the following examples serve to illustrate the invention without restricting it . ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - hci 6 - nle 7 - arg 8 - pro 9 - d - ala 10 - nh 2 the synthesis was carried out according to a solid - phase flow diagram ( peptide synthesis protocol , p . 11 ) with dic / hobt coupling , starting from 3 . 3 g of mbha resin ( loading density 1 . 08 mmol / g ). after hf cleavage from the polymeric support , 3 . 4 g of crude peptide were obtained , which were purified by standard processes of preparative hplc . after subsequent freeze - drying , 1 . 43 g of hplc - uniform product of the empirical formula c 72 h 96 n 17 o 14 cl having correct fab - ms : 1458 . 7 ( m + h + ) ( calc : 1457 . 7 ), and corresponding 1 h - nmr spectrum were obtained . 1 h - nmr ( 500 mhz , d 2 o / dmso - d 6 , δ in ppm ): 8 . 7 to 7 . 2 , several m , arom . h and incompletely exchanged nh ; 6 . 92 and 6 . 58 , 2d , 2 × 2h , arom . h p - cl - phe ; 5 . 2 to 3 . 5 , several m , cα - h and aliph . h ; 3 . 2 to 2 . 6 , several m , aromat . cβ - h 2 . 1 to 0 . 7 , several m , residual aliphat . h ; 1 . 70 , s , 3h , acetyl ; 1 . 20 , d , 3h , cβ - h ala ; 0 . 8 , m , cδ - h leu ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - lys ( b ) 6 - leu 7 - lys ( ipr ) 8 - pro 9 - d - ala 10 - nh 2 the synthesis was carried out according to a flow diagram ( peptide synthesis protocol , p . 11 ) with dic / hobt coupling , starting from 4 . 0 g of mbha resin ( loading density 1 . 11 mmol / g ). after hf cleavage from the polymeric support , 4 . 87 g of crude peptide were obtained , which were purified by standard processes of preparative hplc . after subsequent freeze - drying , 0 . 93 g of hplc - uniform product was obtained , which was reacted with 4 - amidinophenylamino - 4 - oxobutyric acid in the presence of bop as a coupling reagent to give the desired compound . after fresh hplc purification , 148 mg of target compound of the empirical formula c 85 h 112 n 17 o 15 — cl having correct esi - ms : 1647 . 6 ( m + h + ) ( calc : 1645 . 8 ), and corresponding 1 h - nmr spectrum were obtained . 10 . 4 , s , 1h and 9 . 13 , s , 2h , and 8 . 94 , s , 2h , nhs of 4 - amidinoaniline ; 8 . 6 to 7 . 35 , several m , arom . h and nh ; 7 . 22 and 7 . 18 , 2d , 4h , arom . h ( pcl ) phe ; 6 . 95 and 6 . 58 , 2d , 4h , arom . h tyr ; 5 . 2 to 3 . 5 , several m , cα - h and aliphat . h ; 3 . 3 to 2 . 4 , several m , cβ - h and n — ch 3 ; 2 . 1 to 1 . 1 , several m , residual aliphat . h ; 1 . 68 , s , 3h , acetyl ; 1 . 20 , d , 3h , cβ - h ala ; 0 . 83 , dd , 6h , cδ - h leu ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - lys ( b ) 6 - leu 7 - arg 8 - pro 9 - d - ala 10 - nh 2 the synthesis was carried out according to a solid - phase flow diagram ( peptide synthesis protocol , p . 11 ) with dic / hobt coupling , starting from 4 . 0 g of mbha resin ( loading density 0 . 97 mmol / g ). after hf cleavage from the polymeric support , 4 . 0 g of crude peptide were obtained , which were purified by standard processes of preparative hplc . after subsequent freeze - drying , 1 . 39 g of hplc - uniform product were obtained , which were reacted with 4 - amidinophenylamino - 4 - oxobutyric acid in the presence of bop as a coupling reagent to give the desired compound . after fresh hplc purification , 440 mg of target compound of the empirical formula c 82 h 106 n 19 o 15 cl having correct esi - ms : 1632 . 7 ( m + h + ) ( calc : 1631 . 7 ), and corresponding 1 h - nmr spectrum were obtained . 10 . 