CYCLIC CHEMERIN-9 DERIVATIVES

Abstract

The present invention relates to cyclic chemerin-9 derivatives of general formula (I) as described and defined herein, methods of preparing said peptides, and the use of said compounds for the treatment or prophylaxis of diseases, in particular cancer, diabetes, obesity and inflammatory disorders.

Claims

1: A compound of formula (I): ##STR00042## wherein R.sup.1 is absent or represents 6-Carboxytetramethylrhodamine (Tam), ##C(O)R.sup.3, C.sub.8-C.sub.20 fatty acid or the sequence R.sup.4GFLG ##, R.sup.4—C═N—NH-##, R.sup.4—S—S-##, R.sup.4—N═N-##, R-Valin-Citrullin-##, R.sup.4—C(O)O-## or R.sup.4NH—C(O)O-## wherein ## marks the attachment to the terminal amino group of X.sup.1, R.sup.3 represents C.sub.1-C.sub.6-alkylene, aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl or C.sub.3-C.sub.7-heterocycloalkyl, wherein C.sub.1-C.sub.6-alkylene is up to trisubstituted identically or differently by a radical selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, amino and halogen, wherein aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl and C.sub.3-C.sub.7-heterocycloalkyl can be up to trisubstituted identically or differently by a radical selected from the group of C.sub.1-C.sub.4-alkyl, hydroxyl, methoxy, ethoxy, carbonyl, carboxy, amino and halogen, R.sup.4 represents ##STR00043## wherein R.sup.5 represents C.sub.1-C.sub.6-alkylene, aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl or C.sub.3-C.sub.7-heterocycloalkyl, wherein C.sub.1-C.sub.6-alkylene is up to trisubstituted identically or differently by a radical selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, amino and halogen, wherein aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl and C.sub.3-C.sub.7-heterocycloalkyl can be up to trisubstituted identically or differently by a radical selected from the group of C.sub.1-C.sub.4-alkyl, hydroxyl, methoxy, ethoxy, carbonyl, carboxy, amino and halogen, or represents a group of the formula (IIIa) ##STR00044## wherein ** marks the attachment to a nitrogen atom, D.sup.1 is C.sub.1-C.sub.4-alkylene, Y.sup.1 is selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, carboxamide or amino wherein amino might be substituted with 6-carboxytetramethyirhodamine (Tam) via an amide bond, and r represents an integer of from 2 to 15, R.sup.2 represents a group of the formula (II) ##STR00045## or represents a group of the formula (III) ##STR00046## wherein * represents the attachment to the carbonyl atom of the carboxy group of X.sup.3, Z represents a bond or —CH.sub.2—, m represents 1 or 2, n represents 1 or 2, X.sup.1 represents a natural amino acid selected from a list consisting of L, I, F, H, M, W, Y or y or an unnatural amino acid selected from a list consisting of L-Norleucine (Nle), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), 4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluorophenylalanine ((2,5-Difluoro)F), 2-Chlorophenylalanine ((2-Chlor)F), 3-Chlorophenylalanine ((3-Chlor)F), 4-Chlorophenylalanine ((4-Chloro)F), 2-Bromophenylalanine ((2-Bromo)F), 3-Bromophenylalanine ((3-Bromo)F), 4-Bromophenylalanine ((4-Bromo)F), 2-Fluorophenylalanine ((2-Fluoro)F), 3-Fluorophenylalanine ((3-Fluoro)F), 4-Fluorophenylalanine ((4-Fluoro)F), (2,5-difluoro-phenylalanine, 2-Methylphenylalanine ((2-Me)F), 3-Methyl-phenylalanine ((3-Me)F), 4-Methylphenylalanine ((4-Me)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, Phenylglycine (Phg)N-Phenylglycine ((N-Ph)G), 3-Chlorophenylglycine ((3-Chloro-Ph)G), 3-(1,3-Benzothiazol-2-yl)-alanine 1-Benzyl-histidine (H(1-Bn)), 1-Methyl-histidine (H(1-Me)), 3-Methylhistidine (3-Me)H), 2-Pyridylalanine (2-Pal), 3-Pyridylalanine (3-Pal), 4-Pyridylalanine (4-Pal), 3-(Aminomethyl)benzoic acid, 1-Napthylalanine (1-Nal), 2-Napthylalanine (2-Nal), (2R)-Amino-(1-methyl-1H-indazol-5-yl)acetic acid and (2S)-3-(indol-4-yl)-2-(amino)propanoic acid, whereas any natural amino acid and/or unnatural amino acid from that list can be in D- or L-stereoconfiguration, X.sup.2 represents a natural amino acid selected from a list consisting of L, I, F, H, M, W or Y or an unnatural amino acid selected from a list consisting of L-Norleucine (Nle), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), L-4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluoro-L-phenylalanine ((2,5-Difluoro)F), 2-Chloro-L-phenylalanine ((2-Chloro)F), 3-Chloro-L-phenylalanine ((3-Chloro)F), 4-Chloro-L-phenylalanine ((4-Chloro)F), L-2-Bromophenylalanine ((2-Bromo)F), L-3-Bromophenylalanine ((3-Bromo)F), L-4-Bromophenylalanine ((4-Bromo)F), 2-Fluoro-L-phenylalanine ((2-Fluoro)F), 3-Fluoro-L-phenylalanine ((3-Fluoro)F), 4-Fluoro-L-phenylalanine ((4-Fluoro)F), (2,5-difluoro-L-phenylalanine, 2-Methyl-L-phenylalanine ((2-Me)F), 3-Methyl-L-phenylalanine ((3-Me)F), 4-Methyl-L-phenylalanine ((4-Me)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, L-Phenylglycine (Phg)N-Phenylglycine ((N-Ph)G), 3-Chlorophenylglycine ((3-Chloro-Ph)G), 3-(1,3-Benzothiazol-2-yl)-L-alanine 1-Benzyl-L-histidine (H(1-Bn)), 1-Methyl-L-histidine (H(1-Me)), L-3-Methylhistidine (3-Me)H), L-2-Pyridylalanine (2-Pal), L-3-Pyridylalanine (3-Pal), L-4-Pyridylalanine (4-Pal), 3-(Aminomethyl)benzoic acid, L-1-Napthylalanine (1-Nal), L-2-Napthylalanine (2-Nal), (2R)-Amino-(1-methyl-1H-indazol-5-yl)acetic acid and (2S)-3-(indol-4-yl)-2-(amino)propanoic acid, X.sup.3 represents the natural amino acid P, or an unnatural amino acid selected from a list consisting of 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), L-Hydroxyproline (Hyp), (2S,4S)-4-Trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,4S)-4-fluoroproline ((cis-4-Fluoro)P), trans-4-fluoroproline ((trans-4-Fluoro)P), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-trans-3-hydroxyproline ((3S—OH)P, L-Pipecolic acid (Pip), (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (6S)-5-Azaspiro-[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S)-2-Amino-4,4,4-trifluorobutanoic acid, (2S,3aS,6aS)-octahydrocyclopenta[b].sup.¬pyrrole-2-carboxylic acid, trans-4-fluoroproline ((trans-4-Fluoro)P), (2S,4S)-4-fluoroproline ((cis-4-Fluoro)P), L-4,4-difluoroproline ((Difluoro)P), rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 1) and rel-(3R,6R)-1,1-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (enantiomer 2), X.sup.4 represents any natural amino acid or an unnatural amino acid, whereas any natural amino acid and/or unnatural amino acid can be in D- or L-stereoconfiguration, X.sup.5 represents a natural amino acid selected from a list consisting of F, H, W or Y or an unnatural amino acid selected from a list consisting of Cyclohexylalanine (Cha), 2,3,3a,4,5,6,7,7a-Octahydrindole-2-carboxylic acid (Oic), L-4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluoro-L-phenylalanine ((2,5-Difluoro)F), 2-Chlor-L-phenylalanine ((2-Chloro)F), 3-Chloro-L-phenylalanine ((3-Chloro)F), 4-Chloro-L-phenylalanine ((4-Chloro)F), L-2-Bromophenylalanine ((2-Bromo)F), L-3-Bromophenylalanine ((3-Bromo)F), L-4-Bomophenylalanine ((4-Bromo)F), 2-Fluoro-L-phenylalanine ((2-Fluor)F), 3-Fluoro-L-phenylalanine ((3-Fluoro)F), 4-Fluoro-L-phenylalanine ((4-Fluoro)F), (2,5-difluoro-L-phenylalanine, 2-Methyl-L-phenylalanine ((2-Me)F), 3-Methyl-L-phenylalanine ((3-Me)F), 4-Methyl-L-phenylalanine ((4-Me)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, L-Phenylglycine (Phg)N-Phenylglycine ((N-Ph)G), 3-Chlorophenylglycine ((3-Chloro-Ph)G), 3-(1,3-Benzothiazol-2-yl)-L-alanine 1-Benzyl-L-histidine (H(1-Bn)), 1-Methyl-L-histidine (H(1-Me)), L-3-Methylhistidine (3-Me)H), L-2-Pyridylalanine (2-Pal), L-3-Pyridylalanine (3-Pal), L-4-Pyridylalanine (4-Pal), 3-(Aminomethyl)benzoic acid, L-1-Napthylalanine (1-Nal), L-2-Napthylalanine (2-Nal), (2R)-Amino-(1-methyl-1H-indazol-5-yl)acetic acid and (2S)-3-(indol-4-yl)-2-(amino)propanoic acid, X.sup.6 represents any natural amino acid or an unnatural amino acid, whereas any natural amino acid and/or unnatural amino acid can be in D- or L-stereoconfiguration, wherein any natural amino acid or an unnatural amino acid bearing an amino group might be substituted with 6-Carboxytetramethylrhodamine (Tam) or ##C(O)R.sup.3, wherein ## marks the attachment to the terminal amino group of X.sup.1, R.sup.3 represents C.sub.1-C.sub.6-alkylene, aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl or C.sub.3-C.sub.7-heterocycloalkyl,  wherein C.sub.1-C.sub.6-alkylene is up to trisubstituted identically or differently by a radical selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, amino and halogen, wherein aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl and C.sub.3-C.sub.7-heterocycloalkyl can be up to trisubstituted identically or differently by a radical selected from the group of C.sub.1-C.sub.4-alkyl, hydroxyl, methoxy, ethoxy, carbonyl, carboxy, amino and halogen, X.sup.7 represents a natural amino acid selected from a list consisting of F, H, W or Y or an unnatural amino acid selected from a list consisting of Cyclohexylalanine (Cha), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), L-4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluoro-L-phenylalanine ((2,5-Difluoro)F), 2-Chloro-L-phenylalanine ((2-Chloro)F), 3-Chloro-L-phenylalanine ((3-Chloro)F), 4-Chloro-L-phenylalanine ((4-Chloro)F), L-2-Bromophenylalanine ((2-Bromo)F), L-3-Bromophenylalanine ((3-Bromo)F), L-4-Bromophenylalanine ((4-Bromo)F), 2-Fluoro-L-phenylalanine ((2-Fluoro)F), 3-Fluoro-L-phenylalanine ((3-Fluoro)F), 4-Fluoro-L-phenylalanine ((4-Fluoro)F), (2,5-difluoro-L-phenylalanine, 2-Methyl-L-phenylalanine ((2-Me)F), 3-Methyl-L-phenylalanine ((3-Me)F), 4-Methyl-L-phenylalanine ((4-Me)F), (2S)-3-(2,3-difluorophenyl)-2-aminopropanoic acid, L-Phenylglycine (Phg)N-Phenylglycine ((N-Ph)G), 3-Chlorophenylglycine ((3-Chloro-Ph)G), 3-(1,3-Benzothiazol-2-yl)-L-alanine 1-Benzyl-L-histidine (H(1-Bn)), 1-Methyl-L-histidine (H(1-Me)), L-3-Methylhistidine (3-Me)H), L-2-Pyridylalanine (2-Pal), L-3-Pyridylalanine (3-Pal), L-4-Pyridylalanine (4-Pal), 3-(Aminomethyl)benzoic acid, L-1-Naphthylalanine (1-Nal), L-2-Naphthylalanine (2-Nal), (2R)-Amino-(1-methyl-1H-indazol-5-yl)acetic acid and (2S)-3-(indol-4-yl)-2-(amino)propanoic acid, or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt thereof, with the proviso, that compound YFP[cQFAFC] and yFP[xQFAWC], or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt of any of the foregoing are excluded.

