EP1651597A2

EP1651597A2 - N-sulphonylated amino acid derivatives, method for the production and use thereof

Info

Publication number: EP1651597A2
Application number: EP04733311A
Authority: EP
Inventors: Jörg STÜRZEBECHER; Torsten Steinmetzer; Andrea Schweinitz; Anne STÜRZEBECHER; Kerstin Uhland
Original assignee: Curacyte Chemistry GmbH
Current assignee: Medicines Company Leipzig GmbH
Priority date: 2003-05-16
Filing date: 2004-05-17
Publication date: 2006-05-03
Also published as: AU2004238558A1; JP2007500734A; CA2525886A1; WO2004101507A3; EP2316830A3; WO2004101507A2; JP4653100B2; US7772251B2; CA2525886C; EP2316830A2; US20070055065A1; US20110002992A1; DE10322191A1; DE10322191B4

Abstract

The invention relates to N-sulphonylated amino acid derivatives wherein an aryl radical is bound to an amino acid via the sulphonyl groups in the N-terminal and a radical is bound in the C-terminal via the carbonyl group. Said radical contains at least one imino group and at least one other basic group representing an optionally modified amino-, amidino- or guanidino group. The invention also relates to a method for the production of said compounds and to the use thereof, especially as inhibitors of matriptase.

Description

N-sulfonylated amino acid derivatives, process for their preparation and their use

The present invention relates to N-sulfonylated amino acid derivatives, an aryl residue being bonded to the amino acid via the sulfonyl group N-terminal and a residue having at least one imino group and at least one further basic group, which is a C, being bonded via the carbonyl group optionally represents modified amino, amidino or guanidino group. The invention also relates to processes for the preparation of these compounds and their use, preferably as medicaments and in this regard in particular as matriptase inhibitors. ^"

Proteases regulate numerous physiological processes that enable or stimulate the growth and metastasis of tumor cells. This applies in particular to the proteolytic degradation of the extracellular matrix proteins surrounding the tumor cells, which enables the tumor cells which have been deposited by tumors to invade adjacent tissues and the lymphatic or blood system. Proteases are also involved in the activation of growth factors, e.g. stimulate the proliferation of • tumor cells or angiogenesis and thus enable tumor growth. These proteolytic enzymes include various matrix metalloproteases, membrane bound metalloproteases, lysosomal cysteine proteases and a variety of serine proteases such as e.g. Urokinase, plasmin, elastase, thrombin or cathepsin G, and also the type II transmembrane serine protease matriptase or MT-SP1 (Hopper et al, J. Biol. Chem. 276, 857-860, 2001).

There have been numerous attempts to inhibit the growth and metastasis of tumors by using protease inhibitors, but attempts with inhibitors of matrix metalloproteases have so far shown little effect in clinical studies (cous- sens et al., Science 295, 2387-2392, 2002). In the meantime, initial clinical studies with urokinase inhibitors have also been initiated, but no results are yet known about their effectiveness.

Matriptase is a trypsin-like serine protease that was originally isolated from breast cancer cells and preferentially cleaves C-terminal peptide bonds of the basic amino acid arginine. (Shi et al., Cancer Res. 53, 1409-1415, 1993; Lin et al., J. Biol. Chem. 272, 9147-9152, 1997).

In 1998, the gene of matriptase as a putative tumor suppressor was cloned by a subtractive hybridization method using healthy and carcinogenic intestinal tissue (Zhang et al. Cytogenet. Cell Genet. 83, 56-57, 1998).

Matriptase and MT-SP1 (Abbreviation for "membrane-type serine protease 1) (Takeuchi et al., Proc. Natl. Acad. Sci. USA 96, 11054-11061, 1999; Takeuchi et al, J. Biol. Chem. 275 , 26333-26342, 2000) have the same cDNA. However, due to alternative splicing, the protein sequence of the matriptase is shortened by 172 amino acids at the N-terminus compared to MT-SP1. The gene for MT-SP1 was isolated from an epithelial cell line of a prostate tumor.

In the context of the present invention, the term “matriptase” denotes any trypsin-like protein with a molecular weight of 72 to 92 kDa which is derived from the gene sequences with the entry numbers AF118224, AF133086, BANKIt257050 and NM021978 (GenBank / EBI Data Bank) and has already been described earlier (Takeuchi et al., Proc. Natl. Acad. Sei. USA 96, 11054-11061, 1999; Lin et al., J. Biol. Chem. 274, 18231-18236, 1999). In particular, the term "matriptase" denotes both the single and double chain form of the protein. The zymogenic, inactive form of the matriptase is a single chain protein. The double chain form of matriptase is its active form, which has catalytic activity. In the context of the present invention, therefore, in particular, the term “matriptase” denotes the original matriptase described above as well as MT-SP1. The enzyme is anchored in the membrane of epithelial or cancer cells by means of a transmembrane domain, the serine protease domain of the matriptase being located on the cell surface and thus in the extracellular space (Hopper et al., J. Biol. Chem. 276, 857 -860, 2001). It was therefore suspected that matriptase could be involved in the proliferation and metastasis of breast cancer cells through the breakdown or remodeling of extracellular matrix proteins, the activation of latent growth factors and other proteolytic cascades (Shi et al., Cancer Res. 53, 1409-1415, 1993 ; Lin et al., J. Biol. Chem. 272, 9147-9152, 1997).

Matriptase could also be isolated from human milk, but in this case it was almost completely in the form of a proteolytically inactive complex with the endogenous inhibitor HAI-1 (Lin et al., J. Biol. Chem. 274, 18237-18242, 1999). In contrast, the matriptase from breast cancer cells is largely in an uncomplexed and therefore catalytically active form, and only a small part is bound to HAI-1.

The first potential substrates for matriptase have been described. Matriptase is able to activate the hepatocyte growth factor (HGF), which is also referred to as the scattering factor (Lee et al., J. Biol. Chem. 275, 36720-36725, 2000). Pro-HGF is secreted by cancer or stromal cells in inactive form as a single-chain protein and is converted into the active two-chain form (HGF) in the extracellular space by cleavage at the C-terminal of Arg495. Binding of HGF activates the cell surface receptor c-Met and phosphorylates it on certain tyrosine residues. A close correlation between high expression of c-Met, matriptase and HAI-1 and a poor prognosis in breast cancer patients has recently been demonstrated (Kang et al., Cancer Res. 63, 1101-1105, 2003). When examining ovarian tumors, it was also shown that matriptase is expressed to a greater extent. It was found that, in contrast to type I / II tumors, especially in advanced type III / IV tumors, matriptase is expressed almost exclusively without HAI-1. This indicates that in the advanced stage there is an imbalance between the matriptase and the inhibitor HAI-1, which increases the proteolytic activity of the matriptase and therefore probably also the invasive potential of the tumor cells (Oberst et al., Clin. Cancer Res 8, 1101- 1107, 2002).

In addition to activating Pro-HGF, matriptase may also be involved in activating the plasminogen activator cascade. Thus, matriptase is able to activate pro-urokinase to urokinase (uPA) (Lee et al, J. Biol. Chem. 275, 36720-36725, 2000; Takeuchi et al., J. Biol. Chem. 275, 26333 -26342, 2000), which converts plasminogen into plasmin. Plasmin is the principal activator of the matrix metalloproteases, which are involved in the breakdown of extracellular matrix proteins, which is considered a prerequisite for metastasis.

By Ihara et al. (J. Biol. Chem. 277, 16960-16967, 2002) could be shown that gastric cancer cells increasingly express β1-6-N-acetylglucosaminyltransferase (GnT-V), which is able to glycosylate matriptase. This modification makes the matriptase more stable to degradation and is present in an increased concentration in proteolytically active form.

From these findings it can be deduced that with the development of an effective and selective matriptase inhibitor there is a possibility of inhibiting the proliferation of tumors and their metastasis. Although the x-ray structure of the catalytic domain of the matriptase in complex with benzamidine and the bark ankreas trypsin inhibitor has now also been elucidated, only a few inhibitors of the matriptase are known to date (Friedrich et al., J. Biol. Chem. 277, 2160-2168 , 2002).

By Enyedy et al. (J. Med. Chem. 44, 1349-1355, 2001) have described bis-benzamidines, the most effective inhibitor having a K; value of 0.19 μM.

WO 01/97794 describes a method for inhibiting carcinoma progression in which matriptase plays a role. Compounds are used that contain two groups that are capable of being positively charged at a physiological pH. These groups are connected to one another by a chemical structure unit which has a length of 5 to 30, preferably 15 to 24, angstroms. The amino, amidino and guanidino groups and a cyclic group derived from the amidino and guanidino groups are disclosed as positively charged groups. Amino acid derivatives are not mentioned in WO 01/97794, in particular accordingly no sulfonylated amino acid derivatives. Rather, the compounds explicitly disclosed in WO 01/97794 differ fundamentally from the compounds claimed in the context of the present invention.

In WO 02/20475 tripeptide aldehydes with C-terminal arginal are published. After pre-incubation of matriptase with these inhibitors over a period of 30 minutes, IC ₅₀ values of less than 100 nM were determined for the most effective compounds, but exact inhibit constants were not given. These inhibitors presumably bind covalently to matriptase to form a hemiacetal. In the case of the development of inhibitors for other trypsin-like serine proteases, such as thrombin or factor Xa, however, it has been shown that such transition-state-analogous peptide aldehydes are not suitable for the development of an active substance which can be used medically.

By Long et al. (Bioorganic. Med. Chem. Lett. 11, 2515-2519, 2001) described the synthesis of a bicyclic peptide from 14 amino acids, which was originally isolated from the seeds of sunflowers. The peptide inhibits matriptase with an inhibition constant of 0.92 nM, but it can be assumed that these structures are not suitable for the development of an active ingredient.

One of the objects on which the present invention is based was therefore to provide an active ingredient which is also suitable for therapeutic applications and which inhibits matriptase with high activity and specificity.

