang resin).

The reaction is preferably carried out with catalysis (e.g. by adding DMAP - dimethylaminopyridine) or without catalysis.

The excess of linker compounds (occupancy reagent) (e.g. 1.5 equivalents), the number of runs, the temperature range (e.g. 0 ° C), the solvent (e.g. dry pyridine, chloroform) and the base (e.g. pyridine, NaHCO ₃ ) or propene oxide are correct according to the substances used. The substances obtained are cleaned by methods known per se.

The surface-functionalized carrier material obtained according to the general formulas (1) to (16) or (1 ') to (16') is preferably characterized by IR spectroscopy and the occupancy (amount of substance of the linker compound bound to the polymer divided by the mass of substance ) determined by elemental analysis.

The fact that the support material according to the general formulas (1) to (16) or (1 ') to (16') produced in this way and dried in public transport (oil pump vacuum) can be characterized by IR (infrared spectroscopy (compacting technique in potassium bromide)) was not foreseeable due to the complex structure.

The surface-functionalized carrier material obtained according to the general formulas (1) to (16) or (1 ') to (16') is characterized in the IR spectrum by the signals of the carbonyl groups of the linker compound which is linked to the polymeric surface ,

The occupancy of the substances obtained can be determined by elemental analysis (e.g. fluorine, nitrogen and / or sulfur elemental analysis). The linker compounds according to the general formulas (17) to (20) are prepared by directly reacting the corresponding compounds according to the general formulas (43) to (46) with hexafluoroacetone in solution.

The linker compounds of the general formulas (17 ') to (20') are correspondingly prepared by direct reaction of the corresponding compounds of the general formulas (43) to (46) with hexafluorothioacetone in solution.

The linker compounds according to general formulas (21) to (24) are prepared from the compounds according to general formulas (17) to (20) by known methods by converting the carboxy group into an isocyanate group in solution. The linker compounds according to the general formulas (21 ') to (24') are correspondingly prepared from the compounds according to the general formulas (17 ') to (20').

The linker compounds according to general formulas (25) to (28) are prepared from the compounds according to general formulas (17) to (20) by coupling the free carboxy group with a linker in solution to form an ester or amide bond. The linker compounds of the general formulas (25 ') to (28') are correspondingly prepared from the compounds of the general formulas (17 ') to (20').

The linker compounds according to general formulas (29) to (32) are prepared from the compounds according to general formulas (21) to (24) by reaction of the isocyanate function with a linker in solution to form a urea or urethane bond. The linker compounds of the general formulas (29 ') to (32') are correspondingly prepared from the compounds of the general formulas (21 ') to (24').

By installing the hexafluoroacetone or hexafluorothioacetone protective group, the α-amino, α-hydroxy or α-thiol group and selectively the α-carboxy function are protected in one step.

The advantages of the invention can be seen in particular in the following: Since the protection of the amino, hydroxy or thiol function and the activation of the carboxy function in the If new processes are combined into one synthesis step, the application of the new process means that there is no need for synthesis steps. Since the Linkeι ^* compounds are bound to the resin as an activated species, it is guaranteed that every molecule anchored with the resin is also activated. This cannot be guaranteed in the case of subsequent activation that only takes place on the resin. Since a washing process takes place after each reaction step, the dissolved reagents can be used in large excess. This leads to optimal yields. The reaction sequence can be automated.

In the process according to the invention, preference is given to the carboxy group of the compounds of the general formulas (17) to (20) or (17 ') to (20') and of the compounds of the general formulas (25) to (32) or (25 ' ) to (32 ') for the acylation to the ester or amide activated.

The carboxy group of the compounds according to the general formulas (17) to (20) or (17 ') to (20') and (25) to (32) or (25 ') to (32') is activated by processes known per se, preferably by reaction with thionyl chloride or phosphorus pentachloride in an acid chloride, by reaction with DAST (diethylaminosulfur trifluoride) in an acid fluoride or by a coupling reagent, for example TBTU (O- (1 H-bezotriazol-1-yl) -H, N, iV, N'-tetramethyluronium tetrafluoroborate).

The compounds according to the general formulas (25) to (32) or (25 ') to (32') are preferably selected from the compounds according to the general formulas (17) to (24) or (17 ') to (24' ) obtained by attaching the spacer L. The spacer L is linked to the linker compound according to the general formulas (17) to (20) or (17 ') to (20') by acylation to give the ester or amide and the linkage of the spacer L to the linker compound according to general formulas (21) to (24) or (21 ') to (24') by addition to urethane or urea. The spacer L is preferably an amino acid, a peptide or another compound with at least one amino or hydroxy function for the functionalization of the compounds according to the general formulas (17) to (24) or (17 ') to (24 ') and a carboxy function for immobilization on the polymer surface. The linking of the linker compounds of the general formulas (17) to (20) or (17 ") to (20 ') and the general formulas (25) to (32) or (25') to (32 ') with the polymer Surface is preferably carried out by acylation to give the ester or amide and that of the linker compounds of the general formulas (21) to (24) or the general formulas (21 ') to (24') is preferably carried out by addition to the urethane or urea.

When linking the linker compounds of the general formulas (17) to (20) or (17 ') to (20') and the general formulas (25) to (32) or (25 ') to (32') with the polymeric surface by acylation to the ester or amide, a trapping reagent for hydrogen chloride, preferably NaHCO ₃ or pyridine or propene oxide, is preferably used.

The linker compounds according to the general formulas (17) to (32) or (17 ') to (32') are either used once. The process is preferably carried out several times, preferably twice to three times, in order to achieve a quantitative conversion.

The surface-functionalized carrier material according to the invention is preferably produced in a temperature range between -100 ° C. and + 100 ° C., preferably -50 ° C. to 50 ° C., particularly preferably at room temperature.

The reaction is carried out in a solvent or solvent mixture. Chloroform and / or dichloromethane is preferably used as the solvent for base-sensitive substances and pyridine is preferably used for base-insensitive substances.

After the reaction, unreacted substances are removed by washing with a washing liquid. Water, ethyl acetate or another organic solvent or a solvent mixture is preferably used as the washing liquid.

The invention also relates to the use of the carrier material according to the invention for the synthesis of amino acids, peptides, proteins or molecules with at least one peptide structure unit, a first amino acid to be used for the synthesis being covalently bound to the carrier material according to the invention and chain extension by successively attaching further amino acids and / or a chemical modification takes place. The first amino acid used for the synthesis is preferably bound to the support material by a peptide bond between the amino group of the amino acid and the acid function (activated by hexafluoroacetone or hexafluorothioacetone) of the linker compound. The further attachment of amino acids can advantageously take place N-, O- or S-terminal to the amino, hydroxy or thiol group of the linker compound or C-terminal to the carboxy group of the attached amino acid.