2: The compound of formula (I) according to claim 1, wherein R.sup.1 is absent or represents 6-Carboxytetramethylrhodamine (Tam), ##C(O)R.sup.3 or the sequence R.sup.4GFLG##, wherein ## marks the attachment to the terminal amino group of X.sup.1, R.sup.3 represents C.sub.1-C.sub.4-alkylene, wherein C.sub.1-C.sub.4-alkylene is up to trisubstituted identically or differently by a radical selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, amino, fluoro and chloro, R.sup.4 represents ##STR00047## wherein R.sup.5 represents C.sub.1-C.sub.4-alkylene,  wherein C.sub.1-C.sub.4-alkylene is up to trisubstituted identically or differently by a radical selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, amino, chloro and fluoro, or ##STR00048## wherein ** marks the attachment to a nitrogen atom, D.sup.1 is C.sub.1-C.sub.4-alkylene, Y.sup.1 is selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, carboxamide or amino, wherein amino might be substituted with 6-carboxytetramethylrhodamine (Tam) via an amide bond, and r represents an integer of from 2 to 6, R.sup.2 represents a group of the formula (II) ##STR00049## wherein * represents the attachment to the carbonyl atom of the carboxy group of X.sup.3, Z represents a bond or —CH.sub.2—, m represents 1 or 2, n represents 1 or 2, X.sup.1 represents a natural amino acid selected from a list consisting of L, I, F, H, M, W, Y or y or an unnatural amino acid selected from a list consisting of L-Norleucine (Nle), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), 4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluorophenylalanine ((2,5-Difluoro)F), 2-Chlorophenylalanine ((2-Chloro)F), 3-Chlorophenylalanine ((3-Chloro)F), 4-Chlorophenylalanine ((4-Chloro)F), 2-Bromophenylalanine ((2-Bromo)F), 3-Bromophenylalanine ((3-Bromo)F), 4-Bromophenylalanine ((4-Bromo)F), 2-Fluorophenylalanine ((2-Fluoro)F), 3-Fluorophenylalanine ((3-Fluoro)F), 4-Fluorophenylalanine ((4-Fluoro)F), (2,5-difluoro-phenylalanine, 2-Methyl-phenylalanine ((2-Me)F), 3-Methyl-phenylalanine ((3-Me)F), 4-Methylphenylalanine ((4-Me)F), 1-Benzyl-histidine (H(1-Bn)), 1-Methyl-histidine (H(1-Me)), 3-Methylhistidine (3-Me)H), 2-Pyridylalanine (2-Pal), 3-Pyridylalanine (3-Pal), 4-Pyridylalanine (4-Pal), whereas any natural amino acid and/or unnatural amino acid from that list can be in D- or L-stereoconfiguration, X.sup.2 represents a natural amino acid selected from a list consisting of L, I, F, H, M, W or Y or an unnatural amino acid selected from a list consisting of L-Norleucine (Nle), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), L-4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluoro-L-phenylalanine ((2,5-Difluoro)F), 2-Chloro-L-phenylalanine ((2-Chloro)F), 3-Chloro-L-phenylalanine ((3-Chloro)F), 4-Chloro-L-phenylalanine ((4-Chloro)F), L-2-Bromophenylalanine ((2-Bromo)F), L-3-Bromophenylalanine ((3-Bromo)F), L-4-Bromophenylalanine ((4-Bromo)F), 2-Fluoro-L-phenylalanine ((2-Fluoro)F), 3-Fluoro-L-phenylalanine ((3-Fluoro)F), 4-Fluoro-L-phenylalanine ((4-Fluoro)F), (2,5-difluoro-L-phenylalanine, 2-Methyl-L-phenylalanine ((2-Me)F), 3-Methyl-L-phenylalanine ((3-Me)F), 4-Methyl-L-phenylalanine ((4-Me)F), 1-Benzyl-L-histidine (H(1-Bn)), 1-Methyl-L-histidine (H(1-Me)), L-3-Methylhistidine (3-Me)H), L-2-Pyridylalanine (2-Pal), L-3-Pyridylalanine (3-Pal), L-4-Pyridylalanine (4-Pal), X.sup.3 represents the natural amino acid P, or an unnatural amino acid selected from a list consisting of L-Hydroxyproline (Hyp), (2S,4S)-4-Trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,4S)-4-fluoroproline ((cis-4-Fluoro)P), trans-4-fluoroproline ((trans-4-Fluoro)P), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-trans-3-hydroxyproline ((3S—OH)P, (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (6S)-5-Aza.sup.¬spiro.sup.¬[2.4]heptane-6-carboxylic acid, rel-(1R,3R,5R,6R)-6-(trifluoromethyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid, (2S)-2-Amino-4,4,4-trifluorobutanoic acid, (2S,3aS,6aS)-octahydrocyclopenta[b].sup.¬pyrrole-2-carboxylic acid, (2S,4S)-4-fluoroproline ((cis-4-Fluoro)P), L-4,4-difluoroproline ((Difluoro)P), X.sup.4 represents any natural amino acid, whereas any natural amino acid can be in D- or L-stereoconfiguration, X.sup.5 represents a natural amino acid selected from a list consisting of F, H, W or Y or an unnatural amino acid selected from a list consisting of Cyclohexylalanine (Cha), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), L-4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluoro-L-phenylalanine ((2,5-Difluoro)F), 2-Chloro-L-phenylalanine ((2-Chloro)F), 3-Chloro-L-phenylalanine ((3-Chloro)F), 4-Chloro-L-phenylalanine ((4-Chloro)F), L-2-Bromophenylalanine ((2-Bromo)F), L-3-Bromophenylalanine ((3-Bromo)F), L-4-Bromophenylalanine ((4-Bromo)F), 2-Fluoro-L-phenylalanine ((2-Fluoro)F), 3-Fluoro-L-phenylalanine ((3-Fluoro)F), 4-Fluoro-L-phenylalanine ((4-Fluoro)F), (2,5-difluoro-L-phenylalanine, 2-Methyl-L-phenylalanine ((2-Me)F), 3-Methyl-L-phenylalanine ((3-Me)F), 4-Methyl-L-phenylalanine ((4-Me)F), 1-Benzyl-L-histidine (H(1-Bn)), 1-Methyl-L-histidine (H(1-Me)), L-3-Methylhistidine (3-Me)H), L-2-Pyridylalanine (2-Pal), L-3-Pyridylalanine (3-Pal), L-4-Pyridylalanine (4-Pal), X.sup.6 represents any natural amino acid, whereas any natural amino acid can be in D- or L-stereoconfiguration, wherein the amino group of Lysin might be substituted with 6-Carboxytetramethylrhodamine (Tam) or ##C(O)R.sup.3, wherein ## marks the attachment to the terminal amino group of X.sup.1, R.sup.3 represents C.sub.1-C.sub.6-alkylene, aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl or C.sub.3-C.sub.7-heterocycloalkyl,  wherein C.sub.1-C.sub.6-alkylene is up to trisubstituted identically or differently by a radical selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, amino and halogen,  wherein aryl, heteroaryl, C.sub.3-C.sub.8-cycloalkyl and C3-C.sub.7-heterocycloalkyl can be up to trisubstituted identically or differently by a radical selected from the group of C.sub.1-C.sub.4-alkyl, hydroxyl, methoxy, ethoxy, carbonyl, carboxy, amino and halogen, X.sup.7 represents a natural amino acid selected from a list consisting of F, H, W or Y or an unnatural amino acid selected from a list consisting of Cyclohexylalanine (Cha), 2,3,3a,4,5,6,7,7a-Octahydroindole-2-carboxylic acid (Oic), L-4-Bromophenylalanine ((4-Bromo)F), 2,5-Difluoro-L-phenylalanine ((2,5-Difluoro)F), 2-Chloro-L-phenylalanine ((2-Chloro)F), 3-Chloro-L-phenylalanine ((3-Chloro)F), 4-Chloro-L-phenylalanine ((4-Chloro)F), L-2-Bromophenylalanine ((2-Bromo)F), L-3-Bromophenylalanine ((3-Bromo)F), L-4-Bromophenylalanine ((4-Bromo)F), 2-Fluoro-L-phenylalanine ((2-Fluoro)F), 3-Fluoro-L-phenylalanine ((3-Fluoro)F), 4-Fluoro-L-phenylalanine ((4-Fluoro)F), (2,5-difluoro-L-phenylalanine, 2-Methyl-L-phenylalanine ((2-Me)F), 3-Methyl-L-phenylalanine ((3-Me)F), 4-Methyl-L-phenylalanine ((4-Me)F), 1-Benzyl-L-histidine (H(1-Bn)), 1-Methyl-L-histidine (H(1-Me)), L-3-Methylhistidine (3-Me)H), L-2-Pyridylalanine (2-Pal), L-3-Pyridylalanine (3-Pal), L-4-Pyridylalanine (4-Pal), or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt thereof, with the proviso, that compounds YFP[cQFAFC] and yFP[xQFAWC], or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt of any of the foregoing are excluded.

