EP1651597A2 - N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren - Google Patents

N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren

Info

Publication number
EP1651597A2
EP1651597A2 EP04733311A EP04733311A EP1651597A2 EP 1651597 A2 EP1651597 A2 EP 1651597A2 EP 04733311 A EP04733311 A EP 04733311A EP 04733311 A EP04733311 A EP 04733311A EP 1651597 A2 EP1651597 A2 EP 1651597A2
Authority
EP
European Patent Office
Prior art keywords
group
radical
compound
atoms
aryl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP04733311A
Other languages
German (de)
English (en)
French (fr)
Inventor
Jörg STÜRZEBECHER
Torsten Steinmetzer
Andrea Schweinitz
Anne STÜRZEBECHER
Kerstin Uhland
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Medicines Company Leipzig GmbH
Original Assignee
Curacyte Chemistry GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Curacyte Chemistry GmbH filed Critical Curacyte Chemistry GmbH
Priority to EP10013067A priority Critical patent/EP2316830A3/de
Publication of EP1651597A2 publication Critical patent/EP1651597A2/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
    • C07D295/185—Radicals derived from carboxylic acids from aliphatic carboxylic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D205/00—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom
    • C07D205/02—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings
    • C07D205/04—Heterocyclic compounds containing four-membered rings with one nitrogen atom as the only ring hetero atom not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/18—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D211/26—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D211/60—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D211/62—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
    • C07D307/79—Benzo [b] furans; Hydrogenated benzo [b] furans with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring

Definitions

  • 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.
  • proteolytic enzymes include various matrix metalloproteases, membrane bound metalloproteases, lysosomal cysteine proteases and a variety of serine proteases such as e.g.
  • 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).
  • 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.
  • 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).
  • 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.
  • 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).
  • 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.
  • HGF hepatocyte growth factor
  • 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.
  • matriptase may also be involved in activating the plasminogen activator cascade.
  • 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.
  • uPA urokinase
  • 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.
  • gastric cancer cells increasingly express ⁇ 1-6-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.
  • GnT-V ⁇ 1-6-N-acetylglucosaminyltransferase
  • 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.
  • 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.
  • the present invention relates to a compound of the formula (I) or a salt or a prodrug of this compound, wherein
  • Xi and X 2 independently of one another are hydrogen or an alkyl radical having 1, 2 or 3
  • 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
  • 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
  • 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 C-
  • 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):
  • 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:
  • 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. -
  • salts with mineral acids and / or salts with suitable organic acids are preferred.
  • 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.
  • preferred salts of the compounds according to the invention with suitable organic acids are acetates.
  • the compound according to the invention has a structure in which at least one of the radicals Xi and X has a structure (V).
  • 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.
  • m + n is 3 or 4.
  • the residue R_ ⁇ 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.
  • 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
  • alkylene groups having 1, 2, 3, 4 or 5 carbon atoms 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.
  • a 2 is an aryl, heteroaryl or aralkylene group
  • groups are of the structures, for example
  • 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.
  • 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.
  • the cycle according to (V) can contain at least one heteroatom, preference being given here to oxygen, nitrogen or sulfur.
  • 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.
  • none of the methylene groups according to (! ') Is substituted by a hetero atom.
  • 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.
  • 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 ").
  • the present invention also relates to a compound as described above, which compound has the structure (I ")
  • 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 Bonylcuba 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.
  • 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.
  • the cycle of the cyclic imine can be suitably substituted.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • the present invention also relates to a compound as described above, wherein the cycle formed by Xi and X 2 has the following structure
  • Q can be in the 2-, 3- or 4-position relative to the imine nitrogen, the 4-position being particularly preferred.
  • 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.
  • the present invention also describes a compound as described above, which is characterized in that Q is a nitrogen atom.
  • 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.
  • 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.
  • group Ai is not present.
  • the present invention also describes a compound as described above, which is characterized in that Q is a nitrogen atom and ⁇ i is absent.
  • Groups of structures are particularly preferred for T.
  • R 5 is hydrogen.
  • T is a group of the structures
  • T being a functional group of the structure
  • amide or ester groups can be incorporated in both orientations.
  • 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
  • Q is a nitrogen atom
  • 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.
  • a 2 is an aryl, heteroaryl or aralkylene group
  • groups are of the structures, for example
  • 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.
  • 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.
  • 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:
  • 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:
  • R 6 and R 7 are hydrogen.
  • R 4 radicals of the following structures are very particularly preferred:
  • R ⁇ 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
  • 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.
  • 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.
  • group Ai is not present.
  • 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.
  • the functional groups 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.
  • the functional groups T are groups of the structures
  • Groups of structures are particularly preferred for T.
  • R 5 is hydrogen.
  • T is a group of the structures
  • T is very particularly preferably a functional group of the structure
  • amide or ester groups can be incorporated in both orientations.
  • 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
  • the group Ai is not present according to a particularly preferred embodiment of the present invention.
  • the present invention also describes a compound as described above, which has the following structure:
  • 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:
  • 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.
  • a 2 is an aryl, heteroaryl or aralkylene group
  • groups are of the structures, for example
  • 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.
  • 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.
  • halogens preferably fluorine, chlorine or bromine
  • alkyl radicals with preferably 1, 2 or 3 carbon atoms such as methyl, ethyl, n-propyl or isopropyl, particularly preferably methyl and ethyl
  • substituents preferably methyl, ethyl, n-propyl or isopropyl, particularly preferably methyl and ethyl
  • / 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.
  • 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:
  • the present invention therefore also describes compounds, as described above, which are characterized in that they have the following structures:
  • radicals R of the following structures are particularly preferred:
  • N - HR x c 6 further preferably R and R being hydrogen.
  • Re and R are hydrogen.
  • 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
  • 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.
  • 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.
  • 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
  • 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
  • the substituent Rj can generally be positioned at all positions of the aryl radical.
  • the 2-, 3- or 4- position of the R-j radical is accordingly possible, the 3-position of the R ⁇ radical on the phenyl radical being particularly preferred.