4 , s , 1h and 9 . 15 , s , 2h , and 9 . 0 , s , 2h , nhs of 4 - amidinoaniline ; 8 . 60 , m , 2h , arom . h ; 8 . 3 to 7 . 2 , several m , arom . h and nh ; 7 . 27 and 7 . 20 , 2d , 4h , arom . h ( pcl ) phe ; 6 . 96 and 6 . 60 , 2d , 4h , arom . h tyr ; 5 . 2 to 3 . 5 , several m , cα - h and aliphat . h ; 3 . 2 to 2 . 4 , several m , cβ - h and n — ch 3 ; 2 . 1 to 1 . 1 , several m , residual aliphat . h ; 1 . 70 , s , 3h , acetyl ; 1 . 20 , d , 3h , cβ - h ala ; 0 . 85 , dd , 6h , cδ - h leu ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - hci 6 - nle 7 - lys ( ipr ) 8 - pro 9 - d - ala 10 - nh 2 the synthesis was carried out according to a solid - phase flow diagram ( peptide synthesis protocol , p . 11 ) with dic / hobt coupling , starting from 2 . 5 g of mbha resin ( loading density 1 . 08 mmol / g ). after hf cleavage from the polymeric support , 2 . 78 g of crude peptide were obtained , which were purified by standard processes of preparative hplc . after subsequent freeze - drying , 400 mg of hplc - uniform product of the empirical formula c 75 h 102 n 15 o 14 cl having correct esi - ms : 1472 . 6 ( m + h + ) ( calc : 1471 . 7 ), and corresponding 1 h - nmr spectrum were obtained . 1 h - nmr ( 500 mhz , d 2 o / dmso - d 6 , δ in ppm ): 8 . 62 , m , 2h , 8 . 30 , m , 2h , 7 . 80 , m , 4h , 7 . 66 , s , 1h , 7 . 47 , m , 2h , 7 . 36 , d , 1h , aromat . h ; 7 . 25 and 7 . 20 , 2 d , 4h , arom . h ( pcl ) phe ; 6 . 96 and 6 . 63 , 2d , 4h , aromat . h tyr ; 5 . 10 to 4 . 0 , several m , cα - h and aliphat . h ; 3 . 75 to 2 . 65 , several m , cβ - h and n - ch 3 ; 2 . 1 to 1 . 05 , several m , residual aliphat . h ; 1 . 74 , s , 3h , acetyl ; 1 . 23 , d , 3h , cβ - h ala ; 1 . 20 , m , ch 3 isoprop . ; 0 . 8 , m , 3h , cδ - h nle ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - hci 6 - nle 7 - lys ( ipr ) 8 - pro 9 - sar 10 - nh 2 the synthesis was carried out according to a solid - phase flow diagram ( peptide synthesis protocol , p . 11 ) with dic / hobt coupling , starting from 2 . 5 g of mbra resin ( loading density 1 . 08 mmol / g ). after hf cleavage from the polymeric support , 2 . 74 g of crude peptide were obtained , which were purified by standard processes of preparative hplc . after subsequent freeze - drying , 840 mg of hplc - uniform product of the empirical formula c 75 h 102 n 15 o 14 cl having correct esi - ms : 1472 . 6 ( m + h + ) ( calc : 1471 . 7 ), and corresponding 1 h - nmr spectrum were obtained . 1 h - nmr ( 500 mhz , d 2 o / dmso - d 6 , δ in ppm ): 8 . 6 , m , 2h , 8 . 3 , m , 2h , 7 . 85 , m , 2h , 7 . 8 , m , 2h , 7 . 65 , s , 1h , 7 . 46 , m , 2h , 7 . 35 , d , 1h , aromat . h ; 7 . 23 and 7 . 17 , 2 d , 4h , arom . h ( pcl ) phe ; 7 . 0 and 6 . 6 , 2d , 4h , aromat . h tyr ; 5 . 10 to 3 . 8 , several m , cα - h and aliphat . h ; 3 . 75 to 2 . 6 , several m , cβ - h and n - ch 3 ; 2 . 2 to 1 . 05 , several m , residual aliphat . h ; 1 . 70 , s , 3h , acetyl ; 1 . 23 , d , 3h , cβ - h ala ; 1 . 20 , m , ch 3 isoprop . ; 0 . 8 , m , 3h , cδ - h nle 3 -( 4 - fluorophenyl ) propionyl - d - nal ( 1 ) 1 - ser 4 - n - me - tyr 5 - d - lys ( atz ) 6 - leu 7 - arg 8 - pro 9 - d - ala 10 - nh 2 the synthesis was carried out according to a solid - phase flow diagram ( peptide synthesis protocol , p . 11 ) with dic / hobt coupling , starting from 9 . 2 g of mbha resin ( loading density 1 . 08 mmol / g ). after hf cleavage from the polymeric support , 5 . 8 g of crude peptide were obtained , which were purified by standard processes of preparative hplc . after subsequent freeze - drying , 2 . 