3: The compound of formula (I) according to claim 1, wherein R.sup.1 is absent or represents 6-Carboxytetramethylrhodamine (Tam) or the sequence R.sup.4GFLG ##, wherein ## marks the attachment to the terminal amino group of X.sup.1, R.sup.4 represents ##STR00050## wherein R.sup.5 represents methyl or ethyl, or represents a group of the formula (IIIa) ##STR00051## wherein marks the attachment to a nitrogen atom, D.sup.1 is C.sub.1-C.sub.4-alkylene, Y.sup.1 is selected from the group consisting of hydroxyl, methoxy, ethoxy, carboxy, carboxamide or amino, wherein amino might be substituted with 6-carboxytetramethylrhodamine (Tam) via an amide bond, and r represents an integer of from 2 to 4, R.sup.2 represents a group of the formula (II) ##STR00052## wherein * represents the attachment to the carbonyl atom of the carboxy group of X.sup.3, Z represents a bond or —CH.sub.2—, m represents 1 or 2, n represents 1 or 2, X.sup.1 represents a natural amino acid selected from a list consisting of F, H, Y or y, whereas any amino acid from that list can be in D- or L-stereoconfiguration, X.sup.2 represents a natural amino acid selected from a list consisting of F, H, Y or y, whereas any amino acid from that list can be in D- or L-stereoconfiguration, X.sup.3 represents the natural amino acid P, or an unnatural amino acid selected from a list consisting of L-Hydroxyproline (Hyp), (2S,4S)-4-Trifluoromethyl-pyrrolidine-2-carboxylic acid ((4-CF3)P), (2S,4S)-4-fluoroproline ((cis-4-Fluoro)P), trans-4-fluoroproline ((trans-4-Fluoro)P), (2S)-2-amino-4,4,4-trifluorobutanoic acid, L-trans-3-hydroxyproline, (2S,4S)-4-fluoroproline ((cis-4-Fluoro)P), L-4,4-difluoroproline ((Difluoro)P), X.sup.4 represents a natural amino acid selected from a list consisting of Q, A and K, whereas any natural amino acid can be in D- or L-stereoconfiguration, X.sup.5 represents a natural amino acid selected from a list consisting of F, H, W or Y, X.sup.6 represents a natural amino acid selected from a list consisting of Q, A and K, whereas any natural amino acid can be in D- or L-stereoconfiguration, wherein the amino group of K might be substituted with 6-Carboxytetramethylrhodamine (Tam), X.sup.7 represents a natural amino acid selected from a list consisting of F, H, W or Y, or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt, with the proviso, that compounds YFP[cQFAFC] and yFP[xQFAWC], or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt of any of the foregoing are excluded.

4: The compound of formula (I) according to claim 1, wherein R.sup.1 is absent or represents 6-Carboxytetramethylrhodamine (Tam) or the sequence R.sup.4GFLG ##, wherein ## marks the attachment to the terminal amino group of X.sup.1, R.sup.4 represents ##STR00053## wherein R.sup.5 represents methyl, or represents a group of the formula (IIIa) ##STR00054## wherein ** marks the attachment to a nitrogen atom, D.sup.1 is ethylene, Y.sup.1 is amino, wherein amino might be substituted with 6-carboxytetramethylrhodamine (Tam) via an amide bond, and r represents 4, R.sup.2 represents a group of the formula (II) ##STR00055## wherein * represents the attachment to the carbonyl atom of the carboxy group of X.sup.3, Z represents a bond or —CH.sub.2—, m represents 1 or 2, n represents 1 or 2, X.sup.1 represents Y or y, X.sup.2 represents F, X.sup.3 represents P, X.sup.4 represents Q, X.sup.5 represents F, X.sup.6 represents A or K, X.sup.7 represents For W, or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt thereof, with the proviso, that compounds YFP[cQFAFC] and yFP[xQFAWC], or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt of any of the foregoing are excluded.

5: A method for treatment or prophylaxis of a disease, comprising administering to a human or animal in need thereof an effective amount of a compound according to claim 1, or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt thereof.

