DE102009035593A1 - New tetrapeptide derivatives are proprotein convertase inhibitors, useful to treat e.g. viral or bacterial diseases, diabetes, arteriosclerosis, cancer, Alzheimer's disease, obesity, and respiratory tract disease - Google Patents

Abstract

Tetrapeptide derivatives (I) are new. Tetrapeptide derivatives of formula (P5-P4-P3-P2-P1) (I) are new. P1 : arginine mimetic; P2-P4 : amino and/or imino acid radical; and P5 : N-terminal modification of amino or imino acid residue of P4. An independent claim is included for the preparation of (I). ACTIVITY : Virucide; Antibacterial; Antidiabetic; Antiarteriosclerotic; Cytostatic; Neuroprotective; Nootropic; Anorectic; Respiratory-Gen.; Antimicrobial; Nephrotropic. MECHANISM OF ACTION : Proprotein convertase inhibitor. The ability of (I) to inhibit furin was tested using a fluorescence plate reader. The result showed that (S)-6-carbamimidoyl-2-phenylacetylamino-hexanoic acid {(S)-1-[(S)-5-carbamimidoyl-1-(4-carbamimidoyl-benzylcarbamoyl)-pentylcarbamoyl]-2-methyl-propyl}-amide exhibited a inhibition constant (K i) value of 0.0014 mu M.

Description

The The invention relates to N-terminally modified tetrapeptide derivatives with C-terminal arginine mimetic according to claim 1, method for the preparation of the tetrapeptide derivatives according to the invention according to the independent claims 18 and 23, A pharmaceutical composition according to claim 33 and the use the compound of the invention, as well as the inventive Medicament according to claims 35, 36, 37 and 38.

she describes novel peptidomimetics that act as inhibitors of serine protease Furin and other proprotein convertases, such as furin C-terminal from multibasic cleavage sites hydrolyze their substrates, act. In addition, methods for their production are provided.

peptide mimetics are well known. They serve in the fields of pharmacy, Medicine, organic chemistry and biochemistry among others as specific Inhibitors of certain enzymes. It must be every mimetic especially with regard to the particular enzymatic to be influenced Activity can be designed and optimized. Therefore exists a strong dependence of the effectiveness of the peptide mimetic from the respective target enzyme.

The human proprotein convertases belong to the family of serine proteases. According to the current state of knowledge, there are a total of 9 human proprotein convertases of which 7 representatives (furin, PC1 / 3, PC2, PC4, PC5 / 6, PACE4 and PC7) cleave their substrates behind a basic amino acid, while two members (SKI-1 and PCSK9) hydrolyze peptide bonds behind non-basic amino acid residues ( Seidah, NG and Prat, A., J. Mol. Med. 2007, 85, 685-696 ). The proprotein convertases belong to the family of subtilisin-like serine proteases, which differ greatly in their sequence and structure from other serine proteases which also cleave behind basic P1 residues and which include, for example, the trypsin-like serine proteases (the designation P1, P2, used in the application). P3, P4 and P5 correspond to the nomenclature proposed by Schechter and Berger for designating the amino acid positions in protease substrates and substrate analog inhibitors, Schechter & Berger, Biochem. Biophys. Res. Comm., 1967, 27, 157 ).

In contrast to the trypsin-like serine proteases, it is characteristic of representatives of proprotein convertases which cleave behind basic P1 residues that they only efficiently convert substrates if they possess further basic amino acid residues N-terminal of the cleavage site. Furin was the first described member of this group of 7 proteases and is now best characterized ( Thomas, G., Nat. Rev. Mol. Cell. Biol., 2002, 3, 753-766 ). Furin has been found to efficiently cleave substrates especially when in addition to a basic P1 amino acid, they also have other basic P2 and P4 residues. Very similar substrate specificities were also found for PC1 / 3, PC2, PC5 / 6 and PC7 (see MEROPS database http://merops.sanger.ac.uk/ and Remacle AG et al. J. Biol. Chem., 2008, 283, 20897-20906 ), but also substrates of PC4 and PACE4 require basic residues in the P2 and P1 positions.

It There are numerous indications that the proteolytic action of furin very likely the emergence or spread of diseases causes. You may be able to get such effects also taken over by other proprotein convertases in vivo because they have a similar substrate specificity like the furin own. Therefore, the following are made Explanations of the physiological function of furin in principle also for the other above-mentioned proprotein convertases, split the back basic P1 residues.

Thus, furin has been described as an activator of various bacterial toxins, such as the anthrax toxin, the Clostridium septicum α toxin, the Pseudomonas exotoxin, the shiga toxin, the shiga-like toxins or the diphtheria toxin (reviewed in US Pat Thomas, G., Nat. Rev. Mol. Cell. Biol., 2002, 3, 753-766 ). These toxins belong to so-called A / B toxins which contain an active domain (A) and a binding domain (B), both of which are linked together via a furin cleavage sequence. After receptor binding of the toxins they pass through endocytosis into the endosomes of the affected cell and are there split by furin, whereby these toxins can then reach the cytosol of the cells via various mechanisms ( Thomas, G., Nat. Rev. Mol. Cell. Biol., 2002, 3, 753-766 ).

A variety of other bacteria have at least one or more toxins with a furin cleavage site, such. Bacillus cereus, B. anthracis, B. butulinum, Clostridium perfringens, C. difficile, C. spirioforme, Bordetella pertussis, Samonella enterica, some E. coli strains, (e.g., E. coli 0157-H7), identified as EHEC (Enterohaemorrhagic Escherichia coli strains) Hemolytic colitis, the hemolytic uraemic Syndrome, including causing kidney failure. Shigella and E. coli toxins are also referred to as verotoxins because of their similarity.

Furin also plays an essential role in the activation of various pathogenic viruses, for example in highly pathogenic avian influenza A viruses (HPAIV) ( Thomas, G., Nat. Rev. Mol. Cell. Biol., 2002, 3, 753-766 ). They all belong to the H5 or H7 subtypes, which can be differentiated serologically from the other subtypes. So far, they have only one multibasic cleavage site in their glycoprotein spike, hemagglutinin, derived from furin and other proprotein convertases, e.g. B. PC5 / 6, is split. It is to be expected that in the future still further viruses of other subtypes will be identified, which possess Furinspaltstellen in their surface proteins. These highly pathogenic avian influenza A viruses include, for example, the strain H5N1 that appeared in Hong Kong in 1997, while H7N1 had caused considerable losses in poultry in the Netherlands some years ago. The importance of furin or another protein mutant convertase with similar substrate specificity for influenza also became evident when mutinating a furin cleavage site with normally avirulent avian influenza strains possessing only a single arginine at the cleavage site (ie, would not normally be cleaved by furin) which resulted in a very high virulence, although all other virulence factors were present.

Further Virus examples with by Furin and other Proproteinvertvertasen activatable glycoproteins are about 100 significant RNA and DNA viruses, the diseases that are significant in humans and many animal species to lead. These include the RNA negative strand viruses the parainfluenza virus, the measles virus, the mumps virus, the canine distemper virus, the RSV (respiratory syncytial virus) in humans and cattle, the filoviruses (Ebola and Marburg virus, causing severe hemorrhagic fever) and that Newcastle disease virus (virus of non-classical avian influenza). The plus-strand RNA viruses are among those by furin activatable species HIV-1 (Human Immunodeficiency V), HIV-2, HTLV (Human T-Lymphocytic Leukemia V.) And many other retroviruses, coronaviruses (eg the SARS coronavirus), the flaviviruses (yellow fever virus, West Nile virus, dengue virus), they are currently called "emerging viruses" originally narrow areas over wide Spreading parts of the world, but especially in the tropical zones. In our latitudes, the tick-borne encephalitis virus (Engl. "Tick born virus "). Other dreaded viruses are Venezuelan Equine Encephalitis Virus or Rift Valley Fever Virus.

Alles these viruses have a glycoprotein on their surface with a furin cleavage site. After cleavage of this glycoprotein Furin is a peptide sequence unmasked, causing the virus envelope can fuse with membranes of the virus-infected cell and thereby the virus genome is introduced into the cell. In addition, causes the fusion in some viruses that biologically active peptides out be split. By experiments with HIV-1 and cells the no Furin was shown to be possibly at the Activation also other proprotein convertases with similar Substrate specificity take over the function of furin can.

In addition to furin-mediated effects in bacterial and viral diseases, furin or an analogous proprotein convertase may be involved in, enhance or even trigger other diseases. Thus, furin can cleave different pro-forms of growth factors, receptors, adhesion molecules or matrix metalloproteases, triggering various diseases. These conditions may include diabetes, arteriosclerosis, cancer / metastasis, as well as Alzheimer's or obesity (summarized in Bontemps, Y. et al., Med. Res. Rev., 2007, 27, 631-648 ). In neurodegenerative diseases such. For example, in Alzheimer's, furin may be involved in activating the zymogen forms of secretases responsible for the formation of the Aβ peptide ( Bennett, BD et al., J. Biol. Chem., 2000, 275, 37712-37717 ). By the aforementioned furin-mediated cleavage of matrix metalloproteases, adhesion molecules growth factors, growth factor receptors, furin or an analogous proprotein convertase may be directly involved in the development, progression and progression of cancer as well as metastasis and also in the degradation of extracellular matrix proteins.

A review of potential substrate sequences of furin and other proprotein convertases, which require a basic amino acid as P1 residue, is included in the appendix of a recent publication on the substrate specificity of these enzymes ( Supplement to Remacle AG et al. J. Biol. Chem., 2008, 283, 20897-20906 ). However, no suitable inhibitors for these proteases are yet available, although attempts have been made in recent years by various groups to develop furin inhibitors. These include compounds derived from polyarginine peptides. So inhibits, for example, nona-D-arginine amide furin with an inhibition constant of 1.3 nM, while the shorter hexa-D-arg amide has a K _i value of 7.6 nM ( Kacprzak, MM et al., J. Biol. Chem., 2004, 279, 36788-36794 ). An overview of known inhibitors of furin can be found in a recent review ( Basak, A., J. Mol. Med., 2005, 83, 844-855 ). These include pure peptide derivatives derived from the prodomain of furin, but also peptidyl chloromethyl ketones ( Hallenberger, S. et al., Nature, 1992, 360, 358-361 ), Peptidyl aldehydes with C-terminal arginal ( Jean, F. et al., Biochem. J., 1993, 292, 891-899 ), Peptidyl-semicarbazones and also peptides with stabilized pseudopeptide bonds ( Angliker, H., J. Med. Chem., 1995, 38, 4014-4018 ) with which one tried to use binding pockets C-terminal of the cleavage site for the inhibition. Also described were diterpene derivatives which inhibit furin in the micromolar range ( Basak, A. et al., Biochem. J., 1999, 338, 107-113 ). However, these derivatives as well as the other mentioned peptide derivatives with activated P1-carbonyl groups are not suitable for further clinical development. Particularly disadvantageous in these known inhibitors is that they all have a very short half-life in the blood and a high intrinsic reactivity.

