EP1210109A2 - Utilization of an aminopeptidase inhibitor - Google Patents

Utilization of an aminopeptidase inhibitor

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Publication number
EP1210109A2
EP1210109A2 EP01911521A EP01911521A EP1210109A2 EP 1210109 A2 EP1210109 A2 EP 1210109A2 EP 01911521 A EP01911521 A EP 01911521A EP 01911521 A EP01911521 A EP 01911521A EP 1210109 A2 EP1210109 A2 EP 1210109A2
Authority
EP
European Patent Office
Prior art keywords
inhibitor
tumor
immune cells
aminopeptidase
detected
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP01911521A
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German (de)
French (fr)
Inventor
Walter Schubert
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
MPB Meltec Patent und Beteiligungs GmbH
Original Assignee
Meltec Multi-Epitope-Ligand-Technologies GmbH
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Filing date
Publication date
Application filed by Meltec Multi-Epitope-Ligand-Technologies GmbH filed Critical Meltec Multi-Epitope-Ligand-Technologies GmbH
Priority to EP04024180A priority Critical patent/EP1510219A1/en
Publication of EP1210109A2 publication Critical patent/EP1210109A2/en
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5047—Cells of the immune system

Definitions

  • the present invention relates to the use of at least one aminopeptidase inhibitor for the manufacture of a medicament for the treatment of tumor diseases and / or immune diseases, a corresponding pharmaceutical preparation, a method for identifying at least one aminopeptidase inhibitor and a method for identifying at least one further inhibitor which works in combination with the at least one aminopeptidase inhibitor.
  • Aminopeptidases are cell surface enzymes that cleave peptides. They are expressed by different cell types. Their molecular function consists, among other things, in the degradation of biologically active peptides. The further physiological, in particular the cellular functions of the aminopeptidases have not yet been clarified. Recent studies show that aminopeptidase inhibitors can suppress the rate of proliferation and invasion of tumor cells. This suppression of the invasion was generally attributed to the proteolytic activity of cell surface associated aminopeptidases, which cleave the extracellular matrix proteins so that the tumor cells can penetrate into organs and migrate in these organs.
  • Known aminopeptidase inhibitors include actinonin, bestatin, and potent homophthalimide-type inhibitors. Bestatin can be according to J.
  • a particular disadvantage is the fact that the previously existing investigation methods do not reflect the conditions in vivo and therefore often lead to unsatisfactory results, so that the action of the individual aminopeptidase inhibitors cannot be fully elucidated. It remains mostly unclear whether the inhibition of aminopeptidases in tumors in vivo, i.e. on the patient, is effective and whether an adverse effect, consisting in potentiating the invasive behavior in vivo, could not even occur in the case of certain types of tumor by inhibiting the aminopeptidases.
  • the mechanism of action of the aminopeptidase inhibitors with a known effect is completely unknown, so that no new substances can be developed or identified which can only have an extremely specific effect because they affect the cell functions in a known manner. For example, it has not yet been clarified with which other protein interactions occur in the same cell and how these interactions encode complex cell functions. It is therefore also not known whether and which cellular mechanisms that are based on such interactions can be specifically blocked by inhibiting the aminopeptidases, and which new indications result from the clinical use of such inhibitors or from these inhibitors further developed substances could result.
  • One of the objects is achieved by using at least one aminopeptidase inhibitor for the manufacture of a medicament for the treatment of tumor diseases and / or immune diseases, the at least one aminopeptidase inhibitor blocking polarization of invasive human or animal tumor and / or immune cells by modifying at least one surface protein CD13 as a member of a protein network on the surface of the tumor and / or immune cells, the protein network comprising up to 30 surface proteins from a group
  • CD4 1. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
  • CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
  • CD56 12. CD7 13. CD54 14. CD58 15. CD138 16. CD13 17. CD62L 18. CD71 19. CD11b 20. CD36
  • CD2 27. CD20 28. CD10 29. CD44 30. CD80 includes.
  • aminopeptidases control cell surface proteins which do not belong to the class of proteolytic enzymes but to the class of adhesion molecules, these adhesion molecules in a certain combination and spatial Arrangement are crucial for the polarization of the cells.
  • Aminopeptidases are superordinate control proteins in a protein network consisting of up to 30 different protein species on the cell surface, which control the polarization of tumor cells and of other invasive cells, such as immune cells, through targeted interaction with one another and which are listed above.
  • the inhibition of at least one aminopeptidase leads to a reproducible change in surface protein combinations on the cell surface, which always also includes a change in CD13.
  • polarization is to be understood as a process in which a primarily spherical cell changes into an elongated, elongated cell shape via various intermediate stages. This process, which is a shape change controlled by the complex protein network, is the prerequisite for cell migration, i.e. migration, since only cells with an elongated cell shape can migrate. The process of polarization is therefore imperative to all processes of cell migration and thus also to invasion.
  • the invention is based on the knowledge that it is precisely the aminopeptidase inhibitors which alter at least the surface proteins CD13, as a member of the specific protein network defined above, of up to 30 surface proteins, specifically inhibit the first and therefore most important step of the invasion and are therefore used to produce an extremely specific and therefore extremely effective drug for the treatment of tumor diseases and / or immune diseases can be.
  • the at least one aminopeptidase inhibitor can be, for example, an aminopeptidase inhibitor of the homophthalimide type and / or actinonin and / or bestatin and / or an antibody, in particular a monoclonal antibody, against one of the surface proteins. Bestatin in particular counteracts the assumptions set out above by the mechanism of action mentioned and leads to a change in the surface proteins of the protein network which comprises proteins from the group mentioned above. Actinonin, RB 3014 and a monoclonal antibody (clone SJ1 D1) directed against an extracellular domain of CD13 have also proven to be particularly effective.
  • aminopeptidase inhibitors specified above can be used to effectively prevent the polarization of tumor cells as well as the polarization of immune cells, the use of aminopeptidase inhibitors enables effective medicinal products to be used to treat autoimmune diseases or rejection reactions of transplanted organs or allergies, in particular allergies to the respiratory tract , to be provided.
  • At least one further inhibitor can be used to manufacture the medicinal product, which inhibits and / or modifies at least one surface protein that is not an aminopeptidase.
  • inhibition is to be understood as the general inhibition of the function of the at least one surface protein, which can also take place through a change in expression.
  • the blocking of the polarization can be increased enormously by using an inhibitor combination.
  • an antibody against CD45RA can be used as another Inhibitor. This inhibitor in particular enhances the action of an aminopeptidase inhibitor as defined above, so that the polarization can be specifically and particularly effectively inhibited by this inhibitor combination.
  • At least one aminopeptidase inhibitor and / or at least one further inhibitor can, in particular in addition to the change in CD13, bring about a change in at least one further surface protein of the tumor and / or immune cells, which affects adhesion to endothelial cells and / or extracellular structures, in particular to organ-specific ones Endothelial cells and / or to organ-specific extracellular structures.
  • At least one aminopeptidase inhibitor and / or at least one further inhibitor can also cause a change in the adhesive functions of endothelial cells. In this way it can be prevented that the tumor and / or immune cells bind to the endothelial cells, which is essential for polarization.
  • those aminopeptidase inhibitors or those further inhibitors can be used which specifically block the binding to the organ-specific endothelial cells and / or the organ-specific extracellular structures.
  • At least one surface protein in particular an adhesion molecule
  • at least one aminopeptidase inhibitor can be influenced by at least one aminopeptidase inhibitor and / or by at least one further inhibitor.
  • a pharmaceutical preparation which can be produced using at least one aminopeptidase inhibitor and / or a combination of at least one aminopeptidase inhibitor and at least one further inhibitor as described above. Furthermore, one of the tasks listed is achieved by a method for identifying aminopeptidase inhibitors which block the polarization of invasive human or animal tumor and / or immune cells, the method first detecting surface protein combinations of a protein network which are on the surface of the untreated tumor and / or immune cells, the protein network consisting of up to 30 surface proteins from a group
  • CD4 1. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
  • CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
  • CD56 12. CD7 13. CD54 14. CD58 15. CD138
  • CD13 17. CD62L 18. CD71 19. CD11b 20. CD36
  • CD2 27.
  • CD20 28.
  • CD10 29.
  • the or similar tumor and / or immune cells are treated with at least one aminopeptidase inhibitor.
  • the surface protein combinations of the protein network that are located on the surface of the treated tumor and / or immune cells are then detected and compared with the surface protein combinations of the protein network that are located on the surface of the untreated tumor and / or immune cells.
  • the at least one aminopeptidase inhibitor causes the polarization of the tumor and / or immune cells to be blocked.
  • the at least one identified aminopeptidase inhibitor can be added to at least one polarizing tumor and / or immune cell and the further development of the at least one polarizing tumor and / or immune cell can be detected in order to demonstrate the actual blocking of the polarization.
  • the method can further comprise a control step in which the binding of the untreated tumor and / or immune cells to organ-specific endothelial cells and / or to organ-specific extracellular structures is detected, the binding of the tumor cells treated with the at least one identified aminopeptidase inhibitor / or immune cells to the organ-specific endothelial cells and / or the organ-specific extracellular structures is detected and the detected bonds are compared. If a reduced binding is found in the treated tumor and / or immune cells, the polarization is inhibited particularly effectively, since an effective organ-specific adhesion is prevented.
  • One of the above objects is achieved by a method for the identification of inhibitors which, in combination with at least one aminopeptidase inhibitor, block the polarization of invasive human or animal tumor and / or immune cells, in the method first of all surface protein combinations of a protein network which are located on the surface of the untreated tumor and / or immune cells can be detected, the protein network comprising up to 30 surface proteins from a group whose composition has already been listed above.
  • the or similar tumor and / or immune cells are treated with at least one potential inhibitor and the surface protein combinations of the protein network that are on the surface of the treated tumor and / or immune cells are detected.
  • the detected surface protein combinations are then compared, the at least one inhibitor being suitable for blocking the polarization of the tumor and / or immune cells in the event of a deviation due to at least one change in a surface protein.