Accordingly, the present invention relates to a compound of the formula (I) or a salt or a prodrug of this compound, wherein

(a) Xi and X ₂ independently of one another are hydrogen or an alkyl radical having 1, 2 or 3

Are C atoms and at least one of the radicals Xi and X _{2 is} a radical of the structure (I ')

where, if appropriate, at least one of the methylene groups indicated with m or n according to (V) is at least simply substituted with a hydroxyl, a halogen, a pseudo-halogen or a COOR ₂ 'group and R ₂ ' is a linear, branched one or is a cyclic alkyl group with 1 to 10 C atoms, and / or optionally at least one of the C atoms of the methylene groups indicated with m or n according to (T) is replaced by S, N or O and / or optionally at least one of the cycles

is a double bond according to (V), or wherein (b) Xi and X ₂ are bridged to form a cycle such that the compound according to (I) has the structure (I ") where, if appropriate, at least one of the methylene groups according to (I ") indicated with m or n is at least simply substituted with a hydroxyl, a halogen, a pseudo-halogen or a COOR ₂ 'group and R _{2 is} a linear, branched one or is a cyclic alkyl group with 1 to 10 C atoms, and / or optionally at least one of the C atoms which is replaced by m or n indicated methylene groups according to (I ") by S, N or O and / or while maintaining the C- Imino group bonded terminally to the sulfonylated amino acid, if appropriate, at least one of the cycles

bonds forming according to (I ") is a double bond, and wherein

(i) Ri is an optionally partially hydrogenated aryl or heteroaryl group containing at least one of the atoms O, N or S with 5 to 20 C atoms, or a linear, branched or cyclic alkyl group with 1 to 10 C atoms, where Ri is optionally substituted with - at least one halogen and / or pseudohalogen group and / or at least one linear, branched or cyclic alkyl or alkyloxy or alkylthio group with 1 to 10 C atoms, which is optionally at least monosubstituted with a halogen -, Pseudohalogen-, Hydroxy-, Amino-, Cyano-, Amidino-, Guanidino or carboxyl group, where the carboxyl group is optionally esterified with a linear, branched or cyclic alkyl group with 1 to 10 C-atoms, and wherein the linea- right, branched or cyclic alkyl group with 1 to 10 C atoms optionally contains at least one heteroatom selected from the group consisting of O, N and S, and / or at least one aryl or heteroaryl group with 5 to 20 C atoms, wherein this aryl or heteroaryl group is optionally substituted with at least one linear, branched or cyclic alkyl group with 1 to 10 carbon atoms and / or at least one COR ₂ 'and / or COOR ₂ ' group, where R ₂ 'is one is linear, branched or cyclic alkyl group with 1 to 10 C atoms, and / or at least one halogen group and / or at least one pseudohalogen group and / or at least one alkoxy group or an alkylthio group, the alkyl radical in each case having 1 to 10 C atoms, and / or ~ at least one nitro group and / or at least one haloalkyl group with 1 to 10 C atoms, and wherein the aryl or heteroaryl group via an alkylene group with 1 to 3 C atoms or an oxygen atom or via a sulfur atom is bonded to the remainder Ri; - At least one hydroxy, amino, cyano, amidino, guanidino, carboxyl or carboxyalkyl group, where the amino group is optionally acylated and / or wherein the alkyl group of the carboxyalkyl group has 1 to 10 carbon atoms and / or Carboxyl group is optionally esterified or amidated with a linear, branched or cyclic alkyl group having 1 to 10 carbon atoms;

(ii) R _{2 is} an at least monosubstituted aryl group with 1 to 10 C atoms, where at least one of these C atoms is replaced by S, N or O, at least one substituent is a group according to j, R _{2 is} optionally additionally substituted with a hydroxyl, COR ₂ 'or COOR ₂ ' group and R ₂ 'is a linear, branched or cyclic alkyl group having 1 to 10 C atoms; (iii) R _{3 represents} a radical of the following formula (II):

A ₁ TA-R ₄ (II) where

Ai either does not exist or is an alkylene group with 1 to 4 carbon atoms, which is optionally substituted with at least one halogen and / or pseudohalogen group and / or at least one linear, branched or cyclic alkyl group with 1 to 10 carbon atoms and / or at least one aryl or an aralkyl group with 5 to 10 carbon atoms and / or at least one cycloalkyl group with 3 to 10 atoms and / or at least one hydroxyl, cyano, alkyloxy or alkylthio with 1 to 10 carbon atoms, Carboxyl or carboxyalkyl group, the alkyl group of the carboxyalkyl group having 1 to 10 C atoms and / or the carboxyl group optionally being esterified or amidated with a linear, branched or cyclic alkyl radical having 1 to 10 C atoms; - T either does not exist or is one of the following groups:

wherein R _{5 is} hydrogen or an alkyl group with 1 to 10 C atoms or an alkylene group with A ₂ which optionally forms a cycle containing at least one hetero atom and has 1 to 6 C atoms; where the amide or ester bond can be incorporated in both orientations, which also includes the following orientations:

A _{2 is} a linear, branched or cyclic alkylene group with 1 to 10 C atoms or an aryl, heteroaryl or aralkylene group with 1 to 10 C atoms, optionally containing at least one hetero atom, selected from the group consisting of N, S and O , which is optionally substituted with at least one halogen and / or pseudohalogen group and / or at least one linear, branched or cyclic alkyl group with 1 to 10 C atoms and / or ~ at least one aryl or an aralkyl group with 5 to 10 C. -

Atoms and / or at least one cycloalkyl group with 3 to 10 carbon atoms and / or - at least one hydroxyl, cyano, alkyloxy or alkylthio with 1 to 10 carbon atoms, carboxyl or carboxyalkyl group, the alkyl group being the Carboxyalkyl group has 1 to 10 carbon atoms and / or the carboxyl group is optionally esterified or amidated with a linear, branched or cyclic alkyl radical having 1 to 10 carbon atoms; (iv) Rt is one of the following, optionally modified basic groups: where t = 0.1; R ₆ and R independently of one another are hydrogen or an alkyl group having 1 to 6 C atoms or an alkylene group having 1 to 5 C atoms which forms a cycle with A ₂ , or a hydroxyl, amino, alkylamino, acyl or alkyloxycarbonyl group are, wherein the alkylamino, acyl and alkyloxycarbonyl groups independently of one another have 1 to 6 carbon atoms, and wherein R _{8 is} hydrogen or an alkyl group with 1 to 3 carbon atoms or an alkylene group with 1 to 3 carbon atoms which forms a cycle with Rβ;

(v) Q is either a CH group or N;

(vi) j = 0.1.2; = 0, 1, 2, 3; m, n independently of one another = 0, 1, 2, 3, 4, 5, where m + n = 3, 4, 5; and wherein the compound of formula (I) is neither

with s = 0, 1, 2 still

with s = 0.1.

If the above-mentioned compounds according to the invention are present as a salt, salts with mineral acids and / or salts with suitable organic acids are preferred. Among other things, the compounds according to the invention are preferably present as hydrochlorides or as sulfates. Suitable organic acids are, for example, formic acid, acetic acid, methylsulfonic acid, succinic acid, malic acid and trifluoroacetic acid. Among others, preferred salts of the compounds according to the invention with suitable organic acids are acetates.

According to a preferred embodiment, the compound according to the invention has a structure in which at least one of the radicals Xi and X has a structure (V). In the context of this embodiment of the non-cyclic imines bound to the carbonyl group of the amino acid which is medium-sized according to (I), preference is given to compounds in which exactly one of the radicals Xi and X _{2 has} a structure according to (V). The non-cyclic radical is particularly preferably hydrogen, methyl, ethyl or n-propyl, more preferably methyl or ethyl and particularly preferably methyl. With regard to the substituted cyclic radical, embodiments are preferred in which m + n is 3 or 4. The residue R_ι, which is mandatory in residue R _3, can generally be chosen arbitrarily in accordance with the definitions made above. Those radicals R - _t which are not modified and which are selected from the group consisting of are very particularly preferred

The indices m and n of the cycle according to (T) can be chosen such that the radical R _{3 is} basically in the 2-, 3- or, depending on the ring size, also in the 4-position with respect to the nitrogen bonded to the carbonyl group of the medium-sized amino acid can be located. For example, the 3- or the 4-position are preferred, with m = n = 2 the 4-position being particularly preferred.

In the context of the embodiment of the non-cyclic imines bound to the carbonyl group of the amino acid which is medium-sized according to (I), radicals R ₃ in which A _{2 is} not present are further preferred. Also preferred are radicals R ₃ in which the functional group T is either not present or is selected from

Embodiments in which T is not present are very particularly preferred. With regard to group A ₂ , alkylene groups having 1, 2, 3, 4 or 5 carbon atoms, in particular the methylene, ethylene, are within the scope of the non-cyclic imines bonded to the carbonyl group of the amino acid which is medium-sized according to (I). n-Propylene, isopropylene, butylene and pentylene grape are preferred.

In the case where A _{2 is} an aryl, heteroaryl or aralkylene group, groups are of the structures, for example

to be mentioned, where v and w can be independently 0, 1 or 2 and the two alkylene groups can also be positioned in the 1,2 or 1,3 position to one another.

For example, the 1,4-position is preferred. Both the aryl residue and at least one of the two alkylene groups can be suitably substituted as defined above. If A ₂ comprises a heteroaryl group, this preferably has 1 to 3 heteroatoms.

In principle, the cycle according to (V) can contain at least one heteroatom, preference being given here to oxygen, nitrogen or sulfur. If the heteroatom is nitrogen, for example, this nitrogen can have as a further radical, for example, hydrogen or a linear, branched or cyclic alkyl group having 1 to 10 carbon atoms or can form a double bond with an adjacent carbon atom of the cycle. According to a particularly preferred embodiment, none of the methylene groups according to (! ') Is substituted by a hetero atom.

If the cycle according to (V) is additionally substituted, preferred additional substituents include carboxyalkyl groups of the general structure -COOR ₂ , further preferably R ₂ 'being an alkyl group having 1, 2 or 3 carbon atoms and particularly preferably a methyl or Is ethyl group.

According to a preferred embodiment, the compounds according to the invention according to (b) have a cyclic imine bonded to the carbonyl group of the amino acid which is medium-sized according to (I) and thus have a structure according to (I ").

Accordingly, the present invention also relates to a compound as described above, which compound has the structure (I ")

having.

In this regard, the cycle according to (I ") preferably has 5, 6 or 7 ring atoms. Accordingly, it is conceivable that the radical R ₃ in the 2-, 3- or 4-position to the Bonylgruppe of the middle amino acid bound imine nitrogen is located. Preferred embodiments include those in which the cyclic amine cycle has 5 or 6 ring atoms. A six-membered ring is particularly preferred. According to this particularly preferred six-membered ring, the indices m and n can be chosen as desired. Examples of the possible combinations are, for example, m = 0 and n ^, m = 1 and n = 3, m = 2 and n = 2, m = 3 and n = 1, m = 4 and n = 0. According to a very particularly preferred embodiment of the compounds according to the invention, m = n = 2. The group Q according to structure (I) is therefore very particularly preferably in the 4-position relative to the imine nitrogen bonded to the carbonyl group of the medium-sized amino acid.

Accordingly, the present invention also relates to a compound as described above, which is characterized in that m = n = 2.

In the context of the present invention, the cycle of the cyclic imine can be suitably substituted. Among the substituents described above, the COOR ₂ 'group is particularly preferred, wherein R ₂ ' is in turn preferably an alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms and particularly preferably a methyl group or an ethyl group. If the cycle is substituted by a halogen, fluorine, chlorine and bromine are particularly preferred. The hydroxyl group is also a suitable substituent. Furthermore, the cycle can be substituted with two or more identical or different, in particular the substituents mentioned as being preferred.

At least one of the methylene groups indicated in the cycle according to (I ") with m or n may be substituted by a hetero atom, oxygen, nitrogen or sulfur preferably being mentioned here. If the hetero atom is nitrogen, for example, this nitrogen can be a further radical, for example hydrogen or have a linear, branched or cyclic alkyl group with 1 to 10 C atoms or form a double bond with an adjacent C atom of the cycle. In general, the cycle according to (I ") can contain at least one double bond which is either between two heteroatoms, two carbon atoms or a carbon atom and a hetero atom can be formed. Accordingly, within the scope of the present invention Cycles described which have at least one double bond and in which the imine nitrogen bonded to the carbonyl group of the medium-sized amino acid is retained, that is to say bound to the adjacent atoms with three single bonds.

According to a particularly preferred embodiment, none of the methylene groups according to (I ") is substituted by a hetero atom. According to a further particularly preferred embodiment, the methylene groups which form the cycle are not substituted.