Of the The invention is therefore based on the object, for therapeutic Applications to provide suitable active ingredients, the furin or analogous proprotein convertases with high activity and specificity inhibit and useful for the treatment of diseases are involved in which these proteases are involved. In particular, should the active ingredients have a low intrinsic reactivity.

main features The invention are in claim 1, as well as in the independent claims 18, 23, 33, 35, 36, 37 and 38. Embodiments are the subject of claims 2 to 17, 19 to 22, 24 to 32 and 34.

The inhibitors of the invention consist of the 5 segments P5, P4, P3, P2 and P1. P1 is a C-terminal Arginine mimetic, P2, P3 and P4 are amino or imino acid residues and P5 is an N-terminal modification of the amino or imino acid residue P4. P5 is advantageously a variable structure, for example an acyl, sulfonyl or alkyl radical attached to the N-terminal function of the rest P4 is coupled. In addition, the use is the inhibitors according to the invention for the preparation of medicaments for therapy and / or prophylaxis of bacterial and viral diseases. The invention Compounds are also suitable for the production of medicaments, for the therapy and / or prophylaxis of cancer / metastasis, Alzheimer's, Atherosclerosis, diabetes or obesity.

Especially It is advantageous if the inventive Compound a substrate or, most preferably, an inhibitor for the proprotein convertases furin, PC1 / 3, PC2, PC4, PC5 / 6, PACE4 and / or PC7 is.

Surprisingly, it has been found that N-terminally modified tetrapeptide mimetics having a C-terminal arginine mimetic according to the general formula P5-P4-P3-P2-P1 (I)

• – P1 ist ausgewählt aus folgenden Strukturen:
  
  wobei • R₁ ein H, OH, O-CH₃, NH₂, O-CO-CH₃ oder -CO-O-(CH₂)_m-CH₃ mit m eine ganze Zahl von 1 bis 5 ist, und • R₂ ein H oder eine Amidinogruppe
  
  ist, und • n gleich 3, 4 oder 5 ist, und • s gleich 2 oder 3 ist, und
• – P2 eine Amino- oder Iminosäure ausgewählt aus folgenden Strukturen
  
  ist, und wobei • p eine ganze Zahl von 0 bis 5 ist, und • R₃ eine Amino- oder eine Guanidinogruppe sein kann, oder eine -O-Guanidinogruppe wie im Canavaninrest, oder ein einfach oder mehrfach substituierter oder unsubstituierter Aryl- oder Heteroarylrest mit 5 bis 10 Ringatomen, wobei der Heteroarylring bis zu 3 Heteroatome enthalten kann ausgewählt aus N, S oder O, wobei der Substituent am Aryl- oder Heteroarylring ausgewählt ist aus -NH₂, -CH₂-NH₂, -Amidino, -Hydroxyamidino, -Guanidino, -CH₂-Guanidino, -Halogen, -Cl, -Br, -I, -CN, -CF₃, Alkyl mit 1–3 C-Atomen oder Alkoxy mit 1–3 C-Atomen, -COOR₅ mit R₅ ein Wasserstoff oder eine Alkyl mit 1 bis 3 C-Atomen, und • R₄ ein H, OH oder -O-(CH₂)_q-R₆ mit q eine ganze Zahl von 2 bis 4 und R₆ eine Amino- oder Guanidinogruppe ist, und
• – P3 ein beliebiger natürlicher oder unnatürlicher α-Amino- oder α-Iminosäure mit L- oder R-Konfiguration ist, und
• – P4 eine α-Amino- oder α-Iminosäure ausgewählt aus folgenden Strukturen
  
  ist, wobei p, R₃ und R₄ definiert sind wie zuvor, und
• – P5 ausgewählt ist aus • einem -H oder -CO-X-R₇ mit X entweder nicht vorhanden oder X gleich O oder NH, wobei R₇ ein unsubstituierter oder gegebenenfalls einfach oder mehrfach substituierter verzweigter oder unverzweigter, gesättigter oder ungesättigter Alkylrest mit 1 bis 24 C-Atomen und 0 bis 3 Doppelbindungen ist, oder wobei R₇ ein unsubstituierter oder gegebenenfalls einfach oder mehrfach substituierter Aryl-, Heteroaryl, Aralkyl-, Heteroaralkyl oder Cycloalkylrest mit 5 bis 20 Kohlenstoffatomen ist, wobei der Heteroarylring bis zu 3 Heteroatome enthalten kann ausgewählt aus N, S oder O, wobei der oder die gegebenenfalls vorhandenen Substituenten an R₇ unabhängig voneinander ausgewählt werden aus -NH₂, -CH₂-NH₂, -Amidino, -Hydroxyamidino, -Guanidino, -CH₂-Guanidino, -Halogen, -Cl, -Br, -I, -CN, -CF₃, Alkyl mit 1–3 C-Atomen oder Alkoxy mit 1–3 C-Atomen, -COOR₅ mit R₅ ein Wasserstoff oder eine Alkylgruppe mit 1 bis 3 C-Atomen, oder wobei R₇ einen Cholesterolrest darstellt, oder • einem Pyroglutamylrest oder einem anderen Amino- oder Iminosäurerest, oder • einem -SO₂-R₇ wobei R₇ definiert ist wie zuvor, oder • einem unverzweigten oder verzweigten Alkyl mit 1 bis 10-C-Atomen, wobei der Alkylrest gegebenenfalls mit einem Halogen, einer NH₂-, Guanidino-, COOH- oder COOR₅-Gruppe modifiziert und R₅ wie oben definiert ist, oder • ein mit einem bifunktionellen Rest acyliertes Sphingosin bzw. mit einem bifunktionellen Rest acyliertes Sphingosyl-phosphorylcholin folgender Strukturen:
  
  ist und wobei z eine ganze Zahl zwischen 2 und 20 ist.

Very effectively inhibit furin when the residues P1, P2, P3, P4 and P5 are modeled on the Schechter and Berger definition ( Schechter and Berger, Biochem. Biophys. Res. Comm., 1967, 27, 157-162 ) have the following definitions:

• - P1 is selected from the following structures:
  
  where R _{1 is} H, OH, O-CH ₃ , NH ₂ , O-CO-CH ₃ or -CO-O- (CH ₂ ) _m -CH ₃ with m being an integer from 1 to 5, and R _{2 is} an H or an amidino group
  
  is, and • n is 3, 4 or 5, and • s is 2 or 3, and
• P2 is an amino or imino acid selected from the following structures
  
  and wherein • p is an integer from 0 to 5, and • R _{3 can be} an amino or a guanidino group, or an -O-guanidino group as in the canavanine residue, or a mono- or polysubstituted or unsubstituted aryl or heteroaryl residue having 5 to 10 ring atoms, wherein the heteroaryl ring may contain up to 3 heteroatoms selected from N, S or O, wherein the substituent on the aryl or heteroaryl ring is selected from -NH ₂ , -CH ₂ -NH ₂ , -Amidino, -Hydroxyamidino , Guanidino, -CH ₂ -guanidino, -halo, -Cl, -Br, -I, -CN, -CF ₃ , alkyl having 1-3 C atoms or alkoxy having 1-3 C atoms, -COOR ₅ with R _{5 is} a hydrogen or an alkyl having 1 to 3 C atoms, and • R _{4 is} H, OH or -O- (CH ₂ ) _q -R ₆ with q is an integer from 2 to 4 and R _{6 is} an amino or guanidino group, and
• P3 is any natural or unnatural α-amino or α-imino acid of L or R configuration, and
• P4 is an α-amino or α-imino acid selected from the following structures
  
  wherein p, R ₃ and R _{4 are} defined as before, and
• - P5 is selected from • a -H or -CO-XR ₇ with X either not present or X is O or NH, where R _{7 is} an unsubstituted or optionally mono- or polysubstituted branched or unbranched, saturated or unsaturated alkyl radical having 1 to 24 C atoms and 0 to 3 double bonds, or wherein R _{7 is} an unsubstituted or optionally mono- or polysubstituted aryl, heteroaryl, aralkyl, heteroaralkyl or cycloalkyl radical having 5 to 20 carbon atoms, wherein the heteroaryl ring may contain up to 3 heteroatoms selected from N, S or O, wherein the optional substituent (s) on R _{7 are} independently selected from -NH ₂ , -CH ₂ -NH ₂ , -amidino, -hydroxyamidino, -guanidino, -CH ₂ -guanidino, -halogen , -Cl, -Br, -I, -CN, -CF ₃ , alkyl having 1-3 C atoms or alkoxy having 1-3 C atoms, -COOR ₅ with R _{5 is} a hydrogen or an alkyl group having 1 to 3 C atoms, or where R _{7 is} a choles or a -SO ₂ -R ₇ wherein R _{7 is} defined as before, or • an unbranched or branched alkyl having 1 to 10 carbon atoms, wherein the Alkyl radical optionally modified with a halogen, an NH ₂ , guanidino, COOH or COOR ₅ group and R _{5 is} as defined above, or • a sphingosine acylated with a bifunctional radical or sphingosyl-phosphorylcholine acylated with a bifunctional radical structures:
  
  and where z is an integer between 2 and 20.

wobei R₁ wie oben definiert ist und wobei das 4-Amidinobenzylamid-Derivat mit einem Wasserstoffatom als Rest R₁ ganz besonders bevorzugt ist. Um eine verbesserte Membranpermeabilität der Inhibitoren zu erreichen, kann deren Hydrophobizität erhöht werden, indem Prodrugs der 4-Amidinobenzylamid-Derivate eingebaut werden ( Ettmayer et al., J. Med. Chem. 2004, 47, 2393–2404 ), wobei der Rest R₁ in diesen Fällen -OH, -O-CH₃, -NH₂, -O-CO-CH₃ oder -CO-O-(CH₂)_m-CH₃ mit m eine ganze Zahl von 1 bis 5 sein kann. Diese Prodrugs sind auch Gegenstand der vorliegenden Erfindung, Sie werden erst im Organismus in die inhibitorisch wirksamen Verbindungen mit freier Amidinogruppe umgewandelt.According to a preferred embodiment, the compounds according to the invention contain the following structures as P1 radicals:

wherein R _{1 is} as defined above and wherein the 4-Amidinobenzylamid derivative having a hydrogen atom as the radical R _{1 is} very particularly preferred. In order to achieve an improved membrane permeability of the inhibitors, their hydrophobicity can be increased by incorporating prodrugs of the 4-amidinobenzylamide derivatives ( Ettmayer et al., J. Med. Chem. 2004, 47, 2393-2404 ), wherein the radical R ₁ in these cases is -OH, -O-CH ₃ , -NH ₂ , -O-CO-CH ₃ or -CO-O- (CH ₂ ) _m -CH ₃ with m an integer of 1 to 5 can be. These prodrugs are also the subject of the present invention, they are first converted in the organism into the inhibiting compounds with free amidino group.