  • the tumor and / or immune cells of the same type can also be treated with at least one aminopeptidase inhibitor, the combination of the at least one Inhibitor and the at least one aminopeptidase inhibitor in the event of a deviation of the surface protein combinations detected in the two different steps by at least one change in a surface protein CD13 causes the blocking of the polarization of the tumor and / or immune cells.
  • the method can also include a further step in which the at least one identified inhibitor or a combination of the at least one identified inhibitor and the at least one aminopeptide inhibitor is added to at least one polarizing tumor and / or immune cell and further development the at least one polarizing tumor and / or immune cell is detected.
  • the method can advantageously include a control step in which the binding of the untreated tumor and / or immune cells to organ-specific endothelial cells and / or to organ-specific extracellular structures is detected, in which the binding with the at least one identified inhibitor and / or with a combination from the at least one identified inhibitor and the at least one aminopeptidase inhibitor-treated tumor and / or immune cells to the organ-specific endothelial cells and / or the organ-specific extracellular structures is detected and in which the detected bonds are compared.
  • the detection of the surface protein combinations can comprise procedural steps of an automated method for determining molecular classes, molecular groups or molecular parts in a solid or liquid object according to DE 197 09 348 C.
  • control steps listed check whether the polarization is prevented by the at least one aminopeptidase inhibitor and / or the at least one further inhibitor by inhibiting the binding of certain molecules to defined structures.
  • These control steps can be carried out by passing immune cells (lymphocytes) and / or tumor cells in the form of a continuous cell flow in a special device, which is described in DE 199 32 158 A, over at least one sample with the defined structures. While the cells should be bound to the defined structures without treatment with the at least one aminopeptidase inhibitor and / or the at least one further inhibitor, after treatment of the cells with the at least one aminopeptidase inhibitor and / or the at least one further inhibitor takes place no binding or a reduced binding to the structures.
  • the sample can consist, for example, of an organ tissue section.
  • the figure shows, in chronological order, photographs of polarizing cells, untreated and treated with a target inhibitor.
  • the 30 cell surface proteins already listed above could be identified that belong to a specific protein network that controls the early phases of the polarization of tumor cells and immune cells.
  • surface protein combinations of this protein network were untreated by Karpas cells and detected after treatment with the aminopeptidase inhibitor actinonin.
  • two groups V1 and V2 of Karpas cells were formed, the cells of group V1 not being treated, while the cells of group V2 being treated with actinonin.
  • Table 2 lists those surface protein combinations which occur both in the actinonin-treated and in the untreated cells. In this and the other tables 3 and 4, however, only 18 of the 30 proteins are listed by way of example, the detected proteins being identified by 1 and the non-detected proteins being identified by 0.
  • CD2 1. CD3 3. CD4 4. CD8 5. CD16 6. CD56
  • CD57 7. CD26 9. CD38 10. CD71 11. HLA-DR 12. HLA-DQ
  • CD11b 14. CD45RA 15. CD7 16. CD62L 17. CD36 18. CD19
  • Tables 2 to 4 relate to 1000 cells examined in groups V1 and V2.
  • Table 2 shows a total of 203 different protein combinations.
  • Table 3 lists the surface protein combinations that only occur in the untreated Karpas cells and are never found in the Actinonin-treated Karpas cells.
  • the number of protein combinations referred to in Table 3 is 131.
  • Table 4 finally lists those surface protein combinations which only occur in the actinonin-treated Karpas cells. Table 4 contains 60 different protein combinations.
  • Tables 2 to 4 show that when only 18 proteins of the 30 proteins are considered, a total of 394 different surface protein combinations occur, with a total of 334 different combinations being found in the untreated cells and a total of 263 different combinations being detected in the treated cells can.
  • the change in the surface protein combinations detected hereby leads to a specific blocking of the cell polarization.
  • Protein code 1 Proteins [1 - 18], binary number number in
  • Protein code binary code per line 181 1 0 1 1 0 0 1 1 0 1 0 1 0 1 0 1 0 1 1 1 0 0 0.1 0.2
  • Protein code 2 proteins [1 - 1 3], binary number number in
  • 1 protein code binary code per line 87 0 1 0 1 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0.1 0.0
  • Protein code 3 proteins [1 - 1 3], binary number number in
  • 1 protein code binary code per line 0 1 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 1 0 0 0.0 0.1

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Abstract

The invention concerns the utilization of at least one aminopeptidase inhibitor for the production of a medicament used in the treatment of tumor diseases and/or immune diseases, whereby the at least one aminopeptidase inhibitor causes blocking of polarization of invasive human or animal tumor and/or immune cells by modifying at least one surface protein CD13 as member of a protein network on the surface of the tumor and/or immune cell, whereby the protein network comprises up to 30 surface proteins from a defined group. The invention also concerns a pharmaceutical preparation and a method for identifying at least one additional inhibitor acting in combination with the at least one aminopeptidase inhibitor.

Description

Verwendung mindestens eines Aminopeptidasen-Inhibitors Use of at least one aminopeptidase inhibitor
BESCHREIBUNG:DESCRIPTION:
Die vorliegende Erfindung betrifft eine Verwendung mindestens eines Aminopeptidasen-Inhibitors zur Herstellung eines Arzneimittels für die Behandlung von Tumorerkrankungen und/oder Immunkrankheiten, ein entsprechendes pharmazeutisches Präparat, ein Verfahren zur Identifikation mindestens eines Aminopeptidasen-Inhibitors und ein Verfahren zur Identifikation mindestens eines weiteren Inhibitors, der in Kombination mit dem mindestens einen Ami- nopeptidasen-lnhibitor wirkt.The present invention relates to the use of at least one aminopeptidase inhibitor for the manufacture of a medicament for the treatment of tumor diseases and / or immune diseases, a corresponding pharmaceutical preparation, a method for identifying at least one aminopeptidase inhibitor and a method for identifying at least one further inhibitor which works in combination with the at least one aminopeptidase inhibitor.
Aminopeptidasen sind Enzyme der Zelloberfläche, die Peptide spalten. Sie werden von verschiedenen Zelltypen exprimiert. Ihre molekulare Funktion besteht u.a. in der Degradation biologisch aktiver Peptide. Die weiteren physiologischen, insbesondere die zellulären Funktionen der Aminopeptidasen sind bisher nicht geklärt. Jüngste Untersuchungen zeigen, daß Aminopeptidasen- Inhibitoren die Vermehrungsrate und die Invasion von Tumorzellen unterdrük- ken können. Diese Unterdrückung der Invasion wurde allgemein auf die pro- teolytische Aktivität von Zelloberflächen-assoziierten Aminopeptidasen zurückgeführt, die die extrazellulären Matrixproteine spalten, so daß die Tumorzellen in Organe eindringen und in diesen Organen wandern können. Zu den bekannten Aminopeptidasen-Inhibitoren gehören Actinonin, Bestatin und potente Inhibitoren vom Homophtalimid-Typ. Bestatin kann gemäß J. Yoneda et al. Clin. Exp. Metastasis 10, 49-59, 1992 die Degradation des Typ IV Kollagens und damit die Invasion von Tumorzellen verhindern. Der genannten Publikation ist außerdem zu entnehmen, daß Bestatin keinen Einfluß auf die Tumorzelladhäsion und die Migration zu der extra- zellulären Matrix besitzt.Aminopeptidases are cell surface enzymes that cleave peptides. They are expressed by different cell types. Their molecular function consists, among other things, in the degradation of biologically active peptides. The further physiological, in particular the cellular functions of the aminopeptidases have not yet been clarified. Recent studies show that aminopeptidase inhibitors can suppress the rate of proliferation and invasion of tumor cells. This suppression of the invasion was generally attributed to the proteolytic activity of cell surface associated aminopeptidases, which cleave the extracellular matrix proteins so that the tumor cells can penetrate into organs and migrate in these organs. Known aminopeptidase inhibitors include actinonin, bestatin, and potent homophthalimide-type inhibitors. Bestatin can be according to J. Yoneda et al. Clin. Exp. Metastasis 10, 49-59, 1992 prevent the degradation of type IV collagen and thus the invasion of tumor cells. It can also be seen from the publication mentioned that bestatin has no influence on tumor cell adhesion and migration to the extracellular matrix.
Aus einer Veröffentlichung in Biol. Pharm. Bull. 22, 1010-1012, 1999 geht hervor, daß die Hemmung der Invasion durch einen Aminopeptidasen-Inhibitor vom Homophthalimid-Typ PIQ-22 auf eine Hemmung der Ausbildung von Zell- extensionen zurückzuführen ist, wobei die Ursache dieser Hemmung unklar bleibt und eine Inhibition der Aminopeptidase N, im folgenden auch mit CD13, bezeichnet, durch PIQ-22 ausgeschlossen wird. Nach der genannten Veröffentlichung haben die beiden Aminopeptidasen-Inhibitoren Actinonin und Bestatin, die CD13 hemmen, gemäß in vitro Untersuchungen mit einem unspezifi- sehen Matrigel-Analysesystem keine Wirkung auf die Tumorzellinvasion. Für Bestatin konnte auch keine Wirkung auf die Ausbildung von Zellextensionen nachgewiesen werden.A publication in Biol. Pharm. Bull. 22, 1010-1012, 1999 shows that the inhibition of invasion by an aminopeptidase inhibitor of the homophthalimide type PIQ-22 is due to an inhibition of the formation of cell extensions, whereby the cause of this inhibition remains unclear and inhibition of aminopeptidase N, hereinafter also referred to as CD13, is excluded by PIQ-22. According to the publication mentioned, the two aminopeptidase inhibitors actinonin and bestatin, which inhibit CD13, have no effect on tumor cell invasion according to in vitro investigations with a non-specific Matrigel analysis system. No effect on the development of cell extensions could be demonstrated for bestatin.
