EP1268554A2

EP1268554A2 - Diagnostic and medicament for analysing the cell surface proteome of tumour and inflammatory cells and for treating tumorous and inflammatory diseases, preferably using a specific chemokine receptor analysis and the chemokine receptor-ligand interaction

Info

Publication number: EP1268554A2
Application number: EP01923710A
Authority: EP
Inventors: Wolf-Georg Forssmann; Knut Adermann; Aleksandra Heitland; Nikolaj Spodsberg
Original assignee: IPF Pharmaceuticals GmbH
Current assignee: IPF Pharmaceuticals GmbH
Priority date: 2000-03-31
Filing date: 2001-04-02
Publication date: 2003-01-02
Also published as: AU2001250412A1; US20030186889A1; WO2001072830A2; JP2003530325A; WO2001072830A3

Abstract

The invention relates to the production of a medicament and a diagnostic agent, obtained by a proteome analysis, preferably containing at least two different chemokine receptor ligands or chemokine receptor antibodies and in addition, to the use of at least two different chemokine receptor ligands, chemokine receptor antibodies and/or two different chemokine receptors. The medicament preferably contains as inhibitors, ligands or antibodies of at least two chemokine receptors or of the related algorithms of the surface chemokine receptor proteome. The invention also relates to at least one chemokine receptor ligand and/or a chemokine receptor, peptides and antibodies and to the use of the same for diagnosing and treating tumorous and inflammatory diseases. Similarly, clusters of analysed tumour cell surface proteomes, such as ectoproteases, adhesion molecules or different receptor types can be used. The invention relates to a method for diagnostic and therapeutic purposes and for the medicinal and commercial use of chemokines and their corresponding receptors and their antagonists including antibodies, for inhibiting cancer growth and its metastatic spread and for repressing inflammatory and auto-immune diseases. The method is based on the finding that chemokines act on specific tumour and inflammatory cells in an autocrine, paracrine and endocrine manner by means of the disease-specific constellation of chemokine receptor proteome. Primary and secondary tumours and specific inflammatory cells are controlled in their migration and proliferation behaviour. The diagnostic detection of a local increase in expressed and regulated chemokines and of the presence of the chemokine receptor compositions allows the significant repression or complete prevention of cancer growth, tumour metastatic spread and both inflammatory and auto-immune diseases.

Description

Diagnostic and medicinal products for the investigation of the cell surface proteome of tumor and inflammation cells as well as for the treatment of

Do diseases and inflammatory diseases preferably with the help of a specific chemokine receptor analysis and the chemokine receptor

Ligand interaction.

The present invention relates to the preparation of a medicament and a diagnostic agent obtained via a proteome analysis, preferably containing at least two different chemokine receptor ligands or chemokine receptor antibodies, and furthermore the use of at least two different chemokine receptor ligands, chemokine receptor antibodies and / or two different chemokine receptors. The medicament consists in the use of inhibitors, ligands or antibodies of at least two chemokine receptors or the associated algorithms of the surface chemokine receptor proteome as well as the use of at least one chemokine receptor ligand and / or a chemokine receptor, peptides and antibodies and their use for the diagnosis and therapy of tumor diseases and inflammatory diseases. In an analogous manner, clusters of analyzed tumor cell surface proteomes such as ectoproteases, adhesion molecules or various types of receptors can be used.

Recently, it has become increasingly clear that substances of regulation of cell growth and cell migration are of great diagnostic and therapeutic interest and that there are likely to be a large number of unknown factors which control cancer growth, with primary and secondary tumors and the colonized organs with one another to interact. Surprisingly, these factors are of clinically usable importance. These include in particular the chemokines of the CC and the CXC chemokines, preferably, for example, HCC-1 to HCC-3 and SDF-1 (Schulz-Knappe P et al., J Exp ed 183: 295, 1996; Pardigol A et al., Proc Natl Acad Sei USA 95: 6308, 1998; Nagasawa T et al., Proc Natl Acad Sei USA 91: 2305, 1994). The latest work has shown that cell lines from tumor cells have the receptors for the well-known chemokines MCP-1 and RANTES (Wang JM et al., Int J Cancer 75: 900, 1998). These tumor cell lines can thus react to specific chemokines by stimulating the receptors. If the corresponding chemokine peptide is formed in corresponding cells, tissues and organs, an autocrine, paracrine or endocrine reaction can occur, whereby cell migration and cell proliferation are influenced. For example, kidney metastases from tumors that have receptors for MCP-1 and RANTES are said to arise preferentially, since a strong expression of these chemokines is observed in the kidney (Wang JM et al., Int J Cancer 75: 900, 1998). In further new work it was also found that certain chemokines inhibit tumor growth (Wang JM et al., J Interferon Cytokine Res 16: 53, 1996), so that tumor growth can also be negatively influenced by chemokine administration. The role of chemokines in the migration of tumor cells has been observed using breast cancer cell lines (Youngs et al., Int J Cancer 71: 257, 1997). The control of angiogenesis is also influenced positively or negatively by various cytokines. The chemokine receptor CXCR4, via which the chemokine SDF-1 exerts its action, is essential for the vascularization of the gastrointestinal tract (Tachibana K et al., Nature 393: 591, 1998). These factors can therefore also be of crucial importance in the context of the paracrine control of the vascularization of tumors and consequently for tumor maintenance.

The previously known findings regarding the chemokine system and tumors are always based on the observation of individual chemokine receptors. However, the human chemokine system is extremely complex due to over 15 known chemokine receptors with over 40 associated chemokines and is also redundant. Thus, the diagnostic and therapeutic method approaches used up to now are not optimal for carrying out specific diagnostics and therapy based on the chemokine system. With the method according to the invention, a solution to this problem can surprisingly be achieved, which thus allows for the first time reliable diagnosis and therapy of tumor and inflammatory diseases based on the chemokine system or other tumor surface proteome patterns.

A technical problem on which the invention is based is the improvement of the diagnosis of tumors and the presence of inflammatory processes. On Another problem is the improved treatment of tumors and inflammatory diseases.

The problem is solved by a diagnostic agent and a medicament according to the invention.

The diagnostic agent according to the invention contains at least two different ligands of receptors which are involved in a disease process.

The diagnostic agent according to the invention preferably contains at least two different chemokine receptor ligands, such as protein peptide structures, which interact with chemokine receptors (or other tumor cell surface proteins), namely chemokine receptor ligands, chemokine receptor antagonists and / or chemokine receptor antibodies. The chemokine receptor ligands are preferably chemokines, chemokine derivatives, agonists or antagonists of chemokine receptors, antibodies or antibody fragments which at least partially block the binding site of the chemokine receptor and surprisingly result in inhibition of tumor growth.

Chemokines are particularly preferably selected from the group consisting of the natural chemokines C, CC, CXC, CX ₃ C, their analogs, binding proteins and antibodies which bind to the specific receptors in accordance with the chemokines mentioned.

According to the invention the chemokine receptors as a whole Teilproteom and corresponding thereto chemokine receptor ligand in primary and secondary tumors, and circulating single cells are preferably (1) by immunochemical methods (immunohistochemistry using Seπ enschnitten or multiple successive or simultaneous individual cuts, FACS ^analysis') and additionally (2), or alternatively, the expression at the transcription level is recorded and analyzed by molecular biological methods (PCR or Northern analysis, in-situ hybridization). Clusters of analyzed tumor cell surface proteomes such as ectoproteases, adhesion molecules or various types of receptors can also be used according to the invention.

The method according to the invention for the detection of receptors involved in disease processes is closely related to the diagnostic agent according to the invention, expression profiles at the proteome level being examined by means of cell biological or cytochemical methods, in particular by immunochemical methods, immunohistochemistry using serial sections or multiple successive or simultaneous single sections, FACS analysis and / or the expression of receptors at the transcription level by molecular biological methods, in particular PCR, Northern analysis and / or in-situ hybridization processes.