The N-, O- or S- or C-terminal binding of the amino acid to the linker compound is controlled by blocking the amino group or the carboxy group of the amino acid and / or the amino, hydroxyl or thiol group the corresponding linker compound with chemical protecting groups.

If an N-, O- or S-terminal connection is to be made via a coupling reagent (e.g. TBTU), the amino function of the amino acid to be bound must be protected.

In addition, the carboxy function of the amino acid already attached must be protected.

If a C-terminal linkage to the previously linked amino acid is to take place, then at

Activation of the carboxy function by a coupling reagent (e.g. TBTU)

Carboxyfunction of the amino acid to be linked to be protected. In addition, the N, O or S terminus of the linker connection must be protected.

An example of an amino protective group is the Boc protective group (tert-butoxycarbonyl

Protecting group).

The methyl ester represents a protected carboxy function.

An example of a thiol or alcohol protective group is the Bzl protective group (benzyl

Protecting group).

The following schemes and exemplary embodiments are intended to explain the invention in greater detail.

(Scheme 1) In Scheme 1, the gray filled circle divinylbenzene (DVB) means cross-linked styrene

Polymer, [PEGA | has the meaning 2-acrylamidoprop-l-yl- (2-aminoprop-l-yl) polyethylene glycolsoo and dimethylacrylamide mixed with bis 2-acrylamidoprop-l-yl polyethylene glycolsoo OOMe (33) o o -NHCBZ (47)

(Scheme 2)

Scheme 2: Reagents and conditions: i) DMAP, Py, RT (room temperature); ii) H-Lys (Z) - OMe x HCl, DMAP, Py, RT; iii) DMAP, Py, RT; iv) TFA, H ₂ O, RT. Yield (33) → (50) 45%.

(53) (54) (Scheme 3)

Scheme 3: Reagents and conditions: i) 2 HFA, DMF, RT, yield 90%; ii) SOCl ₂ , RT, yield 100%; iii) Trimethylsilylazide, toluene, 0 → 80 ° C, yield 40%.

The ER spectra were recorded on the FTIR ™ Genisis Series from ATI Mattson. The elemental analyzes were measured on the VarioEL V2.6 from Elementar Analysensysteme GmbH.

The NMR spectra were recorded on a VARIAN Gemini 200, 2000 and 300 and a BRUKER DRX 600 spectrometer. The chemical shifts are given in ppm relative to teframethylsilane (TMS, δ = 0 ppm). The coupling constants J are given in Hertz (Hz). The 1H NMR spectra were measured at 200, 300 and 600 MHz and the ¹³ C NMR spectra at 50 and 76 MHz. The ¹⁹ F spectra were measured at 188 and 282 MHz with trifluoroacetic acid (TFA, δ = 0 ppm) as an external standard. The mass spectra were recorded on a Braker Daltronics FT-ICR-MS Apex II and a Fisions VG Autospec.

Example 1:

1 g (0.6 mmol) of Wang resin (Acros, 1% cross-linked with DVB, 0.5-0.6 mmol / g, 200-400 mesh) and a spatula tip of DMAP are placed in 10 ml of dry pyridine. After 5 minutes of treatment in an ultrasound bath and a swelling time of 30 minutes, 270 mg (0.9 mmol) of [5-oxo-2,2-bis (trifluoromethyl) -1I-oxazolidin-3-yl] acetyl chloride are added with stirring. The The reaction mixture is stirred overnight. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-17 ((33) (Scheme 1)) IR (KBr): 1846 cm ^"1 (v lactone) 1745 cm ^{" 1} (v ester) cm-1% T cm-1% T cm-1% T cm -1% T 478.26 73.24 509.11 67.94 532.26 61.72 696.18 43.09 754.03 61.30 792.60 72.99 1099.23 67.43 1166.72 65.21 1226.51 54.18 1276.65 65.54 1288.22 65.77 1448.28 59.60 1506.14 64.26 1606.42 61.65 1644.99 73.25 1733.70 59J2 1816.62 61.64 2310.31 73.61 2366.24 68.31 2848.36 60.17 2919.71 50.96 3023.85 55.07 3056.63 62.76 3174.27 73.47 3210.91 70.34 3234.05 68.71 3261.05 63.75 3289.98 64.66 3349.76 53.01 3357.48 52.92 3388.33 49.10 3399.90 48.76 3436.54 44.26 3446.19 44.64 3455.83 44.79 3477.04 47.41 3517.54 50.08 3544.54 56.67 3586.97 62.93 3608.18 68.96 3619.75 71.26 3793.31 74.62 3806.81 72.87 3826J0 71.57 3845.38 71.12 receiving the IR-spotting trams see Fig. 1

Occupancy (IV elementary analysis): 92% of the max. Occupancy (0.6 mmol / g)

_v x = —Y x. = 0.78% N; -l

Calculation of the occupancy from the percentage of nitrogen: Occupancy: 0.6 mmol / g 0.0078 _- = 0.00055 mol - 1 - 14.07 g - mol error: 6%

^ ^ - = 0.06 x 0.78%

Example 2:

1 g (0.8 mmol) of Wang resin (Novabiochem, 1% cross-linked with DVB, 0.5-1.3 mmol / g, 100-200 mesh) are placed in 10 ml of dry chloroform. After a swelling time of 30 min, 381 mg (1.3 mmol) of (S) - (5-oxo-2,2-bis-trifluomomethyl-oxazolidin-4-yl) acetyl chloride in 1 ml of dry chloroform are added with stirring. After 10 min, 65 mg (0.8 mmol) of dry pyridine in 1 ml of dry chloroform is added dropwise. The reaction mixture is stirred for 49 h. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-77 ((34) (Scheme 1)) IR (KBr): 1817 cm ^'1 (v lactone) 1736 cm ^"1 (v ester) cm-1% T cm-1% T cm-1% T cm -1% T 478.26 73.24 509.11 67.94 532.26 61.72 696.18 43.09 754.03 61.30 792.60 72.99 1099.23 67.43 1166.72 65.21 1226.51 54.18 1276.65 65.54 1288.22 65.77 1448.28 59.60 1506J4 64.26 1606.42 61.65 1644.99 73.25 1733.70 59.12 1816.62 61.64 2310.31 73.61 2366.24 68.31 2848.36 60.17 2919.71 50.96 3023.85 55.07 3056.63 62.76 3174.27 73.47 3210.91 70.34 3234.05 68.71 3261.05 63.75 3289.98 64.66 3349.76 53.01 3357.48 52.92 3388.33 49.10 3399.90 48.76 3436.54 44.26 3446.19 44.64 3455.83 44.79 3477.04 47.41 3517.54 50.08 3544.57.76.6.9 36.9