Especially It is advantageous if P1 is a decarboxylated one Arginine mimetic acts. In particular, there are benzamidine structures Cheap. Compounds having such a P1 radical have one particularly low intrinsic reactivity.

The other mentioned P1 building blocks are known arginine mimetics which have been used for the development of inhibitors of trypsin-like serine protease thrombin (see Review article Steinmetzer and Sturzbecher Current Med. Chem. 2004, 11, 2299-2323 ). The synthesis of these building blocks is known to the person skilled in the art.

In Another preferred embodiment is the remainder P2 is a basic amino acid in the L configuration, especially Preferably, the amino acid residues are arginine, lysine and their longer by a methylene group in the side chain or shorter homo- and nor-derivatives. Likewise, canavanine, 3- and 4-amidinophenylalanine, 3- and 4-aminomethylphenylalanine or but also 3- and 4-guanidinophenylalanine preferred P2 radicals, wherein also all mentioned phenylalanine derivatives as homo- and Norderivate are particularly suitable. As P2 radicals are also pyridylalanine and homopyridylalanine, wherein the nitrogen is in each possible Position of the pyridyl ring can stand.

In Another preferred embodiment is the P3 residue any natural or unnatural α-amino or α-imino acid residue in the L-configuration, as very particularly preferred amino acid residues the proteinogenic amino and imino acid residues are suitable.

In Another preferred embodiment is the remainder P4 a basic amino acid in the L configuration, especially suitable are the amino acid residues arginine and lysine as well their longer by a methylene group in the side chain or shorter homo- and nor-derivatives. Likewise, canavanine, 3- and 4-amidinophenylalanine, 3- and 4-aminomethylphenylalanine or but also 3- and 4-guanidinophenylalanine preferred P4 radicals, wherein also all mentioned phenylalanine derivatives as homo- and Norderivate are particularly suitable. P4 radicals are also pyridylalanine and homopyridylalanine, wherein the nitrogen is in each possible Position of the pyridyl ring can stand.

The P5 residue is very variable and may either be only a hydrogen atom in the simplest case, but it is preferably a hydrophobic saturated or optionally mono- or polyunsaturated acyl radical derived from naturally occurring fatty acids, but also preferably aralkyl-CO radical, or Heteroaralkyl-CO radical or a sulfonyl radical, where all of these radicals may optionally be unsubstituted or substituted with up to three radicals. Particularly suitable substituents are: halogens, in particular chlorine atoms, amino groups, carboxyl groups which may also be present as esters, especially as ethyl esters, amidino-, hydroxyamidino-, guanidino-, -CH ₂ -guanidino-, CF ₃ - and alkyl groups with 1- 3 C atoms or alkoxy groups with 1-3 C atoms.

wobei z eine ganze Zahl von 1 bis 18 ist, ganz besonders bevorzugt ist z gleich 8, 14 oder 16 und auch mit einem bifunktionellen Rest acyliertes Shpingosin bzw. mit einem bifunktionellen Rest acyliertes Shpingosyl-phosphorylcholin folgender Strukturen:

und wobei z eine ganze Zahl zwischen 2 und 20 ist.Particularly suitable P5 groups also have the following structures:

where z is an integer from 1 to 18, very particularly preferably z is 8, 14 or 16 and shpingosine acylated with a bifunctional radical or shpingosyl-phosphorylcholine acylated with a bifunctional radical has the following structures:

and where z is an integer between 2 and 20.

object The invention also provides methods for the synthesis of the described Compounds, the synthesis being characterized by combination of solid phase and solution syntheses or by pure solution synthesis is possible.

• – Synthese des Segments P5-P4-P3-P2 durch Festphasenpeptidsynthese, beispielsweise an einem 2-Chlorotritylchlorid-Harz nach Standardverfahren der Fmoc-Chemie,
• – anschließende Abspaltung des Peptidsegments vom Harz unter geeigneten sauren Bedingungen, wobei Seitenkettenschutzgruppen entweder erhalten oder abgespalten werden,
• – Kopplung eines geeigneten P1-Bausteins mittels racemisierungsfreier Standardverfahren, der gegebenenfalls geschützt oder ungeschützt vorliegen kann, und gegebenenfalls
• – Abspaltung aller eventuell noch vorhandenen Schutzgruppen, dazu gehört beispielsweise die Behandlung mit entsprechenden Säuren, beispielsweise mit Trifluoressigsäure, HBr in Eisessig, HCl in Eisessig, HF, Trifluormethansulfonsäure, wobei die Säuren gegebenenfalls nach Scavanger enthalten können, beispielsweise Wasser, Tri-isopropylsilan, Anisol, Dimethylsulfid oder andere gängige Scavanger mit freien Mercaptogruppen, gegebenenfalls werden noch vorhandene Schutzgruppen durch katalytische Hydrierung mit Wasserstoff und Palladium auf Aktivkohle als Katalysator entfernt, und
• – finale Reinigung durch Kristallisation, Chromatographie, bevorzugt Umkehrphasen- oder Ionenaustauschchromatographie oder durch Gegenstromverteilung, und
• – gegebenenfalls Überführung in eine andere bzw. geeignetere Salzform.

In the synthesis of a compound according to the general formula (I) by means of a combination of solid-phase and solution synthesis, the following steps are carried out:

• Synthesis of the segment P5-P4-P3-P2 by solid phase peptide synthesis, for example on a 2-chlorotrityl chloride resin according to standard methods of Fmoc chemistry,
• Subsequent cleavage of the peptide segment from the resin under suitable acidic conditions, whereby side chain protecting groups are either obtained or cleaved off,
• - Coupling of a suitable P1 block by racemization-free standard method, which may optionally be protected or unprotected, and optionally
• - Removal of any protective groups still present, this includes, for example, the treatment with appropriate acids, such as trifluoroacetic acid, HBr in glacial acetic acid, HCl in glacial acetic acid, HF, trifluoromethanesulfonic acid, the acids may optionally contain after Scavanger, for example water, tri-isopropylsilane, anisole , Dimethylsulfid or other common Scavanger with free mercapto groups, if necessary remaining protecting groups are removed by catalytic hydrogenation with hydrogen and palladium on activated carbon as catalyst, and
• Final purification by crystallization, chromatography, preferably reverse phase or ion exchange chromatography or by countercurrent distribution, and
• - if necessary, conversion into another or more suitable salt form.

Alternatively, for example, for synthesizing compounds having a C-terminal agmatine residue in the first step, a trityl chloride resin is charged with 1,4-diaminobutane by standard methods and successively the remaining building blocks P2, P3, P4 are coupled by Fmoc chemistry and P5 by standard methods Cleavage of the peptide from the resin is carried out with suitable acids or acid-containing cleavage cocktails, either removing all protecting groups or retaining certain protecting groups, such as in the side chain of lysine, followed by final assembly of the guanidino group by reaction of amino groups with pyrazole-carboxamidine.HCl and DIPEA either in DMF or optionally also in organic aqueous solvents containing sodium carbonate or in pure aqueous sodium carbonate solution ( Bernatowicz, MS et al., J. Org. Chem., 1992, 57, 2497-2502 ). Analog can also arbitrary other diamines can be used for the loading of the trityl chloride resin. If, in the end, the reaction with pyrazolecarboxamidine is omitted, the compounds having a C-terminal amino group are obtained.

• a) Synthese eines an der terminalen Amidino- oder Guanidinogruppegeeignet geschützten P1-Bausteins mit freier Aminogruppe, die entweder als Salz oder freie Base vorliegen kann, z. B. von folgender Verbindung:
  