6: The method of claim 5, wherein the disease is a metabolic disorder, cancer or an inflammatory disorder.

7: The method of claim 5, wherein the disease is diabetes mellitus, obesity, an asthmatic disease, an inflammatory disorder or cancer.

8. (canceled)

9: A pharmaceutical composition comprising a compound according to claim 1, or a pharmaceutically acceptable salt, hydrate, solvate or solvate of the salt thereof, in combination with an inert, non-toxic, pharmaceutically suitable auxiliary.

10. (canceled)

11: A method for treatment or prophylaxis of a disease, comprising administering to a human or animal in need thereof an effective amount of a pharmaceutical composition according to claim 9.

12: The method of claim 11, wherein the disease is diabetes mellitus, obesity, an asthmatic disease, an inflammatory disorder or cancer.

13: The method of claim 11, wherein the disease is a metabolic disorder, cancer or inflammatory disorder.

14: The method of claim 5, wherein the compound is administered to a human.

15: The method of claim 11, wherein the pharmaceutical composition is administered to a human.

16: The compound of formula (I) according to claim 1, wherein the compound is [x4,C9]-chemerin-9 YFP[xQFAFC]: ##STR00056##

17: The compound of formula (I) according to claim 1, wherein the compound is [x4,W8,C9]-chemerin-9 YFP[xQFAWC]: ##STR00057##

18: The compound of formula (I) according to claim 1, wherein the compound is EG4-[x4,C9]-chemerin-9 EG(4)-YFP[xQFAFC]: ##STR00058##

Description

SPECIFIC EXAMPLES

Compound List:

[0607]

TABLE-US-00005 Nr Compound name Sequence ($$ 1) Chemerin-9 YFPGQFAFS ($$ 2) Tam-chemerin-9 Tam-YFPGQFAFS ($$ 3) Tam-EG4-chemerin-9 Tam-EG(4)-YFPGQFAFS ($$ 4) [c4,C9]-chemerin-9 YFP[cQFAFC] ($$ 5) Tam-[c4,C9]-chemerin-9 Tam-YFP[cQFAFC] ($$ 6) [c4,C7]-chemerin-9 YFP[cQFC]FS ($$ 7) [c4,C9-TA]-chemerin-9 YFP[c(CH.sub.2)QFAFC] ($$ 8) Tam-[c4,C9-TA]-chemerin-9 Tam-YFP[c(CH.sub.2)QFAFC] ($$ 9) [c4,X9-TA]-chemerin-9 YFP[c(CH.sub.2)QFAFX] ($$ 10) Tam-[c4,X9-TA]-chemerin-9 Tam-YFP[c(CH.sub.2)QFAFX] ($$ 11) [y1,c4,X9-TA]-chemerin-9 yFP[c(CH.sub.2)QFAFX] ($$ 12) [y1,c4,K7(Tam),C9]-chemerin-9 yFP[cQFK(Tam)FC] ($$ 13) [x4,C9]-chemerin-9 YFP[xQFAFC] ($$ 14) [x4,W8,C9]-chemerin-9 YFP[xQFAWC] ($$ 15) [y1,x4,W8,C9]-chemerin-9 yFP[xQFAWC] ($$ 16) EG4-[x4,C9]-chemerin-9 EG(4)-YFP[xQFAFC] ($$ 17) Tam-EG4-[x4,C9]-chemerin-9 Tam-EG(4)-YFP[xQFAFC] ($$ 18) EG4-[x4,W8,C9]-chemerin-9 EG(4)-YFP[xQFAWC] ($$ 19) Tam-EG4-[x4,W8,C9]-chemerin-9 Tam-EG(4)-YFP[xQFAWC] ($$ 20) GFLG-[x4,C9]-chemerin-9 GFLGYFP[xQFAFC] X = Homocysteine, x = D-Homocysteine, TA = thioacetal.

Synthesis of Comparison Examples:

Comparison Example 1: Chemerin-9 ($$ 1)

[0608] ##STR00022##

[0609] After automated synthesis of YFPGQFAFS the peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 30-60% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 5.8 mg (36% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 17.4 min and 8.2 min, respectively. The chemical formula of the peptide is C.sub.54H.sub.66N.sub.10O.sub.3 (monoisotopic mass: 1062.5 Da, average mass: 1063.18 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1063.6 [M+H].sup.+, m/z=1085.6 [M+Na].sup.+ and m/z=1101.5 [M+K].sup.+ were observed. In ESI ion trap, signals at m/z=1063.3 [M+H].sup.+ and m/z=532.3 [M+2H].sup.2+ were observed.

Comparison Example 2: Tam-chemerin-9 ($$ 2)

[0610] ##STR00023##

[0611] After automated synthesis of YFPGQFAFS, the N-terminus of the peptide was modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam. HATU and DIPEA in DMF overnight, the peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 30-80% B in A over 40 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 5.3 mg (24% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 24.4 min and 19.1 min, respectively. The chemical formula of the peptide is C.sub.79H.sub.56N.sub.12O.sub.17 (monoisotopic mass: 1474.6 Da, average mass: 1475.6 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1475.6 [M+H].sup.+, m/z=1497.6 [M+Na].sup.+ and m/z=1513.5 [M+K].sup.+ were observed. In ESI ion trap, signals at m/z=1475.4 [M+H].sup.+ and m/z=738.3 [M+2H].sup.2+ were observed.

Comparison Example 3: Tam-EG4-chemerin-9 ($$3)

[0612] ##STR00024##

[0613] After automated synthesis of YFPGQFAFS, EG(4) was coupled to the N-terminus of the peptide by reaction of 5 equiv of Fmoc-15-amino-4,7,10,13-tetraoxapentadecanoic acid, HOBt and DIC in DMF overnight. The Fmoc-group was cleaved by reaction with 20% piperidine in DMF for 10 min, the reaction was repeated twice. The peptide was N-terminally modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam. HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 30-80% B in A over 40 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 4.8 mg (18.6% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 24.8 min and 16.4 min, respectively. The chemical formula of the peptide is C.sub.90H.sub.107N.sub.3O.sub.22 (monoisotopic mass: 1721.7 Da, average mass: 1722.9 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1722.7 [M+H].sup.+, m/z=1745.7 [M+Na].sup.+ and m/z=1760.7 [M+K].sup.+ were observed. In ESI ion trap, signals at m/z=1722.6 [M+H].sup.+ and m/z=862.1 [M+2H].sup.2+ were observed.

Comparison Example 4: [c4,C9]-chemerin-9 ($$ 4)

[0614] ##STR00025##

[0615] YFPcQFAFC, were synthesized automatically. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Kinetex 5 μm XB-C18 100 Å column employing a gradient of 25 to 55% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 1.9 mg (11.1% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 21.2 min and 11.4 min, respectively. The chemical formula of the peptide is C.sub.55H.sub.66N.sub.10O.sub.12S.sub.2 (monoisotopic mass: 1122.4 Da, average mass: 1123.3 Da). The observed masses were in correspondence to the calculated masses, confirming product identity. In ESI ion trap, signals at m/z=1123.2 [M+H].sup.+ and m/z=562.3 [M+2H].sup.2 were observed.

Comparison Example 5: [c4,C7]-chemerin-9 ($$6)

[0616] ##STR00026##

[0617] YFPcQFCFS was synthesized by automated peptide synthesis. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Kinetex 5 μm XB-C18 100 Å column employing a gradient of 25 to 55% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 1.9 mg (11.1% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 21.0 min and 11.7 min, respectively. The chemical formula of the peptide is C.sub.55H.sub.68N.sub.10O.sub.12S.sub.2 (monoisotopic mass: 1138.4 Da, average mass: 1139.3 Da). The observed masses were in correspondence to the calculated masses, confirming product identity. In ESI ion trap, signals at m/z=1139.2 [M+H].sup.+ and m/z=570.3 [M+2H].sup.2+ were observed.

Synthesis of Examples

Example 1: Tam-[c4,C9]-chemerin-9 ($$ 5)

[0618] ##STR00027##

[0619] After automated synthesis of YFPcQFAFC, the N-terminus of the peptide was modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam, HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Kinetex 5 μm XB-C18 100 Å column employing a gradient of 30 to 65% B in a over 40 min with a flow rate of 15 mL/min. detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 0.7 mg (3% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 27.5 min and 22.3 min, respectively. The chemical formula of the peptide is C.sub.80H.sub.86N.sub.12O.sub.16S.sub.2 (monoisotopic mass: 1534.6 Da, average mass: 1535.8 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, a signal at m/z=1535.6 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1535.3 [M+H].sup.+ and m/z=768.7 [M+2H].sup.2+ were observed.