Besonders nachteilig ist die Tatsache, daß die bisher existierenden Untersu- chungsmethoden nicht die Bedingungen in vivo widerspiegeln und daher häufig zu unbefriedigenden Ergebnissen führen, so daß die Wirkung der einzelnen Aminopeptidasen-Inhibitoren nicht umfassend aufgeklärt werden kann. Meist bleibt unklar, ob die Inhibition der Aminopeptidasen bei Tumoren in vivo, d.h. am Patienten, wirksam ist und ob nicht sogar bei bestimmten Tumorformen durch Inhibition der Aminopeptidasen ein gegenteiliger Effekt, bestehend in einer Potenzierung des Invasionsverhaltens in vivo, eintreten könnte.A particular disadvantage is the fact that the previously existing investigation methods do not reflect the conditions in vivo and therefore often lead to unsatisfactory results, so that the action of the individual aminopeptidase inhibitors cannot be fully elucidated. It remains mostly unclear whether the inhibition of aminopeptidases in tumors in vivo, i.e. on the patient, is effective and whether an adverse effect, consisting in potentiating the invasive behavior in vivo, could not even occur in the case of certain types of tumor by inhibiting the aminopeptidases.
Außerdem ist bei den Aminopeptidasen-Inhibitoren mit bekannter Wirkung der Wirkmechanismus völlig unbekannt, so daß keine neuen Substanzen entwik- kelt oder identifiziert werden können, die nur deshalb äußerst spezifisch wirken können, weil sie in bekannter Art und Weise in die Zellfunktionen eingreifen. Beispielsweise konnte noch nicht geklärt werden, mit welchen anderen Protei- nen in ein- und derselben Zelle Aminopeptidasen Wechselwirkungen eingehen und in welcher Weise diese Wechselwirkungen komplexe Zellfunktionen kodieren. Es ist daher auch nicht bekannt, ob und welche zellulären Mechanismen, die auf solchen Wechselwirkungen beruhen, ggf. durch eine Inhibition der Ami- nopeptidasen gezielt blockiert werden können und welche neuen Indikationen sich hieraus für einen klinischen Einsatz solcher Inhibitoren bzw. der aus diesen Inhibitoren weiter entwickelten Substanzen ergeben könnten.In addition, the mechanism of action of the aminopeptidase inhibitors with a known effect is completely unknown, so that no new substances can be developed or identified which can only have an extremely specific effect because they affect the cell functions in a known manner. For example, it has not yet been clarified with which other protein interactions occur in the same cell and how these interactions encode complex cell functions. It is therefore also not known whether and which cellular mechanisms that are based on such interactions can be specifically blocked by inhibiting the aminopeptidases, and which new indications result from the clinical use of such inhibitors or from these inhibitors further developed substances could result.
Es ist daher Aufgabe der vorliegenden Erfindung einen Aminopeptidasen- Inhibitor mit vordefiniertem und kontrollierbarem Wirkverhalten bereitzustellen, der für die Herstellung eines Arzneimittels zur Behandlung von Tumorerkrankungen und/oder Immunkrankheiten verwendet werden kann. Es ist weiterhin Aufgabe der vorliegenden Erfindung, ein entsprechendes pharmazeutisches Präparat, ein Verfahren zur Identifikation mindestens eines solchen Aminopep- tidasen-lnhibitors und ein Verfahren zur Identifikation mindestens eines weiteren Inhibitors, der in Kombination mit dem mindestens einen Aminopeptidasen- Inhibitor wirkt, bereitzustellen.It is therefore an object of the present invention to provide an aminopeptidase inhibitor with predefined and controllable action which can be used for the production of a medicament for the treatment of tumor diseases and / or immune diseases. It is a further object of the present invention to provide a corresponding pharmaceutical preparation, a method for identifying at least one such aminopeptidease inhibitor and a method for identifying at least one further inhibitor which acts in combination with the at least one aminopeptidase inhibitor.
Eine der Aufgaben wird gelöst durch eine Verwendung mindestens eines Aminopeptidasen-Inhibitors zur Herstellung eines Arzneimittels zur Behandlung von Tumorerkrankungen und/oder Immunkrankheiten, wobei der mindestens eine Aminopeptidasen-Inhibitor eine Blockierung einer Polarisierung von inva- siven menschlichen oder tierischen Tumor- und/oder Immunzellen durch Veränderung mindestens eines Oberflächenproteins CD13 als Mitglied eines Pro- teinnetzwerkes auf der Oberfläche der Tumor- und/oder Immunzellen bewirkt, wobei das Proteinnetzwerk bis zu 30 Oberflächenproteine aus einer Gruppe bestehend ausOne of the objects is achieved by using at least one aminopeptidase inhibitor for the manufacture of a medicament for the treatment of tumor diseases and / or immune diseases, the at least one aminopeptidase inhibitor blocking polarization of invasive human or animal tumor and / or immune cells by modifying at least one surface protein CD13 as a member of a protein network on the surface of the tumor and / or immune cells, the protein network comprising up to 30 surface proteins from a group
1. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD31. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
6. CD26 7. CD38 8. CD45RA 9. CD16 10. CD576. CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
11. CD56 12. CD7 13. CD54 14. CD58 15. CD138 16. CD13 17. CD62L 18. CD71 19. CD11b 20. CD3611. CD56 12. CD7 13. CD54 14. CD58 15. CD138 16. CD13 17. CD62L 18. CD71 19. CD11b 20. CD36
21. CD29 22. CD49d 23. CD18 24. CD49f 25. CD1921. CD29 22. CD49d 23. CD18 24. CD49f 25. CD19
26. CD2 27. CD20 28. CD10 29. CD44 30. CD80 umfaßt.26. CD2 27. CD20 28. CD10 29. CD44 30. CD80 includes.
Mit Hilfe eines Verfahrens der simultanen Detektion einer beliebigen Anzahl von Proteinen der Zelloberfläche wurde gefunden, daß Aminopeptidasen Zell- oberflächenproteine kontrollieren, die nicht zu der Klasse der proteolytischen Enzyme, sondern zu der Klasse der Adhäsionsmoleküle gehören, wobei diese Adhäsionsmoleküle in einer bestimmten Kombination und räumlichen Anordnung entscheidend für die Polarisierung der Zellen sind. Aminopeptidasen sind hiernach übergeordnete Kontrollproteine in einem Proteinnetzwerk bestehend aus bis zu 30 verschiedenen Proteinspezies der Zelloberfläche, welche durch gezielte Wechselwirkung miteinander die Polarisierung von Tumorzellen und von anderen invasiven Zellen, wie Immunzellen, steuern und welche oben aufgeführt sind. Die Inhibition mindestens einer Aminopeptidase führt zu einer reproduzierbaren Veränderung von Oberflächenproteinkombinationen auf der Zelloberfläche, die stets auch eine Veränderung des CD13 umfaßt. Anhand zellbiologischer Experimente an Tumorzellen und an Immunzellen konnte gezeigt werden, daß die Inhibition und die damit verbundene Änderung der Ober- flächenproteinkombinationen zu einer vollständigen Blockierung der Polarisierung der Tumorzellen bzw. der Immunzellen führte. Unter Polarisierung ist hier ein Vorgang zu verstehen, bei dem eine primär sphärische Zelle über verschiedene Zwischenstadien in eine längliche, gestreckte Zellform übergeht. Dieser Vorgang, der eine durch das komplexe Proteinnetzwerk gesteuerte Formveränderung darstellt, ist die Voraussetzung für die Zellwanderung, also die Migration, da nur Zellen mit länglicher Zellform wandern können. Der Vorgang der Polarisierung ist daher allen Vorgängen der Zellwanderung und damit auch der Invasion zwingend vorgeordnet.Using a method for the simultaneous detection of any number of proteins on the cell surface, it was found that aminopeptidases control cell surface proteins which do not belong to the class of proteolytic enzymes but to the class of adhesion molecules, these adhesion molecules in a certain combination and spatial Arrangement are crucial for the polarization of the cells. Aminopeptidases are superordinate control proteins in a protein network consisting of up to 30 different protein species on the cell surface, which control the polarization of tumor cells and of other invasive cells, such as immune cells, through targeted interaction with one another and which are listed above. The inhibition of at least one aminopeptidase leads to a reproducible change in surface protein combinations on the cell surface, which always also includes a change in CD13. On the basis of cell biological experiments on tumor cells and on immune cells it could be shown that the inhibition and the associated change in the surface protein combinations led to a complete blocking of the polarization of the tumor cells or the immune cells. Here, polarization is to be understood as a process in which a primarily spherical cell changes into an elongated, elongated cell shape via various intermediate stages. This process, which is a shape change controlled by the complex protein network, is the prerequisite for cell migration, i.e. migration, since only cells with an elongated cell shape can migrate. The process of polarization is therefore imperative to all processes of cell migration and thus also to invasion.
Demgemäß liegt der Erfindung die Erkenntnis zugrunde, daß genau die Aminopeptidasen-Inhibitoren, die die Veränderung mindestens des Oberflächen- proteins CD13 als Mitglied des spezifischen, oben definierten Proteinnetzwerkes aus bis zu 30 Oberflächenproteinen bewirken, ganz spezifisch den ersten und daher wichtigsten Schritt der Invasion hemmen und daher zur Herstellung eines äußerst spezifisch wirkenden und damit überaus wirksamen Arzneimittels zur Behandlung von Tumorerkrankungen und/oder Immunkrankheiten verwendet werden können.Accordingly, the invention is based on the knowledge that it is precisely the aminopeptidase inhibitors which alter at least the surface proteins CD13, as a member of the specific protein network defined above, of up to 30 surface proteins, specifically inhibit the first and therefore most important step of the invasion and are therefore used to produce an extremely specific and therefore extremely effective drug for the treatment of tumor diseases and / or immune diseases can be.