The present invention thus also relates to the use of at least two different chemokine receptor ligands and / or two different chemokine receptors for the diagnostic characterization of tumors, which is different for each type of tumor and each individual tumor. In particular, the method according to the invention can be used for tumors which originate from the group consisting of colorectal tumors and prostate tumors. However, tumors of other organ systems can also be addressed diagnostically and thus therapeutically using the method according to the invention. In addition, the at least two different chemokine receptors and / or two different chemokine receptor ligands can be used to diagnose inflammatory processes such as organ rejection reactions and to diagnose autoimmune diseases. In particular, tumors, inflammatory diseases and autoimmune diseases of the blood system, the lymphatic system, the cardiovascular system, the nervous system, the respiratory tract, the digestive tract, the endocrine system, the skin including the appendages, the musculoskeletal system and the urogenital tract including the kidney can be diagnosed.

The diagnostic agent according to the invention enables an expansion of the cytological characterization of tumor tissues compared to conventional diagnostic methods, which after diagnosis can also be used to develop a specific therapy. The antagonists / agonists of the associated chemokines and the specific chemokine receptor antibodies are used to influence the cell growth to the chemokine receptors found (and analogously to other tumor cell proteome clusters). Here, an accelerated onset of tumor cell death (apoptosis) is surprisingly observed.

The present invention also relates to a medicament containing at least one inhibitor of at least two chemokine receptors.

The medicament according to the invention preferably contains antagonists of chemokine receptors, antibodies or antibody fragments, which are the binding site block the chemokine receptor at least partially. A chemokine receptor-ligand interaction, preferably a protein-protein (peptide) interaction with non-specific molecules, which are obtained from natural extracts, from synthetic or recombinantly produced binding proteins and from other peptide-protein libraries, is also sufficient for the to bring about a surprising effect of tumor cell apoptosis.

The inhibitors of at least two chemokine receptors which can be used in the medicament according to the invention can be used to produce a medicament for the treatment of tumors, inflammatory diseases, autoimmune diseases of the bone marrow and other organs, rejection reactions in transplants. In particular, tumors, inflammatory diseases and autoimmune diseases of the blood system, the lymphatic system, the cardiovascular system, the nervous system, the respiratory tract, the digestive tract, the endocrine system, the skin including the appendages, the musculoskeletal system and the urogenital tract including the kidney can be treated.

Inhibitors or protein ligands are preferably used which are selected from the group consisting of antagonists of chemokine receptors, antibodies or antibody fragments which at least partially block the binding site of the chemokine receptor.

The tumors that can be treated in particular are selected from the group consisting of colorectal tumors, prostate tumors and other tumor diseases of the blood system, the lymphatic system, the cardiovascular system, the nervous system, the respiratory tract, the digestive tract, the endocrine system stem, the skin including the appendages, the musculoskeletal system and the urogenital tract including the kidney.

The inflammatory processes to be treated in particular are selected from the group consisting of bronchial asthma, chronic inflammatory bowel diseases, organ rejection and other inflammatory processes of the blood system, the lymphatic system, the cardiovascular system, the nervous system, the respiratory tract, the digestive tract, the endocrine system, the skin including the appendages, the musculoskeletal system and the urogenital tract including the kidney. The autoimmune diseases to be treated in particular are selected from the group consisting of rheumatoid arthritis, lupus erythematosus and other chronic inflammations of the blood system, the lymphatic system, the cardiovascular system, the nervous system, the respiratory tract, the digestive tract, the endocrine system, the skin including Appendages, the musculoskeletal system and the urogenital tract including the kidney.

In the diagnosis of organ rejection reactions after organ transplantation, which was carried out in particular on leukocytes obtained from the circulation of patients, it is also possible to use only at least one chemokine receptor ligand and / or one chemokine receptor.

The invention further relates to the use of an inhibitor of a chemokine receptor for the manufacture of a medicament for preventing or alleviating organ rejection reactions after organ transplantation, in particular after transplantation of the heart, liver, kidney and pancreas as well as other organs, tissues and cell systems of the gastrointestinal tract, the respiratory tract and the urogenital tract , Circulatory system, the neuro-endocrinium and the musculoskeletal system as well as the blood and immune system.

In addition to establishing new causalities in a disease event, the method according to the invention can also be used for the purpose of diagnosis and therapy. Using the method according to the invention, for example, the use of chemokines and their corresponding receptors, their antagonists, including antibodies, was found in order to inhibit cancer growth, including metastasis, and to suppress inflammatory and autoimmune diseases. The method according to the invention is based on the finding that chemokines act on specific tumor and inflammatory cells through autocrine, paracrine and endocrine routes via the disease-specific constellation of the chemokine receptor proteome. Primary and secondary tumors as well as specific inflammatory cells are controlled in their migration and proliferation behavior. Diagnostic detection of the locally increased expressed and regulated chemokines and the presence of the chemokine receptor compositions gives rise to the possibility of cancer growth, tumor metastasis and inflammatory as well To decisively suppress or completely prevent autoimmune diseases.

The peptides according to the invention with Seq ID No 1 to 40 can be used as epitopes for generating antibodies. These sequences according to the invention are (ID 1-40):

Amino acid sequences used for the N-terminal domains of human

Chemokine receptors for the production of specific antibodies

1 CXCR1 X-MSNITDPQMWDFDDLNFTGMPPADEDYSPCMLETETLNK-Y

2 CXCR2 X-MEDFNMESDSFEDFWKGEDLSNYSYSSTLPPFLLDAAPCEPESLEINK-Y

3 CXCR3 X-MVLEVSDHQVLNDAEVAALLENFSSSYDYGENESDSCCTSPPCPQDFSLNFDR-Y

4 CXCR4 X-MEGISIYTSDNYTEEMGSGDYDSMKEPCFREENANFNKI-Y

5 CXCR5 X-MNYPLTLEMDLENLEDLFWELDRLDNYNDTSLVENHLCPATGPLMASFKAVFVP-Y

4 CXCR6 X-MAEHDYHEDYGFSSFNDSSQEEHQDFLQFSKV-Y

5 CCR1 X-METPNTTEDYDTTTEFDYGDATPCQKVNERAFGA-Y

6 CCR2 X-MLSTSRSRFIRNTNESGEEVTTFFDYDYGAPCHKFDVKQIGA-Y

7 CCR3 X-MTTSLD1Λ / ETFGTTSYYDDVGLLCEKADTRALMA-Y

8 CCR4 X-MNPTDIADTTLDESIYSNYYLYESIPKPCTKEGIKAFGE-Y

9 CCR5 X-MDYQVSSPIYDINYYTSEPCQKINVKQIAA-Y

10 CCR6a X-MSGESMNFSDVFDSSEDYFVSVNTSYYSVDSEMLLCSLQEVRQFSRL-Y

11 CCR6b X-MNFSDVFDSSEDYFVSVNTSYYSVDSEMLLCSLQEVRQFSRL-Y 12 CCR7 X-MDLGKPMKSVLVVALLVIFQVCLCQDEVTDDYIGDNTTVDYTLFESLCSKKDVRNFKAW-Y

13 CCR8 X-MDYTLDLSVTTVTDYYYPDIFSSPCDAELIQTNGK-Y

14 CCR9a X-MTPTDFTSPIPNMADDYGSESTSSMEDYVNFNFTDFYCEKNNVRQFASH-Y

15 CCR9b X-MADDYGSESTSSMEDYVNFNFTDFYCEKNNVRQFASH-Y

16 CCR10 X-MGTEATEQVSWGHYSGDEEDAYSAEPLPELCYKADVQAFSRAFQPSVSLTVA-Y

17 CCR11 X-MALEQNQSTDYYYEENEMNGTYDYSQYELICIKEDVREFAKV-Y

18 XCR1 X-MESSGNPESTTFFYYDLQSQPCENQAWVFAT-Y

19 CX3CR1 X-MDQFPESVTENFEYDDLAEACYIGDIVVFGT-Y

20 D6 X-MAATASPQPLATEDADSENSSFYYYDYLDEVAFMLCRKDAVVSFGKVFL-Y

Amino acid sequences used for the 2nd extracellular loop domains of human chemokine receptors for the production of specific antibodies