Recording the IR spectrum, see FIG. 2 Illumination (H elemental analysis): 43% of the indicated occupancy (0.8 mmol / g)

_ 1 N x = - Y JC, = 0.50%

Calculation of the occupancy from the percentage of nitrogen: Occupancy: 0.4 mmol / g 0.0050 - = 0.00036 mol- g ^" 1 - 14.07 g - mol Error: 20% s 0.10% 0.20 x 0.50%

Example 3: 1 g (0.6 mmol) of Wang resin (Acros, 1% cross-linked with DVB, 0.5-0.6 mmol / g, 200-400 mesh) are placed in 15 ml of dry chloroform. After a swelling time of 30 min, 271 mg (0.9 mmol) of (R) - (5-oxo-2,2-bis-trifluomomethyl- [1,3] dioxolan-4-yl) acetyl chloride in 1 ml of dry chloroform are stirred added. After 40 min, 76 mg (0.9 mmol) NaHCO _{3 is} added. The reaction mixture is stirred for 25 h. The product is then filtered off, washed with ice water and ethyl acetate and dried in an oil pump vacuum.

TR 5-91 ((35) (Scheme 1)) IR (KBr): 1853 cm ^'1 (v lactone) 1738 cm ^"1 (v ester) cm-1% T cm-1% T cm-1% T cm-1% T 478.26 73.24 509J 1 67.94 532.26 61.72 696.18 43.09 754.03 61.30 792.60 72.99 1099.23 67.43 1166.72 65.21 1226.51 54.18 1276.65 65.54 1288.22 65.760.26.64.42 59.60.26.16.46.45 73.25 1733.70 59.12 1816.62 61.64 2310.31 73.61 2366.24 68.31 2848.36 60.17 2919.71 50.96 3023.85 55.07 3056.63 62.76 3174.27 73.47 3210.91 70.34 3234.05 68.71 3261.05 63.75 3289.98 64.66 3349.76 53.01 3357.48 52.92 3388.33 49.10 3399.90 48.76 3436.54 44.26 3446.19 44.64 3455.83 44.79 3477.04 47.41 3517.54 50.08 3544.54 56.67 3586.97 62.93 3608.18 68.96 3619.75 71.26 3793.31 74.62 3806.81 72.87 3826.10 71.57 3845.38 71.12 For the IR spectrum, see Fig. 3

Occupancy (_E elementary analysis): 63% of the max. Occupancy (0.6 mmol / g)

1 ^N - Y JC, = 4.32% 1-1

Calculation of the occupancy from the percentage of fluorine: Occupancy: 0.4 mmol / g

0.0432 = 0.00038 mol - g 6 J9.00 g - mol ^J error: 1% = ^ = 0.01 x 4.32%

Example 4: 1 g (0.8 mmol) of Wang resin (Νovabiochem, 1% cross-linked with DNB, 0.5-1J mmol / g, 100-200 mesh) are placed in 10 ml of dry chloroform. After a swelling time of 30 min, 389 mg (1.2 mmol) of racemic (5-oxo-2,2-bis-hϊfluoromethyl- [1,3] oxathiolan-4-yl) acetyl chloride in 1 ml of dry chlorofomi are added with stirring. After 10 min 65 mg (0.8 mmol) of dry pyridine in 1 ml of dry chlorofomi is added dropwise. The reaction mixture is stirred for 44 h. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-81 ((36) (Scheme 1)) IR (KBr): 1816 cm ^"1 (v lactone) 1733 cm ^{" 1} (v ester) cm-1% T cm-1% T cm-1% T cm -1% T 478.26 73.24 509.11 67.94 532.26 61.72 696.18 43.09 754.03 61.30 792.60 72.99 1099.23 67.43 1166.72 65.21 1226.51 54.18 1276.65 65.54 1288.22 65.77 1448.28 59.60 1506.14 64.26 1606.42 61.65 1644.99 73.25 1733.70 59J2 1816.62 61.64 2310.31 73.61 2366.24 68.31 2848.36 60.17 2919.71 50.96 3023.85 55.07 3056.63 62.76 3174.27 73.47 3210.91 70.34 3234.05 68.71 3261.05 63.75 3289.98 64.66 3349.76 53.01 3357.48 52.92 3388.33 49.10 3399.90 48.76 3436.54 44.26 3446.19 44.64 3455.83 44.79 3477.04 47.41 3517.54 50.08 3544.54 56.67 3586.97 62.93 3608.18 68.96 3619.75 71.26 3793.31 74.62 3806.81 72.87 3826.10 71.57 3845.38 71.12 receiving the IR spectrum see Fig. 4

Occupancy (S elementary analysis): 51% of the occupancy specified (0.8 mmol / g)

N = - Yx, = 1.33% N ι-l

Calculation of the occupancy from the percentage of sulfur: Occupancy: 0.4 mmol / g 0.0133 = 0.00042 mol g ^" 1 - 32.07 g - mol ^{" 1}

Error: 16%

* _. ^ __ ₌ 0.16 x 133%

Another rule for obtaining the substance is as follows.

0.567 g (0.3 mmol) of Wang resin (Acros, 1% cross-linked with DVB, 0.5-0.6 mmol / g, 200 -400 mesh) are placed in 5 ml of dry chloroform. After a swelling time of 30 min, 129 mg (0.4 mmol) of racemic (5-oxo-2,2-bis-trifluomiethyl- [1,3] oxathiolan-4-yl) acetyl chloride in 71 mg (1.2 mmol) of propene oxide are added with stirring added. Then 0.002 equivalents of DMAP of a 0.008 M solution in chloroform are added with stirring. The reaction mixture is stirred overnight. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-139 ((36) (Scheme 1)) IR (KBr): 1817 cm ^"1 (v lactone) 1734 cm ^{" 1} (v ester) cm-1% T cm-1% T cm-1% T cm -1% T 478.26 73.24 509.11 67.94 532.26 61.72 696.18 43.09 754.03 61.30 792.60 72.99 1099.23 67.43 1166.72 65.21 1226.51 54.18 1276.65 65.54 1288.22 65.77 1448.28 59.60 1506.14 64.26 1606.42 61.65 1644.99 73.25 1733.70 59.12 1816.62 61.64 2310.31 73.61 2366.24 68.31 2848.36 60.17 2919.71 50.96 3023.85 55.07 3056.63 62.76 3174.27 73.47 3210.91 70.34 3234.05 68.71 3261.05 63.75 3289.98 64.66 3349.76 53.01 3357.48 52.92 3388.33 49.10 3399.90 48.76 3436.54 44.26 3446.19 44.64 3455.83 44.79 3477.04 47.41 3517.54 50.08 3544.57.76.67.9 36.9

Recording the IR spectrum, see FIG. 5 Occupancy (_? - elementary analysis): 46% of the maximum occupancy (0.6 mmol / g)