  • the aryl radical can furthermore have at least one further substituent.
  • 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.
  • 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 ⁇ -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.
  • 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.
  • 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.
  • 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.
  • radical Ri is substituted, for example, by at least one halogen, fluorine, chlorine and / or bromine are preferred.
  • radical R 1 is substituted, for example, with at least one esterified carboxyl group, methyl esters and / or ethyl esters are preferred.
  • 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.
  • 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.
  • preferred aryl radicals are phenyl, naphthyl, anthracyl or phenanthryl. These aryl radicals can optionally be present in reduced or / and oxidized form.
  • 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.
  • 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.
  • substituted R 1 radicals are, for example
  • 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.
  • the aryl radical has a substituent, for example, the tert-butyl radical is preferred.
  • 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
  • Atoms can be replaced.
  • 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.
  • the aryl radical is preferably a benzyl or a phenoxy radical
  • the heteroaryl radical is preferably selected from a pyridinylmethyl, pyridinyloxo, pyrimidinyloxo, pyrazinyloxo, pyri
  • the R 3 radical is a guanidinooxyalkyl radical.
  • 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.
  • 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 ')
  • 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.
  • 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
  • this amidino group can again be protected with a suitable protective group, which can be removed again in a suitable process step.
  • 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 )
  • R t ' represents either t or R t , protected with a suitable protective group, or a precursor to t.
  • the C-terminal residue of the middle-sized amino acid according to (I) is built up first.
  • 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.
  • the radical R 4 is an amino group, a guanidino group can also be built up from it here, as described above.
  • 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)
  • 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)
  • Rt ' is a radical R t protected with a protective group
  • the protective group is preferably removed after step (S2 ").
  • an amino group can then be present as the radical R t .
  • 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.
  • W is also a suitable protective group here.
  • 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)
  • Rt ' is a radical R t protected with a protective group
  • the protective group is preferably removed after step (S2 m ).
  • an amino group can then be present as the radical R t .
  • 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.
  • 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.
  • 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.
  • the present invention also relates to a medicament which contains one of the compounds specified above.
  • 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.
  • the compounds according to (I) according to the invention can be used in any form as a medicament.
  • 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.
  • 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.
  • a solution 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.
  • 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.
  • 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.
  • a compound according to the invention according to (I) is used to reduce tumor metastases.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • Analytical HPLC Shimadzu LC-10A system, column: Phenomenex Luna C ⁇ 8 , 5 ⁇ m, 100 ⁇ (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 C ⁇ 8 , 5 ⁇ m, 100 ⁇ (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 ⁇ - Cyano-hydroxycinnamic acid as a matrix, or on an ESI-MS LCQ from Finnigan (Bremen, Germany), measured.
  • 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.
  • 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.
  • 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 ⁇ l (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.
  • 3 x acidic 5% KHSO 4
  • 1 x neutral saturated NaCl solution
  • 3 x basic NaHCO 3 saturated
  • 3 x neutral saturated NaCl Solution
  • 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.
  • 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.
  • 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.
  • 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:
  • BL21 (DE3) cells which contained the vector from the cloning described above, were incubated in LB, 30 ⁇ g / 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 TM 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.
  • Denaturing buffer 6 M guanidinium HCL, 10 mM Tris HCl, 100 mM Na phosphate, pH 8.0
  • Insoluble constituents were centrifuged off (16,000 g, 30 min, 20 ° C.), the supernatant was filtered (0.2 ⁇ m), 10 mM ⁇ -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 ⁇ g / 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).
  • the refolding batch was filtered, concentrated to a concentration of> 300 ⁇ g / 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).
  • the peptide MK (His) 6 at the N-terminus had to be removed to activate the refolded matriptase.
  • the refolding batch was incubated for 2 h at 30 ° C. with 2.5 mU per 50 ⁇ g protein-activated DAPase TM (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
  • 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.
  • 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 ⁇ g Matrigel each, 160,000 DLD-1 cells (Dexter et al, Cancer Research 39: 1020-1025 (1979)) in 100 ⁇ l medium (RPMI 1640, with 2% Ultroser HY serum substitute was applied and the invasive growth was stimulated by adding ProHGF in 400 ⁇ l medium with and without inhibitor (30 ⁇ M). 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.
  • 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.
  • proHGF inactive proform of HGF
  • PC-3 prostate carcinoma cells expressing matriptase were seeded per well of a 96-well plate, incubated overnight in 100 ⁇ l 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 ⁇ M) stimulated by the addition of proHGF. After 6 days the cells were fixed, stained and representative sections were photographed at 100x magnification.
  • the matriptase inhibitors 37 and 54 inhibit the proHGF-induced ⁇ scattering 'of PC-3 cells.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Oncology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Hydrogenated Pyridines (AREA)
  • Plural Heterocyclic Compounds (AREA)
EP04733311A 2003-05-16 2004-05-17 N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren Withdrawn EP1651597A2 (de)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP10013067A EP2316830A3 (de) 2003-05-16 2004-05-17 N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10322191.3A DE10322191B4 (de) 2003-05-16 2003-05-16 N-sulfonylierte Aminosäurederivate, Verfahren zu ihrer Herstellung und ihre Verwendung
PCT/EP2004/005291 WO2004101507A2 (de) 2003-05-16 2004-05-17 N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase-inhibitoren