21 CXC 1 X-RQAYHPNNSSPVCYEVLGNDTAKWR -Y, especially CFRQAYHPNNSSPV

22 CXCR2 X-RRTVYSSNVSPACYEDMGNNTANWR-Y, especially CFRRTvΥSSNVSPA

23 CXCR3 X-LSAHHDERLNATHCQYNFPQVGR-Y, especially CLSAHHDERLNATH

24 CXCR4 X-NVSEADDRYICDRFYPNDLWWVFQ-Y, especially DRFYPNDLWWVFQFC

25 CXCR5 X- VSQGHHNNSLPRCTFSQENQAETHAWFTSR-Y, especially TFSQENQAETHAWFTSRC 26 CXCR6 X-PQIIYGNVFNLDKLICGYHDEAI-Y, in particular

27 CCR1 X-SKTQWEFTHHTCSLHFPHESLREWKL-Y, especially SLHFPHESLREWKLC

28 CCR2 X-TKCQKEDSVYVCGPYFPRGWNNFHTIMR-Y, especially GPYFPRGWNNFHTI RNC

29 CCR3 X-YETEELFEETLCSALYPEDTVYSWRHFHTLRM-Y, especially SALYPEDTVYSWRHFHTLR TC

30 CCR4 X-STCYTERNHTYCKTKYSLNSTTWKVLSSLEI-Y, especially KTKYSLNSTTWKVLSSLEINC

31 CCR5 X-TRSQKEGLHYTCSSHFPYSQYQFWKNFQTLKI-Y, especially SSHFPYSQYQFWKNFQTLKIVC

32 CCR6 X-STFVFNQKYNTQGSDVCEPKYQTVSEPIRW-Y, especially EPKYQTVSEPIRWKC

33 CCR7 X-PELLYSDLQRSSSEQAMRCSLITEHVEA-Y, especially CPELLYSDLQRSSSEQA R

34 CCR8 X-YQVASEDGVLQCYSFYNQQTLKWKIFTNFK -Y, especially YSFYNQQTLKWKIFTNFKMC

35 CCR9 X-PEILYSQIKEESGIAICTMVYPSDESTKL-Y, especially CT VYPSDESTKLK

36 CCR10 X-SQDGQREGQRRCRLIFPEGLTQTV-Y, especially FSQDGQREGQRRC

37 CCR11 X-TVNDNARCIPIFPRYLGTSMKA-Y, especially CIPIFPRYLGTSMKA

38 XCR1 X-HKVLSSGCDYSELTWYLTSVYQH-Y, especially DYSELTWYLTSVYQHC

39 CX3CR1 X-TKQKENECLGDYPEVLQEIWPVLRNVET-Y, in particular LGDYPEVLQEIWPVLRNVET

40 D6 X-QTHENPKGVWNCHADFGGHGTIWKLFLRFQQNL-Y, especially HADFGGHGTIWKLFLRFQQNC

where X represents an amino acid residue or a peptide residue of up to 30 amino acids, where Y represents an amino acid residue or a peptide residue of up to 30 amino acids

To produce the antibodies, the peptides according to the invention (sequences according to ID 1-40), which are normally not immunogenic, are coupled to the protein carrier KLH (keyhole limpet hemocyanin). This coupling takes place using MBS (m-maleimidobenzoyl-N-hydroxysuccinimide ester) via a cysteine terminally integrated in the petid, or directly using carbodiimide.

The antibodies are obtained by conventional methods by immunization, preferably from mice, rabbits, etc. The methods for producing monoclonal antibodies by molecular biological methods, such as recombinant production, can also be used. The antibodies are purified according to the known methods and galenically prepared for use.

Furthermore, cell preparations, cell extracts and in particular membrane isolates from overexpressing, artificially transfected chemokine receptor-carrying cells were used to generate such specific antibodies.

According to the invention, proteins based on known sequences were used as epitopes for generating antibodies. These sequences are known (ID 41-63): CCR1 ACCESSION P32246 355 aa

1 METPNTTEDY DTTTEFDYGD ATPCQKVNER AFGAQLLPPL YSLVFVIGLV GNILWLVLV 61 QYKRLKNMTS IYLLNLAISD LLFLFTLPFW IDYKLKDDWV FGDAMCKILS GFYYTGLYSE 121 IFFIILLTID RYLAIVHAVF ALRARTVTFG VITSIIIWAL AILASMPGLY FSKTQWEFTH 181 HTCSLHFPHE SLREWKLFQA LKLNLFGLVL PLLVMIICYT GIIKILLRRP NEKKSKAVRL 241 IFVIMIIFFL FWTPYNLTIL ISVFQDFLFT HECEQSRHLD LAVQVTEVIA YTHCCVNPVI 301 YAFVGERFRK YLRQLFHRRV AVHLVKWLPF LSVDRLERVS STSPSTGEHE LSAGF

CCR2a ACCESSION P41597 374 aa

1 MLSTSRSRFI RNTNESGEE TTFFDYDYGA PCHKFDVKQI GAQLLPPLYS LVFIFGFVGN 61 MLVVLILINC KKLKCLTDIY LLNLAISDLL FLITLPLWAH SAANEWVFGN AMCKLFTGLY 121 HIGYFGGIFF IILLTIDRYL AIVHAVFALK ARTVTFGVVT SVITWLVAVF ASVPGIIFTK 181 CQKEDSVYVC GPYFPRGWNN FHTIMRNILG LVLPLLIMVI CYSGILKTLL RCRNEKKRHR 241 AVRVIFTIMI VYFLFWTPYN IVILLNTFQE FFGLSNCEST SQLDQATQVT ETLGMTHCCI 301 NPIIYAFVGE KFRSLFHIAL GCRIAPLQKP VCGGPGVRPG KNVKVTTQGL LDGRGKGKSI 361 GRAPEASLQD KEGA

CCR2b ACCESSION NP_000639 360 aa

1 MLSTSRSRFI RNTNESGEEV TTFFDYDYGA PCHKFDVKQI GAQLLPPLYS LVFIFGFVGN 61 MLVVLILINC KKLKCLTDIY LLNLAISDLL FLITLPLWAH SAANEWVFGN AMCKLFTGLY 121 HIGYFGGIFF IILLTIDRYL AIVHAVFALK ARTVTFGVVT SVITWLVAVF ASVPGIIFTK 181 CQKEDSVYVC GPYFPRGWNN FHTIMRNILG LVLPLLIMVI CYSGILKTLL RCRNEKKRHR 241 AVRVIFTIMI VYFLFWTPYN IVILLNTFQE FFGLSNCEST SQLDQATQVT ETLGMTHCCI 301 NPIIYAFVGE KFRRYLSVFF RKHITKRFCK QCPVFYRETV DGVTSTNTPS TGEQEVSAGL

CCR3 ACCESSION P51677 355 aa

1 MTTSLDTVET FGTTSYYDDV GLLCEKADTR ALMAQFVPPL YSLVFTVGLL GNVVVVMILI 61 KYRRLRIMTN IYLLNLAISD LLFLVTLPFW IHYVRGHNWV FGHGMCKLLS GFYHTGLYSE 121 IFFIILLTID RYLAIVHAVF ALRARTVTFG VITSIVTWGL AVLAALPEFI FYETEELFEE 181 TLCSALYPED TVYSWRHFHT LRMTIFCLVL PLLVMAICYT GIIKTLLRCP SKKKYKAIRL 241 IFVIMAVFFI FWTPYNVAIL LSSYQSILFG NDCERSKHLD LVMLVTEVIA YSHCCMNPVI 301 YAFVGERFRK YLRHFFHRHL LMHLGRYIPF LPSEKLERTS SVSPSTAEPE LSIVF