- 1 ^N x = - Tx, ^■ ■ 0.94%

Calculation of the occupancy from the percentage of sulfur occupancy: 0.3 mmol / g

0.0094 - = 0.00029 mol - 1 - 32.07 g - mol error: 10% s _ 0.09% = 0.10 x ^~ 0.94%

Example 5: 1,352 g (0.5 mmol) of HMPA-PEGA resin (Novabiochem, 0.2-0.4 mmol / g) dried in an oil pump vacuum are placed in 20 ml of dry chloroform. After a swelling time of 30 min, 257 mg (0.8 mmol) of racemic (5-oxo-2,2-bis-hifluoromethyl- [1,3] oxathiolan-4-yl) acetyl chloride in 1 ml of dry chlorofomi are added with stirring. After 10 minutes, 43 mg (0.5 mmol) of dry pyridine in 1 ml of dry chloroform is added dropwise. The reaction mixture is stirred for 50 h. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-101 ((40) (Scheme 1)) IR (KBr): 1819 cm ^"1 (v lactone) 1736 cm ^{" 1} (v ester) cm-1% T cm-1% T cm-1% T cm-1 -0 1 512.97 91.85 536.11 83.47 697.14 52.79 754.03 81.47 903.49 90.30 978.70 82.49 1024.98 90.81 1041.37 91.88 1132.01 71.51 1 187.94 77.03 1238.08 58.01 8496.2 90.03 13 14 87.75 1448.28 73.94 1490.71 75.56 1510.96 74.56 1603.53 80.10 1734.66 67.08 1850.37 79.29 1943.90 90.56 2265.96 91.21 2372.99 90.52 2847.39 74.38 2920.68 62.32 3025.78 69.04 3057.60 76.70 3080.74 80.47 3235.02 91.88 3256.23 91.58 3272.62 89.45 3292.87 87.27 3316.01 80.05 3349.76 75.12 3374.83 69.00 3403.76 63.89 3430.76 63.15 3451.01 63.73 3465.47 67.51 3490.54 65.10 3527.18 77.41 3551.29 81.93 3564.79 83.07 3654.46 88.55 3675.68 89.80 3783.67 85.57 3815.49 85.97 3837.67 86.17 3856.95 90.50 3870.45 90.44 Recording the IR spectrum see Fig. 6

Occupancy (S elementary analysis): 46% of the maximum occupancy (0.4 mmol / g)

_v x = - Yx. = 0.60%

Calculation of the occupancy from the percentage of sulfur: Occupancy: 0.2 mmol / g 0.0060 _τ = 0.00019 mol- g ^" 1 -32.07 g - mol ^" Error: 10% s 0.07% = 0.10 x 0.60%

Example 6: 1 g (0.6 mmol) of Wang resin (Acros, 1% cross-linked with DNB, 0.5-0.6 mmol / g, 200-400 mesh) is placed in 10 ml of dry pyridine. After 5 minutes of treatment in an ultrasonic bath and a swelling time of 30 minutes, the mixture is cooled to below 0 ° C. with dry ice and 322 with stirring mg (1.2 mmol) 3-isocyanatomethyl-2,2-bis (ti'ifluormethyl) -l, 3-oxazolidin-5-one added. The reaction mixture is treated for 5 min with ice water cooling in an ultrasonic bath, stirred for 2 h at 4 ° C. (ice water) and overnight at room temperature. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-21 ((37) (Scheme 1)) IR (KBr): 1846 cm ^"1 (v lactone) 1728 cm ^{" 1} (v urethane) cm-1% T cm-1% T cm-1% T cm -1% T 512.97 91.85 536J 1 83.47 697.14 52.79 754.03 81.47 903.49 90.30 978.70 82.49 1024.98 90.81 1041.37 91.88 1132.01 71.51 1187.94 77.03 1238.08 58.01 1296.90 90.03 1448.28 73.94 1318J1 84.27 1371J4 1490.71 75.56 1510.96 87.75 74.56 1603.53 80.10 1734.66 67.08 1850.37 79.29 1943.90 90.56 2265.96 91.21 2372.99 90.52 2847.39 74.38 2920.68 62.32 3025.78 69.04 3057.60 76.70 3080.74 80.47 3235.02 91.88 3256.23 91.58 3272.62 89.45 3292.87 87.27 3316.01 80.05 3349.76 75.12 3374.83 69.00 3403.76 63.89 3430.76 63.15 3451.01 63.73 3465.47 67.51 3490.54 65J0 3527.18 77.41 3551.29 81.93 3564.79 83.07 3654.46 88.55 3675.68 89.80 3783.67 85.57 3815.49 85.97 3837.67 86.17 3856.95 90.50 3870.45 90.44 Recording the IR spectrum see Fig. 7

Occupancy (N-elementary analysis): 93% of the max. Occupancy (0.6 mmol / g)

1 N - Yx. = 1:58%

Calculation of the occupancy from the percentage of nitrogen: Occupancy: 0.6 mmol / g 0.0158 _r = 0.00056 mol- g ^" 2 J4.07 g - mol error: 3%

= - = ^ = 0.03 x 1.58%

Example 7:

656 mg (0.4 mmol) of Wang resin (Acros, 1% cross-linked with DNB, 0.5-0.6 mmol / g, 200-400 mesh) is placed in 10 ml of dry chloroform. After a swelling time of 30 min, 173 mg (0.6 mmol) of (S) -5-isocyanatomethyl-2,2-bis-trifluomomethyl [1,3] dioxolan-4-one are added with stirring. The reaction mixture is stirred for 8 days at room temperature. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-93 ((38) (Scheme 1)) IR (KBr): 1851 cm ^"1 (v lactone) S cm ^ v urethane) cm-1% T cm-1% T cm-1% T cm-1 % T 512.97 91.85 536.11 83.47 697.14 52.79 754.03 81.47 903.49 90.30 978.70 82.49 1024.98 90.81 1041.37 91.88 1132.01 71.51 1187.94 77.03 1238.08 58.01 1296.90 90.03 1318.11 84.27 1371.14 87.75 1448.28 73.94 1490.71 75.56 1510.96 74.56 1603.53 80.10 1734.66 67.08 1850.37 79.29 1943.90 90.56 2265.96 91.21 2372.99 90.52 2847.39 74.38 2920.68 62.32 3025.78 69.04 3057.60 76.70 3080.74 80.47 3235.02 91.88 3256.23 91.58 3272.62 89.45 3292.87 87.27 3316.01 80.05 3349.76 75.12 3374.83 69.00 3403.76 63.89 3430.76 63.15 3451.01 63.73 3465.47 67.51 3490.54 65J0 3527.18 77.41 3551.29 81.93 3564.79 83.07 3654.46 88.55 3675.68 89.80 3783.67 85.57 3815.49 85.97 3837.67 86.17 3856.95 90.50 3870.45 90.44