0 g of hplc - uniform unsubstituted octapeptide were obtained , of which 0 . 4 mmol was reacted with 0 . 5 mmol of 3 - amino - 1 , 2 , 4 - trizole - 5 - carboxylic acid in the presence of pybop as a coupling reagent to give 790 mg of crude product of the desired compound . after fresh hplc purification , 200 mg of target compound of the empirical formula c 64 h 86 n 17 o 12 f having correct fab - ms : 1304 . 6 ( m + h + ) ( calc : 1303 . 6 ) were obtained . 1 h - nmr ( 500 mhz , d 2 o / dmso - d 6 , δ in ppm ): 8 . 14 , m , 1h , 7 . 90 , m , 1h , 7 . 80 , m , 1h , 7 . 50 , m , 2h , 7 . 35 , m , 2h , 7 . 0 , m , 6h , 7 . 63 , m , 2h , aromat . h ; 5 . 0 , m , 1h , 4 . 83 , m , 2h , 4 . 41 , m , 1h , 4 . 30 – 4 . 05 , several m , 4h , cα - h ; 3 . 66 to 2 . 25 , several m , aliphat . and aromat . side - chain h ; 2 . 95 , s , and 2 . 75 , s , n - me ; 2 . 05 to 1 . 1 , several m , residual aliphat . h ; 1 . 20 , d , cβ - h ala ; 0 . 75 , m , 6h , cδ - h leu ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - lys ( b ) 6 - nle 7 - arg 8 - pro 9 - sar 10 - nh 2 the synthesis of the decapeptide was carried out on a polymeric support with a loading density of 0 . 55 mmol / g ( aminomethyl - substituted resin , fmoc protection , type d - 1675 , bachem ). lysine was coupled as fmoc - d - lys ( boc )- oh , and the fmoc protective groups were removed using 20 % piperidine / dmf . after simultaneous removal of all side - chain protective groups and detachment from the polymeric support , the isolated crude peptide was purified by means of preparative hplc . after freeze - drying , 98 . 5 % pure decapeptide was obtained . the substitution on the ε nitrogen of d - lysine with 4 -( 4 - aminophenol ) amino - 1 , 4 - dioxobutyric acid was carried out using pybop in dmf with addition of dipea . the purification of the isolated crude peptide was carried out by means of preparative hplc . the subsequent freeze - drying afforded about 99 % pure product ( trifluoroacetate ) of the empirical formula c82h106cln19o15 having correct fab - ms of 1632 ( m + h ) ( calc : 1631 . 78096 ) ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - lys ( b ) 6 - nle 7 - arg 8 - pro 9 - d - ala 10 - nh 2 the synthesis of the decapeptide was carried out on a polymeric support with a loading density of 0 . 55 mmol / g ( aminomethyl - substituted resin , fmoc protection , type d - 1675 , bachem ). lysine was coupled as fmoc - d - lys ( boc )- oh , and the fmoc protective groups were removed using 20 % piperidine / dmf . after simultaneous removal of all side - chain protective groups and detachment from the polymeric support , the isolated crude peptide with a purity of about 71 % ( hplc ) was reacted further without purification . the side - chain substitution of d - lysine with 4 -( 4 - aminophenol ) amino - 1 , 4 - dioxobutyric acid was carried out using pybop in dmf with addition of dipea . the isolated crude peptide was purified by means of preparative hplc . after subsequent freeze - drying , a 98 . 8 % pure product ( trifluoroacetate ) of the empirical formula c 82 h 106 cln 19 o 15 having correct fab - ms of 1632 ( m + h ) ( calc : 1631 . 78096 ) was obtained . ac - d - nal ( 2 ) 1 - d - cpa 2 - d - pal ( 3 ) 3 - ser 4 - n - me - tyr 5 - d - lys ( b ) 6 - nle 7 - lys ( ipr ) 8 - pro 9 - sar 10 - nh 2 the synthesis of the decapeptide was carried out on a polymeric support with a loading density of 0 . 