CCR4 ACCESSION P51679 360 aa

1 MNPTDIADTT LDESIYSNYY LYESIPKPCT KEGIKAFGEL FLPPLYSLVF VFGLLGNSVV 61 VLVLFKYKRL RSMTDVYLLN LAISDLLFVF SLPFWGYYAA DQWVFGLGLC KMISWMYLVG 121 FYSGIFFVML MSIDRYLAIV HAVFSLRART LTYGVITSLA TWSVAVFASL PGFLFSTCYT 181 ERNHTYCKTK YSLNSTTWKV LSSLEINILG LVIPLGIMLF CYSMIIRTLQ HCKNEKKNKA 241 VKMIFAVVVL FLGFWTPYNI VLFLETLVEL EVLQDCTFER YLDYAIQATE TLAFVHCCLN 301 PIIYFFLGEK FRKYILQLFK TCRGLFVLCQ YCGLLQIYSA DTPSSSYTQS TMDHDLHDAL 361

CCR5 ACCESSION P51681 352 aa

1 MDYQVSSPIY DINYYTSEPC QKINVKQIAA RLLPPLYSLV FIFGFVGNML VILILINCKR 61 LKSMTDIYLL NLAISDLFFL LTVPFWAHYA AAQWDFGNTM CQLLTGLYFI GFFSGIFFII 121 LLTIDRYLAV VHAVFALKAR TVTFGVVTSV ITWVVAVFAS LPGIIFTRSQ KEGLHYTCSS 181 HFPYSQYQFW KNFQTLKIVI LGLVLPLLVM VICYSGILKT LLRCRNEKKR HRAVRLIFTI 241 MIVYFLFWAP YNIVLLLNTF QEFFGLNNCS SSNRLDQAMQ VTETLGMTHC CINPIIYAFV 301 GEKFRNYLLV FFQKHIAKRF CKCCSIFQQE APERASSVYT RSTGEQEISV GL

CCR6a ACCESSION P51684 374 aa

1 MSGESMNFSD VFDSSEDYFV SVNTSYYSVD SEMLLCSLQE VRQFSRLFVP IAYSLICVFG 61 LLGNILVVIT FAFYKKARSM TDVYLLNMAI ADILFVLTLP FWAVSHATGA WVFSNATCKL 121 LKGIYAINFN CGMLLLTCIS MDRYIAIVQA TKSFRLRSRT LPRSKIICLV VWGLSVIISS 181 STFVFNQKYN TQGSDVCEPK YQTVSEPIRW KLLMLGLELL FGFFIPLMFM IFCYTFIVKT 241 LVQAQNSKRH KAIRVIIAVV LVFLACQIPH NMVLLVTAAN LGKMNRSCQS EKLIGYTKTV 301 TEVLAFLHCC LNPVLYAFIG QKFRNYFLKI LKDLWCVRRK YKSSGFSCAG RYSENISRQT 361 SETADNDNAS SFTM

CCR6b 369 aa

1 MNFSDVFDSS EDYFVSVNTS YYSVDSEMLL CSLQEVRQFS RLFVPIAYSL ICVFGLLGNI 61 LVVITFAFYK KARSMTDVYL LNMAIADILF VLTLPFWAVS HATGAWVFSN ATCKLLKGIY 121 AINFNCGMLL LTCISMDRYI AIVQATKSFR LRSRTLPRSK IICLVVWGLS VIISSSTFVF 181 NQKYNTQGSD VCEPKYQTVS EPIRWKLLML GLELLFGFFI PLMFMIFCYT FIVKTLVQAQ 241 NSKRHKAIRV IIAVVLVFLA CQIPHNMVLL VTAANLGKMN RSCQSEKLIG YTKTVTEVLA 301 FLHCCLNPVL YAFIGQKFRN YFLKILKDLW CVRRKYKSSG FSCAGRYSEN ISRQTSETAD 361 NDNASSFTM

CCR7 ACCESSION P32248 378 aa

1 MDLGKPMKSV LVVALLVIFQ VCLCQDEVTD DYIGDNTTVD YTLFESLCSK KDVRNFKAWF 61 LPIMYSIICF VGLLGNGLVV LTYIYFKRLK TMTDTYLLNL AVADILFLLT LPFWAYSAAK 121 SWVFGVHFCK LIFAIYKMSF FSGMLLLLCI SIDRYVAIVQ AVSAHRHRAR VLLISKLSCV 181 GIWILATVLS IPELLYSDLQ RSSSEQAMRC SLITEHVEAF ITIQVAQMVI GFLVPLLAMS 241 FCYLVIIRTL LQARNFERNK AIKVIIAVVV VFIVFQLPYN G VLAQTVAN FNITSSTCEL 301 SKQLNIAYDV TYSLACVRCC VNPFLYAFIG VKFRNDLFKL FKDLGCLSQE QLRQWSSCRH 361 IRRSSMSVEA ETTTTFSP

CCR8 ACCESSION P51685 355 aa

1 MDYTLDLSVT TVTDYYYPDI FSSPCDAELI QTNGKLLLAV FYCLLFVFSL LGNSLVILVL 61 VVCKKLRSIT DVYLLNLALS DLLFVFSFPF QTYYLLDQWV FGTVMCK VS GFYYIGFYSS 121 MFFITLMSVD RYLAWHAVY ALKVRTIRMG TTLCLAVWLT AIMATIPLLV FYQVASEDGV

181 LQCYSFYNQQ TLKWKIFTNF KMNILGLLIP FTIFMFCYIK ILHQLKRCQN HNKTKAIRLV

241 LIVVIASLLF WVPFNVVLFL TSLHSMHILD GCSISQQLTY ATHVTEIISF THCCVNPVIY

301 AFVGEKFKKH LSEIFQKSCS QIFNYLGRQM PRESCEKSSS CQQHSSRSSS VDYIL

CCR9a ACCESSION XP_003251 369 aa

1 MTPTDFTSPI PNMADDYGSE STSSMEDYVN FNFTDFYCEK NNVRQFASHF LPPLYWLVFI 61 VGALGNSLVI LVYWYCTRVK TMTDMFLLNL AIADLLFLVT LPFWAIAAAD QWKFQTFMCK 121 VVNSMYKMNF YSCVLLIMCI SVDRYIAIAQ AMRAHTWREK RLLYSKMVCF TIWVLAAALC 181 IPEILYSQIK EESGIAICTM VYPSDESTKL KSAVLTLKVI LGFFLPFVVM ACCYTIIIHT 241 LIQAKKSSKH KALKVTIT / L TVFVLSQFPY NCILLVQTID AYAMFISNCA VSTNIDICFQ 301 VTQTIAFFHS CLNPVLYVFV GERFRRDLVK TLKNLGCISQ AQWVSFTRRE GSLKLSSMLL 361 ETTSGALSL

CCR9b ACCESSION P51686 357 aa

1 MADDYGSEST SSMEDYVNFN FTDFYCEKNN VRQFASHFLP PLYWLVFIVG ALGNSLVILV 61 YWYCTRVKTM TDMFLLNLAI ADLLFLVTLP FWAIAAADQW KFQTFMCKVV NSMYKMNFYS 121 CVLLIMCISV DRYIAIAQAM RAHTWREKRL LYSKMVCFTI WVLAAALCIP EILYSQIKEE 181 SGIAICTMVY PSDESTKLKS AVLTLKVILG FFLPFVVMAC CYTIIIHTLI QAKKSSKHKA 241 LKVTITVLTV FVLSQFPYNC ILLVQTIDAY AMFISNCAVS TNIDICFQVT QTIAFFHSCL 301 NPVLYVFVGE RFRRDLVKTL KNLGCISQAQ WVSFTRREGS LKLSSMLLET TSGALSL