Accordingly, the present invention also relates to a compound as described above, wherein the cycle formed by Xi and X _{2 has the} following structure

and m, n independently of one another = 0, 1, 2, 3, 4, 5, where m + n = 3, 4, 5. Further preferred are compounds in which m and n are selected such that a six-membered ring is formed becomes. Accordingly, as already described above, Q can be in the 2-, 3- or 4-position relative to the imine nitrogen, the 4-position being particularly preferred. Further particularly preferred is m = n = 2.

In the context of the present invention, Q is a CH group or nitrogen. The present invention also describes compounds in which this CH group carries a suitable substituent instead of hydrogen. Preferred substituents include those with which the methylene groups of the cycle can also be substituted.

Accordingly, the present invention also describes a compound as described above, which is characterized in that Q is a nitrogen atom. In the context of this embodiment, the group Ai which may be present in the radical R ₃ can be chosen as desired within the scope of the definitions described above. Particular preference is given to a group Ai which is a methylene or ethylene or propylene group, in particular a methylene or ethylene group and is optionally substituted, alkyl substituents having 1, 2 or 3 atoms being among the particularly preferred substituents, in particular methyl and ethyl, particularly preferably methyl, and / or halogen, in particular fluorine, chlorine and bromine, and / or cycloalkyl radicals having preferably 5, 6 or 7 carbon atoms and / or carboxyalkyl radicals, the alkyl radical preferably being methyl or ethyl and the Carboxyl group is preferably strengthened or amidated with a methyl or ethyl group.

According to a very particularly preferred embodiment of the compounds according to the invention in which Q is a nitrogen atom, group Ai is not present.

Accordingly, the present invention also describes a compound as described above, which is characterized in that Q is a nitrogen atom and Äi is absent.

With regard to the functional group T which may be contained in radical R ₃ in the context of the present invention, all of the groups described above can be selected. In particular, in the case where Q is a nitrogen atom, functional groups are T groups of the structures

prefers. Groups of structures are particularly preferred for T.

more preferably R _{5 is} hydrogen. According to further particularly preferred embodiments, T is a group of the structures

T being a functional group of the structure

is. The amide or ester groups can be incorporated in both orientations.

Accordingly, the present invention also describes a compound as described above, which is characterized in that Q is a nitrogen atom and T is a functional group of the structure

With regard to the group A _{2 which is} mandatory in the rest R ₃ , there are no restrictions within the definitions described above.

Particularly in the case that Q is a nitrogen atom, and more especially that T is a carbonyl group, ₂ alkylene groups having 1 to 6 carbon atoms are preferred for the group A, the methylene, ethylene, propylene, butylene and particularly preferably Methylene, ethylene and propylene groups are particularly preferred.

to be mentioned, where v and w can independently be 0, 1 or 2 and the two alkylene groups can also be positioned in the 1,2- or 1,3-position to one another. For example, the 1,4-position is preferred. Both the aryl radical and at least one of the two alkylene groups can be suitably substituted. If A ₂ comprises a heteroaryl group, this preferably has 1 to 3 heteroatoms.

The group A ₂ can also be suitably substituted as defined above. Particularly suitable substituents here are halogens, preferably fluorine, chlorine or bromine, and / or alkyl radicals having preferably 1, 2 or 3 carbon atoms, such as methyl, ethyl, n-propyl or isopropyl, particularly preferably methyl and ethyl, and / or cycloalkyl radicals preferably 5, 6 or 7 carbon atoms and / or carboxyalkyl radicals, the alkyl radical preferably being methyl or ethyl and the carboxyl group preferably being esterified or amidated with a methyl or ethyl group.

According to a very particularly preferred embodiment of the present invention, group A _{2 is} not substituted. Accordingly, the present invention describes a compound as described above, which is characterized in that A _{2 is} a methylene, ethylene or propylene group. In particular, the present invention describes a compound as described above, which is characterized in that it has the following structure:

where s = 1, 2, 3.

According to a likewise preferred embodiment, the present invention describes a compound, as described above, which is characterized in that, as functional group T, it is a group of the structure has the following structure:

where s = 1, 2, 3, preferably s = 2.

All of the structures mentioned above can be used as the radical i contained in radical R ₃ . T = 0 is preferred here. Rt residues of the following structures are particularly preferred:

further preferably R ₆ and R ₇ are hydrogen.

R ₄ radicals of the following structures are very particularly preferred:

further preferably R _Ö and R ₇ are hydrogen. Accordingly, the present invention also relates to a compound as described above, which is characterized in that Rj is selected from the group consisting of

According to a likewise preferred embodiment of the present invention, Q is a CH group. Therefore, the present invention also describes compounds as described above which are characterized in that Q is a CH group.

In the context of this embodiment, the group Ai which may be present in the radical R ₃ can be chosen as desired in the same way as described above. Particular preference is given to a group Ai which is a methylene or ethylene or propylene group, in particular a methylene or ethylene group and is optionally substituted, alkyl substituents having 1, 2 or 3 atoms, in particular methyl, being particularly preferred as substituents and ethyl, and / or halogen, in particular fluorine, chlorine and bromine, and / or cycloalkyl radicals with preferably 5, 6 or 7 carbon atoms and / or carboxyalkyl radicals, the alkyl radical being preferably methyl or ethyl and the carboxyl group preferably having a methyl or ethyl group is esterified or amidated.

According to a very particularly preferred embodiment of the compounds according to the invention in which Q is a CH group, group Ai is not present.

Accordingly, the present invention also describes a compound as described above, which is characterized in that Q is a CH group and Ai is not present.

With regard to the functional group T which may be present in radical R ₃ in the context of the present invention, all of the groups described above can to get voted. In particular in the case where Q is a CH group, the functional groups T are groups of the structures

prefers. Groups of structures are particularly preferred for T.

T is very particularly preferably a functional group of the structure

is. The amide or ester groups can be incorporated in both orientations.

Accordingly, the present invention also describes a compound as described above, which is characterized in that Q is a CH group and T is a functional group of the structure

In the event that Q is a CH group and T is one of the preferred functional groups mentioned, the group Ai is not present according to a particularly preferred embodiment of the present invention.

Therefore, the present invention also describes a compound as described above, which has the following structure:

In addition to the 4-position of the radical R ₃ explicitly described in the structural formula above, the 2- or the 3-position are also possible, the 4-position being preferred.

According to a likewise preferred embodiment of the present invention, neither the group Ai nor the functional group T is present.

The present invention therefore also describes a compound, as described above, which has the following structure:

As far as the group A ₂ present in the rest of R _{3 is} concerned, there are, as already discussed above, no restrictions within the definitions described above. Especially in the case that Q is a CH group and further in particular that either T is a group of the structure

is or is not present, the group A _{2 is} alkylene groups having 1 to 6 carbon atoms, in particular the methylene, ethylene, propylene, butylene group, further in particular the methylene, ethylene and propylene group and very particularly the ethylene preferred group.

The group A ₂ can also be suitably substituted as defined above. In particular, halogens, preferably fluorine, chlorine or bromine, and / or alkyl radicals with preferably 1, 2 or 3 carbon atoms, such as methyl, ethyl, n-propyl or isopropyl, particularly preferably methyl and ethyl, are particularly suitable as substituents, and / or cycloalkyl radicals with preferably 5, 6 or 7 carbon atoms and / or carboxyalkyl radicals, the alkyl radical preferably being methyl or ethyl and the carboxyl group preferably being esterified or amidated with a methyl or ethyl group.

According to a very particularly preferred embodiment of the present invention, group A _{2 is} not substituted. Accordingly, the present invention also describes compounds as described above, which are characterized in that they have the following structure:

In addition to the 4-position of the radical R ₃ explicitly described in the structural formulas above, the 2- or the 3-position are also possible, the 4-position being preferred.

Likewise, the present invention therefore also describes compounds, as described above, which are characterized in that they have the following structures:

All of the above-mentioned structures can be used as the radical R- _t contained in radical R ₃ . T = 0 is preferred in this case, radicals R of the following structures are particularly preferred:

NH

N - HR ^x _c 6, further preferably R and R being hydrogen.

Are most preferred radicals R _t of the following structures:

further preferably Re and R are hydrogen.

Accordingly, the present invention also relates to a compound, as described above, which is characterized in that R-i is selected from the group consisting of

In the context of the present invention, particular preference is given to the radical i contained in the radical R ₃ in the structure

With regard to the radical R ₂ , all radicals falling under the definition in (ii) are generally possible. Accordingly, the aryl groups are, for example, the phenyl group or the naphthyl group, it being possible for at least one of the C atoms of this aryl group to be replaced by a heteroatom selected from the group consisting of S, N and O. A phenyl radical is particularly preferred as radical R ₂ . Thienyl and pyridyl are preferred as an aryl radical containing at least one heteroatom.

Accordingly, the present invention relates to a compound as described above, which is characterized in that R _{2 is} an at least monosubstituted phenyl radical, thienyl radical or pyridyl radical.

According to a particularly preferred embodiment, the R _{2 is} an at least monosubstituted phenyl radical.

The aryl radical, particularly preferably the phenyl radical, R ₂ has at least one substituent R as defined above, one of the structures

is preferred.

A substituent R ₄ , which is selected from the group consisting of, is particularly preferred

A radical of the structure is very particularly preferred as the radical R-t

NH

<

N-H

I

H

The substituent Rj can generally be positioned at all positions of the aryl radical. With regard to the particularly preferred phenyl radical, the 2-, 3- or 4- position of the R-j radical is accordingly possible, the 3-position of the RΔ radical on the phenyl radical being particularly preferred.

In addition to the radical _t , the aryl radical can furthermore have at least one further substituent. According to a particularly preferred embodiment, the aryl radical has a single substituent.

The alkylene group with which the radical R _{2 is} bonded to the alpha-C atom of the medium-sized amino acid according to (I) generally has 0 to 3 C atoms. This alkylene group preferably has 1, 2 or 3 carbon atoms, particularly preferably 1 or 2 carbon atoms and very particularly preferably 1 carbon atom.

Accordingly, the present invention describes a compound as described above, which is characterized in that it has the following structure:

The structure shown above can generally be present on the α-C atom of 3-amidinophenylalanine in either the D or the L configuration. The L configuration is particularly preferred in the context of the present invention. In very general terms, the compounds according to the invention according to (I) may be present either in the L or in the D configuration or as a racemate with regard to the medium-sized alpha-amino acid.

Accordingly, the present invention also relates to a compound, as described above, which is characterized in that k = 1 and R _{2 is} a phenyl radical which is meta-substituted with an amidino group, the resulting 3-amidinophenylalanine being present in the L configuration ,

In principle, the compounds according to (I) can be present in the L as well as in the D configuration or S or R configuration if there is another asymmetric C atom or another asymmetry center in addition to the above-mentioned asymmetric carbon atom. Likewise, the compound according to (I) can be present as a racemate. Mixtures of L and D configuration or S and R configuration are also possible, in which the proportion of molecules with D configuration or the proportion of molecules with L configuration or the proportion of molecules with S configuration or the proportion of molecules with the R configuration predominates. Any of the residues Ri described in (i) above can generally be present as the residue Ri.