Recording the IR spectrum, see FIG. 8 Occupancy (ZV elementary analysis): 32% of the max. Occupancy (0.6 mmol / g)

N x = - Yx ,. = 0.27%

Calculation of the occupancy from the percentage of nitrogen: Occupancy: 0.2 mmol / g 0.0027 ^■ = 0.00019 mol - g ^"1 1 - 14.07 g - mol ^{" 1} error: 7%

'= ^^ = 0.07 x 0.27%

Example 8: 500 mg (0.3 mmol) of Wang resin (Acros, 1% cross-linked with DNB, 0.5-0.6 mmol / g, 200-400 mesh) is placed in 10 ml of dry chloroform. After a swelling time of 30 min, 133 mg (0.5 mmol) of racemic 4-isocyanatomethyl-2,2-bis-trifluomomethyl-[1,3] oxathiolan-5-one in 1 ml of dry chloroform are added with stirring. The reaction mixture is stirred for 8 days at room temperature. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-103 ((39) (Scheme 1)) IR (KBr): 1816 cm ^"1 (v lactone) 1734 cm ^{" 1} (urethane) cm-1% T cm-1% T cm-1% T cm-1% T 533.22 77.92 698.10 49.93 753.07 77.78 1099.23 81 .13 1153.23 84.10 1169.62 84.41 1220.72 64.08 1233.26 66.00 1275.68 80.51 1449.25 69.53 1490.71 70.20 151.960.77.61 1815.66 80.70 2848.36 71.85 2920.66 60.26 3000.71 80.92 3023.85 61.97 3056.63 71.46 3079.78 78.27 3307.34 84.87 3365.19 74.76 3383.51 67.92 3398.94 63.37 3416.30 64.25 3436.54 61.85 3465.47 65.66 3549.349

Occupancy (S elementary analysis): 25% of the max. Occupancy (0.6 mmol / g)

1 ^N X - - Yx. = 0.49% N i '-1

Calculation of the occupancy from the percentage of sulfur: Occupancy: 0.2 mmol / g 0.0049: 0.00015 mol- g ^" 1 - 32.07 g - mol error: 16% s _ 0.08% = 0.16 x ^~ 0.49%

Example 9: 0.507 g (0.4 mmol) of Rinkamid-4-methylbenzhydιylamine polymer resin (Acros, 1% cross-linked with DNB, 0.4-0.8 mmol / g, 200-400 mesh) are placed in 5 ml of dry chloroform. After a swelling time of 30 min, 146 mg (0.5 mmol) of (R) - (5-oxo-2,2-bis-trifluoromethyl- [1,3] dioxolan-4-yl) acetyl chloride in 85 mg propene oxide are added with stirring , The The reaction mixture is stirred overnight. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-125 ((41) (Scheme 1)) IR (KBr): 1851 cm ^"1 (v lactone) cm-1% T cm-1% T cm-1% T cm-1% T 696.18 53.56 1033.66 64.92 1124.30 71.58 1172.51 64.32 1207.22 53.33 1238.08 42.55 1288.22 78.24 1448.28 65.88 1500.35 44.96 1606.42 62.09 1673.91 52.06 1681.63 52.14 1727.91 48.04 1851.33 75.26 2915.86 65.08 3025.78 78.19 3307.34 72.84 3320.84 72.39 3345.91 73.30 3367.12 70.46 3409.55 54.59 3440.40 51.46 3490.54 70.61 3754.74 75.57 3783.67 76.50 3912.88 79.15 receiving the IR spectrum see FIG. 10

Occupancy (elementary analysis): 50% of the max. Occupancy (0.8 mmol / g)

1 ^N x = - Y x ,. = 3.95%

0.0395 _τ = 0.00035 mol- 6 - 19.00 g - mol error: 8% s ₌ 0.31% = 0.08 x ^~ 3.95% Example 10: 0.587 g (0.4 mmol) of Rinkamid-4-methylbenzhydrylamine polymer resin (Acros, 1% cross-linked with DVB, 0.4-0.8 mmol / g, 200-400 mesh) are placed in 5 ml of dry chloroform. After a swelling time of 30 min, 138 mg (0.5 mmol) of racemic 4-isocyanatomethyl-2,2-bis-trifluomomethyl [1,3] oxathiolan-5-one in 1 ml of dry chloroform are added with stirring. Then 0.002 equivalents of DMAP of a 0.008 M solution in chloroform are added with stirring. The reaction mixture is stirred for 48 h. The product is then filtered off, washed with ethyl acetate and dried in an oil pump vacuum.

TR 5-149 ((42) (Scheme 1)) IR (KBr): 1817 cm ^'1 (v lactone) cm-1% T cm-1% T cm-1% T cm-1% T 555.40 78.54 694.25 63.24 754.03 84.35 1033.66 68.60 1099.23 75.16 1168.65 66.24 1214.94 55.14 1228.44 54.67 1286.29 71.78 1448.28 71:64 150 & 28 57.14 16Ö6.42 72.40 1673.91 69.94 1727.91 64.33 18K -. 62 81: 49 2370.45.8.8.8.28.28 3023.85 74.19 3056.63 76.49 3160.77 86.17 3270.70 74.40 3280.34 74.26 3342.05 65.69 3378.69 59.07 3428.83 53.04 3504.04 60.33 3733.53 75.64 Recording the IR spectrum see Fig. 11

Occupancy (S elementary analysis): 71% of the max. Occupancy (0.8 mmol / g)

Calculation of the occupancy from the percentage of sulfur: Occupancy: 0.57 mmol / g 0.018 = 0.00057 molg ^"

1 - 32.07 g - mol ^"1 error: 3% = ^ = 0.03 x 1.83%

Example 11:

IR experiments to demonstrate covalent bonding by adding 3-isocyanatomethyl-2,2-bis (trifluoromethyl) -IJ-oxazolidin-5-one to the Wang resin.