CCR10 ACCESSION P46092 362 aa

1 MGTEATEQVS WGHYSGDEED AYSAEPLPEL CYKADVQAFS RAFQPSVSLT VAALGLAGNG 61 LVLATHLAAR RAARSPTSAH LLQLALADLL LALTLPFAAA GALQGWSLGS ATCRTISGLY 121 SASFHAGFLF LACISADRYV AIARALPAGP RPSTPGRAHL VSVIVWLLSL LLALPALLFS 181 QDGQREGQRR CRLIFPEGLT QTVKGASAVA QVALGFALPL GVMVACYALL GRTLLAARGP 241 ERRRALRVVV ALVAAFWLQ LPYSLALLLD TADLLAARER SCPASKRKDV ALLVTSGLAL 301 ARCGLNPVLY AFLGLRFRQD LRRLLRGGSS PSGPQPRRGC PRRPRLSSCS APTETHSLSW 361 DN

CCR11 ACCESSION AAF61299 350 aa

1 MALEQNQSTD YYYEENEMNG TYDYSQYELI CIKEDVREFA KVFLPVFLTI VFVIGLAGNS 61 MVVAIYAYYK KQRTKTDVYI LNLAVADLLL LFTLPFWAVN AVHGWVLGKI MCKITSALYT 121 LNFVSGMQFL ACISIDRYVA VTKVPSQSGV GKPCWIICFC VWMAAILLSI PQLVFYTVND 181 NARCIPIFPR YLGTSMKALI QMLEICIGFV VPFLIMGVCY FITARTLMKM PNIKISRPLK 241 VLLTVVIVFI VTQLPYNIVK FCRAIDIIYS LITSCNMSKR MDIAIQVTES IALFHSCLNP 301 ILYVFMGASF KNYVMKVAKK YGSWRRQRQS VEEFPFDSEG PTEPTSTFSI

CXCR1 ACCESSION P25024 350 aa 1 MSNITDPQMW DFDDLNFTGM PPADEDYSPC MLETETLNKY VVIIAYALVF LLSLLGNSLV

61 MLVILYSRVG RSVTDVYLLN LALADLLFAL TLPIWAASKV NGWIFGTFLC KVVSLLKEVN

121 FYSGILLLAC ISVDRYLAIV HATRTLTQKR HLVKFVCLGC WGLSMNLSLP FFLFRQAYHP

181 NNSSPVCYEV LGNDTAKWRM VLRILPHTFG FIVPLFVMLF CYGFTLRTLF KAHMGQKHRA

241 MRVIFAVVLI FLLCWLPYNL VLLADTLMRT QVIQETCERR NNIGRALDAT EILGFLHSCL

301 NPIIYAFIGQ NFRHGFLKIL AMHGLVSKEF LARHRVTSYT SSSVNVSSNL

56 CXCR2 ACCESSION P25025 360 aa

1 MEDFNMESDS FEDFWKGEDL SNYSYSSTLP PFLLDAAPCE PESLEINKYF VVIIYALVFL 61 LSLLGNSLVM LVILYSRVGR SVTDVYLLNL ALADLLFALT LPIWAASKVN GWIFGTFLCK 121 VVSLLKEVNF YSGILLLACI SVDRYLAIVH ATRTLTQKRY LVKFICLSIW GLSLLLALPV 181 LLFRRTVYSS NVSPACYEDM GNNTANWRML.LRILPQSFGF IVPLLIMLFC YGFTLRTLFK 241 AHMGQKHRAM RVIFAWLIF LLCWLPYNLV LLADTLMRTQ VIQETCERRN HIDRALDATE 301 ILGILHSCLN PLIYAFIGQK FRHGLLKILA IHGLISKDSL PKDSRPSFVG SSSGHTSTTL

57 CXCR3 ACCESSION P49682 368 aa

1 MVLEVSDHQV LNDAEVAALL ENFSSSYDYG ENESDSCCTS PPCPQDFSLN FDRAFLPALY

61 SLLFLLGLLG NGAVAAVLLS RRTALSSTDT FLLHLAVADT LLVLTLPLWA VDAAVQWVFG

121 SGLCKVAGAL FNINFYAGAL LLACISFDRY LNIVHATQLY RRGPPARVTL TCLAVWGLCL

181 LFALPDFIFL SAHHDERLNA THCQYNFPQV GRTALRVLQL VAGFLLPLLV MAYCYAHILA

241 VLLVSRGQRR LRAMRLVVVV VVAFALCWTP YHLVVLVDIL MDLGALARNC GRESRVDVAK

301 SVTSGLGYMH CCLNPLLYAF VGVKFRERMW MLLLRLGCPN QRGLQRQPSS SRRDSSWSET

361 SEASYSGL

58 CXCR4 ACCESSION P30991 352 aa

1 MEGISIYTSD NYTEEMGSGD YDSMKEPCFR EENANFNKIF LPTIYSIIFL TGIVGNGLVI 61 LVMGYQKKLR SMTDKYRLHL SVADLLFVIT LPFWAVDAVA NWYFGNFLCK AVHVIYTVNL 121 YSSVLILAFI SLDRYLAIVH ATNSQRPRKL LAEKWYVGV WIPALLLTIP DFIFANVSEA 181 DDRYICDRFY PNDLWVVVFQ FQHIMVGLIL PGIVILSCYC IIISKLSHSK GHQKRKALKT 241 TVILILAFFA CWLPYYIGIS IDSFILLEII KQGCEFENTV HKWISITEAL AFFHCCLNPI 301 LYAFLGAKFK TSAQHALTSV SRGSSLKILS KGKRGGHSSV STESESSSFH SS

59 CXCR5 ACCESSION P32302 372 aa

1 MNYPLTLEMD LENLEDLFWE LDRLDNYNDT SLVENHLCPA TEGPLMASFK AVFVPVAYSL 61 IFLLGVIGNV LVLVILERHR QTRSSTETFL FHLAVADLLL VFILPFAVAE GSVGWVLGTF 121 LCKTVIALHK VNFYCSSLLL ACIAVDRYLA IVHAVHAYRH RRLLSIHITC GTIWLVGFLL 181 ALPEILFAKV SQGHHNNSLP RCTFSQENQA ETHAWFTSRF LYHVAGFLLP MLVMGWCYVG 241 VVHRLRQAQR RPQRQKAVRV AILVTSIFFL CWSPYHIVIF LDTLARLKAV DNTCKLNGSL 301 PVAITMCEFL GLAHCCLNPM LYTFAGVKFR SDLSRLLTKL GCTGPASLCQ LFPSWRRSSL 361 SESENATSLT TF

60 CXCR6 ACCESSION NP_006555 342 aa

1 MAEHDYHEDY GFSSFNDSSQ EEHQDFLQFS KVFLPCMYLV VFVCGLVGNS LVLVISIFYH 61 KLQSLTDVFL VNLPLADLVF VCTLPFWAYA GIHEWVFGQV MCKSLLGIYT INFYTSMLIL 121 TCITVDRFIV VVKATKAYNQ QAKRMTWGKV TSLLIWVISL LVSLPQIIYG NVFNLDKLIC 181 GYHDEAISTV VLATQMTLGF FLPLLTMIVC YSVIIKTLLH AGGFQKHRSL KIIFLVMAVF 241 LLTQMPFNLM KFIRSTHWEY YAMTSFHYTI MVTEAIAYLR ACLNPVLYAF VSLKFRKNFW 301 KLVKDIGCLP YLGVSHQWKS SEDNSKTFSA SHNVEATSMF QL

61 CX3CR1 ACCESSION P49238 355 aa

1 MDQFPESVTE NFEYDDLAEA CYIGDIVVFG TVFLSIFYSV IFAIGLVGNL L VFALTNSK 61 KPKSVTDIYL LNLALSDLLF VATLPFWTHY LINEKGLHNA MCKFTTAFFF IGFFGSIFFI 121 TVISIDRYLA IVLAANSMNN RTVQHGVTIS LGVWAAAILV AAPQFMFTKQ KENECLGDYP 181 EVLQEIWPVL RNVETNFLGF LLPLLIMSYC YFRIIQTLFS CKNHKKAKAI KLILLVVIVF 241 FLFWTPYNVM IFLETLKLYD FFPSCDMRKD LRLALSVTET VAFSHCCLNP LIYAFAGEKF 301 RRYLYHLYGK CLAVLCGRSV HVDFSSSESQ RSRHGSVLSS NFTYHTSDGD Allll