  (beschrieben in Lila et al., Synth. Commun., 1998, 28, 4419–4429 )
• b) Kopplung einer Nα-Boc oder anderweitig geeignet Nα-urethangeschützten Amino- oder Iminosäure mit geeigneter Seitenkettenschutzgruppe, falls notwendig, an die freie Aminogruppe des P1-Baustein, wobei prinzipiell alle geeigneten Kopplungsverfahren der Peptidchemie mit bekannten Hilfsbasen verwendet werden können, dazu gehören das Mischanhydridverfahren, Carbodiimidverfahren, Aktivesterverfahren, Azidverfahren, Propanphosphonsäureanhydrid-Ver-fahren oder Kopplung mit Triazimoch, TBTU, HBTU, BOP, PyBOP, HATU oder anderer Kopplungsreagentien basierend auf Uronium- oder Phosphoniumsalzen, gefolgt von einer Aufarbeitung des Reaktionsansatzes durch gängige Wasch- und Kristallisationsverfahren sowie gegebenenfalls gefolgt von einem Chromatographieschritt zur Reinigung, und gefolgt von standardmäßiger Abspaltung der Nα-Schutzgruppe unter Erhalt des P2-P1-Segments mit freier Aminogruppe am P2-Baustein, so wird beispielsweise für die Abspaltung der Nα-Boc-Schutzgruppe in der Regel Trifluoressigsäure oder HCl in einem geeigneten organischen Lösungsmittel wie Essigester, Essigsäure, Diethylether oder Dioxan verwendet,
• c) Kopplung einer Nα-Boc oder anderweitig geeignet Nα-urethangeschützten P3-Amino- oder Iminosäure an die freie Aminogruppe des P2-P1-Segments aus Schritt b) analog der im Schritt b) beschriebenen Methoden, gefolgt von der im Schritt b) beschriebenen Aufarbeitung des Reaktionsansatzes und der im Schritt b) beschriebenen Abspaltung der Nα-Schutzgruppe unter Erhalt des P3-P2-P1-Segments mit freier N-terminaler Aminogruppe am P3-Rest,
• d) Kopplung einer Nα-Boc oder anderweitig geeignet Nα-urethangeschützten P4-Amino- oder Iminosäure an die freie Aminogruppe des P3-P2-P1-Segments aus Schritt c) analog der im Schritt b) beschriebenen Methoden, gefolgt von der im Schritt b) beschriebenen Aufarbeitung des Reaktionsansatzes und der im Schritt b) beschriebenen Abspaltung der Nα-Schutzgruppe unter Erhalt des P4-P3-P2-P1-Segments mit freier N-terminaler Aminogruppe am P4-Rest,
• e) Kopplung des P5-Restes an die freie Aminogruppe des P4-P3-P2-P1-Segments aus Schritt d) gegebenenfalls analog der im Schritt b) beschriebenen Methoden, oder Kopplung eines bereits als Säurechlorid oder Aktivester, speziell als Succinimidester, vorliegenden P5-Bausteins in Gegenwart von Hilfsbasen ohne weiteres Kopplungsreagenz.
• f) Abspaltung aller noch vorhandenen Schutzgruppen durch gängige Verfahren, dazu gehört beispielsweise die katalytische Hydrierung mit Wasserstoff und Palladium auf Aktivkohle als Katalysator, oder die Behandlung mit entsprechenden Säuren, beispielsweise mit Trifluoressigsäure, HBr in Eisessig, HCl in Eisessig, HF, Trifluormethansulfonsäure, wobei die Säuren gegebenenfalls nach Scavanger enthalten können, beispielsweise Wasser, Triisopropylsilan, Anisol, Dimethylsulfid oder andere gängige Scavanger mit freien Mercaptogruppen, und
• g) finale Reinigung durch Kristallisation, Chromatographie, bevorzugt Umkehrphasen- oder Ionenaustauschchromatographie oder durch Gegenstromverteilung, und
• h) gegebenenfalls Überführung in eine andere bzw. geeignetere Salzform.

For larger scale synthesis ("upscaling"), methods are particularly suitable in which only pure solution synthesis steps are used. Typically, solution peptide synthesis begins with a suitable C-terminal P1 moiety to which the following Nα-urethane and, if necessary, side-chain protected P2 moiety is coupled, followed by cleavage of the Nα-urethane protecting group and coupling of the following Nα-urethane protected moiety. If necessary, all intermediates occurring in the synthesis can be purified. The Nα-urethane protective group is particularly suitable for the Boc protective group. The synthesis is completed in an analogous way, in the last steps the P5 building block is coupled and then all protecting groups are removed. By means of this pure solution synthesis strategy racemizations can be minimized in the coupling steps, which are often a problem with segment couplings, as can be seen above in the combined solid-phase and solution synthesis. A solution synthesis includes, for example, the following steps:

• a) Synthesis of a terminal amidino or guanidino group suitably protected P1 free amino moiety which may be either salt or free base, e.g. From the following compound:
  
  (described in Lila et al., Synth. Commun., 1998, 28, 4419-4429 )
• b) coupling of an Nα-Boc or otherwise suitably Nα-urethane-protected amino or imino acid with a suitable side chain protecting group, if necessary, to the free amino group of the P1 building block, whereby in principle all suitable coupling methods of peptide chemistry with known auxiliary bases can be used Mixed anhydride method, carbodiimide method, active ester method, azide method, Propanphosphonsäureanhydrid Ver method or coupling with triazimoch, TBTU, HBTU, BOP, PyBOP, HATU or other coupling reagents based on uronium or phosphonium salts, followed by a workup of the reaction mixture by conventional washing and crystallization processes and optionally followed by a purification chromatographic step followed by standard Nα-deprotection to give the P2-P1 free-amino segment on the P2 building block, such as for the Nα-Boc deprotection, for example Trifluoroacetic acid or HCl in a suitable organic solvent such as ethyl acetate, acetic acid, diethyl ether or dioxane,
• c) coupling of an Nα-Boc or otherwise suitably Nα-urethane-protected P3-amino or imino acid to the free amino group of the P2-P1 segment from step b) analogously to the methods described in step b), followed by that described in step b) Working up of the reaction mixture and the cleavage of the Nα protective group described in step b) to obtain the P3-P2-P1 segment with free N-terminal amino group on the P3 radical,
• d) coupling a Nα-Boc or otherwise suitably Nα-urethane-protected P4-amino or imino acid to the free amino group of the P3-P2-P1 segment from step c) analogously to the methods described in step b), followed by the in step b ) and the deprotection of the Nα protective group described in step b) to obtain the P4-P3-P2-P1 segment having a free N-terminal amino group on the P4 radical,
• e) Coupling of the P5 radical to the free amino group of the P4-P3-P2-P1 segment from step d) optionally analogously to the methods described in step b), or coupling of a P5 already present as acid chloride or active ester, especially as succinimide ester Building blocks in the presence of auxiliary bases without further coupling reagent.
• f) cleavage of all remaining protective groups by conventional methods, including, for example, the catalytic hydrogenation with hydrogen and palladium on activated carbon as a catalyst, or treatment with appropriate acids, such as trifluoroacetic acid, HBr in glacial acetic acid, HCl in glacial acetic acid, HF, trifluoromethanesulfonic acid, wherein the acids may optionally contain Scavanger, for example water, triisopropylsilane, anisole, dimethyl sulfide or other common scavangers with free mercapto groups, and
• g) final purification by crystallization, chromatography, preferably reverse phase or ion exchange chromatography or by countercurrent distribution, and
• h) optionally conversion to another or more suitable salt form.

object The invention also relates to methods in which segment couplings in Solution be carried out for such segmental couplings particularly racemization-free method be used, for example, the azide coupling, in which a suitable N-terminal and side chain protected peptide methyl ester converted into the hydrazide by reaction, from which is then prepared by standard method the azide that for coupling to a block or a segment with free amino group is used. On the other hand other coupling methods are used, with particularly weak auxiliary bases, for example, collidine, or suitable additives, For example, 6-Cl-HOBt, HOAt can be used. For the The above methods can all known peptide chemical Synthetic methods, including all amino acid derivatives with suitable protecting groups, suitable coupling reagents, suitable Release reagents used in appropriate solvents be detailed in standard works of peptide chemistry and organic chemistry are described in detail (Houben-Weyl Vol. E22, Synthesis of Peptides).

object The invention also pharmaceutically acceptable or compatible Salts. These salts must be a pharmaceutically acceptable Anion or cation. Suitable pharmaceutically acceptable Acid addition salts of the invention Connections are z. As salts of inorganic acids, such as hydrochloric acid, Hydrobromic, phosphoric, methaphosphoric, nitric, sulfonic and sulfuric acid or organic acids, such as z. Acetic acid, benzenesulfonic, benzoic, citric, ethanesulfonic, Fumaric, gluconic, glycolic, isethionic, lactic, lactobionic, maleic, Methanesulfonic, succinic, p -tuluenesulfonic, tartaric and trifluoroacetic acid. For medical purposes is particularly preferred the chlorine or acetate salt used. Suitable pharmaceutically acceptable basic salts are z. Ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth salts (such as magnesium and calcium salts).

salts with a non-pharmaceutically acceptable anion also within the scope of the invention as useful intermediates more pharmaceutically acceptable for the preparation or purification Salts and / or for use in non-therapeutic, for example in vitro applications.

The term "physiologically functional derivative" used below denotes any physiologically acceptable derivative of a compound of the formula I, z. An ester which, when administered to a mammal, such as. Human, is able to form (directly or indirectly) a compound of formula I or an active metabolite thereof. The physiologically functional derivatives also include prodrugs of the compound according to the invention. Such prodrugs can be metabolized in vivo to a compound of the invention. These prodrugs may or may not be effective. Examples of suitable prodrugs for benzamidine groups are compounds in which the amidine is present as hydroxyamidine, methoxyamidine, acetylhydroxyamidine, alkyloxycarbonylamidine, especially as ethyloxycarbonylamidine or hexyloxycarbonylamidine ( Ettmayer. et al., J. Med. Chem. 2004, 47, 2393-2404 ).

The Compounds according to the invention can also in different stereoisomeric forms but also in polymorphic ones Shapes are present, for. B. as amorphous and crystalline polymorphic forms.

following all references refer to compounds according to formula (I) as described above, as well as their salts, solvates and physiologically functional derivatives.

The The present invention also relates to the use of the compounds of the formula (I) as a medicament for the treatment and prophylaxis of bacterial and viral diseases, as well as diabetes, arteriosclerosis, cancer / metastasis, but also from Alzheimer's or overweight training, in which are furin or analogous proprotein convertases that are behind basic P1 residues split, are involved. Also a pharmaceutical composition containing a compound of formula (I) is the subject of this Invention. The amount of compound according to formula (I), which is required to produce the desired biological Effect is dependent on a number of factors z. As the selected specific compound, the intended Use, route of administration and clinical condition of the patient. For the prophylaxis or therapy of the above conditions can the compounds according to formula (I) themselves are used as a compound, preferably However, with a carrier or excipient in the form of a pharmaceutical composition. The carrier or excipient must of course be compatible, in the sense that he is compatible with the other components of the composition and not harmful to the patient is.

The carrier may be a solid or a liquid, or both, and is preferably formulated with the compound as a single dose, for example, as a tablet, which may contain from 0.05% to 95% by weight of the active ingredient. Other pharmaceutically active substances may also be present, including further compounds of the formula (I). The pharmaceutical compositions according to the invention can be prepared by one of the known pharmaceutical methods, which consist essentially in that the ingredients are mixed with pharmacologically acceptable carriers and / or excipients.

invention Pharmaceutical compositions are especially those which for oral, rectal, topical, peroral (eg sublingual) and parental (e.g., subcutaneous, intramuscular, intradermal, or intravenous) administration, although the most suitable mode of administration in each individual case of the kind and severity of the condition to be treated and the nature of each used compound according to formula (I) is. Aerosols are also suitable pharmaceutical compositions for the compounds according to the invention, which also in the form of aerosol sprays especially for the treatment and prophylaxis of diseases of the respiratory tract suitable are. Also coated formulations and coated slow release formulations belong to the scope of the invention. Preference is given to acid and enteric formulations. Suitable enteric-coated Coatings include cellulose acetate phthalate, polyvinyl acetate phthalate, Hydroxypropylmethylcellulose phthalate and anionic polymers of methacrylic acid and methacrylic acid methyl ester.

suitable Pharmaceutical compounds for oral administration can be in separate units, such as Capsules, cachets, lozenges or tablets, each a certain amount of the compound according to formula (I) included; as a powder or granules; as a solution or Suspension in an aqueous or non-aqueous Liquid; or as an oil-in-water or Water-in-oil emulsion. These compositions can, as already mentioned, after any suitable pharmaceutical Method to be prepared, which includes a step in which the Active substance and the carrier (consisting of one or more additional Constituents can be) brought into contact. In general the compositions are uniform and homogeneously mixing the active ingredient with a liquid and / or finely divided solid support, after which the product is molded if necessary.