- IR (KBr) Wang resin (Acros, 1% cross-linked with DNB, 0.5-0.6 mmol / g, 200-400 mesh): cm-1% T cm-1% T cm-1% T cm-1% T 526.47 86.51 566.97 93.89 692.32 84.78 750.17 90.08 817.67 91.67 1012.45 92.53 1116.58 9456 1164.80 93.74 1226.51 91.19 1369.22 93.74 1444.43 90.02 1492.64 90.49 1600.63 91.90 1864.83 94.41 1941.97 94.70 2175.32 94.72 2318.03 94.22 2364.31 92.68 2846.43 87.07 2919.71 84.23 3029.64 89.55 3072.06 91.70 3340.12 93.30 3369.05 94.16 3403.76 91.54 3430.76 91.24 3469.33 91.49 3500.18 91.98 3540.68 94.34 Recording the IR spectrum see Fig. 12

IR (KBr) 3-isocyanatomethyl-2,2-bis (trifluoromethyl) -l, 3-oxazolidin-5-one: cm-1% T cm-1% T cm-1% T cm-1% T 966.16 28J4 1095.37 10.71 1230.36 1.48 1294.00 27.82 1376.93 66.98 1849.41 1 J3 2256.32 6.87 2279.46 10.07 3386.40 59.99 3415.33 57.14 3424.97 57.01 3436.54 57.31 Recording the IR spectrum see Fig. 13

IR (KBr) of a mixture of 3-isocyanatomethyl-2,2-bis (trifluoromethyl) -lJ-oxazolidin-5-one and TR 5-21 ((37) (Scheme 1)): cm-1% T cm-1 % T cm-1% T cm-1% T 690.39 71.78 744.39 74.02 960.38 77.51 1089.59 44.66 1170.58 26.26 1222.65 8.52 1292.08 45.27 1446.35 36.95 1517.71 65.12 1724.06 39.42 1843.62 19.85 2262.10 52.38 2630.44 53.57.24.29.24.24 3046.99 66.67 3089.42 73.73 3282.27 7110 3309.27 66.85 3345.91 68.03 3388.33 57.11 3444.26 68.28 3473.19 44.18 3517.54 72.19 3594.68 61.73 3864.67 76.25 Recording of the IR spectrum see Fig. 14 IR (KBr) TR 5-21 ((37) (Scheme 1)): cm-1% T cm-1% T cm-1% T cm-1% T 541.SO 85.06 694.25 54.66 754.03 64.07 821.53 85.02 869.74 84.29 958.45 78.53 1087.68 65.57 1166.72 62.57 1230.36 56.75 1297.86 74.37 1448.28 66.61 1509.99 65.37 1604.49 76.24 1727.91 62.03 1845.55 56.68 2848.36 66.08 2919.71 50.91 3023.85 57.85 3060.49 68.37 349.5

IR (KBr):

By comparing the characteristic signals (see table above) of 3 - isocyanatomethyl-2,2-bis (tiifluoπnethyl) -lJ-oxazolidin-5-one, a mixture of 3-isocyanatomethyl-2,2-bis (trifluomomethyl) -l , 3-oxazolidin-5-one and TR 5-21 (Scheme 1) and TR 5-21 (Scheme 1) show that in TR 5-21 (Scheme 1) a covalent bond is obtained by adding 3-isocyanatomethyl-2 , 2-bis (trifluoromethyl) -l, 3-oxazolidin-5-one is present on the Wang resin. Which was to be proved.

Example 12: Synthesis on the surface-functionalized support material (33) according to the invention (Scheme 2) Scheme 2 shows reagents and conditions for the synthesis: i) means DMAP, Py, RT (room temperature), ii) means H-Lys (Z) -OMe x HCl, DMAP, Py, RT. iii) stands for DMAP, Py, RT. iv) means TFA, H ₂ O, RT. The yield of reaction (33) → (50) is 45%.

Obtaining the compound (50): 178 mg (0.6 mmol) of H-Lys (Z) -0CH ₃ xHCl (Ne-CBZ- (E) -LysimethylesterhydiOchlorid) are treated with 2 mL dry pyridine in an ultrasonic bath for 15 min. Then 73 mg (0.6 mmol) DMAP are added. The reaction mixture is treated in an ultrasound bath for 15 min (precipitation).

5 mL dry pyridine are added to 0.5 g (0.28 mmol) (33). After 30 min, the reaction mixture with the H-Lys (Z) -OCH _{3 is} added. The reaction mixture is stirred for 44 h. Then 10 mL water is added and the mixture is stirred for 24 h. The product is then filtered off, washed with pyridine, dichloroethane and water and dried in an oil pump vacuum.

4 mL dry pyridine and a spatula tip DMAP are added to 0.37 g (0.22 mmol) (47). After 30 min, 65 mg (0.34 mmol) of 4- (trifluoromethyl) benzoyl chloride (48) are added at 0 ° C. The reaction mixture is stirred for 23 hours at room temperature. The product is then filtered off, washed with ethyl acetate and water and dried in an oil pump vacuum.

2 ml of CH ₂ C1 _{2 are} added to 0.30 g (0J8 mmol) (49). After 30 min, 2 ml of trifluoroacetic acid (99%) and after 5 min 0.05 ml of water are added. The reaction mixture is stirred for 4 hours at room temperature. The resin is then filtered off and washed with trifluoroacetic acid (99%), dichloromethane and methanol. The filtrate is concentrated in vacuo with water, treated in an ultrasound bath for 5 min and freeze-dried. The crude product is purified by flash column chromatography (solvent: methanol / chloroform = 10/2). Yield (50) 43 mg (45%). 1H NMR (CD ₃ OD): δ = 1J9-1.52 (m, 4H, CH ₂ CH ₂ CH ₂ NCOO, CH ₂ CH ₂ NCOO), 1.60-1.86 (m, 2H, CH ₂ CH), 2.96-3.05 (m, 2H, CH ₂ NCOO), 3.60 / 3.63 (s, 3Η, OCH ₃ ), 3.77 (broad signal, NH), 3.94 (broad singlet, CH ₂ ), 4J0 (broad singlet, CH ₂ ), 4.28 / 4.37 (m, CH), 4.95 (m, CH ₂ C ₆ Η ₅ ), 7J7-7.27 (m, 5H, CeHs), 7.59-7.67 (m, 5Η, C ₆ H ₄ CF ₃ ). ¹³ C NMR (CD ₃ OD): δ = 24.47 / 24.65 (CΗ ₂ CΗ ₂ CΗ ₂ NΗCOO), 30.76 (CH ₂ CH ₂ NHCOO), 32.22 / 32.77 (CH ₂ CH), 41.88 / 41.96 (CH ₂ NHCOO) , 53.30 (OCH ₃ ), 53.51 (CH ₂ ), 54.18 / 54.51 (CH), 55.60 (CH ₂ ), 67.84 / 67.92 (CH ₂ C ₆ H ₅ ), 125.73 (q, J = 271Hz, CF ₃ ), 126.56 / 126.62 (2xCH from C ^ CFs), 129.23 (2xCH from C _Ö I ^ CF _S , 2x H from C ₆ H ₅ ) _. 129.44 (CH from C ₆ H ₅ ), 129.95 (2xCH from C ₆ H ₅ ), 133.52 (m, CCF ₃ ), 138.90 (C from C ₆ H ₅ ), 140.87 / 141.16 (C from CeHjCFs), 159.48 (NHCOO ), 171.76 (CO), 172.04 (CO), 173.82 (CON), 174.53 / 174.70 (COOCH ₃ ). ¹⁹ F NMR (CD ₃ OD): δ = 13.35 (s, CE ₃ ). COZY. MS (ΕSI) m / z 582.20524 (582.20578) [M + H] ⁺ , 604.18751 (604.18772) [M + Naf, 620.15234 [M + K] ⁺ , 1185.38537 [2M + Na 1201.35247 [2M + K] ⁺ . Example 13:

Process for the production of linker compounds (53) and (54) (Scheme 3)

Scheme 3 shows the reagents and conditions: i) means 2 HFA, DMF, RT, where the

Yield is 90%, ii) means SOCl ₂ , RT, the yield being questioned 100%, iii) means trimethylsilyl azide, toluene, 0 → 80 ° C., the yield being 40%.