Example 2: [c4,C9-TA]-chemerin-9 ($$ 7)

[0620] ##STR00028##

[0621] After automated synthesis of YFPcQFAFC the peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 20-70% B in A over 40 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield of the linear peptide was 5.3 mg (31% of theory). 0.6 mg of the peptide was dissolved in THF/H.sub.2O (1:2) in the presence of 3 equiv K.sub.2CO.sub.3, 3 equiv TCEP, and 20 equiv Et.sub.3N. This solution was added stepwise to a solution of 20 equiv CH.sub.2I.sub.2 in THF. The reaction was completed after shaking at rt for 12 h. The peptide was purified on a semipreparative a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 30-60% B in A over 30 min with a flow rate of 5 mL/min. The pure yield of the cyclic peptide was 0.5 mg (83% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 employing a linear gradient of 20-70% B in A over 40 min with a flow rate of 1.0 mL/min. The peptide showed over 95% purity as determined by the absorption at 220 nm, with a retention time of 20.4 min. The chemical formula of the peptide is C.sub.56H.sub.68N.sub.10O.sub.12S.sub.2 (monoisotopic mass: 1136.5 Da, average mass: 1137.3 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1137.5 [M+H].sup.+, 1159.5 [M+Na].sup.+, and 1175.4 [M+K].sup.+ were observed. In ESI ion trap, signals at m/z=1137.3 [M+H].sup.+ and m/z=569.3 [M+2H].sup.2+ were observed.

Example 3: Tam-[c4,C9-TA]-chemerin-9 ($$ 8)

[0622] ##STR00029##

[0623] After automated synthesis of YFcGQFAFC, the N-terminus of the peptide was modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam, HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 30-80% B in A over 40 min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield of the linear peptide was 2.3 mg (10% of theory). The peptide was dissolved in THF/H.sub.2O (1:2) in the presence of 3 equiv K.sub.2CO.sub.3, 3 equiv TCEP, and 20 equiv Et.sub.3N. This solution was added stepwise to a solution of 20 equiv Ch.sub.2I.sub.2 in THF. The reaction was completed after shaking at rt for 12 h. The peptide was purified on a semi-preparative a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 30-60% B in A over 30 min with a flow rate of 5 mL/min. The pure yield of the cyclic peptide was 0.73 mg (31% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 26.9 min and 22.6 min, respectively. The chemical formula of the peptide is C.sub.81H.sub.88N.sub.12O.sub.16S.sub.2 (monoisotopic mass: 1548.6 Da, average mass: 1549.8 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, a signal at m/z=1549.6 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1549.4 [M+H].sup.+ and m/z=775.3 [M+2H].sup.2+ were observed.

Example 4: [c4,X9-TA]-chemerin-9 ($$ 9)

[0624] ##STR00030##

[0625] A 2-chlortrityl chloride resin was loaded with Fmoc-Homocysteine by reaction of the resin with 1.5 equiv of the amino acid and 5 equiv of DIPEA overnight. Loading was determined by cleaving the Fmoc-group off a defined amount of resin using piperidine and measuring the absorption of the piperidin-Fmoc adduct at λ=301 nm. After subsequent automated synthesis of YFPcQFAF, the peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 30-65% B in A over 35 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield of the linear peptide was 1.3 mg (7.5% of theory). The peptide was dissolved in THF/H.sub.2O (1:2) in the presence of 3 equiv K.sub.2CO.sub.3, 3 equiv TCEP, and 20 equiv Et.sub.3N. This solution was added stepwise to a solution of 20 equiv CH.sub.2I.sub.2 in THF. The reaction was completed after shaking at rt for 12 h. The peptide was purified on a semi-preparative a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 20-70% B in A over 40 min with a flow rate of 5 mL/min. The pure yield of the cyclic peptide was 0.7 mg (53% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm XB-C18 100 Å column employing linear gradients of 20-70% B in A over 40 min with flow rates of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm, with a retention time of 20.9 min and 13.9 min, respectively. The chemical formula of the peptide is C.sub.57H.sub.70N.sub.10O.sub.12S.sub.2 (monoisotopic mass: 1150.5 Da, average mass: 1151.4 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1151.5 [M+H].sup.+, 1173.5 [M+Na].sup.+, and 1189.5 [M+K].sup.+ were observed. In ESI ion trap, signals at m/z=1151.3 [M+H].sup.+ and m/z=576.2 [M+2H].sup.2+ were observed.

Example 5: Tam-[c4,X9-TA]-chemerin-9 ($$ 10)

[0626] ##STR00031##

[0627] A 2-chlortrityl chloride resin was loaded with Fmoc-Homocysteine by reaction of the resin with 1.5 equiv of the amino acid and 5 equiv of DIPEA overnight. Loading was determined by cleaving the Fmoc-group off a defined amount of resin using piperidine and measuring the absorption of the pieridine-Fmoc adduct at λ=301 nm. After subsequent automated synthesis of YFPcQFAF, the N-terminus of the peptide was modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam. HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 30-80% B in A over 40 min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield of the linear peptide was 2.3 mg (10% of theory). The peptide was dissolved in THF/H.sub.2O (1:2) in the presence of 3 equiv K.sub.2CO.sub.3, 3 equiv TCEP, and 20 equiv Et.sub.3N. This solution was added stepwise to a solution of 20 equiv Ch.sub.2I.sub.2 in THF. The reaction was completed after shaking at rt for 12 h. The peptide was purified on a semi-preparative a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 30-60% B in A over 30 min with a flow rate of 5 mL/min. The pure yield of the cyclic peptide was 0.73 mg (31% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 26.9 min and 22.6 min. respectively.

Example 6: [y1,c4,X9-TA]-chemerin-9 ($$ 11)

[0628] ##STR00032##

[0629] A 2-chlortrityl chloride resin was loaded with Fmoc-Homocysteine by reaction of the resin with 1.5 equiv of the amino acid and 5 equiv of DIPEA overnight. Loading was determined by cleaving the Fmoc-group off a defined amount of resin using piperidine and measuring the absorption of the pieridine-Fmoc adduct at λ=301 nm. After subsequent automated synthesis of YFPcQFAF, the peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 30-65% B in A over 35 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield of the linear peptide was 2.5 mg (11% of theory). The peptide was dissolved in THF/H.sub.2O (1:2) in the presence of 3 equiv K.sub.2CO.sub.3, 3 equiv TCEP, and 20 equiv Et.sub.3N. This solution was added stepwise to a solution of 20 equiv CH.sub.2I.sub.2 in THF. The reaction was completed after shaking at rt for 12 h. The peptide was purified on a semi-preparative a Kinetex 5 μm XB-C18 100 Å column employing a linear gradient of 20-70% B in A over 40 min with a flow rate of 5 mL/min. The pure yield of the cyclic peptide was 0.8 mg (32% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm, with retention times of 22.3 min and 12.9 min, respectively. The chemical formula of the peptide is C.sub.57H.sub.70N.sub.10O.sub.12S.sub.2 (monoisotopic mass: 1150.5 Da, average mass: 1151.4 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, a signal at m/z=1151.4 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1151.3 [M+H].sup.+ and m/z=576.3 [M+2H].sup.2+ were observed.

Example 7: [y1,c4,K7(Tam),C9]-chemerin-9 ($$12)

[0630] ##STR00033##

[0631] The peptide was synthesized incorporating a Dde-protected lysine at position 7 to allow selective modification of the peptide at the lysine said chain. After automated synthesis of yFPcQFK(Dde)FC, the Dde protecting group was cleaved by repeated reaction with 2% hydrazine in DMF, the reaction was monitored by measuring the UV absorbance of the Dde-hydrazine adduct at λ=300 nm. The peptide was modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam, HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a linear gradient of 30-60% B in A over 30 min, detecting the peptide by measuring the absorption at λ=220 nm. The yield of the pure peptide was 1.2 mg (5% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm, with retention times of 24.1 min and 15.9 min, respectively. The chemical formula of the peptide is C.sub.83H.sub.93N.sub.13O.sub.16S.sub.2 (monoisotopic mass: 1591.6 Da, average mass: 1592.9 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1592.7 [M+H].sup.+, m/z=1614.7 [M+Na].sup.+, and at m/z=1630.6 [M+K].sup.+ was observed. In ESI ion trap, signals at m/z=797.0 [M+2H].sup.2+ and m/z=531.7 [M+3H].sup.3+ were observed.