Publications (1)

Publication Number Publication Date
EP1651597A2 true EP1651597A2 (de) 2006-05-03

Family

ID=33440914

Family Applications (2)

Application Number Title Priority Date Filing Date
EP04733311A Withdrawn EP1651597A2 (de) 2003-05-16 2004-05-17 N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren
EP10013067A Withdrawn EP2316830A3 (de) 2003-05-16 2004-05-17 N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren

Family Applications After (1)

Application Number Title Priority Date Filing Date
EP10013067A Withdrawn EP2316830A3 (de) 2003-05-16 2004-05-17 N-sulfonylierte aminosäurederivate und ihre verwendung als matriptase- inhibitoren

Country Status (7)

Country Link
US (2) US7772251B2 (enExample)
EP (2) EP1651597A2 (enExample)
JP (1) JP4653100B2 (enExample)
AU (1) AU2004238558A1 (enExample)
CA (1) CA2525886C (enExample)
DE (1) DE10322191B4 (enExample)
WO (1) WO2004101507A2 (enExample)

Families Citing this family (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE399175T1 (de) * 2002-03-11 2008-07-15 Curacyte Ag Hemmstoffe der urokinase, ihre herstellung und verwendung
DE10301300B4 (de) 2003-01-15 2009-07-16 Curacyte Chemistry Gmbh Verwendung von acylierten 4-Amidino- und 4-Guanidinobenzylaminen zur Inhibierung von Plasmakallikrein
DE10322191B4 (de) * 2003-05-16 2014-02-27 The Medicines Company (Leipzig) Gmbh N-sulfonylierte Aminosäurederivate, Verfahren zu ihrer Herstellung und ihre Verwendung
DE10342108A1 (de) * 2003-09-11 2005-04-14 Curacyte Chemistry Gmbh Basisch-substituierte Benzylaminanaloga als Inhibitoren des Gerinnungsfaktors Xa, ihre Herstellung und Verwendung
JP2008510726A (ja) * 2004-08-20 2008-04-10 エントレメッド インコーポレイテッド プロテイナーゼ活性化受容体アンタゴニストを含む組成物および方法
DE102005044319A1 (de) * 2005-09-16 2007-03-22 Curacyte Chemistry Gmbh 2-(Aminomethyl)-5-Chlor-Benzylamid-Derivate und ihre Verwendung als Hemmstoffe des Gerinnungsfaktors Xa
JP5406722B2 (ja) 2006-10-16 2014-02-05 グリュネンタール・ゲゼルシャフト・ミト・ベシュレンクテル・ハフツング ブラジキニン１受容体モジュレータとしての置換されたスルホンアミド誘導体
DE102006050672A1 (de) * 2006-10-24 2008-04-30 Curacyte Discovery Gmbh Hemmstoffe des Plasmins und des Plasmakallikreins
DE102007010815B3 (de) 2007-03-06 2008-07-31 Curacyte Discovery Gmbh Meta-substituierte Phenylsulfonylamide sekundärer Aminosäureamide, ihre Herstellung und Verwendung als Hemmstoffe der Matriptase
JP5260976B2 (ja) * 2008-02-08 2013-08-14 東ソー株式会社 血液凝固第ｖｉｉｉ因子ｃ２ドメインタンパク質の製造方法
US8912176B2 (en) 2009-02-02 2014-12-16 Evotec Ag Azetidines as histamine H3 receptor antagonists
KR20120058468A (ko) 2009-05-18 2012-06-07 오리온 코포레이션 프로테아제 억제제
US8921319B2 (en) 2010-07-07 2014-12-30 The Medicines Company (Leipzig) Gmbh Serine protease inhibitors
EP2655399B1 (en) 2010-12-21 2017-09-27 The Medicines Company (Leipzig) GmbH Trypsin-like serine protease inhibitors, their preparation and use as selective inhibitors of the clotting factors iia and xa
WO2012162828A1 (en) * 2011-06-02 2012-12-06 Socpra Sciences Santé Et Humaines S.E.C. Matriptase inhibitors and uses thereof against orthomyxoviridae infections
US11130780B2 (en) 2015-03-09 2021-09-28 Washington University Inhibitors of growth factor activation enzymes
KR102704242B1 (ko) 2017-06-20 2024-09-09 임브리아 파마슈티칼스, 인크. 심장 대사의 효율을 증가시키기 위한 조성물 및 방법
EP3654968B1 (en) 2017-07-17 2023-08-16 Merck Sharp & Dohme LLC Metallo-beta-lactamase inhibitors and methods of use thereof
WO2020081361A1 (en) 2018-10-17 2020-04-23 Imbria Pharmaceuticals, Inc. Methods of treating rheumatic diseases using trimetazidine-based compounds
US11780811B2 (en) 2020-06-30 2023-10-10 Imbria Pharmaceuticals, Inc. Methods of synthesizing 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate
US11530184B2 (en) 2020-06-30 2022-12-20 Imbria Pharmaceuticals, Inc. Crystal forms of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate
US11883396B2 (en) 2021-05-03 2024-01-30 Imbria Pharmaceuticals, Inc. Methods of treating kidney conditions using modified forms of trimetazidine

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3942114C2 (de) * 1988-12-26 1997-09-04 Hiroyoshi Hidaka Verbindungen mit einer Wirkung für eine gleichmäßige Gefäßmuskelentspannung und pharmazeutische Zusammensetzung, die diese enthält
WO1992008709A1 (de) 1990-11-15 1992-05-29 Pentapharm Ag Meta-substituierte phenylalanin-derivate
US5607937A (en) * 1993-02-10 1997-03-04 Pentapharm Ag Piperazides of substituted phenylalanine derivatives as thrombin inhibitors
CH689611A5 (de) 1995-03-01 1999-07-15 Pentapharm Ag Urokinase-Inhibitoren.
EP1264828B1 (de) * 1998-07-20 2004-04-14 Wilex AG Urokinase-Inhibitoren
EP1114024B1 (de) 1998-09-18 2002-11-27 Pentapharm A.G. Urokinase-inhibitoren
US6797504B1 (en) 2000-09-08 2004-09-28 Dendreon San Diego Llc Inhibitors of serine protease activity of matriptase or MTSP1
EP1355637A2 (en) 2000-06-21 2003-10-29 Georgetown University Medical Center Inhibitors of matriptase for the treatment of cancer
AU2003206945A1 (en) * 2002-02-22 2003-09-09 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Use of proteinase inhibitors in the treatment of autoimmune diseases
DE10322191B4 (de) * 2003-05-16 2014-02-27 The Medicines Company (Leipzig) Gmbh N-sulfonylierte Aminosäurederivate, Verfahren zu ihrer Herstellung und ihre Verwendung