Obtaining the compound (52):

Hexafluoroacetone is slowly introduced into a slurry of 4.97 g (263 mmol) (51) and 20 ml of dry DMF (ultrasound) with vigorous stirring. After the reaction has ended (hexafluoroacetone backflow in a dry ice cooler, bubble counter, ¹⁹ F-NMR, clear solution), the solution is concentrated in an oil pump vacuum, mixed with ice water (ultrasound) and freeze-dried. The residue is taken up in chloroform, filtered, concentrated in vacuo, mixed with ice water (ultrasound) and freeze-dried. No further cleaning steps are necessary. Yield: 30.36 g (90%), colorless crystals, mp 79-80 ° C. IR (KBr): v = 3352, 1818, 1707 cm ^"1. 1H NMR (CDC1 ₃ ): δ = 1.30-1.80 (m, 7H), 1.88 (m, br, 1H), 2.37 (t, J = 7 Hz, 2H), 3.00 (d, J = 7 Hz, 1H), 3.94 (m, 1H). ¹³ C NMR (CDC1 ₃ ): δ = 24.3, 24.9, 28.5, 32.6, 33.8, 54.6, 88.4 (sept, J = 34 Hz), 120.3 (q, J = 286 Hz), 121.3 (q, J = 288 Hz), 171.3, 179.7. ¹⁹ F NMR (CDC1 ₃ ): δ = -3.82 - -4.19 (m, CF ₃ ). MS (ESI) m / z = 338.08212 (338.08215) [M + H] ⁺ . Obtaining compound (53):

11.5 ml (119.0 mmol) of thionyl chloride are added to 4.06 g (12.0 mmol) (52) (cool with dry ice below 0 ° C) while stirring. The solution is stirred at RT for 48 hours, concentrated in vacuo and dried in an oil pump vacuum. No further cleaning steps are necessary. Yield: 4.27 g (100% o); bp 140 ° C / 0.2 ton-. IR (film): v = 3377, 1826 cm ^"1. 1H NMR (CDC1 ₃ ): δ = 1.33-1.47 (m, 3H), 1.50 (m, 1H), 1.60-1.78 (m, 3H), 1.86 ( m, 1H), 2.89 (t, J = 7 Hz, 2H), 3.27 (br.s, 1H), 3.95 (m, 1H). ¹³ C NMR (CDC1 ₃ ): δ = 24.7, 24.8, 27.9, 32.5 , 46.9, 54.5, 88.4 (sept, J = 34 Hz), 120.3 (q, J = 286 Hz), 121.3 (q, J = 289 Hz), 171.4, 174.0. ¹⁹ F NMR (CDC1 ₃ ): δ = - 2.95 (m, CF ₃ ), -2.79 (m, CF ₃ ). MS (EI) m / z = 319 (40) [M-HC1] ⁺ , 301 (20), 273 (8), 246 (14) , 222 (50), 206 (9), 178 (16), 152 (13), 140 (12), 109 (39), 81 (66), 73 (100). Obtaining compound (54):

1.28 ml (9.4 mmol) of trimethylsilyl azide (97%) are added to 2.72 g (8J mmol) (53) in 12 ml of dry toluene (cool with dry ice below 0 ° C). The solution will be Stirred for 24 hours at 3 ° C. and 46 hours at room temperature, then heated to 80 ° C. until the reaction had ended (bubble counter) and then concentrated in an oil pump vacuum (rotary evaporator).

The purification is carried out by flash column chromatography (0 == 2cm, h = 32cm) with CHC1 ₂ . Yield: 1.00 g (40%). IR (film): v = 3375, 2280, 1828 cm ^"1. 1H NMR (CDC1 ₃ ): δ = 1.35-2.00 (m, 7H), 1.88 (m, IH), 3.07 (d, J = 7 Hz, IH), 3.31 (t, J = 7 Hz, 2H), 3.95 (dd, J = 7 Hz, J = 12 Hz, IH). ¹³ C NMR (CDC1 ₃ ): δ = 24.6, 26.1, 30.9, 32.6, 42.8, 54.5, 88.4 (sept, J = 34 Hz), 120.3 (q, J = 287 Hz), 121.3 (q, J - 289 Hz), 122.0 (NCO), 171.3. ¹⁹ F NMR (CDC1 ₃ ): δ = -2.72 (m, CF ₃ ), -2.84 (m, CF ₃ ).

Claims

P atent claims

1. Surface-functionalized carrier material with a polymeric surface and at least one linker compound covalently bound thereto according to the general formulas (1) to (16),

d) (2) (3) (4)

in which P means the polymeric surface, n has the meaning 1 to 12, Ri and R ₂ independently mean H or an alkyl group and L means a spacer which links the linker compound to the polymeric surface.

2. Surface-functionalized carrier material with a polymeric surface and at least one linker compound covalently bound thereto according to the general formulas (1 ⁵ ) to (16 '),

(V) (2') (3') (4')

in which P means the polymeric surface, n has the meaning 1 to 12, RI and R2 independently mean H or an alkyl group and L means a spacer which links the linker compound to the polymeric surface.

3. Surface-functionalized carrier material according to points 1 or 2, characterized in that the polymeric surface and / or the carrier material is an organic polymer.

4. Surface-functionalized carrier material according to claim 3, characterized in that the organic polymer is polypropylene, polyethylene, polysulfone, polystyrene, polyvinyl chloride, polyacrylonitrile, cellulose, amylose, agarose, polyamide, polyimide, polytefrafluoroethylene, polivinylidene difluoride, polyester, polycarbonate, polyacrylate, polyacrylamide or a Is a derivative of these or a copolymer or a blend of these.

5. Surface-functionalized carrier material according to points 1 or 2, characterized in that the carrier material is an inorganic and / or mineral material.

6. Surface-functionalized carrier material according to address 5, characterized in that the carrier material is a glass, a silicate, a ceramic material or a metal.