Example 8: [x4,C9]-chemerin-9 ($$ 13)

[0632] ##STR00034##

[0633] After automated synthesis of QFAFC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of YFP. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Kinetex 5 μm XB-C18 100 Å column employing a gradient of 25 to 55% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220n. The pure yield was 1.9 mg (11.1% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 20.5 min and 11.3 min, respectively. The chemical formula of the peptide is C.sub.56H.sub.68N.sub.10O.sub.12S.sub.2 (monoisotopic mass: 1136.5 Da, average mass: 1137.3 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1137.5 [M+H].sup.+, at m/z=1159.5 [M+Na].sup.+, and m/z=1175.5 [M+K].sup.+ were observed. In ESI ion trap, signals at m/z=1137.2 [M+H].sup.+ and m/z=569.2 [M+2H].sup.2+ were observed.

Example 9: [x4,W8,C9]-chemerin-9 ($$ 14)

[0634] ##STR00035##

[0635] After automated synthesis of QFAWC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of YFP. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 column employing a gradient of 30 to 60% B in A over 30 min with a flow rate of 15 mL/min. detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 2.0 mg (11.3% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Kinetex 5 μm biphenyl 100 Å column employing linear gradients of 20-70% B in A over 40 min with a flow rate of 1.0 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 20.6 min and 12.1 min, respectively. The chemical formula of the peptide is C.sub.58H.sub.69N.sub.11O.sub.12S.sub.2 (monoisotopic mass: 1175.5 Da, average mass: 1176.4 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF, signals at m/z=1176.5 [M+H].sup.+, and at m/z=1198.5 [M+Na].sup.+ were observed. In ESI ion trap, signals at m/z=1176.2 [M+H].sup.+ and m/z=588.8 [M+2H].sup.2+ were observed.

Example 10: [y1,x4,W8,C9]-chemerin-9 ($$ 15)

[0636] ##STR00036##

[0637] After automated synthesis of QFAWC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of yFP. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a gradient of 20 to 50% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 1.8 mg (10.2% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 0.6 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 22.7 min and 14.2 min, respectively. The chemical formula of the peptide is C.sub.58H.sub.69N.sub.11O.sub.12S.sub.2 (monoisotopic mass: 1175.46 Da; average mass: 1176.38 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF a signal at m/z=1176.5 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1176.2 [M+H].sup.+ and m/z=588.8 [M+2H].sup.2+ were observed.

Example 11: EG4-[x4,C9]-chemerin-9 ($$ 16)

[0638] ##STR00037##

[0639] After automated synthesis of QFAFC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of YFP. EG(4) was coupled to the N-terminus of the peptide by reaction of 5 equiv of Fmoc-15-amino-4,7,10,13-tetraoxapentadecanoic acid, HOBt and DIC in DMF overnight. The Fmoc-group was cleaved by reaction with 20% piperidine in DMF for 10 min, the reaction was repeated twice. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a gradient of 20 to 50% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 4.5 mg (21.6% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 0.6 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 21.9 min and 13.9 min, respectively. The chemical formula of the peptide is C.sub.67H.sub.89N.sub.11O.sub.17S.sub.2 (monoisotopic mass: 1383.59 Da; average mass: 1384.63 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF a signal at m/z=1384.6 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1384.3 [M+H].sup.+ and m/z=692.9 [M+2H].sup.2+ were observed.

Example 12: Tam-EG4-[x4,C9]-chemerin-9 ($$ 17)

[0640] ##STR00038##

[0641] After automated synthesis of QFAFC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of YFP. EG(4) was coupled to the N-terminus of the peptide by reaction of 5 equiv of Fmoc-15-amino-4,7,10,13-tetraoxapentadecanoic acid, HOBt and DIC in DMF overnight. The Fmoc-group was cleaved by reaction with 20% piperidine in DMF for 10 min, the reaction was repeated twice. The peptide was N-terminally modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam. HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a gradient of 20 to 50% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 1.8 mg (6.7% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 0.6 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 22.9 min and 14.9 min, respectively. The chemical formula of the peptide is C.sub.92H.sub.109N.sub.13O.sub.21S.sub.2 (monoisotopic mass: 1795.73 Da; average mass: 1797.07 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF a signal at m/z=1796.7 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1797.3 [M+H].sup.+ and m/z=599.7 [M+2H].sup.2+ were observed.

Example 13: EG4-[x4,W8,C9]-chemerin-9 ($$ 18)

[0642] ##STR00039##

[0643] After automated synthesis of QFAWC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of YFP. EG(4) was coupled to the N-terminus of the peptide by reaction of 5 equiv of Fmoc-15-amino-4,7,10,13-tetraoxapentadecanoic acid, HOBt and DIC in DMF overnight. The Fmoc-group was cleaved by reaction with 20% piperidine in DMF for 10 min, the reaction was repeated twice. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a gradient of 20 to 50% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 4.6 mg (22.2% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 0.6 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 21.4 min and 13.4 min, respectively. The chemical formula of the peptide is C.sub.67H.sub.89N.sub.11O.sub.17S.sub.2 (monoisotopic mass: 1383.59 Da; average mass: 1384.63 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF a signal at m/z=1384.6 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1384.3 [M+H].sup.+ and m/z=692.9 [M+2H].sup.2+ were observed.

Example 14: Tam-EG4-[x4,W8,C9]-chemerin-9 ($$19)

[0644] ##STR00040##

[0645] After automated synthesis of QFAFC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of YFP. EG(4) was coupled to the N-terminus of the peptide by reaction of 5 equiv of Fmoc-15-amino-4,7,10,13-tetraoxapentadecanoic acid, HOBt and DIC in DMF overnight. The Fmoc-group was cleaved by reaction with 20% piperidine in DMF for 10 min, the reaction was repeated twice. The peptide was N-terminally modified with 6-carboxytetramethylrhodamine (Tam) by reaction with 2 equiv of Tam, HATU and DIPEA in DMF overnight. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et.sub.2O/hexane (1:3). The precipitate was washed with Et.sub.2O and the peptide was incubated in TBS, 20% ACN. pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a gradient of 20 to 50% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 1.9 mg (6.69% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 0.6 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 22.4 min and 14.4 min, respectively. The chemical formula of the peptide is C.sub.92H.sub.109N.sub.13O.sub.21S.sub.2 (monoisotopic mass: 1795.73 Da; average mass: 1797.07 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF a signal at m/z=1796.7 [M+H].sup.+ was observed. In ESI ion trap, signals at m/z=1797.3 [M+H].sup.+ and m/z=599.7 [M+2H].sup.2+ were observed.

Example 15: GFLG-[x4,C9]-chemerin-9 ($$ 20)

[0646] ##STR00041##

[0647] After automated synthesis of QFAFC, D-homocysteine (x) was coupled by reaction of 5 equiv of HOBt, DIC and Fmoc-D-Homocysteine(Trt)-OH in DMF overnight, followed by automated synthesis of GFLGYFP. The N terminus of the peptide was acetylated on resin with Ac.sub.2O and DIPEA in DCM for 15 min. The peptide was treated with 90% TFA, 7% thioanisol, 3% ethanedithiol for 3 h to deprotect all side chains and cleave the peptide from the resin, followed by precipitation in ice-cold Et2O/hexane (1:3). The precipitate was washed with Et2O and the peptide was incubated in TBS, 20% ACN, pH 7.8 for 48 h to promote the formation of the intramolecular disulfide bond. The peptide was purified by RP-HPLC on a Aeris 3.6 μm 100 Å XB-C18 column employing a gradient of 20 to 50% B in A over 30 min with a flow rate of 15 mL/min, detecting the peptide by measuring the absorption at λ=220 nm. The pure yield was 1.7 mg (7.3% of theory). Purity was determined by RP-HPLC on a Jupiter 4 μm Proteo 90 Å C12 and on a Aeris 3.6 μm 100 Å XB-C18 column employing linear gradients of 20-70% B in A over 40 min with flow rates of 0.6 and 1.55 mL/min, respectively. The peptide showed over 95% purity as determined by the absorption at 220 nm on both columns, with retention times of 22.5 min and 14.1 min, respectively. The chemical formula of the peptide is C.sub.77H.sub.96N.sub.14O.sub.17S.sub.2 (monoisotopic mass: 1552.65 Da; average mass: 1553.82 Da). The observed masses were in correspondence to the calculated masses, confirming product identity: In MALDI-ToF a signal at m/z=1575.6 [M+Na].sup.+ was observed. In ESI ion trap, signals at m/z=1554.6 [M+H]+ and m/z=777.3 [M+2H]2+ were observed.