Der mindestens eine Aminopeptidasen-Inhibitor kann beispielsweise ein Ami- nopeptidasen-lnhibitor vom Homophtalimid-Typ und/oder Actinonin und/oder Bestatin und/oder ein Antikörper, insbesondere ein monoklonaler Antikörper, gegen eines der Oberflächenproteine sein. Insbesondere Bestatin wirkt entgegen den oben dargelegten Annahmen durch den genannten Wirkmechanismus und führt zu einer Veränderung der Oberflächenproteine des Proteinnetzwerks, das Proteine aus der oben genannten Gruppe umfaßt. Als besonders wir- kungsvoll haben sich weiterhin das Actinonin, RB 3014 und ein gegen eine extrazelluläre Domäne des CD13 gerichteter, monoklonaler Antikörper (Klon SJ1 D1) erwiesen.The at least one aminopeptidase inhibitor can be, for example, an aminopeptidase inhibitor of the homophthalimide type and / or actinonin and / or bestatin and / or an antibody, in particular a monoclonal antibody, against one of the surface proteins. Bestatin in particular counteracts the assumptions set out above by the mechanism of action mentioned and leads to a change in the surface proteins of the protein network which comprises proteins from the group mentioned above. Actinonin, RB 3014 and a monoclonal antibody (clone SJ1 D1) directed against an extracellular domain of CD13 have also proven to be particularly effective.
Da durch die oben spezifizierten Aminopeptidasen-Inhibitoren neben der Pola- risierung von Tumorzellen auch die Polarisierung von Immunzellen wirkungsvoll verhindert werden kann, können durch Verwendung der Aminopeptidasen- Inhibitoren wirkungsvolle Arzneimittel zur Behandlung von Autoimmunkrankheiten oder Abstoßungsreaktionen von transplantierten Organen oder Allergien, insbesondere Allergien der Atemwege, bereitgestellt werden.Since the aminopeptidase inhibitors specified above can be used to effectively prevent the polarization of tumor cells as well as the polarization of immune cells, the use of aminopeptidase inhibitors enables effective medicinal products to be used to treat autoimmune diseases or rejection reactions of transplanted organs or allergies, in particular allergies to the respiratory tract , to be provided.
Vorteilhafterweise kann zur Herstellung des Arzneimittels mindestens ein weiterer Inhibitor eingesetzt wird, der mindestens ein Oberflächenprotein verändert und/oder inhibiert, das keine Aminopeptidase ist. Unter Inhibition ist in diesem Fall die allgemeine Hemmung der Funktion des mindestens einen Oberflä- chenproteins zu verstehen, die auch durch eine Expressionsänderung erfolgen kann. Durch den Einsatz einer Inhibitorkombination kann die Blockierung der Polarisierung enorm verstärkt werden. Beispielsweise kann als ein weiterer Inhibitor ein Antikörper gegen CD45RA eingesetzt werden. Gerade dieser Inhibitor verstärkt die Wirkung eines Aminopeptidasen-Inhibitor nach der obigen Definition besonders, so daß die Polarisierung durch diese Inhibitorkombination gezielt und besonders effektiv gehemmt werden kann.Advantageously, at least one further inhibitor can be used to manufacture the medicinal product, which inhibits and / or modifies at least one surface protein that is not an aminopeptidase. In this case, inhibition is to be understood as the general inhibition of the function of the at least one surface protein, which can also take place through a change in expression. The blocking of the polarization can be increased enormously by using an inhibitor combination. For example, as another Inhibitor an antibody against CD45RA can be used. This inhibitor in particular enhances the action of an aminopeptidase inhibitor as defined above, so that the polarization can be specifically and particularly effectively inhibited by this inhibitor combination.
Mindestens ein Aminopeptidasen-Inhibitor und/oder mindestens ein weiterer Inhibitor kann insbesondere neben der Veränderung des CD13 eine Veränderung mindestens eines weiteren Oberflächenproteins der Tumor- und/oder Immunzellen bewirken, das die Adhäsion an Endothelzellen und/oder extrazellulä- re Strukturen, insbesondere an organspezifische Endothelzellen und/oder an organspezifische extrazelluläre Strukturen, vermittelt. Mindestens ein Aminopeptidasen-Inhibitor und/oder mindestens ein weiterer Inhibitor kann auch eine Veränderung der adhäsiven Funktionen von Endothelzellen bewirken. Auf diese Weise kann verhindern werden, daß die Tumor- und/oder Immunzellen an die Endothelzellen binden, was für die Polarisierung unerläßlich ist. Um eine Invasion in ein bestimmtes Organ oder die Migration innerhalb dieses Organs gezielt zu verhindern, können diejenigen Aminopeptidase-Inhibitoren bzw. diejenigen weiteren Inhibitoren eingesetzt werden, die spezifisch die Bindung an die organspezifische Endothelzellen und/oder die organspezifischen extrazellu- lären Strukturen blockieren.At least one aminopeptidase inhibitor and / or at least one further inhibitor can, in particular in addition to the change in CD13, bring about a change in at least one further surface protein of the tumor and / or immune cells, which affects adhesion to endothelial cells and / or extracellular structures, in particular to organ-specific ones Endothelial cells and / or to organ-specific extracellular structures. At least one aminopeptidase inhibitor and / or at least one further inhibitor can also cause a change in the adhesive functions of endothelial cells. In this way it can be prevented that the tumor and / or immune cells bind to the endothelial cells, which is essential for polarization. In order to specifically prevent an invasion into a certain organ or the migration within this organ, those aminopeptidase inhibitors or those further inhibitors can be used which specifically block the binding to the organ-specific endothelial cells and / or the organ-specific extracellular structures.
Besonders vorteilhaft ist es weiterhin, wenn durch mindestens einen Aminopeptidasen-Inhibitor und/oder durch mindestens einen weiteren Inhibitor die Expression mindestens eines Oberflächenproteins, insbesondere eines Adhäsionsmoleküls, beeinflußbar ist.It is furthermore particularly advantageous if the expression of at least one surface protein, in particular an adhesion molecule, can be influenced by at least one aminopeptidase inhibitor and / or by at least one further inhibitor.
Eine der obigen Aufgaben wird durch ein pharmazeutisches Präparat gelöst, das unter einer Verwendung mindestens eines Aminopeptidasen-Inhibitors und/oder einer Kombination von mindestens einem Aminopeptidasen-Inhibitor und mindestens einem weiteren Inhibitor nach den obigen Ausführungen herstellbar ist. Weiterhin wird eine der aufgeführten Aufgaben durch ein Verfahren zur Identifikation von Aminopeptidasen-Inhibitoren gelöst, die eine Blockierung einer Polarisierung von invasiven menschlichen oder tierischen Tumor- und/oder Immunzellen bewirken, wobei bei dem Verfahren zunächst Oberflächenproteinkombi- nationen eines Proteinnetzwerkes detektiert werden, die sich auf der Oberfläche der unbehandelten Tumor- und/oder Immunzellen befinden, wobei das Proteinnetzwerk bis zu 30 Oberflächenproteine aus einer Gruppe bestehend ausOne of the above objects is achieved by a pharmaceutical preparation which can be produced using at least one aminopeptidase inhibitor and / or a combination of at least one aminopeptidase inhibitor and at least one further inhibitor as described above. Furthermore, one of the tasks listed is achieved by a method for identifying aminopeptidase inhibitors which block the polarization of invasive human or animal tumor and / or immune cells, the method first detecting surface protein combinations of a protein network which are on the surface of the untreated tumor and / or immune cells, the protein network consisting of up to 30 surface proteins from a group
1. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD31. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
6. CD26 7. CD38 8. CD45RA 9. CD16 10. CD576. CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
11. CD56 12. CD7 13. CD54 14. CD58 15. CD13811. CD56 12. CD7 13. CD54 14. CD58 15. CD138
16. CD13 17. CD62L 18. CD71 19. CD11b 20. CD3616. CD13 17. CD62L 18. CD71 19. CD11b 20. CD36
21. CD29 22. CD49d 23. CD18 24. CD49f 25. CD1921. CD29 22. CD49d 23. CD18 24. CD49f 25. CD19
26. CD2 27. CD20 28. CD10 29. CD44 30. CD8026. CD2 27. CD20 28. CD10 29. CD44 30. CD80
umfaßt. In einem nächsten Schritt werden die oder gleichartige Tumor- und/oder Immunzellen mit mindestens einem Aminopeptidasen-Inhibitor behandelt. Anschließend werden die Oberflächenproteinkombinationen des Proteinnetzwerkes, die sich auf der Oberfläche der behandelten Tumor- und/oder Immunzellen befinden, detektiert und mit den Oberflächenproteinkombinatio- nen des Proteinnetzwerks, die sich auf der Oberfläche der unbehandelten Tumor- und/oder Immunzellen befinden, verglichen. Der mindestens eine Aminopeptidasen-Inhibitor bewirkt bei einer Abweichung durch mindestens eine Veränderung des Oberflächenproteins CD13 die Blockierung der Polarisierung der Tumor- und/oder Immunzellen.includes. In a next step, the or similar tumor and / or immune cells are treated with at least one aminopeptidase inhibitor. The surface protein combinations of the protein network that are located on the surface of the treated tumor and / or immune cells are then detected and compared with the surface protein combinations of the protein network that are located on the surface of the untreated tumor and / or immune cells. In the event of a deviation due to at least one change in the surface protein CD13, the at least one aminopeptidase inhibitor causes the polarization of the tumor and / or immune cells to be blocked.