7. Surface-functionalized carrier material according to one of claims 3 to 6, characterized in that the carrier material is a composite of at least one inorganic and / or mineral material and at least one organic polymer.

8. Surface-functionalized carrier material according to one of the preceding claims, characterized in that the carrier material is in the form of a membrane, a film, a plate, a microtiter plate, a reaction vessel, a microscope slide, a fiber, a hollow fiber, a fleece, a fabric, a powder , a granulate or particles is present and is porous or non-porous.

9. Surface-functionalized carrier material according to point 8, characterized in that the carrier material is in the form of a membrane with a symmetrical or asymmetrical pore structure.

10. Surface-functionalized carrier material according to claim 8 or 9, characterized in that a pore size of 1 nm to 10 μ is required.

1. Process for producing a surface-functionalized carrier material with a polymeric surface and at least one linker compound covalently bound thereto according to the general formulas (1) to (16),

d) (2) (3) (4)

where P means the polymeric surface of the support material, n has the meaning 1 to 12, Ri and R ₂ independently mean H or an alkyl group and L means a spacer which links the linker compound to the polymeric surface, by converting at least one linker compound of the general formulas (17) to

(32),

(17) (18) (19) (20)

(21) (22) (23) (24)

(25) (26) (27) (28)

in which n, Ri and R ₂ have the above meaning and L means a spacer which is intended to link the linker connections to the polymeric surface,

with a carrier material that contains functional groups that can be linked.

2. Process for producing a surface-functionalized carrier material with a polymeric surface and at least one linker compound covalently bound thereto according to the general formulas (1') to (16'),

(1') (2') (3') (4')

where P means the polymeric surface of the carrier material, n has the meaning 1 to 12, Ri and R ₂ independently of one another mean H or an alkyl group and L means a spacer which links the linker compound to the polymeric surface by reacting at least one linker compound of the general one Forms (17') to (32')

(25') (26") (27') (28')

in which n, Ri and R ₂ have the above meaning and L means a spacer which is intended to link the linker compounds to the polymeric surface, with a carrier material which contains functional groups capable of being linked.

13. The method according to claim 11, characterized in that for the production of at least one of the compounds according to the general formulas (1) to (4), (9) to (16) and (25) to (28) the carboxy group of corresponding compound according to the general formulas (17) to (20) and (25) to (32) is activated before the reaction with the carrier material.

14. The method according to claim 12, characterized in that for the production of at least one of the compounds according to the general formulas (1 ') to (4'), (9') to (16') and (25') to (28') the carboxy group of the corresponding compound according to the general formulas (17') to (20') and (25') to (32') before the reaction with the support material is activated.

15. The method according to claim 13 or 14, characterized in that the activation of the carboxy group of at least one of the compounds according to the general formulas (17) to (20) and (25) to (32) or the compounds according to the general formulas ( 17') to (20') and (25') to (32') by reaction of the carboxy group with a thionyl chloride or phosphorus pentachloride to form an acid chloride.

16. The method according to claim 11 or 12, characterized in that at least one of the compounds according to the general formulas (25) to (32) from the corresponding compound according to the general formulas (17) to (24) or at least one of the compounds according to general formulas (25') to (32') can be obtained from the corresponding compound according to the general formulas (17') to (24') by attaching the spacer L.

17. The method according to claim 16, characterized in that the spacer L is linked to at least one of the linker compounds according to the general formulas (17) to (20) or according to the general formulas (17 ') to (20') by acylation to the ester or amide or to at least one of the linker compounds according to the general formulas (21) to (24) or (21 ') to (24') by addition to the urethane or urea.

18. The method according to claim 16 or 17, characterized in that the spacer L is an amino acid, a peptide or another compound with at least one amino or hydroxy function for binding to the compounds according to the general formulas (17) to ( 24) or (17') to (24') and a carboxy function for immobilization to the polymeric surface.

19. The method according to any one of claims 11 to 18, characterized in that the linking of at least one linker compound of the general formulas (17) to (20), (17 ') to (20'), (25) to (32) and ( 25 ') to (32') with the polymeric surface by acylation to the ester or amide or the linkage of at least one Linker compound of the general formulas (21) to (24) or (17 ') to (24 J is carried out by addition to the urethane or urea.

20. The method according to claim 19, characterized in that when linking the linker compounds of the general formulas (17) to (20), (17 ') to (20'), (25) to (32) and (25') to (32') a scavenging reagent for hydrogen chloride is used with the polymeric surface by acylation to the ester or amide.

21. The method according to claim 20, characterized in that NaHCO ₃ or pyridine or propene oxide is preferably used as the scavenging reagent for hydrogen chloride.

22. Process according to points 11 or 12, characterized in that the linker compounds according to the general formulas (17) to (32) are used once or the process is repeated several times.

23. The method according to claim 11 or 12, characterized in that a solvent or solvent mixture and a temperature range between -100 ° C and + 100 ° C are used to obtain the surface-functionalized carrier material.

24. The method according to claim 23, characterized in that chloroform and/or dichloromethane is used as the solvent for base-sensitive substances and pyridine is used for base-insensitive substances.

25. The method according to claim 23, characterized in that the reaction takes place at room temperature.

26. Method according to paragraph 11 or 12, characterized in that after the reaction, unreacted substances are removed by washing with a washing liquid.

27. The method according to claim 26, characterized in that water, ethyl acetate or an organic solvent or a solvent mixture is used as the washing liquid.

28. The method according to any one of claims 11 to 27, characterized in that a Wang resin, a PEGA resin, is used as the polymeric surface (P) and/or carrier material Rinkamid resin or a material mentioned in claims 3 to 10 for the carrier material is used.

29. Method according to one of the statements 11 to 28, characterized in that the obtained surface-functionalized carrier material is characterized according to the general formulas (1) to (16) or (1 ') to (16') by infrared spectroscopy and / or the occupancy is determined by elementary analysis.

30. Use of a carrier material according to one of claims 1 to 10 for the synthesis of amino acids, peptides, proteins or molecules with at least one peptide structural unit, wherein a first amino acid to be used for the synthesis is covalently bound to a surface-functionalized carrier material according to one of claims 1 to 10 and chain extension occurs through successive attachment of further amino acids and/or chemical modification.

31. Use according to claim 30, characterized in that the first amino acid used for the synthesis is bound to the carrier material according to one of claims 1 to 10 by a peptide bond between the amino group of the amino acid and the activated acid function of the linker compound and then further bonds of amino acids optionally N-, O- or S- or C-terminal.

32. Use according to claim 30 or 31, characterized in that the N-, O- or S- or C-terminal connection of the amino acid to the linker connection is controlled by blocking the amino group or the carboxy group of the amino acid and / or the amino, hydroxy or thiol group of the corresponding link compound with chemical protecting groups.