If the radical Ri is substituted, for example, by at least one halogen, fluorine, chlorine and / or bromine are preferred.

If the radical R 1 is substituted, for example, with at least one esterified carboxyl group, methyl esters and / or ethyl esters are preferred.

If the R 1 radical is substituted, for example, with at least one amino group, this amino group can be acylated, the acetyl group being particularly preferred.

For example, particularly preferred are mono-, bi- or tricyclic aryl radicals and heteroaryl radicals, in which at least one double bond is optionally hydrogenated and / or which have at least one heteroatom selected from O, S and N, where a heteroaryl radical R 1 is also two or more identical or can contain different heteroatoms. Examples of preferred aryl radicals are phenyl, naphthyl, anthracyl or phenanthryl. These aryl radicals can optionally be present in reduced or / and oxidized form. With regard to the naphthyl radical, for example a 1,2-dihydronaphthyl, a 1,4-dihydronaphthyl or even a 1,2,3,4-tetrahydronaphthyl radical is possible. The oxidized form of the naphthyl radical can be present, for example, as a 1,4-naphthoquinoyl radical. The anthraquinoyl radical can be present in oxidized form, for example as 1,4- or 9,10-anthraquinoyl radical or 1,4- or 9,10-anthraquinroloyl radical, and the phenanthryl radical as phenanthrene-quinoyl radical. Examples of heteroaryl radicals are, for example, pyrrolyl, furanyl, thiophenyl, pyridyl, pyrimidyl, pyrazyl, triazyl, imidazolyl, thiazolyl, oxazolyl, indolyl, purinyl, pyronyl, pyridonyl, quinolyl, isoquinolyl. Ri radicals such as indenyl or tetrahydroindenyl are also included.

The radical R 1 can furthermore be suitably substituted, preference being given, for example, to linear, branched or cyclic alkyl radicals having 1 to 10 carbon atoms. Linear or branched alkyl radicals with 1, 2, 3 or 4 carbon atoms are particularly preferred. Atoms. For example, isopropyl and tert-butyl are particularly preferred as substituents. Cyclic alkyl radicals are likewise preferred as substituents, cyclic alkyl radicals having 5, 6 or 7 carbon atoms and in particular 6 carbon atoms being particularly preferred. Aryl or heteroaryl groups may also be mentioned as substituents, where the heteroaryl groups contain a heteroatom selected from N, S and O and can contain two or more identical or different heteroatoms. The substituents of the radical Ri can in turn be suitably substituted.

Both heteroaryl and aryl as well as alkyl radicals can be bound to the radical Ri by a sulfur bridge atom or an oxygen bridge atom or via an alkylene chain with 1-3 C atoms. Accordingly, the radical RI can be substituted, for example, with an alkyloxy, alkylthio, aryloxy, arylthio, heteroaryloxy or heteroarylthio group.

Examples of substituted R 1 radicals are, for example

^tf AX AΑ ^ a

An aryl radical which is preferably present can in principle also carry more than one substituent. Embodiments in which the aryl radical has no, one, two or three substituents are particularly preferred. If, for example, the aryl radical has three substituents, alkyl radicals having 1, 2 or 3 carbon atoms are preferred. Alkyl radicals having 2 or 3 carbon atoms are particularly preferred, particularly preferably alkyl radicals having 3 carbon atoms, and particularly preferably isopropyl. The 2,4,6-triisopropylphenyl radical, for example, is particularly preferred here.

If the aryl radical has a substituent, for example, the tert-butyl radical is preferred.

In a further suitable embodiment, the R 1 radical is an aryl radical, preferably a phenyl radical, which is in turn substituted with an aryl, heteroaryl or alkyl radical with a further aryl radical or heteroaryl radical via an oxygen bridge atom or via an alkylene chain with 1-3 C atoms. The aryl or heteroaryl radical, for example a pyridine, may be unsubstituted or likewise substituted at a suitable position, in the ortho, meta or para position, for example with at least one alkyl group, for example a methyl group and / or with at least one a halogen atom, preferably with one chlorine atom or two chlorine atoms or one of the two fluorine atoms and / or with at least one trihalomethyl group, preferably with one or two of a trifluoromethyl group (s) and / or with

with at least one alkoxy group, preferably a methoxy group.

Examples of such residues Ri are about

the oxygen connecting the two aryls through an alkylene chain with 1-3 C-

Atoms can be replaced.

According to a particularly preferred embodiment, the Ri radical is selected from tert-butylphenyl, cyclohexylphenyl, 5,6,7,8-tetrahydronaphthyl, naphthyl, anthracyl, anthraquinoyl and anthrahydroquinoyl, pyridyloxyphenyl, phenyloxypyridyl, pyridyl-alkylphenyl with a C ₁ -C ₃ -alkyl , The alkylene group which connects the radical R 1 and the sulfonyl group generally has no, one or two carbon atoms. It preferably has no or one carbon atom, particularly preferably it has no carbon atom and is therefore not present.

An alternative embodiment of the invention are compounds of the formula I "

wherein Ri, R ₂ , Q, j and k are defined as described above, m = n = 2 and R _{3 is} an aryl or heteroaryl radical, and wherein the aryl radical is preferably a benzyl or a phenoxy radical and the heteroaryl radical is preferably selected from a pyridinylmethyl, pyridinyloxo, pyrimidinyloxo, pyrazinyloxo, pyridinylthio radical and the aryl or heteroaryl radical is unsubstituted or substituted with at least one halogen, preferably fluorine or chlorine, at least one alkoxy radical, preferably methoxy radical and / or at least one trifluoromethyl radical.

In a further preferred embodiment, the R ₃ radical is a guanidinooxyalkyl radical.

Examples of such connections are connections in which the building block in formula F 'is replaced by

which are commercially available from Array Bipharma, Bolder Colorado, U.S.A ..

Also included are compounds in which an amino acid, preferably glycine, is incorporated between the sulfonyl group and the 3-amidinophenylalanine group of the formula I.

The compounds described above can generally be prepared according to any suitable method. The compounds according to the invention are preferably prepared by processes in which a sulfonic acid chloride is reacted with an amino acid or an amino acid derivative in a first step. Accordingly, the present invention relates to a process for the preparation of a compound as described above, which comprises the following step (S1): (S1) reaction of a compound of the general structure (El ')

with a compound of the general structure (El ")

to obtain a compound of the general structure (ZP1)

where R ₂ "is the aryl radical R ₂ , substituted either with Ri or with i, protected with a suitable protective group, or with a substituent which is a precursor to i.

If, for example, the aryl radical R ₂ is preferably substituted with an amidino group i, the substituent which is a precursor to the amidino group, for example a cyano group, is converted to an amidino group in one, preferably two or more suitable process steps by reaction with hydroxylamine and subsequent hydrogenation can be. In the following, this amidino group can again be protected with a suitable protective group, which can be removed again in a suitable process step. If compounds of the general structure (I ") according to the invention are prepared in which the group Q is nitrogen and the alkylene group Ai is not present, production processes are preferred in which, starting from a compound of the structure (ZPl), these are initially in one Step (S2 ') with a cyclic compound of the general structure (E2')

implemented, where W is a suitable protecting group. A connection of the general structure (ZP2 ¹ )

obtained, from which the protective group W is preferably split off in a subsequent step (S3 ¹ ). In a next step (S4 '), the compound obtained according to (S3 ¹ ) is combined with a compound of the general structure (E2 ")

HO-TA ₂ -R; (E2 ") implemented, a compound of the general structure (Pl)

is obtained, wherein R4 'is either R _t or R _t , protected with a suitable protective group, or a precursor to _t . If _t 'is a radical Rt protected with a protective group, the protective group is preferably removed after step (S4'). According to the invention, an amino group can then be present as residue t. In a next step, a guanidino group can be built up from this in accordance with a process known to the person skilled in the art, for example by reaction with pyrazole carboxamidine.

A likewise preferred process for the preparation of compound according to (I "), in which the group Q is nitrogen and the alkylene group Ai is not present, comprises a step (S2 ') ₅ in which a compound of the general structure (E2 ^m )

where W is a suitable protective group, with a compound of the general structure (E2 ")

HO-TA -R; _(E2 ») is implemented, whereby a connection of the general structure (ZP2")

is obtained in which R _t 'represents either _t or R _t , protected with a suitable protective group, or a precursor to t. In this process, the C-terminal residue of the middle-sized amino acid according to (I) is built up first. In a next step (S3 ') the protective group W is preferably split off, and in a next step (S4') the compound obtained according to (S3 ') with a compound of the general structure (ZPl) to obtain a compound of the general structure (Pl ) implemented. If the radical R _{4 is} an amino group, a guanidino group can also be built up from it here, as described above. If compounds of the general structure (I ") according to the invention are prepared in which the group Q is CH, production processes are preferred in which starting from compounds according to (ZPl) these are carried out in a step (S2") with a compound of the general structure ( E3)

are implemented, wherein R _t 'is either R _t or R _t , protected with a suitable protective group, or a precursor to _t . This preferred implementation results in a compound of the general structure (P2)

receive. If Rt 'is a radical R _t protected with a protective group, the protective group is preferably removed after step (S2 "). According to the invention, an amino group can then be present as the radical R _t . In a next step, this can be used, according to a process known to the person skilled in the art, for example, by reaction with pyrazolecarboxamide, a guanidino group can be built up.

If compounds of the general structure (I ") according to the invention are prepared in which T is - (C = O) -NH-, Q is CH and Ai is not present, preference is given to preparation processes in which (S2 '") first compounds of the general structure (E3 ') with a connection of the general structure (E3 ") be implemented, whereby a connection of the general structure (ZP3)

is obtained. The rest Rt 'has the meaning described in the further production processes described above. W is also a suitable protective group here. In a next step, the protective group W is preferably removed in a step (S3 '") according to a suitable method. This is preferably followed by a step (S4'") in which the compound obtained according to (S3 '") with a compound is implemented according to (ZPl), a compound of the general structure (P3)

is obtained. If Rt 'is a radical R _t protected with a protective group, the protective group is preferably removed after step (S2 ^m ). According to the invention, an amino group can then be present as the radical R _t . In a next step, a guanidino group can be built up from this in a method known to the person skilled in the art, for example by reaction with pyrazole carboxamidine. According to a preferred embodiment of this last-described method variant, a compound having the general structure (E3 "), for example a compound which has a radical _t 'as a precursor to a radical Rt according to the invention, this precursor being a -CN group. In one or more suitable steps, this -CN group is further preferably built up to an amidino group in accordance with a process known to the person skilled in the art, for example by reaction with a hydroxylamine, subsequent reaction with acetic anhydride and subsequent hydrogenation Hydroxyamidino derivative can be converted directly into an amidine derivative by hydrogenation, the amidino group optionally being protected again intermediately with a suitable protective group. The reaction with hydroxylamine is very particularly preferably carried out after step (S2 '") and before the Sc step (S3 '"). The reaction with acetic anhydride is also particularly preferably carried out before step (S3"'). The hydrogenation to the amidino group is preferably carried out after step (S4 ^m ).

The final purification of the compounds prepared in this way is very particularly preferably carried out by preparative, reversed-phase HPLC or by crystallization from a suitable solvent or solvent mixture or by countercurrent distribution.

In addition to the compounds described above per se and methods for their preparation, the present invention also relates to a medicament which contains one of the compounds specified above.