So For example, a tablet can be prepared by a Powder or granules of the compound is pressed or shaped, optionally with one or more additional ingredients. Pressed tablets can be made by tableting the compound in free-flowing form, such as a powder or granules, optionally mixed with a binder, lubricants, inert diluent and / or one (more) surface-active / dispersing agent Means be made in a suitable machine. molded Tablets can be made by molding the powdery, moistened with an inert liquid diluent Connection can be made in a suitable machine.

pharmaceutical Compositions suitable for peroral (sublingual) Administration include lozenges containing a A compound according to formula (I) with a flavor usually, sucrose or gum arabic or tragacanth, and lozenges that allow the administration of an inert Base like gelatin and glycerine or sucrose and gum arabic include.

suitable pharmaceutical compositions for parenteral administration preferably comprise sterile aqueous preparations of a Compound according to formula (I), preferably isotonic with the blood of the intended recipient. These preparations are preferably administered intravenously, although administration is also subcutaneous, intramuscular or intradermally as injection.

These Preparations may preferably be made by: the compound is mixed with water and the resulting solution sterile and isotonic with the blood. Injectable invention Compositions generally contain from 0.1 to 5% by weight the active compound.

In terms of The further formulation is based on common manuals directed.

The The invention also relates to methods for the production of pharmaceutical Compositions in which one or more compounds of the general formula (I) with suitable carriers and excipients (see above) mixed.

The compound according to the invention or a medicament according to the invention can be used with particular advantage for the therapy or prophylaxis of a virally or bacterially-caused disease as well as for the therapy and prophylaxis of diabetes, arteriosclerosis, cancer, Alzheimer's disease or obesity in oral, subcutaneous, intravenous or transdermal form or in the form of a spray for the treatment of respiratory diseases.

Advantageous may be the compound of the invention or a Medicament according to the invention in this case for therapy or prophylaxis of diseases caused by bacterial Pathogen, in particular caused by pathogens from the group Pseudomonas spec., Shigella spec., Corynebacterium diphtheriae, Bacillus cereus, Bacillus anthracis, Bacillus butulinum, Clostridium perfringens, Clostridium difficile, Clostridium septicum, Clostridium sprioforme, Bordetella pertussis, Samonella enterica or by selected Escherichia coli strains, especially E. coli strains which hemolytic colitis, the hemolytic uraemic Syndrome and / or kidney failure, especially E. coli 0157-H7.

• • hochpathogene aviäre Influenza A-Viren, vor allem der H5- bzw. H7-Subtypen,
• • RNA-Negativstrangviren wie den Parainfluenzaviren, das Masern-Virus, das Mumpsvirus, das Hundestaupevirus, das Respiratorische-Syncytial-Virus, Filoviren wie das Ebola- und Marburg-Virus und das Newcastle-Disease-Virus, sowie
• • Plusstrang-RNA-Viren, wie HIV-1, HIV-2, HTLV und andere Retroviren, insbesondere Coronaviren wie das SARS-Coronavirus, die Flaviviren, Gelbfieber-Viren, West-Nile-Virus., Dengue-Virus. oder das Frühsommer-Meningo-Encephalitis-Virus, das Venezuelan equine encephalitis Virus oder das Rift Valley Fever Virus verwendet werden.

In the therapy or prophylaxis of diseases caused by viruses, the compound or the drug, in particular in diseases caused by viruses of the group

• Highly pathogenic avian influenza A viruses, especially the H5 or H7 subtypes,
• RNA negative strand viruses such as parainfluenza viruses, measles virus, mumps virus, canine distemper virus, respiratory syncytial virus, filoviruses such as Ebola and Marburg virus and Newcastle disease virus, as well as
• • Positive-strand RNA viruses such as HIV-1, HIV-2, HTLV and other retroviruses, especially coronaviruses such as SARS coronavirus, flaviviruses, yellow fever viruses, West Nile virus, dengue virus. or the tick-borne encephalitis virus, the Venezuelan equine encephalitis virus, or the Rift Valley Fever virus.

you recognizes that the compound of the invention or the pharmaceutical according to the invention especially advantageous for inhibiting the proprotein convertases furin, PC1 / 3, PC2, PC4, PC5 / 6, PACE4 and / or PC7.

Further Features, details and advantages of the invention will become apparent the wording of the claims and the description below of embodiments with reference to the drawings. Show it:

1 the inhibition of HA cleavage of avian influenza virus by varying concentrations of inhibitor 1 and the reference inhibitor Dec-Arg-Val-Lys-Arg-CMK

2 the multicyclic propagation of FPV in cell culture in the presence of inhibitor 1 and without inhibitor.

A detailed description of the figures can be found in the description Example 7 and 8.

embodiments

Methods for analysis of the compounds

Analytical HPLC

to Analytical reversed-phase HPLC was a LC-10A HPLC system Company Shimadzu, consisting of subsystems CTO-10A column oven, LC-10ATvp pumps (2x), DGU-14A degasser, SIL-10Axl autoinjector, SCL-10Avp system controller, SPD-M10Avp photodiode array detector and a column 250 / 4,6 Nucleodur 100-5 C18 ec of the company Macherey-Nagel (Düren, Germany), using the associated software Shimadzu CLASS-VP, Version 7.2.1, used. The detection was carried out at 220 nm. Serving as eluent Water with 0.1% TFA (A) and acetonitrile with 0.1% TFA (B) at one Flow rate of 1 mL / min and a linear gradient (increase of 1% B / min). The respective starting conditions (% B) are given in the syntheses.

Preparative HPLC

Preparative RP-HPLC was carried out on a Varian HPLC system consisting of Varian PrepStar Model 218 preparative pumps (2x), Varian ProStar Model 320 UV-Vis detector, Varian fraction collector Model 701, and a VP 250/32 column Nucleodur 100-5 C8 ec from Macherey-Nagel (Düren, Germany), using the associated Star software V. 6.0. The detection succeed The eluent was also water with 0.1% TFA (A) and acetonitrile with 0.1% TFA (B) at a flow rate of 20 mL / min and a suitable gradient.

mass spectroscopy

The Spectra were taken with a device from Applied Biosystems (QTrap 2000) or a company Autospec spectrometer Micromass added.

TLC

For thin layer chromatography, silica gel precast Adamant UV ₂₅₄ from Macherey-Nagel was used. The eluent used was a mixture of n-butanol, glacial acetic acid and water (4: 1: 1). The compounds were detected by UV absorption at 254 nm. In addition, a ninhydrin solution (300 mg of ninhydrin dissolved in 100 ml of n-butanol and 3 ml of glacial acetic acid) were used as spray reagents and, after incubation of the TLC plate in a chlorine atmosphere, an o-tolidine solution (150 mg o-tolidine and 2.1 g KI dissolved in 2 mL glacial acetic acid and 148 mL water).

NMR

The ¹ H and ¹³ C NMR spectra were recorded with an ECX-400 NMR spectrometer from Jeol Inc. USA ( ¹ H NMR: 400 MHz, ¹³ C NMR: 100 MHz). The samples were dissolved in commercially available deuterated solvents of commercial grade. The chemical shifts were reported as δ values in parts per million (ppm), and the solvent signals were used as a reference.

used abbreviations

• Ac

  acetyl

  Amb

  Amidobenzyl

  Boc

  tert-butyloxycarbonyl

  Bzl

  benzyl

  Cbz

  benzyloxycarbonyl

  DIPEA

  diisopropylethylamine

  DCM

  dichloromethane

  DMF

  N, N-dimethylformamide

  Fmoc

  fluorenylmethyloxycarbonyl

  hArg

  homoarginine

  HBTU

  O-benzotriazole-N, N, N ', N'-tetramethyl-uronium-hexafluoro-phosphate

  HPLC

  High performance liquid chromatography

  i. V.

  in a vacuum

  LM

  solvent

  MS

  mass spectroscopy

  NMM

  N-methylmorpholine

  NMP

  N-methylpyrrolidone

  NMR

  Nuclear Magnetic Resonance Spectroscopy

  phac

  phenylacetyl

  PyBOP

  Benzotriazol-1-yl-N-oxy-tris (pyrrolidino) phosphonium hexafluorophosphate

  RT

  room temperature

  Bu

  tert-butyl

  TFA

  trifluoroacetic

  Tfa

  trifluoroacetyl

  THF

  tetrahydrofuran

bzw. als ausführliche Struktur

(im Folgenden werden die Standardaminosäuren immer im Drei-Buchstabencode angegeben). Schritt a) 4-Amidinobenzylamin × 2HCl

All of the Fmoc amino acids, resins, coupling reagents and other reagents used in the synthesis were purchased from Orpegen, Iris Biotech, Novabiochem, Bachem, Aldrich, Fluka or Acros. Example 1 Phac-Arg-Val-Arg-4-amidinobenzylamide × 3 TFA = Inhibitor 1

or as a detailed structure

(In the following, the standard amino acids are always given in three-letter code). Step a) 4-amidinobenzylamine × 2HCl

10 g (32.5 mmol) of Boc-4-acetylhydroxyamidinobenzylamide (prepared analogously Schweinitz, A. et al., Med. Chem., 2006, 2, 349-361 ) were dissolved in 300 ml of 90% acetic acid, treated with 650 mg of catalyst (10% Pd / C) and hydrogenated with hydrogen for 6 h at about 35 ° C and 36 h at RT. The catalyst was filtered off and the LM i. V. removed. The remaining residue was dissolved in 150 ml of water, admixed with 40 ml of concentrated HCl and stirred for 1.5 h. The solution was i. Concentrated, the residue was redissolved in water and the LM i. V. concentrated. Redissolution in water and lyophilization yields the product (off-white solid, 6.8 g (30.6 mmol)).