55 mmol / g ( aminomethyl - substituted resin , fmoc protection , type d - 1675 , bachem ). lysine was coupled as fmoc - d - lys ( boc )- oh , and the fmoc protective groups were removed using 20 % piperidine / dmf . after simultaneous removal of all side - chain protective groups and detachment from the polymeric support , the isolated crude peptide ( concentration about 59 %, hplc ) was purified by means of preparative hplc . after freeze - drying , 95 % pure decapeptide was obtained . the side - chain substitution of d - lysine with 4 -( 4 - aminophenol ) amino - 1 , 4 - dioxobutyric acid was carried out using pybop in dmf with addition of dipea . the isolated crude peptide was purified by means of preparative hplc . after subsequent freeze - drying , a 96 . 6 % pure product ( trifluoroacetate ) of the empirical formula c 85 h 112 cln17o 15 having correct fab - ms of 1646 ( m + h ) ( calc : 1645 . 8218 ) was obtained . the compounds according to formula i according to the invention were investigated for their receptor binding . the process closely followed the process described in beckers et al ., eur . j . biochem . 231 , 535 – 543 ( 1995 ). cetrorelix obtained according to the synthesis disclosed above was iodinated with [ 125 i ] ( amersham ; specific activity 80 . 5 bq / fmol ) using the iodogen reagent ( pierce ). the reaction mixture was purified by reverse - phase high - performance liquid chromatography , monoiodinated cetrorelix being obtained without unlabelled peptide . in each case , about 80 % of the [ 125 i ]- cetrorelix and the unlabelled compound according to the invention were suitable for the specific receptor association . the compounds according to the invention can be tested for their in - vitro action using the following methods 1 and 2 , the binding affinities in the binding assay being determined with [ 125 i ]- cetrorelix ( method 1 ) and the functional activities being determined with triptorelin as an agonist stimulus ( method 2 ). receptor binding assay according to beckers , t ., marheineke , k ., reiländer , h ., hilgard p . ( 1995 ) “ selection and characterization of mammalian cell lines with stable overexpression of human pituitary receptors for gonadoliberin ( gnrh )” eur . j . biochem . 0 . 231 , 535 – 543 . for investigation of the receptor binding , cetrorelix was iodinated using the iodogen reagent ( pierce ) with [ 125 i ] ( amersham ; 80 . 5 bq / fmol specific activity ). the reaction mixture was purified by high - performance liquid chromatography with exchanged phases , monoiodinated cetrorelix being obtained without unlabelled peptide . about 80 % of the [ 125 i ] cetrorelix was capable of specific receptor association . the receptor binding assay was carried out under physiological conditions as described ( beckers et al ., 1995 ) using intact cells . subconfluent cultures of stably transfected ltk − cells , which express the human lhrh receptor , were separated off by incubation in nacl / p i ( 137 mm nacl , 2 . 7 mm kcl , 8 . 1 mm na 2 hpo 4 , 11 . 47 mm kh 2 po 4 )/ 1 mm edta and collected by centrifugation . the cell pellet was resuspended in binding buffer ( dmem without h 2 co 3 , with 4 . 5 g / l of glucose , 10 mm hepes ph 7 . 5 , 0 . 5 % ( mass / volume ) bsa , 1 g / l bacitracin , 0 . 1 g / l sbti , 0 . 1 % ( mass / volume ) nan 3 ). for displacement assays , 0 . 