Accordingly, the present invention also relates to a medicament containing at least one compound according to (I) or a salt of this compound and optionally pharmaceutically suitable auxiliaries and / or additives.

The present invention likewise describes a compound according to (I) for use as a medicament, the medicament optionally additionally containing pharmaceutically suitable auxiliaries and / or additives. Suitable auxiliaries and / or additives, which serve, for example, to stabilize and / or preserve the medicament, are disclosed, for example, in H. Sucker et al., Pharmaceutical Technology, 2nd Edition, Georg Thieme Verlag, Stuttgart (1991), the relevant one Disclosure is incorporated by reference into the context of the present invention. The pharmaceutically suitable auxiliaries and / or additives include, for example, physiological saline solutions, Ringer's dextrose, Ringer's lactate, demineralized water, stabilizers, antioxidants, complexing agents, antimicrobial compounds, proteinase inhibitors and / or inert gases.

In general, the compounds according to (I) according to the invention can be used in any form as a medicament. In possible embodiments of the invention, the medicament is administered, for example, in the form of a tablet, a dragee, a cap, a pellet, suppository, a solution, in particular a solution for injection or infusion, eye, nose and ear drops, a juice, a Capsule, an emulsion or suspension, a globule, styli, aerosol, powder, a paste, cream or ointment.

The present invention therefore also relates to a medicament, as described above, which is characterized in that it is in the form of a tablet, a dragee, a capsule, a pellet, a suppository, a solution, in particular a solution for injection or infusion, from the eye. , Nasal or ear pots, a juice, an emulsion or suspension, a globule, a styli, an aerosol, a powder, a paste, a cream or an ointment.

Likewise, the present invention therefore also describes a compound according to (I) for use as a medicament, the medicament in the form of a tablet, a coated tablet, a capsule, a pellet, a suppository, a solution, in particular a solution for injection or infusion, from eyes -, nose or ear drops, a juice, an emulsion or suspension, a globule, a styli, an aerosol, a powder, a paste, a cream or an ointment. According to a particularly preferred embodiment of the present invention, the above-mentioned compounds according to (I) and / or their salts or the above-mentioned medicaments containing these compounds according to (I) and / or their salts and optionally at least one pharmaceutically suitable auxiliary and / or additive used to diagnose and / or treat a tumor. The prophylaxis of a tumor is also additionally or alternatively possible, the compounds being particularly useful for preventing and / or reducing the metastasis of tumors.

A compound according to the invention according to (I) or its salt or a medicament as described above can also be used very generally, for example, in parenteral use, in particular in intraartial, intravenous, intramuscular or subcutaneous form, in enteral use, in particular for oral or rectal use, or in topical use Application form, in particular as a dermatological, can be used. Intravenous or subcutaneous applications are preferred.

These forms of use are particularly suitable for diagnosis and / or therapy and / or prophylaxis of a tumor.

The present invention therefore also relates to the use of a compound according to (I) or a salt of this compound or a medicament as described above for the diagnosis, therapy or prophylaxis of a tumor and for preventing and / or reducing the metastasis of a tumor, in particular in the oral, subcutaneous region - ner, intravenous or transdermal form.

The present invention also describes the use of a compound according to (I) or a salt of this compound for producing a medicament for the diagnosis, therapy or prophylaxis of a tumor and for preventing and / or reducing the metastasis of a tumor. The present invention further describes this use for the production of an orally, subcutaneously, intravenously or transdermally applicable medicament.

According to a particularly preferred embodiment of the present invention, a compound according to the invention according to (I) is used to reduce tumor metastases.

Accordingly, the present invention relates to the above-described use of the compound according to (I) or a salt of this compound or a medicament as described above, the formation of tumor metastases being reduced.

The present invention also describes a method, as described above, for producing a medicament for reducing the formation of tumor metastases.

Likewise, the present invention therefore relates in particular to the use of a compound according to (I) or a salt of this compound or a medicament containing the compound according to (I) or a salt of this compound for inhibiting the matriptase.

In particular, the present invention also relates to the use of a compound according to (I) or a salt of this compound or a medicament containing the compound according to (I) or a salt of this compound for inhibiting the matriptase, the matriptase being MT-SP1.

Accordingly, the present invention also describes a process for the preparation of a medicament containing a compound according to (I) or a salt of this compound for inhibiting matriptase.

The present invention also describes a method for producing a medicament, containing a compound according to (I) or a salt of this compound, for inhibiting the matriptase, the matriptase being MT-SP1. The compounds described above can also be present in form of prodrugs, for example by modifying the amidino group with a hydroxyl or a C ₁ -C ₆ -alkyloxycarbonyl group, which only spontaneously after being absorbed into the organism and or by one or more endogenous enzymes in the inhibitory active species can be converted, which can improve the bioavailability and the pharmacokinetic properties of the compounds.

The invention is explained in more detail in the following examples and figures.

Description of the figures

Figure 1 shows the inhibition of invasive growth by the matriptase inhibitors 37 and 54 from Example 9.

FIG. 2 shows the inhibition of proHGF-induced scattering of PC-3 cells by the matriptase inhibitors 37 and 54 from example 10.

Examples

methods

Analytical HPLC: Shimadzu LC-10A system, column: Phenomenex Luna Cι ₈ , 5 μm, 100 Ä (250 x 4.6 mm) solvent A: 0.1% TFA in water, B: 0.1% TFA in ACN, Gradient: 10% B to 70% B in 60 min, 1 ml / min flow, detection at 220 or 215 nm.

Preparative HPLC: Shimadzu LC-8A system, column: Phenomenex Luna Cι ₈ , 5 μm, 100 Ä (250 x 30 mm) solvent A: 0.1% TFA in water, B: 0.1% TFA in ACN, gradient: 10% B to 55% B in 120 min, 10 ml / min flow, detection at 220 nm.

Mass spectroscopy: The mass spectra were recorded on a compact sample from Kratos (Manchester, England) with a time-of-flight measurement detector and α- Cyano-hydroxycinnamic acid as a matrix, or on an ESI-MS LCQ from Finnigan (Bremen, Germany), measured.

used abbreviations

Ac Acetyl

AcOH acetic acid

ACN acetonitrile ß-Ala ß-alanine

Boc tert-butyloxycarbonyl

CKIBE isobutyl chlorocarbonate

DAE 1,2-diaminoethane

DCM dichloromethane

DIEA diisopropylethylamine

DMF N, N-dimethylformamide iNip isonipecotinic acid

Kj inhibition constant

NMM N-methylmorpholine

Phe (3-AcOxam) 3 - (acetyloxamidino) phenylalanine

Phe (3-Am) 3 - amidinophenylalanine

Phe (3-CN) 3 - (cyano) phenylalanine

Phe (3-oxam) 3 - (oxamidino) phenylalanine

PyBop benzotriazole-1-n-N-oxytris (ynolidino) phosphonium hexafluorophosphate

Pzd piperazide

RT room temperature

TFA trifluoroacetic acid

THF tetrahydrofuran

Tips 2,4,6- (triisopropyl) phenylsulfonyl

Z benzyloxycarbonyl Example 1: Synthesis of Anthracensulfonyl-Phe (3-Am) -Pzd-ß-Ala x 2 TFA

(Compound 10 from Table 1)

la) Boc-Pzd-ßAla-Z

2 g (8.96 mmol) of Z-βAla-OH were dissolved in 20 ml of THF and 0.99 ml (8.96 mmol) of NMM and 1.17 ml (8.96 mmol) of CKIBE were added at -15 ° C , The mixture was stirred at -15 ° C. for 10 min, then 1.67 g (8.96 mmol) of Boc-piperazide (Fluka) and an additional 400 μl (3.6 mmol) of NMM were added. The mixture was stirred for a further hour at -15 ° C. and further overnight at room temperature. The solvent was then removed in vacuo, the residue taken up in ethyl acetate, in each case

Washed 3 times with 5% KHSO ₄ , saturated NaHCO ₃ solution and NaCl-saturated water and then dried over Na SO ₄ . The solvent was removed in vacuo. A pale oil remained as a residue, which crystallized in the refrigerator overnight.

Yield: 3.2 g (8.17 mmol), HPLC: 51.69% B

lb) H-Pzd-βAla-Z x HC1

3.2 g (8.17 mmol) of Boc-Pzd-βAla-Z were dissolved in glacial acetic acid, mixed with 50 ml of IN HC1 in glacial acetic acid and left to stand at room temperature with occasional shaking. The solvent was partially removed in vacuo and the product was precipitated by adding diethyl ether, suction filtered, washed again with diethyl ether and dried in vacuo. Yield: 2.13 g (6.5 mmol) white solid, HPLC: 28.19% B

lc) Boc-Phe (3-CN) -OH

2.5 g (13.1 mmol) H-Phe (3-CN) -OH were dissolved in 100 ml dioxane and at 0 ° C with 13 ml (13 mmol) IN NaOH and 3.16 g (14.5 mmol ) Boc pyrocarbonate added. The mixture was stirred at 0 ° C. for 20 min and then at room temperature for 4 h, a total of 7 ml (7 mmol) of IN NaOH were added in portions, as a result of which the pH was kept constant at 8-8.5. The solvent was removed in vacuo, the residue was taken up in ethyl acetate, washed 3 times with 5% KHSO ₄ and 3 times with NaCl-saturated water and then dried over Na ₂ SO ₄ . The solvent was removed in vacuo. A white solid was obtained as a residue.

Yield: 2.11 g (7.3 mmol) white solid, HPLC: 45.93% B

ld) Boc-Phe (3-AcOxam) -OH

2.11 g (7.3 mmol) of Boc-Phe (3-CN) -OH were dissolved in 100 ml of methanol and with 760 mg (10.95 mmol) of hydroxylamine x HC1 and 1.9 ml (10.95 mmol) DIEA transferred. The mixture was stirred under reflux for 6 h. Another 266 mg (3.84 mmol) of hydroxylamine x HCl and 665 μl (3.84 mmol) of DIEA were added and the mixture was stirred under reflux for a further 3 h and then at room temperature overnight. The solvent was then removed in vacuo. A pale oil remained as a residue, which was dissolved in 50 ml of glacial acetic acid and 2 ml (22 mmol) of acetic anhydride were added. The mixture is stirred for 30 minutes at room temperature. The solvent was removed in vacuo, the residue was taken up in ethyl acetate, washed 3 × with 5% KHSO ₄ and NaCl-saturated water and then dried over Na ₂ SO ₄ . The solvent was removed in vacuo.

Yield: 3.31 g (colorless oil), HPLC: 26.59% B le) Boc-Phe (3-AcOxam) -Pzd-ßAla-Z

0.92 g (2.8 mmol) H-Pzd-ßAla-Z x HC1 and 1.02 g (2.8 mmol) Boc-Phe (3-AcOxam) - OH were dissolved in 40 ml DMF and at 0 ° C with 1.46 g (2.8 mmol) PyBop and 1.46 ml (8.4 mmol) DIEA. The mixture was stirred at 0 ° C. for 20 min and at room temperature for a further 2 h. The solvent was then removed in vacuo, the residue was taken up in ethyl acetate, washed 3 × with 5% KHSO ₄ , saturated NaHCO ₃ solution and NaCl-saturated water and then dried over Na ₂ SO ₄ . The solvent was removed in vacuo.