62 XCR1 ACCESSION P46094 333 aa

1 MESSG PEST TFFYYDLQSQ PCENQAWVFA TLATTVLYCL VFLLSLVGNS LVLWVLVKYE 61 SLESLTNIFI LNLCLSDLVF ACLLPVWISP YHWGWVLGDF LCKLLNMIFS ISLYSSIFFL 121 TIMTIHRYLS VVSPLSTLRV PTLRCRVLVT MAVWVASILS SILDTIFHKV LSSGCDYSEL 181 TWYLTSVYQH NLFFLLSLGI ILFCYVEILR TLFRSRSKRR HRTVKLIFAI VVAYFLSWGP 241 YNFTLFLQTL FRTQIIRSCE AKQQLEYALL ICRNLAFSHC CFNPVLYVFV GVKFRTHLKH 301 VLRQFWFCRL QAPSPASIPH SPGAFAYEGA SFY

63 D6 ACCESSION XP_003126 384 aa

1 MAATASPQPL ATEDADSENS SFYYYDYLDE VAFMLCRKDA VVSFGKVFLP VFYSLIFVLG 61 LSGNLLLLMV LLRYVPRRRM VEIYLLNLAI SNLLFLVTLP FWGISVAWHW VFGSFLCKMV 121 STLYTINFYS GIFFISCMSL DKYLEIVHAQ PYHRLRTRAK SLLLATIVWA VSLAVSIPDM 181 VFVQTHENPK GVWNCHADFG GHGTIWKLFL RFQQNLLGFL LPLLAMIFFY SRIGCVLVRL 241 RPAGQGRALK IAAALVVAFF VLWFPYNLTL FLHTLLDLQV FGNCEVSQHL DYALQVTESI 301 AFLHCCFSPI LYAFSSHRFR QYLKAFLAAV LGWHLAPGTA QASLSSCSES SILTAQEEMT 361 GMNDLGERQS ENYPNKEDVG NKSA The pharmaceutical compositions of the invention can in suitable pharmaceutical application forms, in particular in lyophilized form ingested with mannitol or similar sugars in sterile ampoules for dissolution in physiological saline and / or infusion solution for repeated single injection and / or continuous infusion in amounts of 300 mg to 30 mg of pure chemokine receptor ligands, in particular chemokines, chemokine agonists or antagonists and chemokine and chemokine receptor antibodies are administered per therapy unit. The medicament according to the invention is preferably administered systemically or locally in a galenical application form, in which the medicament is used in biocompatible microspheres, and via aerosol, intravenous or subcutaneous application.

According to the invention, this method can also be used as follows. The tumor cells whose receptor composition is to be investigated are grown in parallel in vitro, and the cells obtained therefrom are also examined for their receptor composition and treated with chemokines, preferably of the types HCC-1, HCC-2, MCP-1, RANTES or SDF-1 , In addition, the known analogues were also used. With the modified Boyden migration chamber method, it can be determined that the tumor cells react chemotactically when agonists that bind to the corresponding chemokine receptors are administered. The inhibition of their migration is confirmed by prior incubation with antagonists or receptor antibodies.

When using highly purified antibodies, it was found that these are capable of causing the apoptosis of tumor cells in in vitro and in vivo models. If cell lines or removed tumor cells are grown using the usual cell culture methods, their survival time in vitro is greatly reduced by adding chemokine antibodies, the corresponding receptors of which have previously been detected on the cell surface of these cell types. An apoptosis of a large number of these cultured cells can be observed.

Surprisingly, a decrease in tumor cells due to apoptosis can also be found in in vivo models.

Since nude mice have a deficient immune system, the metastatic behavior in a host body can be examined in a nude mouse model without the known immune reactions between species taking place and the foreign cells being rejected. Nude mice are vaccinated in a manner known per se with tumor cells or tumor cell lines whose chemokine receptor distribution pattern had been analyzed and the metastasis by these cells is checked when treated with chemokines and when treated with chemokine antagonists and / or receptor antibodies. Surprisingly, it is found that chemokine antagonists and receptor antibodies belonging to the found receptors significantly inhibit or prevent metastasis, while the addition of chemokines leads to a modulation of tumor growth. Surprisingly, it also emerges that the preparations analyzed by immunohistochemistry show a specific distribution of chemokines and chemokine receptors in the tumor and surrounding tissue. This identified additional targeted intervention options.

This intervention in accordance with the method according to the invention consists in additionally expanding the proteome analysis of the chemokines and their receptors by analyzing antitumor peptides / proteins in the same method. This results in the expansion of the diagnostic and therapeutic approach, in particular to use antagonists directed against further clusters of the tumor cell surface proteome. These effects can be enhanced by a combination with chemokine receptor antagonists and antibodies.

To confirm these findings, tumor cell lines (e.g. preferably LNCaP, PC-3, DU-145, HT-29, Caco-2, T-84) can also be stably transfected with one or more of the chemokine receptors. After subsequent injection of these cells into animals in which a chemokine corresponding to the receptor is overexpressed in the liver, such tumor cells settle. Such modified cells thus preferably form metastases in the liver.

When using the method in samples from effector cells in inflammatory (acute rejection in kidney transplantation) and autoimmune diseases (rheumatoid arthritis), a specific composition of the chemokine / chemokine receptor proteome, in particular the chemokine receptors CXCR4, and CCR5 and / or CXCR3, can also be used in the rejection of Kidney transplants can be demonstrated. In vitro experiments have also shown that a mixture of antibodies and antagonists directed at these receptors strongly inhibits the migration of disease-specific effector cells.

The invention is explained in more detail below with the aid of examples. Example 1 :

Generation of specific antibodies against chemokine receptors (preferably CCR and CXCR) and other clusters of the tumor cell surface proteome.

For the production of specific antibodies against chemokine receptors it has surprisingly been found that the use of specific amino acid sequences (see Tables 1 and 2) in particular against the (1) N-terminal extracellular and the (2) second extracellular loop domain of this 7-TM D receptors (seven transmembrane domains) are particularly suitable for immunization if the synthesized peptides are coupled to carrier molecules according to the usual methods and are injected into mice. Furthermore, multiple antigenic peptides (MAP), which are linked to larger molecules by means of lysine, or chemokine receptor-transfected cell lines (sequences according to Table 3) are also suitable for producing these antibodies. As a further method, the use of immunogens from stably transfected, chemokine receptor-bearing cells has proven surprisingly well, membrane isolates, cell extracts with complete or fragmented receptors or even lyophilized total preparations being used.

The mice (type NZW X NZB) are used to produce monoclonal antibodies, which is carried out using the routine methods of IPF PharmaCeuticals GmbH. The antibodies checked by Western blot and ELISA can be used for the above-mentioned diagnostic and therapeutic purposes if they are highly purified using the laboratory methods of IPF PharmaCeuticals GmbH. The sequences used for the chemokine receptors must be selected in accordance with sequence ID 1-63.

Example 2:

Detection of chemokines and chemokine receptors in primary tumors, tumor metastases and single tumor cells.