• ¹ H-NMR (400 MHz, D ₂ O): δ [ppm] = 4.46 (s, 2H, H-1), 7.80-8.01 (2d, 4H, arom.). ¹³ C-NMR (100 MHz, D ₂ O): δ [ppm] = 42.64 (2H, H-1), 128.65 (2H, H-3), 129.26 (1H, H-5) , 129.61 (2H, H-4), 138.69 (1H, H-2), 166.47 (1H, H-6).

Step b) Phac-Arg (Pbf) -Val-Arg (Pbf) -OH

The loading of the 2-chloro-trityl chloride resin (1.5 g) with Fmoc-Arg (Pbf) -OH was carried out in a glass vessel with frit bottom analogous to a standard procedure of the manufacturer with equimolar amounts of Fmoc-Arg (Pbf) -OH (1 , 51 g, 2.325 mmol) and 4 eq. DIPEA (9.3 mmol, 1.55 mL) based on the indicated resin loading (1.55 mmol / g) in dry DCM over a period of 2 h. Thereafter, the reaction solution was sucked off, the resin washed 3 × with DCM / methanol / DIPEA (85/10/5 (v / v)), 3 × with DCM, 2 × with DMF and 2 × with DCM and dried. Subsequently, the remaining building blocks were manually coupled using a standard Fmoc synthesis protocol using phenylacetic acid instead of an Fmoc amino acid in the last step. After completion of the synthesis, the resin was washed several times with DMF and DCM. The cleavage from the resin was carried out with 1% TFA in DCM (v / v) in 30 min with shaking. The cleavage solution was squeezed out and i. V., then the cleavage of the resin was under identical conditions repeated 2 ×. The product was lyophilized with 80% t-butanol and used directly for the next step in the synthesis (white powder, HPLC: start at 30% B, elution at 35.57 min, MS over 1051.5, MS Ft 1074.3 (m.p. M + Na) ⁺ ).

Step c) Phac-Arg-Val-Arg-4-amidinobenzylamide × 3 TFA = inhibitor 1

105 mg (0.1 mmol) of Phac-Arg (Pbf) -Val-Arg (Pbf) -OH as crude product, 50.9 mg (0.3 mmol) of 6-Cl-HOBt, 23 mg (0.1 mmol) 4-Amidinobenzylamide × 2 HCl and 57.3 mg (0.11 mmol) of PyBOP were dissolved in 500 μL of DMF and treated with 51.4 μL (0.3 mmol) of DIPEA. The batch was stirred for 2 h at RT. The LM was i. V. and the remaining oily residue with 2 ml of a mixture of 95% TFA, 2.5% triisopropylsilane and 2.5% water (v / v) added. After shaking for 3 h at RT, the product was precipitated by addition of diethyl ether, centrifuged off and the precipitate was washed twice with diethyl ether. The purification was carried out by preparative HPLC, the product-containing fractions were combined and lyophilized (white powder 60 mg, (HPLC: start at 1% B, elution at 24.17 min, MS calc. 678.4, MS gef. 679.4 (cf. M + H) ⁺ ) Example 2 Phac-Arg-Val-Arg-Agmatine × 3 TFA = Inhibitor 2

Step a) Phac-Arg-Val-Arg-NH- (CH ₂ ) ₄ -NH ₂ × 3 TFA = Inhibitor 2a

100 mg of trityl chloride resin was 4-fold excess 1,4-diaminobutane (0.6 mmol) based on the indicated resin loading (1.5 mmol / g) of the manufacturer in the reaction vessels of the peptide synthesizer (Syro 2000, Multisyntech GmbH, Witten, Germany) with dry THF as LM over a period of time loaded for 2 hours. Thereafter, the LM was sucked off and the resin with 3 × DCM / methanol / DIPEA (85/10/5 (v / v)), 3x with DCM, 2x with DMF and washed 2x with DCM.

The The following synthetic steps were carried out by means of a multiple peptide synthesizer according to a standard Fmoc protocol in DMF / NMP with double coupling and 4-fold excesses of Fmoc-amino acid, HOBt and HBTU in the presence of 8 equivalents of DIPEA. For the Fmoc cleavage was in each case a mixture of piperidine / DMF / NMP (1: 1: 1). In the last synthesis cycle, phenylacetic acid became coupled like a normal Fmoc amino acid. After completion In the synthesis, the resin was washed several times with DMF and DCM.

The reaction vessels were removed from the machine and 2 mL of a mixture of 95% TFA, 2.5% triisopropylsilane and 2.5% water (v / v) was added to the resin. After shaking for 2 h at RT, the cleavage solution was squeezed out and the product was precipitated by addition of diethyl ether and centrifuged off. The resulting precipitate was washed twice with diethyl ether and then purified by preparative HPLC, the product-containing fractions combined and lyophilized (white powder, HPLC: start at 1% B, elution at 22.3 min, MS calc. 617.79, MS Found 309.68 (M + 2H) ²⁺ / 2).

Step b) Phac-Arg-Val-Arg-Agmatine × 3 TFA = inhibitor 2

Schritt a) N-Boc-(4-Aminomethyl)-Piperidid

48 mg (about 0.05 mmol) of Phac-Arg-Val-Arg-NH- (CH ₂ ) ₄ -NH ₂ × 3 TFA were dissolved in a mixture of 1 mL DMF and 1 mL 1 M Na ₂ CO ₃ - Solution with 0.15 mmol Pyrazolcarboxamidin × HCl and 0.2 mmol DIPEA added. The batch was stirred at RT for 36 h, the LM i. V. and the product was purified by preparative HPLC and lyophilized (white powder, HPLC: start at 1% B, elution at 23.3 min, MS calc. 659.8, MS F, 660.4 (M + H) ⁺ ). Example 3 Phac-Arg-Val-Arg-4-amidomethyl-N-amidinopiperidide × 3 TFA = inhibitor 3

Step a) N-Boc- (4-aminomethyl) piperidide

210.4 mg (1 mmol) of 4- (Tfa-amidomethyl) -piperidine ( Steinmetzer et al., J. Enz. Inhib., 1999, 14, 203-216 ) were dissolved with 174 μL (1 mmol) DIPEA in 2.5 mL DMF. Under ice-cooling, 240 mg (1.1 mmol) Boc ₂ O were added, the mixture was stirred for 1 h under ice-cooling and overnight at RT, then the LM i. V., the residue taken up in EA, 3 × with 5% KHSO ₄ solution. and 3 times with sat. NaCl solution and dried with Na ₂ SO ₄ . The LM was again i. V. removed (oily residue, which crystallizes slowly at 4 ° C). The intermediate was dissolved with 3 mL dioxane and 3 mL 1 M NaOH and stirred at 40 ° C for 3 h. Subsequently, 10 mL of water were added and the product was extracted 3 × with DCM. The combined DCM phases were dried with Na ₂ SO ₄ and then the LM i. V. (white solid, HPLC: start at 1% B, elution at 26.6 min).

Step b) Phac-Arg-Val-Arg-4- (amidomethyl) -piperidine × 3 TFA = inhibitor 3a

The coupling of N-Boc- (4-aminomethyl) piperidide to Phac-Arg (Pbf) -Val-Arg (Pbf) -OH (obtained from step b of Synthesis Example 1) was carried out analogously to step c from Synthesis Example 1. Nach Cleavage of the Pbf and Boc protecting groups remaining on the peptide with a mixture of 95% TFA, 2.5% triisopropylsilane and 2.5% water (v / v) over a period of 2 h at RT gave the intermediate product contained in the cleavage solution precipitated by addition of diethyl ether and centrifuged off, the precipitate washed twice with diethyl ether and i. V. dried. The product was purified by preparative HPLC, the product-containing fractions were combined and lyophilized (white powder, HPLC: start at 1% B, elution at 22.7 min, MS calc. 643.43, MS mL. 644.5 (M + H) ⁺ .

Step c) Phac-Arg-Val-Arg-4- (amidomethyl) -N-amidinopiperide × 3 TFA = inhibitor 3

The construction of the guanidino group was carried out by reacting the Phac-Arg-Val-Arg-4- (amidomethyl) -piperidine × 3 TFA with pyrazolecarboxamidine × HCl and DIPEA analogously to the procedure described in step b of Example 2. The product was purified by preparative HPLC and lyophilized (white powder, HPLC: start at 1% B, elution at 24.0 min, MS above 685.5, MS F, 686.6 (M + H) ⁺ .

Example 4

Analogous of the synthetic strategy for inhibitors 2 and 2a made further connections. For the first synthesis step, the loading of the trityl chloride resin (about 100 mg each with an initial charge of 1.5 mmol / g) in each case the corresponding diamine derivative was used, for example, 1,3-diaminopropane for the inhibitors 4 and 4a, or p-diaminoxylol for the compounds 6 and 6a. The structures of the inhibitors are summarized in Table 1, all compounds were prepared by preparative HPLC as TFA salts receive.

Table: 1 Inhibitors of the general structure Phac-Arg-Val-Arg-P1.

Example 5

The Inhibitors 8-11 were prepared according to the following synthetic strategy prepared as described above lysine Drivat Fmoc-Lys (Cbz) -OH was used for solid-phase peptide synthesis.

Step a) Synthesis of Phac-Arg (Pbf) -Val-Lys (Cbz) -NH- (CH ₂ ) ₄ -NH-trityl resin

The Synthesis was carried out analogously to the step a) in Synthesis Example 2 below Use of trityl chloride resin and Fmoc-Lys (Cbz) -OH as building block, however, without cleavage of the protecting groups and the peptide of Resin.

Step b) Phac-Arg-Val-Lys (Cbz) -NH- (CH ₂ ) ₄ -NH ₂ as 2x TFA salt = inhibitor 8

The cleavage of the Pbf protecting group and the peptide from the Phac-Arg (Pbf) -Val-Lys (Cbz) -NH- (CH ₂ ) ₄ -NH-trityl resin was carried out with a mixture of 95% TFA, 2.5%. Triisopropylsilane and 2.5% water (v / v) for 1.5 h at RT to give Phac-Arg-Val-Lys (Cbz) -NH- (CH ₂ ) ₄ -NH ₂ as 2x TFA -Salt. The product was precipitated with diethyl ether, washed twice with diethyl ether and purified by preparative HPLC (inhibitor 8, white lyophilised powder by preparative HPLC, HPLC: start at 1% B, elution at 36.2 min, MS calc. 723.4, MS found 724.6 (M + H) ⁺ ).