Yield: 2.49 g (light oil), HPLC: 48.13% B

lf) H-Phe (3-AcOxam) -Pzd-βAla-Z x HC1

2.49 g of Boc-Phe (3-AcOxam) -Pzd-ßAla-Z (crude product) were dissolved in glacial acetic acid, mixed with 30 ml of IN HC1 in glacial acetic acid and left standing at room temperature with occasional shaking. The solvent was partially removed in vacuo and the product was precipitated by adding diethyl ether, suction filtered, washed again with diethyl ether and dried in vacuo.

Yield: 1.32 g (2.3 mmol) white solid, HPLC: 32.89% B

lg) Anthracensuflonyl-Phe (3-AcOxam) -Pzd-ßAla-Z

At 0 ° C and 207.7 mg (0.361 mmol) H-Phe (3-AcOxam) -Pzd-ßAla-Z, 127 μl (0.73 mmol) DIEA and 100 mg (0.361 mmol) anthracensulfonyl chloride (Fluka) in 10 ml of DMF dissolved. The mixture was stirred at 0 ° C. for 20 min and then at room temperature overnight. The solvent was removed in vacuo, the residue was taken up in ethyl acetate, in each case 3 × with 5% KHSO ₄ , saturated NaHCO ₃ solution and NaCl-saturated water and then dried over Na ₂ SO ₄ . The solvent was removed in vacuo. A pale oil remained as residue, which was used directly for the next synthesis step without further purification. HPLC: 57.65% B

lh) Anthracensuflonyl-Phe (3-Am) -Pzd-ßAla

The crude product Ig was dissolved in 50 ml of 90% acetic acid and 5 ml of 1N HCl and 30 mg of catalyst (10% Pd / C) were added. The mixture was hydrogenated at 40 ° C. and normal pressure overnight with hydrogen. The catalyst was then filtered off and the solvent was concentrated in vacuo. Part of the crude product was cleaned with preparative reversed-phase HPLC.

HPLC: 34.11% B MS: calculated 586.24 (monoisotopic), found 587.79 [M + H] ⁺

Example 2: Synthesis of Anthracensulfonyl-Phe (3-Am) -Pzd-CO-CH -CH - Guanidino x 2 TFA (Compound 11 from Table 1)

2a) Anthracensulfonyl-Phe (3-Am) -Pzd-CO-CH ₂ -CH ₂ -Guanidmo

Approximately 115 mg of crude product of anthracensuflonyl-Phe (3-Am) -Pzd-ßAla (1 h) was dissolved in 10 ml of DMF and 90.3 mg (0.616 mmol) of pyrazole carboxamidine x HCl and 107 μl (0.616 mmol) of DIEA were added. The mixture was stirred overnight and then the solvent removed in vacuo. The remaining residue was dried in vacuo and purified without preparative reversed-phase HPLC.

HPLC: 35.09% B

MS: calculated 628.26 (monoisotopic), found 629.4 [M + H] ⁺

Example 3: Synthesis of 2,4,6-triisopropylphenylsulfonyl-Phe (3-Am) - iNip-DAE-H x 2 TFA (Compound 3 from Table 1)

3a) Tips-Phe (3-CN) -OH

1 To a solution of 1.05 eq. HL-Phe (3-CN) -OH (3 g, 15.8 mmol) in dioxane and 2.1 eq. In NaOH (31.5 ml), a solution of 1 eq. Tips-Cl (97%, 4.69 g, 15 mmol) added dropwise in dioxane. The pH of the solution was checked and kept at pH 8-9 with 1M NaOH. After 4 h, the LM was removed in vacuo, the residue was taken up in ethyl acetate, washed 3 times acid (5% KHSO ₄ ) and 3 times neutral (saturated NaCl solution). The organic phase was dried over Na ₂ SO ₄ , the LM was removed in vacuo and the product obtained was recrystallized from ethyl acetate / hexane (yellowish, crystalline compound).

Yield: 6.6 g (96.3%) HPLC: 71.1% B 3b) Tips-Phe (3-OAm) -OH

A solution of 1 eq. Tips-Phe (3-CN) -OH (3 g, 6.6 mmol), 1.5 eq. Hydroxylamine x HCl (685 mg, 9.9 mmol) and 3 eq. DIEA (3.4 ml, 19.8 mmol) were dissolved in absolute ethanol, boiled under reflux for 4 h and then stirred at RT while adding hydroxylamine x HCl and base (DIEA, pH 8-9) until no starting material was found in the HPLC more was found. After removing the LM, the residue was taken up in ethyl acetate, washed 3 times acid (5% KHSO) and 3 times neutral (saturated NaCl solution). The organic phase was dried over Na ₂ SO ₄ , the LM was removed in vacuo and the product obtained was recrystallized from ethyl acetate / hexane.

Yield: 1.63 g (white crystals; 50.4%), HPLC: 51.0% B

3c) Tips-Phe (3-AcOAm) -OH

1 eq. Tips-Phe (3-OAm) -OH (2.5 g, 5.1 mmol) was dissolved in 100 ml glacial acetic acid, then 1.5 eq. Acetic anhydride (724 ul, 7.6 mmol) added and stirred for 15 min. After removing the LM in a vacuum, the product is obtained as a white powder.

Yield: 2.7 g (99.4%) HPLC: 64.5% B

3d) H-iNip-DAE-Z x HCl

497 mg (2.17 mmol) of Boc-isonipecotic acid were dissolved in 250 ml (2.27 mmol) of NMM in 10 ml of dried THF. At -15 ° C 296 μl (2.27 mmol) of isobutyl chloroformate were added and the mixture was stirred for a further 10 min. 500 mg (2.17 mmol) of NZl, 2-diaminoethane × HCl and 250 μl (2.27 mmol) of NMM were then added, the mixture was stirred for a further 1 h at -15 ° C. and then for 4 h at RT. The LM was removed in vacuo, the residue was taken up in ethyl acetate and 3 x acidic (5% KHSO ₄ ), 1 x neutral (saturated NaCl solution), 3 x basic (NaHCO ₃ saturated) and 3 x neutral (saturated NaCl solution ) washed. The ethyl acetate phase was then dried over Na ₂ SO ₄ and the LM was removed in vacuo, the product being obtained as an amorphous substance (HPLC: 50.3% B). The crude product was dissolved in 20 ml IN hydrogen chloride in glacial acetic acid and left to stand at RT for 1 hour. The LM was then removed in vacuo and the product was lyophilized.

Yield: 722 mg HPLC: 26.4% B

3e) Tips-Phe (3-Am) -iNip-DAE-H x 2 TFA

150 mg (0.28 mmol) Tips-Phe (3-AcOAm) -OH were dissolved with 97 mg (0.28 mmol) H-iNip-DAE-Z x HCl in 5 ml DMF and with stirring in an ice bath to 0 ° C cooled. 122 μl (0.70 mmol) DIEA and 154 mg (0.29 mmol) PyBOP were added to the cooled solution. After 15 min, the ice bath was removed and the mixture was stirred at RT for a further 2 h.

The LM was then removed in vacuo and the residue was taken up in ethyl acetate and 3 x acidic (5% KHSO ₄ ), 1 x neutral (saturated NaCl solution), 3 x basic (NaHCO ₃ saturated) and 3 x neutral (saturated NaCl Solution) washed. The

The ethyl acetate phase was then dried over Na ₂ SO ₄ and the LM was removed in vacuo. The residue was dissolved in 90% glacial acetic acid at 10% by weight

Catalyst (10% Pd / C) added and hydrogenated with hydrogen at RT overnight. The

Catalyst was filtered off and the LM was removed to dryness in vacuo and the residue was purified by preparative reversed-phase HPLC.

Yield: 92 mg HPLC: 43.3% B MS: calculated 626.36 (monoisotopic), found 628.1 [M + H] ⁺ Example 4: Synthesis of 2,4,6-triisopropylphenylsulfonyl-Phe (3Am) -iNip-NH-CH ₂ -CH ₂ -Guanidino x 2 TFA (Compound 34 from Table 1)

4a) Tips-L-Phe (3-Am) -iNip-NH-CH ₂ -CH ₂ -Guanidino x 2 TFA

85 mg (0.1 mmol) Tips-Phe (3-Am) -iNip-DAE-H x 2 TFA were dissolved in 5 ml DMF and with 30 mg (0.2 mmol) pyrazole carboxamidine and 55 μl (0.3 mmol ) DIEA relocated. The mixture was stirred at RT overnight. The solvent was then removed in vacuo and the residue was purified by preparative reversed-phase HPLC.

HPLC: 43.7% B MS: calculated 668.38 (monoisotopic), found 669.8 [M + H] ⁺

Example 5: Synthesis of 2-Nas-Phe (3-Am) -4 (aminoethyl) piperidide x 2 TFA

(Compound 36 from Table 1)

5a) 2-Nas-Phe (3-CN) -OH

To 1.9 g (10 mmol) of H-Phe (3CN) -OH, dissolved in 100 ml of dioxane / water mixture and 22 ml of 1 N NaOH solution, 2.49 g (11 mmol) were added at 0 ° C -Nas chloride (dissolved in dioxane) was added dropwise over a period of 30 min. The mixture was stirred at 0 ° C. for 1 h and further at room temperature overnight. The solvent was then removed in vacuo and the residue was dissolved in water (adjusted to pH 8-9 with NaOH). The water phase was extracted twice with diethyl ether and then the pH was adjusted to 3-4 with 1N HCl. The product was extracted 3 times with ethyl acetate and the ethyl acetate phase was washed 3 times with 5% KHSO ₄ and NaCl-saturated water and then dried over Na ₂ SO ₄ . The solvent was removed in vacuo. A light oil remained, which crystallized out in the refrigerator.

Yield: 3.51 g (9 mmol), HPLC: 51.02% B

5b) 2-Nas-Phe (3-AcOxam) -OH

3.5 g (9 mmol) of 2-Nas-Phe (3CN) -OH were dissolved in 100 ml of methanol, and 1.04 g (15 mmol) of hydroxylamine × HCl and 2.61 ml (15 mmol) of DIEA were added. The mixture was stirred under reflux for 6 h. Then another 700 mg (10 mmol) hydroxylamine x HCl and 1.74 ml (10 mmol) DIEA were added. The mixture was stirred under reflux for a further 4 h and then at room temperature overnight. On- finally the solvent was removed in vacuo. A pale oil remained as a residue, which was dissolved in 50 ml of glacial acetic acid and mixed with 2.83 ml (30 mmol) of acetic anhydride. The mixture was stirred at room temperature for 1 h. The solvent was removed in vacuo, the residue was taken up in ethyl acetate, washed 1 × with 5% KHSO ₄ solution and 3 × with NaCl-saturated water and then dried over Na SO ₄ . The solvent was almost removed in vacuo. The product slowly crystallized out and was suctioned off.