In the surgical treatment of tumors (eg colon, prostate) and their metastases, which were removed from the liver or lymph nodes, for example, the tumor cells could be obtained and analyzed by immunohistochemical methods and other molecular biological methods. In the immunochemical and molecular biological analyzes it was found that the primary tumor cells, the metastases and circulating single cells (obtained from the blood of patients) contain a specific composition of chemokines and chemokine receptors. The algorithm of this composition is individually and tumor-dependent of high specificity, which surprisingly enables targeted tumor treatment based on the diagnosed proteome clusters using the method according to the invention. Surprisingly, the algorithms of the chemokine receptors are preferably suitable here. According to the invention, these algorithms derived from the tests are defined as follows:

X = new chemokine receptors to be identified n = 0 to co other chemokine receptors or chemokine receptors to be newly identified

CCR1 + CCR2 + n CCR2 + CCR5 + n CCR3 + CCR8 + n

CCR1 + CCR3 + n CCR2 + CCR6 + n CCR3 + CCR9 + n

CCR1 + CCR4 + n CCR2 + CCR7 + n CCR3 + CCR10 + n

CCR1 + CCR5 + n CCR2 + CCR8 + n CCR3 + CCR11 + n

CCR1 + CCR6 + n CCR2 + CCR9 + n CCR3 + CXCR1 + n

CCR1 + CCR7 + n CCR2 + CCR10 + n CCR3 + CXCR2 + n

CCR1 + CCR8 + n CCR2 + CCR11 + n CCR3 + CXCR3 + n

CCR1 + CCR9 + n CCR2 + CXCR1 + n CCR3 + CXCR4 + n

CCR1 + CCR10 + n CCR2 + CXCR2 + n CCR3 + CXCR5 + n

CCR1 + CCR11 + n CCR2 + CXCR3 + n CCR3 + CXCR6 + n

CCR1 + CXCR1 + n CCR2 + CXCR4 + n CCR3 + XCR1 + n

CCR1 + CXCR2 + n CCR2 + CXCR5 + n CCR3 + CX3CR1 + n

CCR1 + CXCR3 + n CCR2 + CXCR6 + n CCR3 + D6 + n

CCR1 + CXCR4 + n CCR2 + XCR1 + n CCR3 + X + n

CCR1 + CXCR5 + n CCR2 + CX3CR1 + n

CCR1 + CXCR6 + n CCR2 + D6 + n CCR4 + CCR1 + n

CCR1 + XCR1 + n CCR2 + X + n CCR4 + CCR2 + n

CCR1 + CX3CR1 + n CCR4 + CCR3 + n

CCR1 + D6 + n CCR3 + CCR1 + n CCR4 + CCR5 + n

CCR1 + X + n CCR3 + CCR2 + n CCR4 + CCR6 + n

CCR3 + CCR4 + n CCR4 + CCR7 + n

CCR2 + CCR1 + n CCR3 + CCR5 + n CCR4 + CCR8 + n

CCR2 + CCR3 + n CCR3 + CCR6 + n CCR4 + CCR9 + n

CCR2 + CCR4 + n CCR3 + CCR7 + n CCR4 + CCR10 + n CCR4 + CCR11 + n CCR6 + CCR9 + n CCR8 + CCR6 + n

CCR4 + CXCR1 + n CCR6 + CCR10 + n CCR8 + CCR7 + n

CCR4 + CXCR2 + n CCR6 + CCR11 + n CCR8 + CCR9 + n

CCR4 + CXCR3 + n CCR6 + CXCR1 + n CCR8 + CCR10 + n

CCR4 + CXCR4 + n CCR6 + CXCR2 + n CCR8 + CCR11 + n

CCR4 + CXCR5 + n CCR6 + CXCR3 + n CCR8 + CXCR1 + n

CCR4 + CXCR6 + n CCR6 + CXCR4 + n CCR8 + CXCR2 + n

CCR4 + XCR1 + n CCR6 + CXCR5 + n CCR8 + CXCR3 + n

CCR4 + CX3CR1 + n CCR6 + CXCR6 + n CCR8 + CXCR4 + n

CCR4 + D6 + n CCR6 + XCR1 + n CCR8 + CXCR5 + n

CCR4 + X + n CCR6 + CX3CR1 + n CCR8 + CXCR6 + n

CCR6 + D6 + n CCR8 + XCR1 + n

CCR5 + CCR1 + n CCR6 + X + n CCR8 + CX3CR1 + n

CCR5 + CCR2 + n CCR8 + D6 + n

CCR5 + CCR3 + n CCR7 + CCR1 + n CCR8 + X + n

CCR5 + CCR4 + n CCR7 + CCR2 + n

CCR5 + CCR6 + n CCR7 + CCR3 + n CCR9 + CCR1 + n

CCR5 + CCR7 + n CCR7 + CCR4 + n CCR9 + CCR2 + n

CCR5 + CCR8 + n CCR7 + CCR5 + n CCR9 + CCR3 + n

CCR5 + CCR9 + n CCR7 + CCR6 + n CCR9 + CCR4 + n

CCR5 + CCR10 + n CCR7 + CCR8 + n CCR9 + CCR5 + n

CCR5 + CCR11 + n CCR7 + CCR9 + n CCR9 + CCR6 + n

CCR5 + CXCR1 + n CCR7 + CCR10 + n CCR9 + CCR7 + n

CCR5 + CXCR2 + n CCR7 + CCR11 + n CCR9 + CCR8 + n

CCR5 + CXCR3 + n CCR7 + CXCR1 + n CCR9 + CCR10 + n

CCR5 + CXCR4 + n CCR7 + CXCR2 + n CCR9 + CCR11 + n

CCR5 + CXCR5 + n CCR7 + CXCR3 + n CCR9 + CXCR1 + n

CCR5 + CXCR6 + n CCR7 + CXCR4 + n CCR9 + CXCR2 + n

CCR5 + XCR1 + n CCR7 + CXCR5 + n CCR9 + CXCR3 + n

CCR5 + CX3CR1 + n CCR7 + CXCR6 + n CCR9 + CXCR4 + n

CCR5 + D6 + n CCR7 + XCR1 + n CCR9 + CXCR5 + n

CCR5 + X + n CCR7 + CX3CR1 + n CCR9 + CXCR6 + n

CCR7 + D6 + n CCR9 + XCR1 + n

CCR6 + CCR1 + n CCR7 + X + n CCR9 + CX3CR1 + n

CCR6 + CCR2 + n CCR9 + D6 + n

CCR6 + CCR3 + n CCR8 + CCR1 + n CCR9 + X + n

CCR6 + CCR4 + n CCR8 + CCR2 + n

CCR6 + CCR5 + n CCR8 + CCR3 + n CCR10 + CCR1 + n

CCR6 + CCR7 + n CCR8 + CCR4 + n CCR10 + CCR2 + n

CCR6 + CCR8 + n CCR8 + CCR5 + n CCR 10+ CCR3 + n

n X n X Xn n X n X n X n X o X n X n X n X n X n X Xn Xn n X n n n o X X X n n n n n o o n o n n o n n n n r> n n n n n n n n n n X X X X X X X X X X X X X X X X X X X X o

73 73 73 73 73 73 n

73 73 73 73 n 73 n 73 7 73 n 73 n 73 n 73 o 73 n n 73 n 73 r 73> n n n n

73 73 73 73 7n3 n 73 n 73 n 73 n 73 n 73 n 73 n 73 n 73 n 73 n 73 o 73 7) NJ ro r ro NJ MN) NJ M ro MK> M NJ M ro l- l- f - l- H- ⁱ l- ⁱ l- ⁱ l- li l- l- h- ⁱ l- ⁱ l- ⁱ l- ⁱ h- ⁱ H- ⁱ l- ⁱ h-

+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

X α n X

X n n X n X Xn Xn π X nn on on nn nn on nn nn nn nn nn X n n n O cα X cn n n X n X n X X X n X n n 7n3 n n n O n o O n

+ 73 7n

+ co n 73 nnnnn 73 73 73 73 73 73 73 73 73 73 3 n 73 n 73 n 73 7n3 7n3 n 73 7 l- 73 73 73 73 l- l- ⁱ ÜD 00 j cn cπ 4 LO NJ h- ⁱ + + LA) n 73 l- 7n3 n 73 7n3 n 73 73 H- h- ⁱ KΩ co J cn U1. LO r

3 3 73 + cn π 4 ^ ω l- l- O + + + + + + + + + + 3 3 73 + cn Ol ω ro l- ⁱ o + + + + + + + +