25 × 10 6 cells / 100 μl were incubated with approximately 225 pm of the [ 125 i ]- cetrorelix ( specific activity 5 – 10 × 10 5 dpm / pmol ) and various concentrations of unlabelled compound according to the invention as competitor . the cell suspension in 100 μl of binding medium was layered in 400 μl assay tubes over 200 μl of 84 % by volume silicone oil ( merck type 550 )/ 16 % by volume paraffin oil . after incubation for 1 h at 37 ° c . with slow , continuous shaking , the cells were separated from the incubation medium by centrifugation for 2 min at 9000 rpm ( rotor type hta13 . 8 ; heraeus sepatec , osterode / germany ). the tips of the tubes which contained the cell pellet were cut off . cell pellet and supernatants were then analysed by counting the γ radiation . the amount of non - specifically bound material was determined at a final concentration of 1 μm with inclusion of unlabelled cetrorelix and was typically ≦ 10 % of the total bound material . the analysis of the binding data was carried out using the ebda / ligand analysis programme ( biosoft v3 . 0 ). the assay was carried out , provided with some modifications , as described in beckers , t ., reiländer , h ., hilgard , p . ( 1997 ) “ characterization of gonadotropin - releasing hormone analogs based on a sensitive cellular luciferase reporter gene assay ”, analyt . biochem . 251 , 17 – 23 ( beckers et al ., 1997 ). 10 , 000 cells per well , which express the human lhrh receptor and a luciferase reporter gene , were cultured for 24 h in microtitre plates using dmem with additives and 1 % ( v : v ) fcs i . the cells were then stimulated with 1 nm [ d - trp 6 ] lhrh for 6 h . antagonistic compounds according to the invention were added before the stimulation and the cells were lysed at the end for the quantification of the cellular luc activity . the calculation of the ic 50 values from dose - effect curves was carried out by non - linear regression analysis using the hill model ( programme edx 2 . 0 from c . grunwald , arzneimittelwerk dresden ). the quantification of the luc activity was carried out in duplicate essentially as described ( promega technical bulletins # 101 / 161 ) using the respective luciferase assay system ( promega e4030 ). owing to addition of coenzyme a ( coa ), an oxidation of luciferyl - coa takes place with advantageous kinetics . after the removal of the culture medium from the microtitre plate , the cells were lysed by addition of 100 μl of lysis buffer ( 25 mm tris - phosphate ph 7 . 8 , 2 mm dithiothreitol , 2 mm 1 , 2 - diaminocyclohexane - n , n , n ′, n ′- tetraacetic acid ( cdta ), 10 % ( v : v ) glycerol , 1 % ( v : v ) triton x - 100 ). after incubation at room temperature for 15 min , 10 μl of cell lysate were transferred into a white microtitre plate suitable for luminometric detection ( dynatech ). the enzymatic reaction was initiated by addition of 50 μl of assay buffer ( 20 mm tricine ph 7 . 8 , 1 . 07 mm ( mgco 3 ) 4 mg ( oh ) 2 , 2 . 67 mm mgso 4 , 0 . 1 mm ethylenediaminetetraacetic acid ( edta ), 33 . 3 mm dithiothreitol , 270 μm coenzyme a , 470 μm glow - worm ( photinus pyralis ) luciferin , 530 μm ratpna 2 ). after one minute , the luminescence was determined for a total time of one second with a signal half - life of five minutes using the eg & amp ; g berthold microlumat lb 96 p . in this way , the following in - vitro data were obtained , k d being the binding affinities and ic 50 being the functional activity and pm being picomoles per liter :
the invention relates to peptides which contain n - methylated amino acid units and have improved water solubility . medicaments in which the peptides according to the invention are contained can be used for the treatment of hormone - dependent tumors and hormone - influenced non - malignant disorders .