Yield: 3.02 g (6.64 mmol), pale solid, HPLC: 44.04% B

5c) 2 ~ Nas-Phe (3-AcOxam) -4 (aminoethyl) piperidide x HCl

100 mg (0.22 mmol) of 2-Nas-Phe (3-AcOxam) -OH and 50 mg (0.22 mmol) of 4- (2-Boc-aminoethyl) piperidine (Tyger Scientific Inc., Princeton, NY) dissolved in 10 ml DMF and 115 mg (0.22 mmol) PyBop and 115 μl (0.22 mmol) DIEA were added at 0 ° C. The mixture was stirred at 0 ° C. for 20 min and at room temperature for a further 3 h. The solvent was then removed in vacuo. The residue was taken up in ethyl acetate, 2 x with 5% KHSO ₄ solution, 1 x with NaCl-saturated water, 2 x with sat. NaHCO ₃ solution and washed 3 times with NaCl-saturated water and then dried over Na ₂ SO ₄ . The solvent was removed in vacuo.

The crude product was dissolved in glacial acetic acid, mixed with 5 ml IN HCl in glacial acetic acid and left at room temperature with occasional shaking. The solvent was removed in vacuo, a light oil remained.

Yield: 105 mg oil, HPLC: 35.66% B

5d) 2-Nas-Phe (3-Am) -4 (aminoethyl) piperidide x 2 TFA

The crude product 5c was dissolved in 50 ml of 90% acetic acid and 15 mg of catalyst (10% Pd / C) were added. The reaction was carried out at 40 ° C. and normal pressure overnight Hydrogen hydrogenates. The catalyst was then filtered off and the solvent was concentrated in vacuo. A third of the crude product was purified using preparative reversed-phase HPLC.

Yield: 17.6 mg HPLC: 29.03% B

MS: calculated 507.23 (monoisotopic), found 508.4 [M + H] ⁺

Example 6: Synthesis of 2 ~ Nas-Phe (3-Am) -4 (guanidinoethyl) piperidide x 2 TFA

(Compound 37 from Table 1)

6a) 2-Nas-Phe (3-Am) -4 (guanidinoethyl) piperidide x 2 TFA

Approximately 80 mg of crude product of 2-Nas-Phe (3-AcOxam) -4 (aminoethyl) piperidide x HCl (5d) was dissolved in 5 ml of DMF and with 65 mg (0.45 mmol) of pyrazole carboxamidine x HCl and 105 μl (0, 6 mmol) DIEA added. After 3 h, a further 21.5 mg (0.15 mmol) pyrazolecarboxamidine x HCl and 35 μl (0.15 mmol) DIEA were added and the mixture was further stirred overnight. The solvent was removed in vacuo and the remaining residue was purified by preparative reversed-phase HPLC.

Yield: 42 mg HPLC: 31.19% B

MS: calculated 549.25 (monoisotopic), found 550.4 [M + H] ⁴ Example 7: Preparation of the catalytic domain of matriptase

Cloning: The catalytic domain of matriptase was amplified by PCR with the following primer pair:

Sense primer:

5 '- GGCAATTCCATATGAAACATCACCATCATCACCATGrroTTGGGGGCiCGG ^ rGCG -3'

Antisense primer: 5 '- GCATGAATTCTTATACCCCAGTGTTCTCTTTGATCCA -3

Sense primers and antisense primers were chosen so that at the 5 'end before the protease domain (italic) an Ndel interface (bold) followed by the peptide sequence Met Lys (His) ₆ or at the 3' end of matriptase (italic) an EcoRI interface (bold) was inserted. The PCR product was cloned via Ndel and EcoRI in pET24 (Novagen), a vector for expression in Escherichia coli.

The catalytic domain of matriptase was expressed in inactive and insoluble form in Escherichia coli, purified, refolded and then activated. The steps were:

Expression and Purification: BL21 (DE3) cells (Novagen), which contained the vector from the cloning described above, were incubated in LB, 30 μg / ml kanamycin at 37 ° C. and 250 rpm. Expression was induced at an OD ₆₀ o of 0.6 by adding 1 mM IPTG and the incubation was continued for one hour. Then, the cells were pelleted, digested with 5 mL of Bug Buster ™ Protein Extraction Reagent (Novagen) and the DNA with 25 U / ml per 1 g cell pellet Benzonase ^® nuclease (Novagen) was digested. The protein aggregates were washed and denatured with 5 ml denaturing buffer (6 M guanidinium HCL, 10 mM Tris HCl, 100 mM Na phosphate, pH 8.0) per 1 g pellet. Insoluble constituents were centrifuged off (16,000 g, 30 min, 20 ° C.), the supernatant was filtered (0.2 μm), 10 mM β-mercaptoethanol was added, and the mixture was subsequently purified on a metal to purify the catalytic domain from matriptase Chelate chromatography column (1 ml NiNTA (Qiagen) per 10 ml supernatant). The column was washed (8 M urea, 10 mM Tris HCl, 100 mM Na phosphate, pH 6.3) and the purified protein eluted with 8 M urea, 10 mM Tris HCl, 100 mM Na phosphate, pH 4.5.

Refolding: The fractions containing matriptase were pooled, derivatized with glutathione and then refolded to a refinement concentration of 50 μg / ml in refolding buffer (50 mM Tris HCl, 0.5 M L-arginine, 20 mM CaCl ₂ , 1 mM EDTA, 0 , 1 M NaCl, pH 7.5). After 3 days of incubation at room temperature, the refolding batch was filtered, concentrated to a concentration of> 300 μg / ml (Centricon Plus-80, Amicon) and by gel filtration (PD 10 columns, Pharmacia) in activation buffer (20 mM Na phosphate, 150 mM NaCl, pH 7.0).

Activation: Since a correctly processed N-terminus is a prerequisite for the activity of serine proteases, the peptide MK (His) ₆ at the N-terminus had to be removed to activate the refolded matriptase. For this purpose, the refolding batch was incubated for 2 h at 30 ° C. with 2.5 mU per 50 μg protein-activated DAPase ™ (Qiagen) and the activated matriptase was separated from the non-activated matriptase and the DAPase via metal chelate chromatography.

The yield of active matriptase was approximately 0.9 mg / l bacterial culture. The proteolytic activity was demonstrated by cleavage of the chromogenic substrate Pefachrome tPA (Pentapharm).

Example 8: Determination of the inhibitory effect of matriptase with the inhibitors listed in Table 1

To determine the inhibitory effect, 200 μl Tris buffer (0.05 M, 0.154 M NaCl, 5% ethanol, pH 8.0; contains the inhibitor), 25 μl substrate (CH ₃ SO ₂ -D-HHT-Gly - Arg-pNA; 2 and 1 mM) and 50 μl Matriptase (0.5 μg / ml) incubated at 25 ° C. After 3 min, the reaction was interrupted by adding 25 μl of acetic acid (50%) and the absorption at 405 nm is determined using a microplate reader (Dynatech MR 5000). The Kj values were determined according to Dixon (Biochem. J. 55, 170-171, 1953) by linear regression using a computer program. The Kj values (Table 1) are the average of at least three determinations.

Table 1: Determination of the Kj values for the inhibition of matriptase

Example 9: Inhibition of Invasive Growth by Matriptase Inhibitors (Matrigel Assay)

A common test system for invasive growth at a more cellular level is the Matrigel invasion assay. Cells are placed on an artificial extracellular matrix and examined how many cells pass through them within a certain period of time.

It is shown here as an example for the matriptase inhibitors 37 and 54 that the invasive growth is influenced and the migration of matriptase-expressing colon carcinoma cell line DLD-1 through the matrigel is inhibited:

The wells of a 'trans well' plate were coated with 10 μg Matrigel each, 160,000 DLD-1 cells (Dexter et al, Cancer Research 39: 1020-1025 (1979)) in 100 μl medium (RPMI 1640, with 2% Ultroser HY serum substitute was applied and the invasive growth was stimulated by adding ProHGF in 400 μl medium with and without inhibitor (30 μM). After 48 hours of incubation at 37 ° C and 5% CO _2, the cells that had migrated through the matrix were fixed, stained and at 100- times magnification photographed.

As Figure 1 shows, the invasion of DLD-1 cells by the extracellular matrix is stimulated by the addition of proHGF. This indicates that the zymogen is activated on the cell surface. This effect is significantly inhibited by the matriptase inhibitors.

Example 10: Inhibition of cell scattering by matriptase inhibitors

Because of its ability to induce the detachment of cells from the island-like bandage and the spread of the cells, HGF is also referred to as a 'scatter factor'. This function can be demonstrated at the cellular level using the so-called 'scatter assay'. Cells are sown and their spread stimulated by the addition of HGF is documented after a certain point in time. HGF can be formed by activating the inactive proform of HGF (proHGF). In the following it is shown as an example for the matriptase inhibitors 37 and 54 that the activation of proHGF by cellular matriptase is prevented and the "scattering" of proHGF stimulated cells is reduced.

For this purpose 500 prostate carcinoma cells (PC-3) expressing matriptase were seeded per well of a 96-well plate, incubated overnight in 100 μl medium without fetal calf serum (Nut Mix. F-12, 2% Ultroser HY) (37 ° C, 5% CO ₂ ) and then the 'scattering' with and without matriptase inhibitors (3 μM) stimulated by the addition of proHGF. After 6 days the cells were fixed, stained and representative sections were photographed at 100x magnification.

As FIG. 2 shows, the matriptase inhibitors 37 and 54 inhibit the proHGF-induced ^ scattering 'of PC-3 cells.

In the concentrations used, neither proHGF nor the inhibitors have an influence on the proliferation of PC-3 cells. This suggests that the one shown Effect based on a changed 'scatter' behavior and not a changed doubling time of the cells.

Claims

claims

Compound according to formula (I)

or a salt or a prodrug of this compound, wherein

(a) Xi and X _{2 are} independently hydrogen or an alkyl radical having 1, 2 or 3 carbon atoms and at least one of the radicals Xi and X _{2 is} a radical of the structure (V)

where, if appropriate, at least one of the methylene groups indicated with m or n according to (P) is at least simply substituted with a hydroxyl, a halogen, a pseudohalogen or a COOR ₂ 'group and R ₂ ' is a linear, branched or cyclic Is alkyl group with 1 to 10 C atoms, and / or optionally at least one of the C atoms of the methylene groups indicated with m or n according to (T) is replaced by S, N or O and / or optionally at least one of the cycles is a double bond according to (V), or where (b) X and X ₂ are bridged to form a cycle such that the compound according to (I) has the structure (I ")

has, where appropriate at least one of the m or n indexed methylene groups according to (I ") is at least simply substituted with a hydroxyl, a halogen, a pseudohalogen or a COOR ₂ 'group and R ₂ ' is a linear, branched or is cyclic alkyl group with 1 to 10 C atoms, and / or optionally at least one of the C atoms which is replaced with m or n indicated methylene groups according to (I ") by S, N or O and / or while maintaining the C-terminal to the sulfonylated amino acid-bound imino group, optionally at least one of the cycles

bonds forming according to (I ") is a double bond, and wherein

(i) Ri is an optionally partially hydrogenated aryl or heteroaryl group containing at least one of the atoms O, N or S, with 5 to 20 carbon atoms. Atoms, or a linear, branched or cyclic alkyl group with 1 to

10 carbon atoms, wherein R 1 is optionally substituted with at least one halogen and / or pseudohalogen group and / or at least one linear, branched or cyclic alkyl or alkyloxy or alkylthio group with 1 to 10 carbon atoms, which is optionally substituted at least once is with a halogen, pseudohalogen, hydroxyl, amino, cyano, amidino, guanidino or carboxyl group, the carboxyl group optionally being esterified with a linear, branched or cyclic alkyl group having 1 to 10 C atoms, and the linear, branched or cyclic