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2004101507A2 *

Also Published As

Publication number Publication date
US7772251B2 (en) 2010-08-10
WO2004101507A3 (de) 2006-03-23
CA2525886C (en) 2013-01-08
JP2007500734A (ja) 2007-01-18
EP2316830A2 (de) 2011-05-04
JP4653100B2 (ja) 2011-03-16
US20110002992A1 (en) 2011-01-06
WO2004101507A2 (de) 2004-11-25
DE10322191B4 (de) 2014-02-27
AU2004238558A1 (en) 2004-11-25
EP2316830A3 (de) 2011-08-17
US20070055065A1 (en) 2007-03-08
CA2525886A1 (en) 2004-11-25
DE10322191A1 (de) 2004-12-09

Similar Documents

Publication Publication Date Title
DE10322191B4 (de) N-sulfonylierte Aminosäurederivate, Verfahren zu ihrer Herstellung und ihre Verwendung
DE69830410T2 (de) Meta-benzamidinderivate als serinprotease-inhibitoren
EP1294742B1 (de) Urokinase-hemmstoffe
EP0511347A1 (de) Meta-substituierte phenylalanin-derivate
WO1999054294A1 (de) Neue substituierte amide, deren herstellung und anwendung
WO1992016549A1 (de) Para-substituierte phenylalanin-derivate
US4873253A (en) Phenylalanine derivative and proteinase inhibitor
DE69921994T2 (de) Heterozyklische verbindungen mit faktor xa hemmender wirkung
EP3297992B1 (en) Heterocyclicalkyl derivative compounds as selective histone deacetylase inhibitors and pharmaceutical compositions comprising the same
EP0508220B1 (de) Amidinophenylalaninderivate, Verfahren zu deren Herstellung, deren Verwendung und diese enthaltende Mittel
EP1114024B1 (de) Urokinase-inhibitoren
EP0574808A1 (de) Amidin-Biphenylderivate, diese Verbindungen enthaltende Arzneimittel und Verfahren zu ihrer Herstellung
EP1455790B1 (de) Verwendung von pyridin-2,4-dicarbons urediamiden und pyrimid in-4,6-dicarbons urediamiden zur selektiven inhibierung von kollagenasen
EP1664089B1 (de) Basisch-substituierte benzylaminanaloga als inhibitoren des gerinnungsfaktors xa, ihre herstellung und verwendung
EP2132174B1 (de) Meta-substituierte phenylsulfonylamide sekundärer aminosäureamide als hemmstoffe der matriptase zur hemmung des tumorwachstums und/oder der metastasierung von tumoren
EP1373234B1 (de) Urokinase-inhibitoren
DE10251019A1 (de) Neue Pyrimidin-4,6-dicarbonsäurediamide zur selektiven Inhibierung von Kollagenasen
EP1511721A2 (de) Guanidinophenylalanin-verbindungen als urokinase-inhibitoren
DE60010344T2 (de) Diketopiperazinderivate als thrombininhibitoren
DE10209299A1 (de) Cyclische N-substituierte alpha-Iminocarbonsäuren zur selektiven Inhibierung von Kollagenase
DE10254092A1 (de) Neue Pyrimidin-4,6-dicarbonsäurediamide zur selektiven Inhibierung von Kollagenasen

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

PUAK Availability of information related to the publication of the international search report

Free format text: ORIGINAL CODE: 0009015

17P Request for examination filed

Effective date: 20051202

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LI LU MC NL PL PT RO SE SI SK TR

AX Request for extension of the european patent

Extension state: AL HR LT LV MK

DAX Request for extension of the european patent (deleted)
RIC1 Information provided on ipc code assigned before grant

Ipc: C07D 295/18 20060101AFI20061116BHEP

17Q First examination report despatched

Effective date: 20070731

EL Fr: translation of claims filed
RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: THE MEDICINES COMPANY (LEIPZIG) GMBH

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20140722