In einem zusätzlichen Schritt kann der mindestens eine identifizierte Aminopeptidasen-Inhibitor zu mindestens einer polarisierenden Tumor- und/oder Immunzelle zugegeben werden und die weitere Entwicklung der mindestens einen polarisierenden Tumor- und/oder Immunzelle detektiert werden, um so die tatsächliche Blockierung der Polarisierung nachzuweisen. Das Verfahren kann weiterhin einen Kontrollschritt umfassen, bei dem die Bindung der unbehandelten Tumor- und/oder Immunzellen an organspezifische Endothelzellen und/oder an organspezifische extrazelluläre Strukturen detek- tiert wird, die Bindung der mit dem mindestens einen identifizierten Aminopeptidasen-Inhibitor behandelten Tumor- und/oder Immunzellen an die organspezifischen Endothelzellen und/oder die organspezifischen extrazellulären Strukturen detektiert wird und die detektierten Bindungen verglichen werden. Wird bei den behandelten Tumor- und/oder Immunzellen eine verminderte Bindung festgestellt, wird die Polarisierung besonders effektiv gehemmt, da eine wirkungsvolle organspezifische Adhäsion verhindert wird.In an additional step, the at least one identified aminopeptidase inhibitor can be added to at least one polarizing tumor and / or immune cell and the further development of the at least one polarizing tumor and / or immune cell can be detected in order to demonstrate the actual blocking of the polarization. The method can further comprise a control step in which the binding of the untreated tumor and / or immune cells to organ-specific endothelial cells and / or to organ-specific extracellular structures is detected, the binding of the tumor cells treated with the at least one identified aminopeptidase inhibitor / or immune cells to the organ-specific endothelial cells and / or the organ-specific extracellular structures is detected and the detected bonds are compared. If a reduced binding is found in the treated tumor and / or immune cells, the polarization is inhibited particularly effectively, since an effective organ-specific adhesion is prevented.
Eine der obigen Aufgaben wird durch ein Verfahren zur Identifikation von Inhibitoren gelöst, die in Kombination mit mindestens einem Aminopeptidasen- Inhibitor eine Blockierung einer Polarisierung von invasiven menschlichen oder tierischen Tumor- und/oder Immunzellen bewirken, wobei in dem Verfahren zunächst Oberflächenproteinkombinationen eines Proteinnetzwerkes, die sich auf der Oberfläche der unbehandelten Tumor- und/oder Immunzellen befinden, detektiert werden, wobei das Proteinnetzwerk bis zu 30 Oberflächenproteine aus einer Gruppe umfaßt, deren Zusammensetzung bereits oben aufgeführt ist. Die oder gleichartige Tumor- und/oder Immunzellen werden mit mindestens einem potentiellen Inhibitor behandelt und die Oberflächenproteinkombinatio- nen des Proteinnetzwerkes, die sich auf der Oberfläche der behandelten Tumor- und/oder Immunzellen befinden, werden detektiert. Daraufhin werden die detektierten Oberflächenproteinkombinationen verglichen, wobei der mindestens eine Inhibitor bei einer Abweichung durch mindestens eine Veränderung eines Oberflächenproteins zur Blockierung der Polarisierung der Tumor- und/oder Immunzellen geeignet ist.One of the above objects is achieved by a method for the identification of inhibitors which, in combination with at least one aminopeptidase inhibitor, block the polarization of invasive human or animal tumor and / or immune cells, in the method first of all surface protein combinations of a protein network which are located on the surface of the untreated tumor and / or immune cells can be detected, the protein network comprising up to 30 surface proteins from a group whose composition has already been listed above. The or similar tumor and / or immune cells are treated with at least one potential inhibitor and the surface protein combinations of the protein network that are on the surface of the treated tumor and / or immune cells are detected. The detected surface protein combinations are then compared, the at least one inhibitor being suitable for blocking the polarization of the tumor and / or immune cells in the event of a deviation due to at least one change in a surface protein.
Die oder die gleichartigen Tumor- und/oder Immunzellen können neben dem mindestens einen Inhibitor auch mit mindestens einem Aminopeptidasen- Inhibitor behandelt werden, wobei die Kombination von dem mindestens einen Inhibitor und dem mindestens einen Aminopeptidasen-Inhibitor bei einer Abweichung der in den zwei verschiedenen Schritten detektierten Oberflächenproteinkombinationen durch mindestens eine Veränderung eines Oberflächenproteins CD13 die Blockierung der Polarisierung der Tumor- und/oder Immunzellen bewirkt.In addition to the at least one inhibitor, the tumor and / or immune cells of the same type can also be treated with at least one aminopeptidase inhibitor, the combination of the at least one Inhibitor and the at least one aminopeptidase inhibitor in the event of a deviation of the surface protein combinations detected in the two different steps by at least one change in a surface protein CD13 causes the blocking of the polarization of the tumor and / or immune cells.
Das Verfahren kann außerdem einen weiteren Schritt umfassen, bei dem der mindestens eine identifizierte Inhibitor oder eine Kombination aus dem mindestens einen identifizierten Inhibitor und dem mindestens einem Aminopeptida- sen-lnhibitor zu mindestens einer polarisierenden Tumor- und/oder Immunzelle zugegeben wird und die weitere Entwicklung der mindestens einen polarisierenden Tumor- und/oder Immunzelle detektiert wird.The method can also include a further step in which the at least one identified inhibitor or a combination of the at least one identified inhibitor and the at least one aminopeptide inhibitor is added to at least one polarizing tumor and / or immune cell and further development the at least one polarizing tumor and / or immune cell is detected.
Vorteilhafterweise kann das Verfahren einen Kontrollschritt umfassen, in dem die Bindung der unbehandelten Tumor- und/oder Immunzellen an organspezifische Endothelzellen und/oder an organspezifische extrazelluläre Strukturen detektiert wird, in dem die Bindung der mit dem mindestens einen identifizierten Inhibitor und/oder mit einer Kombination aus dem mindestens einen identifizierten Inhibitor und dem mindestens einen Aminopeptidasen-Inhibitor behandelten Tumor- und/oder Immunzellen an die organspezifischen Endothelzellen und/oder die organspezifischen extrazellulären Strukturen detektiert wird und in dem die detektierten Bindungen verglichen werden.The method can advantageously include a control step in which the binding of the untreated tumor and / or immune cells to organ-specific endothelial cells and / or to organ-specific extracellular structures is detected, in which the binding with the at least one identified inhibitor and / or with a combination from the at least one identified inhibitor and the at least one aminopeptidase inhibitor-treated tumor and / or immune cells to the organ-specific endothelial cells and / or the organ-specific extracellular structures is detected and in which the detected bonds are compared.
Grundsätzlich kann das Detektieren der Oberflächenproteinkombinationen Ver- fahrensschritte eines automatisierten Verfahrens zur Bestimmung von Molekülklassen, Molekülgruppen oder Molekülteilen in einem festen oder flüssigen Objekt gemäß der DE 197 09 348 C umfassen. Bei diesen Verfahrensschritten können in ein- und demselben Objekt, nämlich beispielsweise in einer Probe von Immunzellen und/oder Tumorzellen, Oberflächenproteine mittels sequen- tieller Aufbringung von Reagenzlösungen Yn (n = 1,2,3...N) mittels einer automatischen Vorrichtung untersucht und gemessen werden, wobei die Verfahrensschritte umfassen: I. Aufnahme einer ersten Reagenzlösung Y1 aus einem Behälter, der die Reagenzlösung enthält,In principle, the detection of the surface protein combinations can comprise procedural steps of an automated method for determining molecular classes, molecular groups or molecular parts in a solid or liquid object according to DE 197 09 348 C. In these process steps, surface proteins can be examined in one and the same object, namely, for example, in a sample of immune cells and / or tumor cells, by means of sequential application of reagent solutions Yn (n = 1,2,3 ... N) by means of an automatic device and measured, the process steps comprising: I. receiving a first reagent solution Y1 from a container containing the reagent solution,
II. Aufbringen der Reagenzlösung Y1 auf das Objekt, das sich auf einer objekttragenden Vorrichtung befindet, III. Einwirken der Reagenzlösung über einen automatisch eingestellten Zeitraum,II. Applying the reagent solution Y1 to the object, which is located on an object-carrying device, III. Exposure of the reagent solution over an automatically set period of time,
IV. Aufnahme von mindestens einem Einzelmarkierungsmuster des zuvor mit der ersten Reagenzlösung Y1 markierten Objekts,IV. Recording at least one individual marking pattern of the object previously marked with the first reagent solution Y1,
V. Wiederholung der Schritte l-IV durch Aufbringen der ersten Reagenzlösung Y1 oder einer zweiten Reagenzlösung Y2 oder eines Gemisches aus erster und zweiter Reagenzlösung, undV. repetition of steps I-IV by applying the first reagent solution Y1 or a second reagent solution Y2 or a mixture of first and second reagent solutions, and
VI. Wiederholung der Schritte I bis V mit weiteren Reagenzlösungen Yn (n = 2,3... N) oder einem Gemisch davon und wobei die in jedem Verfahrenszyklus erhaltenen Markierungsverteilungsmuster durch computergestützte Bildüberlagerung in ein komplexes molekulares Kombinationsmuster des zu untersuchenden Objekts überführt werden.VI. Repetition of steps I to V with further reagent solutions Yn (n = 2.3 ... N) or a mixture thereof and wherein the marker distribution patterns obtained in each process cycle are converted into a complex molecular combination pattern of the object to be examined by computer-aided image superimposition.
Aus diesem Kombinationsmuster können Informationen über das Vorhandensein der oben genannten Proteine gewonnen werden und damit auch die Oberflächenproteinkombinationen detektiert werden, wenn die verwendeten Reagenzlösungen markierte Substanzen enthalten, die gegen die entsprechenden Proteine gerichtet sind.Information about the presence of the abovementioned proteins can be obtained from this combination pattern and thus also the surface protein combinations can be detected if the reagent solutions used contain labeled substances which are directed against the corresponding proteins.
Bei den aufgeführten Kontrollschritten wird überprüft, ob die Polarisierung durch den mindestens einen Aminopeptidasen-Inhibitor und/oder den mindestens einen weiteren Inhibitor verhindert wird, indem die Bindung bestimmter Moleküle an definierte Strukturen gehemmt wird. Diese Kontrollschritte können durchgeführt werden, indem Immunzellen (Lymphozyten) und/oder Tumorzellen in Form eines kontinuierlichen Zellflusses in einem speziellen Gerät, das in der DE 199 32 158 A beschrieben wird, über mindestens eine Probe mit den definierten Strukturen geleitet werden. Während ohne Behandlung mit dem mindestens einen Aminopeptidasen- Inhibitor und/oder dem mindestens einen weiteren Inhibitor eine Bindung der Zellen an die definierten Strukturen erfolgen sollte, findet nach der Behandlung der Zellen mit dem mindestens einen Aminopeptidasen-Inhibitor und/oder dem mindestens einen weiteren Inhibitor keine Bindung oder eine verminderte Bindung an die Strukturen statt. Die Probe kann beispielsweise aus einem Organgewebeschnitt bestehen.The control steps listed check whether the polarization is prevented by the at least one aminopeptidase inhibitor and / or the at least one further inhibitor by inhibiting the binding of certain molecules to defined structures. These control steps can be carried out by passing immune cells (lymphocytes) and / or tumor cells in the form of a continuous cell flow in a special device, which is described in DE 199 32 158 A, over at least one sample with the defined structures. While the cells should be bound to the defined structures without treatment with the at least one aminopeptidase inhibitor and / or the at least one further inhibitor, after treatment of the cells with the at least one aminopeptidase inhibitor and / or the at least one further inhibitor takes place no binding or a reduced binding to the structures. The sample can consist, for example, of an organ tissue section.