Step c) Phac-Arg-Val-Lys (Cbz) -agmatine × 2 TFA = inhibitor 9

Phac-Arg-Val-Lys (Cbz) -NH- (CH ₂ ) ₄ -NH ₂ × 2 TFA (inhibitor 8) was reacted with 3-fold excess of pyrazolecarboxamidine × HCl and 4-fold excess of DIPEA in DMF. The product was purified by preparative HPLC (inhibitor 9, white lyophilized powder after preparative HPLC, HPLC: start at 1% B, elution at 36.8 min, MS calc. 765.5, MS F 766.5 (M + H ) ⁺ .

Step d) Phac-Arg-Val-Lys-Agmatine × 3 TFA = inhibitor 10

Phac-Arg-Val-Lys (Cbz) -agmatine × 2 TFA (inhibitor 9) was hydrogenated with H ₂ and 10% by mass Pd / C as catalyst in 90% acetic acid for 12 h at RT and the product was purified by preparative HPLC (inhibitor 10 , white lyophilized powder by preparative HPLC, HPLC: start at 1% B, elution at 23.1 min, MS calc. 631.4, MS F, 316.7 (M + 2H) ²⁺ / 2).

Step e) Phac-Arg-Val-hArg-Agmatine × 3 TFA = inhibitor 11

Phac-Arg-Val-Lys-agmatine × 3 TFA (inhibitor 10) was mixed with a mixture of equal volumes of DMF and 1M Na ₂ CO ₃ solution containing ca. 3 eq. Pyrazolecarboxamidine × HCl and 4 eq. DIPEA reacted analogously to step b of Example 2 (inhibitor 11, white lyophilized powder by preparative HPLC, HPLC: start at 1% B, elution at 24.3 min, MS calc. 673.4, MS F. 674.4 (M + H) ⁺ .

Example 6

Enzyme kinetic studies for determination the inhibitory effect on furin

The determination of the inhibitory effect was carried out using a fluorescence plate reader Safire ² from Tecan (λ _Ex = 380 nm, λ _Em = 460 nm) and Pyr-Arg-Thr-Lys-Arg-AMC (Bachem, No. I-1650). performed as a substrate. Recombinantly produced human furin ( Kacprzak, MM et al., J. Biol. Chem., 2004, 279, 36788-36794 ) was used in the measurement batch in a concentration of about 0.95 nM.

Tabelle 2: K_i-Werte (μM) ausgewählter Inhibitoren für die Hemmung von Furin Inhibitor K_i-Wert (μM) 1 0,0014 2 0,077 2a 25 3 0,068 3a 10 4 0,097 4a 1,3 5 1,6 10 0,049 11 0,047 For the determination of the inhibition constant, the inhibitor concentration was varied at a constant substrate concentration (12.5 μM in the batch) over at least the range of one order of magnitude, and the steady-state velocity was determined in each case. Parallel to each inhibitor measurement, K _m and V _{max were} determined by means of a v / S characteristic. The K _i values were calculated by adjusting the determined rates as a function of the inhibitor concentration to the rate equation for competitively reversible binding inhibitors:

Table 2: K _i values (μM) of selected inhibitors of inhibition of furin inhibitor K _i value (μM) 1 0.0014 2 0.077 2a 25 3 0,068 3a 10 4 0.097 4a 1.3 5 1.6 10 0,049 11 0.047

Example 7

Inhibition of proteolytic cleavage of the Hemagglutinin (HA) of the bird flu virus (FPV)

The HA surface protein of avian influenza virus (belongs to to the highly pathogenic influenza viruses of subtypes H5 and H7) produced as precursor protein HA0.

HA0 is at a multibasic cleavage site by the protease furin or hydrolyzed by a furin-like enzyme, these Proteases occur in virtually all cells. This creates the Subunit HA1 and the membrane-bound subunit HA2. These Cleavage is a prerequisite for the formation of infectious viruses, so that they can fuse with the endosomal membrane of the target cell.

If the proteolytic cleavage of HA in HA1 and HA2 is inhibited, the fusion and thus also the further multiplication of the viruses are inhibited. Cells infected with avian influenza viruses were incubated with different concentrations of inhibitor 1 and a previously described control inhibitor ( Decanoyl-Arg-Val-Lys-Arg-chloromethyl ketone, Hallenberger, S. et al., Nature, 1992, 360, 358-361 ) and examined the cleavage of the HA.

In 1 For example, the inhibition of avian influenza virus HA cleavage by different concentrations of inhibitor 1 and the reference inhibitor Dec-Arg-Val-Lys-Arg-CMK is shown.

The Gelbild A shows the results of the following experiment: Confluent MDCK cell cultures were incubated with influenza virus A / FPV / Rostock / 34 (H7N1) infected at an MOI of 10 per cell. 16 hours after infection the cells were lysed and the inhibition of HA cleavage (PSKKRKKR ↓ GLFG) depending on the inhibitor concentration by means of SDS-PAGE and Western blotting. The viral proteins became immunochemical with a rabbit anti-FPV serum using the ECL test Pierce: HA0 (82 kDa), nucleoprotein NP (56 kDa) and HA1 (~ 50 kDa), with HA2 (32 kDa) with the antiserum used is not detectable.

The statistical evaluation is in picture B of the 1 shown. One recognizes the quantification of the inhibition of HA cleavage. In each case, three independent experiments were carried out for both inhibitors, the quantification was carried out by means of a second monoclonal antibody, which is labeled with an IR dye and detectable with the LI-COR Odyssey Image system. The maximum amount of HA0 at 50 μM Dec-Arg-Val-Lys-Arg-CMK was set as 100% inhibition of cleavage. The intensities of the HA0 bands at the different inhibitor concentrations were normalized with the intensity of the bands for the nucleoprotein (NP at 56 kDa).

Example 8

Inhibition of multicyclic replication of the FPV in the presence of the inhibitor 1

In another experiment, the long-term effects of inhibitor 1 on multicyclic Ver increase in bird flu virus (FPV) ( See Garden et al., Virology 1989, 172, 25-31 ). MDCK cells were infected with FPV at low multiplicity of infection (MOI) of 10 ^-5 and incubated in the presence of a 25 μM concentration of inhibitor 1 over a period of 72 hours. As a control, the infected cells were incubated without inhibitor. The multiplication of the FPV was investigated by virus titration in the cell culture supernatants with a standard hemagglutination test ( Garten et al., Virology 1989, 172, 25-31, Figure 3 ). The release of virus in the group treated with the inhibitor is delayed by approximately 24 h compared to the untreated control group. The delayed virus replication in the presence of the inhibitor also makes it clear that the vitality of the cells is not affected by the inhibitor

2 shows the multicyclic proliferation of FPV in cell culture in the presence (filled bars) of inhibitor 1 (25 μM) and without inhibitor (unfilled bars). MDCK cell cultures were incubated with influenza strain A / FPV / Rostock / 34 (H7N1) at an MOI of 10 ^-5 per cell at 37 ° C. At the indicated time points (10, 18, 32, 48, 60, and 72 h after infection), the amount of virus in the medium was determined by the hemagglutination test. The mean values of four independent experiments are shown.

Example 9

• *Start der HPLC bei 30% B, bei allen anderen Verbindungen Start der HPLC bei 1% B.

According to the methods described above, the Ki values for the inhibition of furin were also determined for the synthesized inhibitors shown in Table 3. Table 3: Structures of further synthesized inhibitors and K _i values for the inhibition of furin for selected inhibitors (nb = not determined).

• * Start HPLC at 30% B, for all other compounds start HPLC at 1% B.

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

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Claims (38)

Compound according to formula (I) P5-P4-P3-P2-P1 (I) wherein P5-P4-P3-P2-P1 is a tetrapeptide derivative and wherein P1 is an arginine mimetic, P2, P3, P4 are each an amino and / or imino acid residue and P5 is an N-terminal modification of the amino or imino acid residue P4. Compound characterized in that it is a salt or a prodrug of the compound according to formula (I) is. A compound according to any one of claims 1 or 2, characterized in that the compound is an inhibitor for the proprotein convertases furin, PC1 / 3, PC2, PC4, PC5 / 6, PACE4 and / or PC7 is. Compound according to one of claims 1 to 3, characterized in that P1 is selected from the following structures:

wherein • R _{1 is} selected from the group H, OH, O-CH ₃ , NH ₂ , O-CO-CH ₃ or -CO-O- (CH ₂ ) _m -CH ₃ where m is an integer from 1 to 5 is and • R _{2 is} H or amidino, and • n is 3, 4 or 5, and • s is 2 or 3. Compound according to claim 4, characterized in that P1 is preferably selected from the following structures

particularly preferred

Compound according to one of Claims 1 to 5, characterized in that P2 and P4 are each an amino or imino acid radical selected from the following structures

and wherein • p is an integer from 0 to 5, and • R _{3 can be} an amino or a guanidino group, or an -O-guanidino group, or a mono- or polysubstituted or unsubstituted aryl or heteroaryl radical having 5 to 10 ring atoms in which the heteroaryl ring may contain up to 3 heteroatoms selected from N, S or O, where the substituent on the aryl or heteroaryl ring is selected from -NH ₂ , -CH ₂ -NH ₂ , -amidino, -hydroxyamidino, -guanidino, - CH ₂ -guanidino, -halogen, -Cl, -Br, -I, -CN, -CF ₃ , -alkyl having 1-3 C-atoms or -alkoxy with 1-3 C-atoms, -COOR ₅ with R ₅ is a hydrogen or an alkyl having 1 to 3 C atoms, and R _{4 is} -H, -OH or -O- (CH ₂ ) _q -R ₆ is q is an integer from 2 to 4 and R _{6 is} an amino - or guanidino group. Compound according to claim 6, characterized in that P2 is in L-configuration and is preferably selected from the structures