III Experimental Section—Biological Assays

Test Systems and Methods

Cell Culture

[0648] COS-7 and HEK293 cells were cultivated in DMEM supplemented with 10% FBS or DMEM/Ham's F12 supplemented with 15% FBS, respectively. All cells were maintained in T75 cell culture flasks at 37° C., 95% humidity and 5% CO.sub.2 (standard conditions).

1. Plasma Stability Assay

[0649] Investigation of peptide stability in blood plasma was carried out as described previously. (Hoppenz, Els-Heindl et al., A Selective Carborane-Functionalized Gastrin-Releasing Peptide Receptor Agonist as Boron Delivery Agent for Boron Neutron Capture Therapy. J Org Chem, 2020, 85(3): 1446-1457) Tam-labeled peptides were dissolved in human blood plasma at a concentration of 10.sup.−5 M and incubated at 37° C. and 250 rpm. Samples taken at the respective time points were added to a solution of 0.1% SDS in ACN/EtOH (1:1). After incubation at −20° C. for 20 min, the supernatant was transferred to a new tube and incubated again at −20° C. for at least 3 h. The solution was filtered by centrifugation using Costar Spin-X tubes (0.22 μm) and the filtrate was analyzed by RP-HPLC on a VariTide RPC. 6 μm, 200 Å column (Agilent technologies, Santa Clara, USA) employing a linear gradient of 15-65% (v/v) A in B over 40 min. The fluorescence of the peptide was detected at λ=573 nm. Peaks were integrated and the peak containing the intact peptide was normalized to the sample taken at t=0 min (100%). Plasma half-life was determined using one-phase decay in GraphPad Prism 5.03.

2. Calcium Mobilization Assay

[0650] COS-7 cells were transfected in 75 cm.sup.2 cell culture flasks with 12 μg of the hCMKLR1_eYFP_G.sub.αΔ6qi4myr_pV2 plasmid overnight using Metafectene Pro. Transfected cells were seeded in 96 well plates (100 μL cell suspension in DMEM+10% FBS/well) and incubated overnight. The following day, the Ca.sup.2+-mobilization was performed as described previously. (Hoppenz, Els-Heindl et al., A Selective Carborane-Functionalized Gastrin-Releasing Peptide Receptor Agonist as Boron Delivery Agent for Boron Neutron Capture Therapy. J Org Chem, 2020, 85(3): 1446-1457) Briefly, cells were incubated with Fluo-2-AM solution (2.3 μM Fluo-2-AM, 0.06% (v/v) Pluronic-F127 in assay buffer). After 1 h, the Fluo-2-am solution was replaced with assay buffer (20 mM HEPES, 2.5 mM Probenecid in HBSS, pH 7.5) and the basal Ca.sup.2-level was measured for 20 s with a Flexstation 3 (λ.sub.ex=485 nm, λ.sub.em=525 nm). The ligand was added, and Ca.sup.2+-response was measured for 40 s. The resulting maximum over basal values were calculated for each well, and normalized to the top and bottom values of the control curve (chemerin-9 $$1). All experiments were performed in triplicates, each experiment was repeated at least twice. Nonlinear regression was calculated using GraphPad Prism 5.

3. Bioluminescence Resonance Energy Transfer (BRET)

[0651] HEK293 cells were transiently transfected using 75 cm.sup.2 cell culture flasks with cell monolayers (confluency of ˜80%). Plasmid DNA of the C-terminally eYFP tagged human CMKLR1 and the chimeric G protein G.sub.αΔ6qi4myr in a pVitro2 vector (7.8 μg) and Renilla-luciferase 8-tagged Arrestin 3 in pcDNA3 (0.2 μg) and 24 μl MetafectenePro were separately added to 900 μl DMEM/Ham's F12 and incubated for 10 min before unification and incubation at RT for 20 min. 6 ml DMEM/Ham's F12 with 15% FCS at 37° C. was added on the cell monolayer. The plasmid solution was added and cells were incubated overnight before seeding. Cell seeding was carried out in white 96-well polystyrene cell culture microplates, coated with poly-D lysine. Transfected cells were detached with 1 ml trypsin/EDTA, 21 ml DMEM/Ham's F12 with 15% FCS was added and 100,000-200.000 cells in 100 μl per well were seeded. Afterwards, the cells were incubated overnight at 37° C. The assay was performed under unsterile conditions. First, medium was displaced with 100 μl BRET buffer (HBSS, 25 mM HEPES. pH 7.3) and 50 μl of luciferase substrate coelenterazine-h (final concentration of 4.2 μM) was added. Afterwards, the cells were stimulated with the peptides in different concentrations (10.sup.−5 to 10.sup.−12 M) dissolved in BRET buffer. 50 μl of the peptide dilution were used for cell stimulation. Buffer without peptide was used as a negative control. BRET effect was measured 15 min after agonist addition with a Tecan infinite plate reader using two filter sets at 37° C. (luminescence filter 400 nm-470 nm and fluorescence filter 505 nm-590 nm) and plotted as a function of fluorescence/luminescence ratio. The values of the negative control were subtracted, non-linear regression was calculated using GraphPad Prism. The curves were normalized to the positive control wild type chemerin 9 ($$1). All measurements were performed in four technical replicates, all experiments were repeated at least twice.

4. Fluorescence Microscopy

[0652] Cellular arrestin 3 recruitment was verified and CMKLR1 receptor uptake was tested in HEK293 cells. Ibidi 15μ-slides were coated with poly D-lysine before cell seeding. Cells were washed with DPBS prior to detachment with 1 ml trypsin/EDTA. A Neubauer chamber was used to count the amount of cells/ml medium after addition of 9 ml DMEM/Ham's F12 with 15% FCS. The cell suspension was diluted to 140,000 cells/200 μl, which were seeded. Incubation was carried out overnight at 37° C. Afterwards, cells were transiently transfected. Plasmid DNA of hCMKLR1_eYFP_G.sub.αΔ6qi4myr_pV2 (0.9 μg) and mCherry_Arr3_pcDNA3 (0.1 μg) and 8 μl Lipofectamine were separately added to 100 μl DMEM/Ham's F12 and incubated for 10 min before unification and incubation at RT for 20 min. Incubation was carried out overnight at 37° C. Then, the cells were starved with 200 μl OptiMEM and 1 μl of Hoechst 33342 for 30 min. The medium was replaced by 200 μl of OptiMEM and the to status was documented. The OptiMEM was then replaced by 200 μl of 1 μM peptide in OptiMEM. Fluorophore excitation was analyzed using different filters, depending on the emission wavelength of the fluorophore and the time of exposure was adjusted to each fluorophore individually. All images were processed identically with the AxioVision software (Carl Zeiss AG, Oberkochen, Germany).

TABLE-US-00006 TABLE 1 Filter sets (Zeiss) used for fluorophore detection. Fluorophore Used for labeling Excitation [nm] Emission [nm] Hoechst33342 cell nucleus 352 455 mCherry arrestin 3 549 577 YFP hCMKLR1 514 526 Hoechst33342: 2′-(4-Ethoxyphenyl)-6-(4-methyl-1-piperazinyl)-1H,3′H-2,5′-bibenzimidazole; YFP: yellow fluorescent protein; hCMKLR1: chemokine like receptor 1;

Results

Activity Studies in Ca.SUP.2+ Mobilization Assay

[0653] To test the ability of the synthesized peptides to induce G protein-mediated signaling at the CMKLR1, a Ca.sup.2+ mobilization assay was performed. All tested cyclic chemerin-9 variants showed G protein-signaling to various extends (Table 2). The most active cyclic derivate Example 8: [x4,C9]-chemerin-9 ($$ 13) showed a 2-fold higher activity than linear chemerin-9 (comparison example 1). Introducing a tryptophan in position 8 increased the activity even more ($$14), but combining modifications Trp.sup.8 and D-Tyr.sup.1 in one peptide led to a significantly decreased activity ($$15).