+ 3 + + + + + + + 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3

3 3 3 3 3 3 3 + 3 + + + + + + + 3

3 3 3 3 3 3 3

3 3

CXCR4 + D6 + n CXCR6 + XCR1 + n CX3CR1 + CXCR4 + n

CXCR4 + X + n CXCR6 + CX3CR1 + n CX3CR1 + CXCR5 + n

CXCR6 + D6 + n CX3CR1 + CXCR6 + n

CXCR5 + CCR1 + n CXCR6 + X + n CX3CR1 + XCR1 + n

CXCR5 + CCR2 + n CX3CR1 + D6 + n

CXCR5 + CCR3 + n XCR1 + CCR1 + n CX3CR1 + X + n

CXCR5 + CCR4 + n XCR1 + CCR2 + n

CXCR5 + CCR5 + n XCR1 + CCR3 + n

CXCR5 + CCR6 + n XCR1 + CCR4 + n

CXCR5 + CCR7 + n XCR1 + CCR5 + n

CXCR5 + CCR8 + n XCR1 + CCR6 + n

CXCR5 + CCR9 + n XCR1 + CCR7 + n

CXCR5 + CCR10 + n XCR1 + CCR8 + n

CXCR5 + CCR11 + n XCR1 + CCR9 + n

CXCR5 + CXCR1 + n XCR1 + CCR10 + n

CXCR5 + CXCR2 + n XCR1 + CCR11 + n

CXCR5 + CXCR3 + n XCR1 + CXCR1 + n

CXCR5 + CXCR4 + n XCR1 + CXCR2 + n

CXCR5 + CXCR6 + n XCR1 + CXCR3 + n

CXCR5 + XCR1 + n XCR1 + CXCR4 + n

CXCR5 + CX3CR1 + n XCR1 + CXCR5 + n

CXCR5 + D6 + n XCR1 + CXCR6 + n

CXCR5 + X + n XCR1 + CX3CR1 + n XCR1 + D6 + n

CXCR6 + CCR1 + n XCR1 + X + n

CXCR6 + CCR2 + n

CXCR6 + CCR3 + n CX3CR1 + CCR1 + n

CXCR6 + CCR4 + n CX3CR1 + CCR2 + n

CXCR6 + CCR5 + n CX3CR1 + CCR3 + n

CXCR6 + CCR6 + n CX3CR1 + CCR4 + n

CXCR6 + CCR7 + n CX3CR1 + CCR5 + n

CXCR6 + CCR8 + n CX3CR1 + CCR6 + n

CXCR6 + CCR9 + n CX3CR1 + CCR7 + n

CXCR6 + CCR10 + n CX3CR1 + CCR8 + n

CXCR6 + CCR11 + n CX3CR1 + CCR9 + n

CXCR6 + CXCR1 + n CX3CR1 + CCR10 + n

CXCR6 + CXCR2 + n CX3CR1 + CCR11 + n

CXCR6 + CXCR3 + n CX3CR1 + CXCR1 + n

CXCR6 + CXCR4 + n CX3CR1 + CXCR2 + n

CXCR6 + CXCR5 + n CX3CR1 + CXCR3 + n

Claims

claims

1. Diagnostic agent containing at least two different ligands of receptors that are involved in a disease process.

2. Diagnostic agent containing at least two different chemokine receptor ligands.

3. Diagnostic agent according to claim 2, wherein the chemokine receptor ligands are chemokines, chemokine derivatives, agonists or antagonists of chemokine receptors, agonists and antagonists of chemokine receptors, antibodies or antibody fragments that at least partially block the binding site of the chemokine receptor.

4. Diagnostic agent according to claim 2 and 3, wherein the chemokines are selected from the group consisting of C, CC, CXC, CX ₃ C chemokines their pharmacolagic analogs, binding proteins and antibodies that bind to the specific receptors according to the said chemokines.

5. Use of at least two different ligands of receptors that are involved in a disease process for the diagnosis of diseases.

6. Use according to claim 5, wherein at least two chemokine receptor ligands and / or two different chemokine receptors are used for the diagnosis of tumors.

7. Use according to claim 6, wherein the tumors are selected from the group consisting of colorectal tumors, prostate tumors.

8. Use of at least two different chemokine receptor ligands and / or two different chemokine receptors for the diagnosis of organ rejection reactions.

9. Use of at least two different chemokine receptor ligands and / or two different chemokine receptors for the diagnosis of inflammatory processes.

10. Use of at least two different chemokine receptor ligands and / or two different chemokine receptors for the diagnosis of autoimmune diseases.

11. Medicament containing at least one inhibitor of at least two different ligands of receptors which are involved in a disease process.

12. Medicament according to claim 11, comprising at least one inhibitor of at least two chemokine receptors.

13. Medicament according to claim 12 containing antagonists of chemokine receptors, antibodies or antibody fragments which at least partially block the binding site of the chemokine receptor.

14. Use of inhibitors of at least two different ligands of receptors which are involved in a disease process for the treatment of these diseases.

15. Use according to claim 14, wherein at least two chemokine receptors for the manufacture of a medicament for the treatment of tumors, inflammatory processes, autoimmune diseases, diseases of the bone marrow.

16. Use according to claim 14, wherein tumors, inflammatory processes, autoimmune diseases of the blood vessel system, the lymphatic system, the respiratory tract, the digestive tract and the urogenital tract, including the kidney, are affected.

17. Use according to claim 15 and 16, wherein the inhibitors are selected from the group consisting of antagonists of chemokine receptors, antibodies or antibody fragments which at least partially block the binding sites of the chemokine receptor and thus modulate their function.

18. Use according to any one of claims 15 to 17, wherein the tumors are selected from the group consisting of colorectal tumors, prostate tumors, and other tumor diseases of the blood system, the lymphatic system, the cardiovascular system, the nervous system, and the respiratory tract, of the digestive tract, the endocrine system, the skin including appendages, the musculoskeletal system and the urogenital tract including the kidney.

19. Use according to any one of claims 15 to 17, wherein the inflammatory processes are selected from the group consisting of bronchial asthma, chronic inflammatory bowel diseases, organ rejection, and further inflammatory processes of the blood system, the lymphatic system, the cardiovascular system, the nervous system, of the respiratory tract, the digestive tract, the endocrine system, the skin including appendages, the musculoskeletal system and the urogenital tract including the kidney.

20. Use according to one of claims 15 to 17, wherein the autoimmune diseases are selected from the group consisting of rheumatoid arthritis, lupus erythematosus, and further chronic inflammation of the blood system, the lymphatic system, the cardiovascular system, the nervous system, the respiratory tract, of the digestive tract, the endocrine system, the skin including appendages, the musculoskeletal system and the urogenital tract including the kidney.

21. Use of at least one chemokine receptor ligand and / or a chemokine receptor for the diagnosis of organ rejection reactions after organ transplantation.

22. Use of a chemokine receptor inhibitor in the manufacture of a medicament for preventing or ameliorating organ rejection reactions after organ transplantation.

23. Use according to claim 22, after transplants of the liver, kidney, pancreas, small intestine and other organs, tissues and cell systems of the gastrointestinal tract, the respiratory tract, the urogenital tract, the circulatory system, the neuro-endocrine and the musculoskeletal system as well as the blood and immune system ,

24. Peptides with Seq ID No 1 to 40.

25. Use of the peptides according to claim 24 for the production of antibodies against a chemokine receptor.

26. A method for recognizing receptors involved in disease processes, wherein expression profiles are examined at the proteome level by means of cell biological or cytochemical methods, in particular by immunochemical methods, immunohistochemistry by means of serial sections or multiple successive or simultaneous single sections, FACS analysis and / or the expression of receptors Transcription level through molecular biological methods, in particular PCR, Northern analysis and / or in situ hybridization methods.

27. The method according to claim 26, wherein a diagnostic agent according to one of claims 1 to 4 is used.