Weitere Merkmale und Vorteile der Erfindung gehen aus den im folgenden aufgeführten Untersuchungsergebnissen hervor, die unter anderem anhand von einer Figur beschrieben werden.Further features and advantages of the invention emerge from the test results listed below, which are described, inter alia, using a figure.
Die Figur zeigt in zeitlicher Abfolge Fotografien von polarisierenden Zel- len, unbehandelt und mit einem Targetinhibitor behandelt.The figure shows, in chronological order, photographs of polarizing cells, untreated and treated with a target inhibitor.
Mit Hilfe zahlreicher Untersuchungen konnten die bereits oben aufgeführten 30 Zeiloberflächenproteine identifiziert werden, die zu einem spezifischen Proteinnetzwerk gehören, das die frühen Phasen der Polarisierung von Tumorzellen und Immunzellen kontrolliert. In einer weiteren Untersuchung wurden nun Oberflächenproteinkombinationen dieses Proteinnetzwerkes von Karpas-Zellen unbehandelt und nach der Behandlung mit dem Aminopeptidasen-Inhibitor Actinonin detektiert. Zu diesem Zweck wurden zwei Gruppen V1 und V2 von Karpas-Zellen gebildet, wobei die Zellen der Gruppe V1 nicht behandelt wurden, während die Zellen der Gruppe V2 mit Actinonin behandelt wurden. In Tabelle 2 sind diejenigen Oberflächenproteinkombinationen zusammengestellt, die sowohl bei den mit Actinonin behandelten als auch bei den unbehandelten Zellen vorkommen. In dieser und den weiteren Tabellen 3 und 4 sind jedoch beispielhaft nur 18 der 30 Proteine aufgeführt, wobei die detektierten Proteine mit 1 gekennzeichnet sind und die nicht-detektierten Proteine mit 0 gekennzeichnet sind.With the help of numerous studies, the 30 cell surface proteins already listed above could be identified that belong to a specific protein network that controls the early phases of the polarization of tumor cells and immune cells. In a further investigation, surface protein combinations of this protein network were untreated by Karpas cells and detected after treatment with the aminopeptidase inhibitor actinonin. For this purpose, two groups V1 and V2 of Karpas cells were formed, the cells of group V1 not being treated, while the cells of group V2 being treated with actinonin. Table 2 lists those surface protein combinations which occur both in the actinonin-treated and in the untreated cells. In this and the other tables 3 and 4, however, only 18 of the 30 proteins are listed by way of example, the detected proteins being identified by 1 and the non-detected proteins being identified by 0.
Die Proteine sind von 1 bis 18 durchnumeriert, wobei die Nomenklatur aus der Tabelle 1 hervorgeht. Tabelle 1The proteins are numbered from 1 to 18, with the nomenclature shown in Table 1. Table 1
1. CD2 2. CD3 3. CD4 4. CD8 5. CD16 6. CD561. CD2 2. CD3 3. CD4 4. CD8 5. CD16 6. CD56
7. CD57 8. CD26 9. CD38 10. CD71 11. HLA-DR 12. HLA-DQ7. CD57 8. CD26 9. CD38 10. CD71 11. HLA-DR 12. HLA-DQ
13. CD11b 14. CD45RA 15. CD7 16. CD62L 17. CD36 18. CD1913. CD11b 14. CD45RA 15. CD7 16. CD62L 17. CD36 18. CD19
Die in den Tabellen 2 bis 4 angegebenen Zellanzahlen beziehen sich auf jeweils 1000 untersuchte Zellen in den Gruppen V1 und V2. In der Tabelle 2 ist eine Gesamtanzahl von 203 unterschiedlichen Proteinkombinationen aufgeführt.The cell numbers given in Tables 2 to 4 relate to 1000 cells examined in groups V1 and V2. Table 2 shows a total of 203 different protein combinations.
In der Tabelle 3 sind die Oberflächenproteinkombinationen aufgeführt, die nur bei den unbehandelten Karpas-Zellen vorkommen und nie bei den Actinonin- behandelten Karpas-Zellen zu finden sind. Die Anzahl der in dieser Tabelle 3 bezeichneten Proteinkombinationen beträgt 131.Table 3 lists the surface protein combinations that only occur in the untreated Karpas cells and are never found in the Actinonin-treated Karpas cells. The number of protein combinations referred to in Table 3 is 131.
In der Tabelle 4 sind schließlich diejenigen Oberflächenproteinkombinationen aufgeführt, die ausschließlich bei den Actinonin-behandelten Karpas-Zellen vorkommen. Die Tabelle 4 enthält 60 verschiedene Proteinkombinationen.Table 4 finally lists those surface protein combinations which only occur in the actinonin-treated Karpas cells. Table 4 contains 60 different protein combinations.
Aus den Tabellen 2 bis 4 geht hervor, daß bei Betrachtung von lediglich 18 Proteinen der 30 Proteine insgesamt 394 unterschiedliche Oberflächenprotein- kombinationen auftreten, wobei bei den unbehandelten Zellen insgesamt 334 unterschiedliche Kombinationen zu finden sind und bei den behandelten Zellen insgesamt 263 unterschiedliche Kombinationen detektiert werden können. Die hiermit nachgewiesene Änderung der Oberflächenproteinkombinationen führt zu einer spezifischen Blockierung der Zellpolarisierung.Tables 2 to 4 show that when only 18 proteins of the 30 proteins are considered, a total of 394 different surface protein combinations occur, with a total of 334 different combinations being found in the untreated cells and a total of 263 different combinations being detected in the treated cells can. The change in the surface protein combinations detected hereby leads to a specific blocking of the cell polarization.
Eine weitere Untersuchung wird anhand der Figur erläutert. In (I) ist der normale zelluläre Prozeß der Polarisierung einer Tumorzelle gezeigt. Durch in vitro life imaging wird erfaßt, wie sich eine Sarkom-Zelle aus einer primär sphärischen Zellform heraus über die Bildung von 3 Zellextensionen (tripolare Zellform) und anschließende gezielte Rückbildung nur einer der drei Extensionen (weißer Pfeil bei 360 min) polarisiert. Die Definition einer Längsachse ist Voraussetzung für die darauf folgende Zellmigration. In (II) ist nachgewiesen, daß die Applikation eines selektiven Targetinhibitiors, hier ein monoklonaler Antikörper gegen eine extrazelluläre Domäne des CD13 (schwarzer Pfeil), die Zell- polarisierung vollständig verhindert. Die Zelle wird sphärisch und hoch adhäsiv, was aus dem Vergleich zwischen einer Fotografie der inhibierten Zelle (II) nach 480 min und einer Fotografie der nicht-inhibierten Zelle (I) nach 480 min (I) hervorgeht.A further investigation is explained using the figure. The normal cellular process of polarization of a tumor cell is shown in (I). In vitro life imaging captures how a sarcoma cell emerges from a primarily spherical cell shape via the formation of 3 cell extensions (tripolar cell shape) and subsequent targeted regression of only one of the three extensions (white arrow at 360 min) polarized. The definition of a longitudinal axis is a prerequisite for the subsequent cell migration. In (II) it has been demonstrated that the application of a selective target inhibitor, here a monoclonal antibody against an extracellular domain of CD13 (black arrow), completely prevents cell polarization. The cell becomes spherical and highly adhesive, which is evident from the comparison between a photograph of the inhibited cell (II) after 480 min and a photograph of the uninhibited cell (I) after 480 min (I).
Ähnliche Ergebnisse erhält man bei dem Einsatz von Actinonin oder Bestatin als Targetinhibitor. Similar results are obtained when using actinonin or bestatin as a target inhibitor.