particularly preferred

Compound according to one of claims 6 or 7, characterized in that R3 is preferably • an amino, guanidino or -O-guanidino group or • is a substituted aryl radical having 6 ring atoms or an unsubstituted pyridyl radical, wherein the nitrogen in the pyridyl ring at any position and optionally also present as N-oxide, and wherein the substituent on the aryl ring is selected from -NH ₂ , -CH ₂ -NH ₂ , -Amidino, -Hydroxyamidino, -Guanidino, or -CH ₂ -Guanidino. A compound according to any one of claims 6 to 8, characterized in that P4 is preferably in L configuration wherein P4 is more preferably an amino acid residue in L configuration is. A compound according to any one of claims 1 to 9, characterized in that P3 is any natural one or unnatural α-amino or α-imino acid residue with L or D configuration. Connection according to claim 10, characterized in that that P3 is a non-basic α-amino or α-imino acid residue in L configuration, the remainder of a proteinogenic α-amino or α-imino acid. Connection according to claim 10, characterized in that that P3 is a basic α-amino acid residue in D configuration is. Connection according to claim 12, characterized in that that P3 is a D-arginine, D-homoarginine, D-norarginine, D-lysine, D-homolysin, D-norlysine, D-3-amidinophenylalanine, D-4-amidinophenylalanine, a D-pyridylalanine or a D-homopyridylalanine, wherein the nitrogen is in the pyridyl ring anywhere stands. Compound according to one of claims 1 to 13, characterized in that P5 is selected from one of the groups • -H, -CO-R ₇ or -CO-XR ₇ , or • pyroglutamyl radical or another amino or imino acid radical, or • SO ₂ -R ₇ , or • unbranched or branched alkyl radical having 1 to 10 carbon atoms, wherein the alkyl radical optionally with a halogen, an NH ₂ , guanidino, COOH or COO-R ₅ group, or • a shpingosine acylated with a bifunctional residue or shpingosyl-phosphorylcholine acylated with a bifunctional residue of the following structures

in which • z is an integer between 2 and 20, • X is O or NH, • R _{5 is} a hydrogen or an alkyl having 1 to 3 carbon atoms, and • R _{7 is} i) unsubstituted or optionally mono- or polysubstituted branched or is unbranched, saturated or unsaturated alkyl radical having 1 to 24 C atoms and 0 to 3 double bonds, or ii) an unsubstituted or optionally mono- or polysubstituted aryl, heteroaryl, aralkyl, heteroaralkyl or cycloalk radical having 5 to 20 carbon atoms wherein the heteroaryl ring may contain up to 3 heteroatoms selected from N, S or O, wherein the optionally present substituent (s) on R _{7 are} independently selected from -NH ₂ , -CH ₂ -NH ₂ , -Amidino, -Hydroxyamidino , Guanidino, -CH ₂ -guanidino, -halo, -Cl, -Br, -I, -CN, -CF ₃ , -alkyl having 1-3 C atoms or alkoxy having 1 to 3 C atoms, -COO -R ₅ with or iii) is a cholesterol residue. A compound according to claim 14, characterized in that P5 is preferably selected from the group -CO-R ₇ , -CO-XR ₇ or SO ₂ -R ₇ . Compound according to one of claims 14 or 15, characterized in that P5 is more preferably selected from

following structures,

where z is an integer from 1 to 18, more preferably 8, 14 or 16. A compound according to any one of claims 1 to 16, characterized in that • P1 is a 4-amidinobenzylamide radical, • P2 is an arginyl, lysyl or canavanine radical in L configuration, • P3 is a non-basic α-amino acid or α-imino acid radical in L configuration, • P4 is an arginyl, lysyl or canavanine residue in the L-configuration, and • P5 is a phenylacetyl or another acyl residue, the compounds, including their salts,

are very particularly preferred. A process for the preparation of a compound according to any one of claims 1 to 17 by a combination of solid phase and solution synthesis by means of the following steps: • synthesis of the segment P5-P4-P3-P2 by solid-phase peptide synthesis, • cleavage of the peptide segment from the resin under appropriate acidic conditions to either conserve or cleave side-chain protecting groups, • coupling of a suitable P1 building block using standard racemization-free methods, where the P1 building block is either protected or unprotected, • purification by crystallization, chromatography, preferably reverse phase or ion exchange chromatography or by countercurrent distribution, A method according to claim 18, characterized characterized in that after the coupling of a suitable P1 block the step is performed • Spin-off all remaining protective groups. A method according to claim 19, characterized characterized in that the cleavage of the protecting groups by • the Treatment with acids, preferably with trifluoroacetic acid, HBr in glacial acetic acid, HCl in glacial acetic acid, HF or trifluoromethanesulfonic acid, or • by catalytic hydrogenation with hydrogen and palladium on activated carbon as catalyst removed, or • by another method suitable for the particular protecting group he follows. Method according to claim 20, characterized in that that the acids contain scavengers, preferably water, Triisopropylsilane, anisole, dimethyl sulfide or other common ones Scavenger with free mercapto groups. Method according to one of the claims 18 to 21, characterized in that after cleaning the step is carried out • overpass in a different salt form. A process for the preparation of a compound according to any one of claims 1 to 17 by solution synthesis comprising the following steps: a) synthesis of a P1 with free amino group suitably protected at the terminal amidino or guanidino group, wherein the amino group is present either as salt or free base, preferably synthesis of following compound:

b) coupling of a Nα-Boc or otherwise suitably Nα-urethane protected amino or imino acid with or without a suitable side chain protecting group to the free amino group of the P1 block, c) workup of the reaction mixture by conventional washing and crystallization dations d) cleavage of the Nα-protecting group to obtain the P2-P1 segment having a free amino group on the P2 component, e) coupling the Nα-Boc or otherwise suitably Nα-urethane-protected P3-amino or imino acid to the free amino group of the P2-P1 segment, f) work-up g) Cleavage of the Nα-protecting group to give the P3-P2-P1 segment with free N-terminal amino group on the P3 radical, h) coupling of the Nα-Boc or otherwise suitable Nα-urethane-protected P4 Amino or imino acid to the free amino group of the P3-P2-P1 segment from step h) i) work-up of the reaction batch by customary washing and crystallization processes j) Cleavage of the Nα-protecting group to give the P4-P3-P2-P1 segment with free N-terminal amino group at the P4 residue, k) coupling of the P5 residue to the free amino group of the P4-P3-P2-P1 segment Step j) l) removal of all protective groups still present, m) purification by crystallization and / or chromatography, Method according to claim 23, characterized as coupling method, all coupling methods of peptide chemistry can be used with known auxiliary bases. Method according to claim 23 or 24, characterized in that the mixed anhydride method, Carbodiimide method, active ester method, azide method, propanephosphonic anhydride method or the coupling with triazimoch, TBTU, HBTU, BOP, PyBOP, HATU or other coupling reagents based on uronium or phosphonium salts is applied. Method according to one of claims 23 to 25, characterized in that for the cleavage of the Nα-Boc protective group Trifluoroacetic acid or HCl in a suitable organic Solvent, preferably ethyl acetate, acetic acid, Diethyl ether or dioxane is used. Method according to one of claims 23 to 26, characterized in that after the work-up accordingly Steps c), f) and i) each a chromatography step for Cleaning is performed. Method according to one of claims 23 to 27, characterized in that P5 as acid chloride or is present as an active ester and the coupling of the P5 according to step k) takes place in the presence of auxiliary bases without further coupling reagent. Method according to one of claims 23 to 28, characterized in that the cleavage according to step l) • the catalytic hydrogenation with hydrogen and palladium on activated carbon as a catalyst, or by • the Treatment with appropriate acids, preferably with trifluoroacetic acid, HBr in glacial acetic acid, HCl in glacial acetic acid, HF, trifluoromethanesulfonic acid he follows. Method according to claim 29, characterized that the acids scavenger, preferably water, triisopropylsilane, Anisole, dimethyl sulfide or other common scavengers with contain free mercapto groups. Method according to one of claims 23 to 30, characterized in that the cleaning according to step m) preferably by reverse phase or ion exchange chromatography or by countercurrent distribution. Method according to one of claims 23 to 31, characterized in that after the cleaning accordingly Step m) the transfer to another salt form he follows. Medicament containing at least one compound according to one of claims 1 to 17. Medicament according to Claim 33, characterized that it is in the form of a tablet, a dragee, a capsule, one Pellets, a suppository, a solution, in particular an injection or infusion solution, ophthalmic, nasal or ear drops, a juice, an emulsion or suspension, a globule, a styli, an aerosol, an aerosol spray, a powder, paste, cream or ointment. Use of a compound according to a of claims 1 to 17 or of a medicament according to claim 33 or 34 for the treatment or prophylaxis of a viral or bacterial conditional diseases, as well as for the therapy and prophylaxis of diabetes, Arteriosclerosis, cancer, Alzheimer's or overweight, in particular in oral, subcutaneous, intravenous or transdermal Form or in the form of a spray for the treatment of diseases in the Respiratory tract. Use of a compound according to a of claims 1 to 17 or of a medicament according to claim 33 to 34 caused for the therapy or prophylaxis of diseases by bacterial agents, in particular caused by pathogens from the group Pseudomonas spec., Shigella spec., Corynebacterium diphtheriae, Bacillus cereus, Bacillus anthracis, Bacillus butulinum, Clostridium perfringens, Clostridium difficile, Clostridium septicum, Clostridium sprioforme, Bordetella pertussis, Samonella enterica or by selected Escherichia coli strains, in particular E. coli strains, the hemolytic Colitis, the Hemolytic Uraemic Syndrome and / or Cause kidney failure, especially E. coli 0157-H7. Use of a compound according to any one of claims 1 to 17 or a medicament according to claim 33 to 34 for the therapy or prophylaxis of diseases caused by viruses, in particular caused by viruses of the group • highly pathogenic avian influenza A viruses, especially the H5 or H7 Subtypes, • RNA negative strand viruses such as the parainfluenza viruses, the measles virus, the mumps virus, the canine distemper virus, respiratory syncytial virus, filoviruses such as the Ebola and Marburg virus and the Newcastle disease virus; and • positive-strand RNA viruses such as HIV-1, HIV-2, HTLV and other retroviruses, in particular coronaviruses such as the SARS coronavirus, the flaviviruses, yellow fever viruses, West Nile virus, dengue virus or the early summer meningocephalitis virus, the Venezuelan equine encephalitis virus or the Rift Valley Fever virus. Use of a compound according to a of claims 1 to 17 or of a medicament according to claim 33 to 34 for the inhibition of the proprotein convertases furin, PC1 / 3, PC2, PC4, PC5 / 6, PACE4 and / or PC7.