Alkyl group with 1 to 10 carbon atoms optionally contains at least one heteroatom selected from the group consisting of O, N and S, and / or at least one aryl or heteroaryl group with 5 to 20 carbon atoms, this aryl or heteroaryl group is optionally substituted with at least one linear, branched or cyclic alkyl group with 1 to 10 C atoms and / or at least one COR ₂ 'and / or COOR ₂ ' group, where R ₂ 'is a linear, branched or cyclic alkyl group with 1 until 10

Is C atoms, and / or at least one halogen group and / or at least one pseudohalogen group and / or at least one alkoxy group or an alkylthio group, the alkyl radical each having 1 to 10 C atoms, and / or at least one nitro group and / or at least one Haloalkyl group with 1 to 10 C atoms, and wherein the aryl or heteroaryl group is bonded to the radical R 1 via an alkylene group with 1 to 3 C atoms or an oxygen atom or via a sulfur atom; at least one hydroxy, amino, cyano, amidino, guanidino,

Carboxyl or carboxyalkyl group, where the amino group is optionally acylated and / or wherein the alkyl group of the carboxyalkyl group has 1 to 10 C atoms and / or the carboxyl group is optionally esterified or amidated with a linear, branched or cyclic alkyl group with 1 to 10 C atoms is; (ii) R _{2 is} an at least monosubstituted aryl group having 1 to 10 C atoms, where at least one of these C atoms is replaced by S, N or O, at least one substituent is a group according to R ₄ , R ₂ optionally additionally is substituted by a hydroxyl, COR ₂ 'or COOR ₂ ' group and R ₂ 'is a linear, branched or cyclic alkyl group having 1 to 10 C atoms; (iii) R _{3 represents} a radical of the following formula (II):

A-, TA- R ₄ (II) where

Ai either does not exist or is an alkylene group with 1 to 4 carbon atoms, which is optionally substituted with at least one halogen and / or pseudohalogen group and / or at least one linear, branched or cyclic alkyl group with 1 to 10 carbon atoms and / or at least one aryl or an aralkyl group with 5 to 10 C atoms and / or ~ at least one cycloalkyl group with 3 to 10 atoms and / or - at least one hydroxy, cyano, alkyloxy or alkylthio with 1 to 10 C atoms , Carboxyl or carboxyalkyl group, the alkyl group of the carboxyalkyl group having 1 to 10 carbon atoms and / or the carboxyl group, if appropriate is esterified or amidated with a linear, branched or cyclic alkyl radical having 1 to 10 carbon atoms;

T either does not exist or is one of the following groups:

wherein R _{5 is} hydrogen or an alkyl group with 1 to 10 C atoms or an alkylene group with A which may contain at least one hetero atom and has an alkylene group with 1 to 6 C atoms;

A _{2 is} a linear, branched or cyclic alkylene group with 1 to 10 C atoms or an aryl, heteroaryl or aralkylene group with 1 to 10 C atoms, optionally containing at least one hetero atom, selected from the group consisting of N, S and O , which is optionally substituted with at least one halogen and / or pseudohalogen group and / or at least one linear, branched or cyclic alkyl group with 1 to 10 C atoms and / or at least one aryl or an aralkyl group with 5 to 10 carbon atoms and / or ~ at least one cycloalkyl group with 3 to 10 carbon atoms and / or at least one hydroxy, cyano, alkyloxy or alkylthio with 1 to 10 carbon atoms, carboxyl or carboxyalkyl group, the Alkyl group of the carboxyalkyl group has 1 to 10 carbon atoms and / or the carboxyl group optionally is esterified or amidated with a linear, branched or cyclic alkyl radical having 1 to 10 carbon atoms; (iv) _{t is} one of the following, optionally modified basic groups:

where t = 0.1; and R ₇ independently of one another, hydrogen or an alkyl group having 1 to 6 C atoms or an alkylene group having 1 to 5 C atoms which forms a cycle with A ₂ , or a hydroxyl, amino, alkylamino, acyl or alkyloxycarbonyl group are, where the alkylamino, acyl and alkyloxycarbonyl groups independently of one another have 1 to 6 carbon atoms, and wherein R _{8 is} hydrogen or an alkyl group with 1 to 3 carbon atoms or an alkylene group with 1 to 3 carbon atoms, which forms a cycle with R _Ö ;

(v) Q is either a CH group or N;

(vi) j = 0.1.2; k = 0, 1, 2, 3; m, n independently of one another = 0, 1, 2, 3, 4, 5, where m + n = 3, 4, 5; and wherein the compound of formula (I) is neither

with s = 0, 1, 2 still

with s = 0.1.

2. A compound according to claim 1, characterized in that T is present and T is one of the groups as defined in claim 1, it being possible for the amide and ester groups to be incorporated in both orientations.

3. A compound according to claim 1 or 2, characterized in that it has the structure (I ").

4. A compound according to claims 1 to 3, wherein j = 0 and Ri is an aryl radical which is substituted at most twice.

5. A compound according to claim 4, wherein the aryl group is substituted by an oxygen bridge atom or a sulfur bridge atom or by a C ₁ -C ₃ alkylene chain with a further aryl group or a heteroaryl group.

6. A compound according to claim 5, wherein the aryl radical which is substituted by an oxygen bridge atom with a further aryl radical or a heteroaryl radical is a phenyl radical or a pyridyl radical.

7. A compound according to claim 3 or 4, wherein the aryl radical or the heteroaryl radical carries at least one substituent selected from the group consisting of chlorine, fluorine,

Trifluoromethyl, methyl, methoxy.

8. A compound according to claim 4, wherein the aryl radical is substituted with at least one alkoxy group.

9. A compound according to any one of claims 1 to 4, wherein Ri is selected from the group consisting of tert-butylphenyl, cyclohexylphenyl, 5,6,7,8-tetrahydronaphthyl, naphthyl, anthracyl, anthraquinoyl and anthrahydroquinoyl, pyridyloxyphenyl, phenyloxypyridyl, Pyridylalkylphenyl with a Ci-C ₃ alkyl.

10. A compound according to any one of claims 1 to 9, wherein R _{2 is} an at least monosubstituted phenyl radical, thienyl radical or pyridyl radical.

11. A compound according to any one of claims 1 to 10, wherein k = 1 and R _{2 is} a phenyl radical which is meta-substituted with an amidino group, the resulting 3-amidinophenylalanine being present in the L configuration.

12. A compound according to any one of claims 1 to 11, wherein m = n = 2.

13. A compound according to any one of claims 1 to 12, wherein the cycle formed by Xi and X has the following structure:

14. A compound according to any one of claims 1 to 13, wherein Ai is absent and T is.

15. A compound according to any one of claims 1 to 14, wherein A _{2 is} a methylene, ethylene or propylene group and Rt is selected from

16. A compound according to any one of claims 2 to 15, wherein m = n = 2 and R _{3 is} a guanidinooxyalkyl radical or an aryl or heteroaryl radical, and wherein the aryl radical is preferably a benzyl or phenoxy radical and the heteroaryl radical is preferably selected is from a pyridinylmethylene, pyridinyloxo, pyrimidinyloxo, pyrazinyloxo, pyridinylthio radical and the aryl or heteroaryl radical is unsubstituted or substituted with at least one halogen, at least one methoxy radical and / or at least one trifluoromethyl radical.

17. A compound according to any one of the preceding claims, characterized in that an amino acid, preferably a glycine, is incorporated between the sulfenyl radical and the 3-amidinophenylalanine radical of the formula I.

18. A process for the preparation of a compound according to one of claims 1 to 17 or a compound (AI) or (A2), comprising the step (S1): (S1) reacting a compound of the general structure (El ¹ ) with a compound of the general structure (El ")

maintaining a connection of the general structure (ZPl)

where R ₂ "is the aryl radical R ₂ , substituted either with R _t or with R _t , protected with a suitable protective group, or with a substituent which is a precursor to R *.

19. The method according to claim 18, wherein Q = N and Ai is not present, additionally comprising the steps (S2 '), (S3 *) and (S4'):

(S2 ') implementation of the compound (ZPl) with a compound of the general structure (E2')

where W is a suitable protective group, to give a compound of the general structure (ZP2 ')

(S3 ') deprotection W;

(S4 ') reaction of the compound obtained according to (S3') with a compound of the general structure (E2 ")

HO-TA ₂ -R ₄ '(E2 ») to obtain a compound of the general structure (Pl)

where R- _t 'is either R ₄ or R ₄ , protected with a suitable protecting group, or a precursor to R ₄ ; or (S2 ') implementation of a compound of the general structure (E2 ^m )

HO-TA ₂ -R _(E2 » ₎ to obtain a compound of the general structure (ZP2") H ₂

WN; : Q - T - A ₂ - R (ZP2 ")

H ₂ where R represents either Rt or Rt, protected with a suitable protecting group, or a precursor to _t ;

(S3 ¹ ) deprotection W;

(S4 ') reaction of the compound obtained according to (S3') with a compound of the general structure (ZPl) to obtain a compound of the general structure (Pl).

20. The method according to claim 18 wherein Q = CH, additionally comprising the step (S2 "): (S2") reaction of the compound according to (ZPl) with a compound of the general structure (E3)

where _t 'is either t or R _t , protected with a suitable protective group, or a precursor to R _t , to give a compound of the general structure (P2)

21. The method according to claim 18, wherein T = - (C = O) -NH-, Q = CH and Ai are not present, additionally comprising the steps (S2 ^m ), (S3 ^{, M} ) and (S4 '"): ( S2 '") implementation of a compound of the general structure (E3')

with a connection of the general structure (E3 ")

H ₂ NA ₂ -R ₄ '(E3 ") to obtain a compound of the general structure (ZP3)

(S3 ^{, M} ) removal of the protective group W;

(S4 ^m ) reaction of the compound obtained according to (S3 ^{, π} ) with a compound according to (ZPl) to obtain a compound of the general structure (P3)

22. The method according to claim 18, additionally comprising the step of converting the connection (ZPl) with a connection of the general structure (E2 ')

where Q = N, m = n = 2 and R3 is as defined in claim 15.

23. Medicament containing at least one compound according to one of claims 1 to 17 or a compound (AI) or (A2) or a salt of these compounds.

24. Medicament according to spoke 23, characterized in that it is in the form of a tablet, a dragee, a capsule, a pellet, a suppository, a solution, in particular a solution for injection or infusion, eye, nose or ear drops, a juice, a Emulsion or suspension, a globule, a styli, an aerosol, a powder, a paste, a cream or an ointment is used.

25. Use of a compound according to one of claims 1 to 17 or a compound (AI) or (A2) or a salt of these compounds or a medicament according to claim 23 or 24 for the diagnosis, therapy or prophylaxis of a tumor, in particular in oral, subcutaneous, intravenous or transdermal form.

26. Use according to claim 25, wherein the formation of tumor metastases is reduced.

27. Use of a compound according to any one of claims 1 to 17 or one

Compound (AI) or (A2) or a salt of these compounds or one

Medicament according to claim 23 or 24 for inhibiting matriptase.

28. Use according to spoke 27, characterized in that the matriptase is MT-SP1.