Tabelle 2Table 2
V1 V2 ZellanZellan¬V1 V2 cell cell cell
Proteincode1 Proteine [1 - 18], binär zahl zahl in inProtein code 1 Proteins [1 - 18], binary number number in
Nr. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1/1000 1/1000No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1/1000 1/1000
1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 136.4 153.21 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 136.4 153.2
2 0 0 1 0 0 0 0 1 0 1 0 1 0 0 1 0 0 0 78.2 91.92 0 0 1 0 0 0 0 1 0 1 0 1 0 0 1 0 0 0 78.2 91.9
3 0 0 1 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 71.9 57.63 0 0 1 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 71.9 57.6
4 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 40.0 62.24 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 40.0 62.2
5 0 0 1 0 0 0 0 1 0 0 0 1 0 0 1 0 0 0 50.7 37.95 0 0 1 0 0 0 0 1 0 0 0 1 0 0 1 0 0 0 50.7 37.9
6 0 0 1 0 0 0 0 1 0 1 0 1 0 0 0 0 0 0 43.9 44.36 0 0 1 0 0 0 0 1 0 1 0 1 0 0 0 0 0 0 43.9 44.3
7 0 0 1 1 0 0 0 1 0 1 0 1 0 0 1 0 0 0 43.0 37.67 0 0 1 1 0 0 0 1 0 1 0 1 0 0 1 0 0 0 43.0 37.6
8 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 26.5 41.88 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 26.5 41.8
9 0 0 1 0 0 0 0 1 0 1 1 0 0 1 0 0 0 33.2 31.49 0 0 1 0 0 0 0 1 0 1 1 0 0 1 0 0 0 33.2 31.4
10 0 0 1 1 0 0 0 1 0 1 1 0 1 1 0 0 0 23.5 29.710 0 0 1 1 0 0 0 1 0 1 1 0 1 1 0 0 0 23.5 29.7
11 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 20.6 29.311 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 20.6 29.3
12 0 0 1 1 0 0 0 1 0 1 1 0 0 1 0 0 0 21.5 24.212 0 0 1 1 0 0 0 1 0 1 1 0 0 1 0 0 0 21.5 24.2
13 0 0 1 0 0 0 0 1 0 0 1 0 0 1 0 0 0 29.4 15.413 0 0 1 0 0 0 0 1 0 0 1 0 0 1 0 0 0 29.4 15.4
14 0 0 0 0 0 0 0 1 0 1 0 1 0 0 0 0 0 0 13.2 22.314 0 0 0 0 0 0 0 1 0 1 0 1 0 0 0 0 0 0 13.2 22.3
15 1 0 1 1 0 0 0 1 0 1 1 0 1 1 0 0 0 15.1 17.315 1 0 1 1 0 0 0 1 0 1 1 0 1 1 0 0 0 15.1 17.3
16 0 0 1 0 0 0 0 1 0 1 1 0 1 1 0 0 0 17.1 15.016 0 0 1 0 0 0 0 1 0 1 1 0 1 1 0 0 0 17.1 15.0
17 0 0 1 0 0 0 0 1 0 1 0 1 0 1 1 0 0 0 15.7 15.517 0 0 1 0 0 0 0 1 0 1 0 1 0 1 1 0 0 0 15.7 15.5
18 0 0 1 1 0 0 0 1 0 1 0 1 0 1 1 0 0 0 14.6 9.718 0 0 1 1 0 0 0 1 0 1 0 1 0 1 1 0 0 0 14.6 9.7
19 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 0 0 0 9.8 13.419 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 0 0 0 9.8 13.4
20 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 3.7 14.520 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 3.7 14.5
21 0 0 1 1 0 0 0 1 0 0 0 1 0 0 1 0 0 0 11.1 6.321 0 0 1 1 0 0 0 1 0 0 0 1 0 0 1 0 0 0 11.1 6.3
22 0 0 1 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 9.2 6.722 0 0 1 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 9.2 6.7
23 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 7.5 7.423 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 7.5 7.4
24 0 0 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 8.6 5.724 0 0 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 8.6 5.7
25 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 6.9 7.325 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 6.9 7.3
26 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 3.1 11.026 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 3.1 11.0
27 0 1 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 5.9 7.527 0 1 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 5.9 7.5
28 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 4.4 8.928 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 4.4 8.9
29 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 0 0 0 4.9 7.029 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 0 0 0 4.9 7.0
30 0 0 1 1 0 0 0 1 0 0 1 1 0 0 1 0 0 0 7.7 4.030 0 0 1 1 0 0 0 1 0 0 1 1 0 0 1 0 0 0 7.7 4.0
31 1 0 1 0 0 0 0 1 0 1 1 1 0 1 1 0 0 0 6.8 2.831 1 0 1 0 0 0 0 1 0 1 1 1 0 1 1 0 0 0 6.8 2.8
32 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 3.5 6.132 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 3.5 6.1
33 0 0 1 0 0 0 0 1 0 1 1 1 0 0 0 0 0 0 4.2 5.333 0 0 1 0 0 0 0 1 0 1 1 1 0 0 0 0 0 0 4.2 5.3
34 0 0 1 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 6.2 3.234 0 0 1 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 6.2 3.2
35 0 0 1 0 0 0 0 1 0 0 0 1 0 1 1 0 0 0 4.5 4.835 0 0 1 0 0 0 0 1 0 0 0 1 0 1 1 0 0 0 4.5 4.8
36 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 3.0 5.436 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 3.0 5.4
37 0 1 0 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 4.2 3.937 0 1 0 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 4.2 3.9
38 0 0 1 0 0 0 0 1 1 1 0 1 0 0 1 0 0 0 3.8 4.338 0 0 1 0 0 0 0 1 1 1 0 1 0 0 1 0 0 0 3.8 4.3
39 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 5.5 2.539 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 5.5 2.5
Proteincode = Binärcode pro Zeile 181 1 0 1 1 0 0 1 1 0 1 0 1 0 1 1 0 0 0 0.1 0.2Protein code = binary code per line 181 1 0 1 1 0 0 1 1 0 1 0 1 0 1 1 0 0 0 0.1 0.2
182 0 1 0 0 0 0 0 0 1 1 0 0 0 1 1 0 0 0 0.1 0.2182 0 1 0 0 0 0 0 0 1 1 0 0 0 1 1 0 0 0 0.1 0.2
183 0 0 1 1 0 0 0 1 0 1 0 0 0 0 1 0 0 0 0.1 0.2183 0 0 1 1 0 0 0 1 0 1 0 0 0 0 1 0 0 0 0.1 0.2
184 0 0 1 0 0 0 0 1 1 0 1 1 0 1 1 0 0 0 0.1 0.2184 0 0 1 0 0 0 0 1 1 0 1 1 0 1 1 0 0 0 0.1 0.2
185 0 0 1 0 0 0 0 1 1 0 1 1 0 0 0 0 0 0 0.1 0.2185 0 0 1 0 0 0 0 1 1 0 1 1 0 0 0 0 0 0 0.1 0.2
186 0 0 0 0 0 0 0 1 1 0 1 1 0 0 0 0 0 0 0.1 0.2186 0 0 0 0 0 0 0 1 1 0 1 1 0 0 0 0 0 0 0.1 0.2
187 1 1 1 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0.1 0.1187 1 1 1 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0.1 0.1
188 1 0 1 0 0 0 1 1 0 1 1 1 0 1 1 1 0 0 0.1 0.1188 1 0 1 0 0 0 1 1 0 1 1 1 0 1 1 1 0 0 0.1 0.1
189 0 1 1 1 0 0 0 1 1 1 0 0 0 0 1 1 0 0 0.1 0.1189 0 1 1 1 0 0 0 1 1 1 0 0 0 0 1 1 0 0 0.1 0.1
190 0 1 1 1 0 0 0 1 1 0 0 1 0 0 1 0 0 0 0.1 0.1190 0 1 1 1 0 0 0 1 1 0 0 1 0 0 1 0 0 0 0.1 0.1
191 0 1 0 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.1 0.1191 0 1 0 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.1 0.1
192 0 0 1 1 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.1 0.1192 0 0 1 1 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.1 0.1
193 0 0 1 1 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0.1 0.1193 0 0 1 1 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0.1 0.1
194 0 0 1 0 0 0 1 1 0 0 1 1 0 1 1 0 0 0 0.1 0.1194 0 0 1 0 0 0 1 1 0 0 1 1 0 1 1 0 0 0 0.1 0.1
195 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 1 0 0 0.1 0.1195 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 1 0 0 0.1 0.1
196 0 0 1 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0.1 0.1196 0 0 1 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0.1 0.1
197 0 0 1 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0.1 0.1197 0 0 1 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0.1 0.1
198 0 0 0 1 0 0 0 0 0 1 1 1 0 0 1 0 0 0 0.1 0.1198 0 0 0 1 0 0 0 0 0 1 1 1 0 0 1 0 0 0 0.1 0.1
199 0 0 0 1 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0.1 0.1199 0 0 0 1 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0.1 0.1
200 0 0 0 1 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0.1 0.1200 0 0 0 1 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0.1 0.1
201 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0.1 0.1201 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0.1 0.1
202 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 0.1 0.1202 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 0.1 0.1
203 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 1 0 0 0.1 0.1 gesamt 971.3 990.9 203 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 1 0 0 0.1 0.1 total 971.3 990.9
Tabelle 3Table 3
V1 V2 ZellanZellan¬V1 V2 cell cell cell
Proteincode2 Proteine [1 - 1 3], binär zahl zahl in inProtein code 2 proteins [1 - 1 3], binary number number in
Nr. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1/1000 1/1000No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1/1000 1/1000
1 0 0 1 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 1.7 0.01 0 0 1 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 1.7 0.0