TABLE-US-00007 TABLE 2 Activity data of chemerin-9 derivates at the CMKLR1 in a Ca.sup.2+ mobilization assay. Cpd Code EC.sub.50 [nM] pEC50 ± SEM $$ 1 Comparison Example 1: Chemerin-9 ($$ 1) 10 8.021 ± 0.039 $$ 4 Comparison Example 4: [c4,C9]-chemerin-9 ($$ 4) 64 7.192 ± 0.147 $$ 6 Comparison Example 5: [c4,C7]-chemerin-9 ($$6) 63 7.204 ± 0.106 $$ 7 Example 2: [c4,C9-TA]-chemerin-9 ($$ 7) 13 7.888 ± 0.130 $$ 9 Example 4: [c4,X9-TA]-chemerin-9 ($$ 9) 37 7.429 ± 0.110 $$ 11 Example 6: [y1,c4,X9-TA]-chemerin-9 ($$ 11) 28 7.553 ± 0.067 $$ 13 Example 8: [x4,C9]-chemerin-9 ($$ 13) 5 8.259 ± 0.074 $$ 14 Example 9: [x4,W8,C9]-chemerin-9 ($$ 14) 3 8.499 ± 0.058 $$ 15 Example 10: [y1,x4,W8,C9]-chemerin-9 ($$ 15) 207 6.683 ± 0.101 $$ 16 Example 11: EG4-[x4,C9]-chemerin-9 ($$16) 17 7.766 ± 0.226 $$ 18 Example 13: EG4-[x4,W8,C9]-chemerin-9 ($$ 18) 193 6.714 ± 0.088 $$ 20 Example 15: GFLG-[x4,C9]-chemerin-9 ($$ 20) 21 7.676 ± 0.221 X = Homocysteine, x = D-Homocysteine, TA = thioacetal.

Plasma Stability Studies

[0654] The stability of the different peptides in blood plasma was investigated for Tam-modified derivates to enable monitoring the degradation of the peptides in RP-HPLC. All N-terminally Tam-labeled cyclic chemerin-9 variants ($$5, $$8, $$9) were completely stable in blood plasma over a time period of 24 h. Introducing the Tam-label at a side chain while simultaneously inducing an N-terminal D-Tyr gave the equally stable Example 7: [y1,c4,K7(Tam),C9]-chemerin-9 ($$12). This demonstrates that all cyclic variants with an N-terminal stabilization can be expected to be stable in blood plasma. This was verified by testing the chemerin-9 variants bearing an N-terminal ethylene glycol linker ($$17, $$19), both were completely stable for at least 48 h).

TABLE-US-00008 TABLE 3 Plasma stability of Tam-modified chemerin-9 derivates. Cpd Code t.sub.1/2 [h] $$ 2 Comparison Example 2: Tam-chemerin-9 ($$ 2) <0.2 $$ 5 Example 1: Tam-[c4,C9]-chemerin-9 ($$ 5) >24 $$ 8 Example 3: Tam-[c4,C9-TA]-chemerin-9 ($$ 8) >24 $$ 10 Example 5: Tam-[c4,X9-TA]-chemerin-9 ($$ 10) >24 $$ 12 Example 7: [y1,c4,K7(Tam),C9]-chemerin-9 ($$ 12) >24 $$ 17 Example 12: Tam-EG4-[x4,C9]-chemerin-9 ($$ 17) >48 h $$ 19 Example 14: Tam-EG4-[x4,W8,C9]-chemerin-9 ($$ 19) >48 h

Internalization Studies in BRET and Microscopy

[0655] Arrestin recruitment is the first step in the internalization process of GPCR that follows activation of the G protein pathway. The bioluminescence resonance energy transfer (BRET) assay was used to determine the potency of cyclic chemerin variants to recruit arrestin 3 to the CMKLR1 receptor after stimulation. HEK293 cells were transiently transfected with CMKLR1 receptor fused with eYFP fluorophore and Arrestin 3 tagged with Rluc8 luciferase. The transfected cells were seeded, incubated with the luciferase substrate coelenterazine-h and stimulated with different peptide concentrations, resulting in measurable BRET signals. The sigmoidal concentration-response-curves were normalized to the positive control WT chemerin 9 ($$1). In the BRET assays, it could be shown that all but one of the tested peptides are still able to induce arrestin 3 recruitment to the activated CMKLR1 receptor with different potencies (Table 1).

TABLE-US-00009 TABLE 1 Arrestin recruitment data of selected peptides obtained in a BRET based assay. All data was obtained from at least two individual experiments and normalized to the control compound chemerin-9 ($$ 1). EC.sub.50 pEC.sub.50 ± E.sub.max Cpd Code [nM] SEM [%] ± SEM ($$ 1) Comparison Example 1: Chemerin-9 ($$ 1) 37  7.4 ± 0.05 96 ± 2 ($$ 13) Example 8: [x4,C9]-chemerin-9 ($$ 13) 78 7.1 ± 0.1 86.8 ± 4.sup.  ($$ 14) Example 9: [x4,W8,C9]-chemerin-9 ($$14) 158 6.8 ± 0.2  97 ± 10 ($$ 15) Example 10: [y1,x4,W8,C9]-chemerin-9 ($$ 15) — — — ($$ 16) Example 11: EG4-[x4,C9]-chemerin-9 ($$ 16) 101  6.8 ± 0.08 102 ± 5  ($$ 18) Example 13: EG4-[x4,W8,C9]-chemerin-9 ($$18) 418 6.4 ± 0.2 69 ± 8 ($$ 20) Example 15: GFLG-[x4,C9]-chemerin-9 ($$ 20) 100 7.0 ± 0.2 89 ± 5 x = D-homocysteine

[0656] Cyclization of chemerin 9 by a disulfide bond ($$13) leads to only slightly shifted EC.sub.50 and E.sub.max values compared with wild type chemerin 9. An additional exchange at position 8 to tryptophan is also accepted ($$14). Interestingly, a further modification with D-tyrosine at position 1 leads to a peptide that is no longer able to induce arrestin 3-recruitment, despite its activity in G protein activation in a Ca.sup.2+ assay ($$15), making it a biased ligand. Elongation of the N-terminus of cyclic peptides with an ethylene glycol linker ($$16, $$18) or short peptide linker ($$20) is accepted with respect to arrestin-recruitment.

[0657] FIG. 1: Arrestin 3 recruitment to the CMKLR1 after stimulation with chemerin-9 variants. Impact of cyclization and amino acid substitutions on the activity chemerin-9

[0658] Impact of cyclization and amino acid substitutions on the activity chemerin-9 is shown in FIG. 1. A) Cyclization ($$13) leads to a slight loss of activity compared to linear chemerin-9 ($$1). Exchanging position 8 for a tryptophan ($$14) has no impact, while changing position 8 for tryptophan and position 1 for D-tyrosine completely abolishes arrestin recruitment ($$15). B) N-terminal elongation of cyclic peptides with either polyethylene glycol or peptide linker has no impact ($$16, $$ 20), unless a tryptophane is present in position 8 ($$18).

[0659] To verify the arrestin-recruitment and analyze the internalization of the CMKLR1 receptor itself, HEK293 cells were used and transiently transfected with fluorescent labeled variants of the two molecules. These cells express the human CMKLR1 fused to a C terminally yellow fluorescent protein (YFP) and arrestin 3 with a red fluorescent mCherry protein.

[0660] FIG. 2: Internalization of CMKLR1 and arrestin 3 recruitment. HEK293 cells expressing the hCMKLR1 (green) and arrestin 3 (Arr3; red) due to transient transfection were used for the internalization studies. Inter-nalization of the receptor was observed for 30 min stimulation with 1 μM via fluorescence microscopy after nuclei staining (blue). Representative pic-tures for time point 15 min were chosen and processed identically with the AxioVision software; scale bar: 15 μm; n≥2;

[0661] Without stimulation, the receptor was located at the cell membrane and the arrestin was distributed in the cytosol (FIG. 2, 0 min). After stimulation with chemerin-9 (comparison example 1, $$1), arrestin 3 was recruited to the CMKLR1 receptor followed by internalization of the CMKLR1-arrestin complex. A similar behavior was demonstrated for the cyclic variant Example 8: [x4,C9]-chemerin-9 ($$13), Example 9: [x4,W8,C9]-chemerin-9 ($$14) and Example 11: EG4-[x4,C9]-chemerin-9 ($$16). Example 13: EG4-[x4,W8,C9]-chemerin-9 ($$18) shows good arrestin 3 recruitment, but slightly lower receptor internalization compared to chemerin-9 (comparison example 1, $$1). In contrast, neither internalization of the hCMKLR1 nor arrestin 3 recruitment could be detected for Example 10: [y1,x4,W8,C9]-chemerin-9 ($$15). Thus, the bias behavior of this compound was verified. These results obtained in fluorescence microscopy confirm the results obtained in the BRET-based assay for all peptides.