2 0 0 0 1 0 0 0 1 0 1 0 1 0 0 0 0 0 0 1.2 0.02 0 0 0 1 0 0 0 1 0 1 0 1 0 0 0 0 0 0 1.2 0.0
3 0 1 1 1 0 0 0 1 1 1 0 1 0 1 1 1 0 0 0.8 0.03 0 1 1 1 0 0 0 1 1 1 0 1 0 1 1 1 0 0 0.8 0.0
4 1 0 1 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 0.7 0.04 1 0 1 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 0.7 0.0
5 0 0 1 0 0 0 1 1 0 1 0 1 0 1 1 0 0 0 0.7 0.05 0 0 1 0 0 0 1 1 0 1 0 1 0 1 1 0 0 0 0.7 0.0
6 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0.7 0.06 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0.7 0.0
7 1 0 1 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0.6 0.07 1 0 1 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0.6 0.0
8 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0.6 0.08 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0.6 0.0
9 1 1 1 1 0 0 0 1 1 0 1 0 1 1 1 0 0 0.5 0.09 1 1 1 1 0 0 0 1 1 0 1 0 1 1 1 0 0 0.5 0.0
10 0 0 1 1 0 0 0 1 1 0 1 0 0 0 0 0 0 0.5 0.010 0 0 1 1 0 0 0 1 1 0 1 0 0 0 0 0 0 0.5 0.0
11 0 0 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.5 0.011 0 0 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.5 0.0
12 1 1 1 0 0 0 0 1 1 0 1 0 1 1 1 0 0 0.4 0.012 1 1 1 0 0 0 0 1 1 0 1 0 1 1 1 0 0 0.4 0.0
13 1 1 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0.4 0.013 1 1 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0.4 0.0
14 1 0 1 1 0 0 0 1 1 1 1 0 0 1 0 0 0 0.4 0.014 1 0 1 1 0 0 0 1 1 1 1 0 0 1 0 0 0 0.4 0.0
15 1 0 1 0 0 0 0 1 0 0 0 1 0 1 1 0 0 0 0.4 0.015 1 0 1 0 0 0 0 1 0 0 0 1 0 1 1 0 0 0 0.4 0.0
16 1 0 0 1 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0.4 0.016 1 0 0 1 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0.4 0.0
17 1 0 0 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0.4 0.017 1 0 0 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0.4 0.0
18 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0.4 0.018 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0.4 0.0
19 0 1 1 0 0 0 0 1 1 1 0 1 0 1 1 1 0 0 0.4 0.019 0 1 1 0 0 0 0 1 1 1 0 1 0 1 1 1 0 0 0.4 0.0
20 0 1 1 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0.4 0.020 0 1 1 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0.4 0.0
21 0 1 0 0 0 0 0 0 1 1 0 1 0 0 1 1 0 0 0.4 0.021 0 1 0 0 0 0 0 0 1 1 0 1 0 0 1 1 0 0 0.4 0.0
22 0 1 0 0 0 0 0 0 1 1 0 0 0 1 1 1 0 0 0.4 0.022 0 1 0 0 0 0 0 0 1 1 0 0 0 1 1 1 0 0 0.4 0.0
23 0 0 1 1 0 0 0 1 1 0 1 1 0 0 1 0 0 0 0.4 0.023 0 0 1 1 0 0 0 1 1 0 1 1 0 0 1 0 0 0 0.4 0.0
24 0 0 1 1 0 0 0 1 1 0 0 1 0 0 1 0 0 0 0.4 0.024 0 0 1 1 0 0 0 1 1 0 0 1 0 0 1 0 0 0 0.4 0.0
25 0 0 1 0 0 0 0 1 0 1 0 1 1 0 1 0 0 0 0.4 0.025 0 0 1 0 0 0 0 1 0 1 0 1 1 0 1 0 0 0 0.4 0.0
26 0 0 1 0 0 0 0 1 0 0 1 0 0 0 1 0 0 0 0.4 0.026 0 0 1 0 0 0 0 1 0 0 1 0 0 0 1 0 0 0 0.4 0.0
27 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0.4 0.027 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0.4 0.0
28 1 1 1 0 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.2 0.028 1 1 1 0 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.2 0.0
29 1 1 0 0 0 0 0 1 1 1 0 0 0 1 1 1 0 0 0.2 0.029 1 1 0 0 0 0 0 1 1 1 0 0 0 1 1 1 0 0 0.2 0.0
30 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0.2 0.030 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0.2 0.0
31 1 0 1 0 0 0 1 1 1 1 1 1 0 1 1 0 0 0 0.2 0.031 1 0 1 0 0 0 1 1 1 1 1 1 0 1 1 0 0 0 0.2 0.0
32 0 1 1 1 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.2 0.032 0 1 1 1 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.2 0.0
33 0 1 1 1 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0.2 0.033 0 1 1 1 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0.2 0.0
34 0 1 1 1 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0.2 0.034 0 1 1 1 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0.2 0.0
35 0 0 1 1 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0.2 0.035 0 0 1 1 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0.2 0.0
36 0 0 1 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0.2 0.036 0 0 1 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0.2 0.0
37 0 0 1 0 0 0 0 1 0 1 1 1 0 1 1 1 0 0 0.2 0.037 0 0 1 0 0 0 0 1 0 1 1 1 0 1 1 1 0 0 0.2 0.0
38 0 0 1 0 0 0 0 1 0 1 1 0 0 0 1 0 0 0 0.2 0.038 0 0 1 0 0 0 0 1 0 1 1 0 0 0 1 0 0 0 0.2 0.0
39 0 0 1 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0.2 0.039 0 0 1 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0.2 0.0
1 Proteincode = Binärcode pro Zeile 87 0 1 0 1 0 0 0 0 1 0 0 0 0 1 0 0 0 0.1 0.0 1 protein code = binary code per line 87 0 1 0 1 0 0 0 0 1 0 0 0 0 1 0 0 0 0.1 0.0
88 0 1 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.1 0.088 0 1 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.1 0.0
89 0 1 0 0 0 0 0 1 1 0 1 0 1 1 0 0 0 0.1 0.089 0 1 0 0 0 0 0 1 1 0 1 0 1 1 0 0 0 0.1 0.0
90 0 1 0 0 0 0 0 1 1 0 1 0 0 0 1 0 0 0.1 0.090 0 1 0 0 0 0 0 1 1 0 1 0 0 0 1 0 0 0.1 0.0
91 0 1 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0.1 0.091 0 1 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0.1 0.0
92 0 0 1 1 0 0 0 1 1 0 1 0 1 1 1 0 0 0.1 0.092 0 0 1 1 0 0 0 1 1 0 1 0 1 1 1 0 0 0.1 0.0
93 0 0 1 1 0 0 0 1 0 1 1 1 1 1 1 0 0 0 0.1 0.093 0 0 1 1 0 0 0 1 0 1 1 1 1 1 1 0 0 0 0.1 0.0
94 0 0 1 1 0 0 0 1 0 1 1 1 1 0 1 1 0 0 0.1 0.094 0 0 1 1 0 0 0 1 0 1 1 1 1 0 1 1 0 0 0.1 0.0
95 0 0 1 1 0 0 0 1 0 1 0 0 0 1 1 0 0 0 0.1 0.095 0 0 1 1 0 0 0 1 0 1 0 0 0 1 1 0 0 0 0.1 0.0
96 0 0 1 1 0 0 0 1 0 0 1 1 0 0 1 1 0 0 0.1 0.096 0 0 1 1 0 0 0 1 0 0 1 1 0 0 1 1 0 0 0.1 0.0
97 0 0 1 1 0 0 0 1 0 0 0 1 0 0 1 1 0 0 0.1 0.097 0 0 1 1 0 0 0 1 0 0 0 1 0 0 1 1 0 0 0.1 0.0
98 0 0 1 1 0 0 0 0 1 1 0 1 0 0 1 0 0 0 0.1 0.098 0 0 1 1 0 0 0 0 1 1 0 1 0 0 1 0 0 0 0.1 0.0
99 0 0 1 1 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0.1 0.099 0 0 1 1 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0.1 0.0
100 0 0 1 0 0 0 1 1 1 1 1 1 0 1 1 0 0 0 0.1 0.0100 0 0 1 0 0 0 1 1 1 1 1 1 0 1 1 0 0 0 0.1 0.0
101 0 0 1 0 0 0 1 1 1 1 0 1 0 1 1 0 0 0 0.1 0.0101 0 0 1 0 0 0 1 1 1 1 0 1 0 1 1 0 0 0 0.1 0.0
102 0 0 1 0 0 0 1 1 1 0 1 1 0 1 1 0 0 0 0.1 0.0102 0 0 1 0 0 0 1 1 1 0 1 1 0 1 1 0 0 0 0.1 0.0
103 0 0 1 0 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.1 0.0103 0 0 1 0 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.1 0.0
104 0 0 1 0 0 0 0 1 1 1 0 1 1 0 1 0 0 0 0.1 0.0104 0 0 1 0 0 0 0 1 1 1 0 1 1 0 1 0 0 0 0.1 0.0
105 0 0 1 0 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.1 0.0105 0 0 1 0 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.1 0.0
106 0 0 1 0 0 0 0 1 1 0 0 1 0 0 0 1 0 0 0.1 0.0106 0 0 1 0 0 0 0 1 1 0 0 1 0 0 0 1 0 0 0.1 0.0
107 0 0 1 0 0 0 0 1 0 1 1 1 1 0 1 0 0 0 0.1 0.0107 0 0 1 0 0 0 0 1 0 1 1 1 1 0 1 0 0 0 0.1 0.0
108 0 0 1 0 0 0 0 1 0 1 0 1 0 0 0 1 0 0 0.1 0.0108 0 0 1 0 0 0 0 1 0 1 0 1 0 0 0 1 0 0 0.1 0.0
109 0 0 1 0 0 0 0 1 0 1 0 0 0 1 1 0 0 0 0.1 0.0109 0 0 1 0 0 0 0 1 0 1 0 0 0 1 1 0 0 0 0.1 0.0
110 0 0 1 0 0 0 0 1 0 0 1 1 0 1 0 0 0 0 0.1 0.0110 0 0 1 0 0 0 0 1 0 0 1 1 0 1 0 0 0 0 0.1 0.0
111 0 0 1 0 0 0 0 1 0 0 0 1 0 1 1 1 0 0 0.1 0.0111 0 0 1 0 0 0 0 1 0 0 0 1 0 1 1 1 0 0 0.1 0.0
112 0 0 1 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.1 0.0112 0 0 1 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0.1 0.0
113 0 0 1 0 0 0 0 0 1 0 1 1 0 0 1 0 0 0 0.1 0.0113 0 0 1 0 0 0 0 0 1 0 1 1 0 0 1 0 0 0 0.1 0.0
114 0 0 1 0 0 0 0 0 1 0 0 1 0 0 1 0 0 0 0.1 0.0114 0 0 1 0 0 0 0 0 1 0 0 1 0 0 1 0 0 0 0.1 0.0
115 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0.1 0.0115 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0.1 0.0
116 0 0 1 0 0 0 0 0 0 1 1 0 0 0 1 0 0 0 0.1 0.0116 0 0 1 0 0 0 0 0 0 1 1 0 0 0 1 0 0 0 0.1 0.0
117 0 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0.1 0.0117 0 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0.1 0.0
118 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0.1 0.0118 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0.1 0.1
119 0 0 0 1 0 0 0 1 0 0 0 1 0 1 1 0 0 0 0.1 0.0119 0 0 0 1 0 0 0 1 0 0 0 1 0 1 1 0 0 0 0.1 0.0
120 0 0 0 1 0 0 0 0 1 0 0 1 0 0 1 0 0 0 0.1 0.0120 0 0 0 1 0 0 0 0 1 0 0 1 0 0 1 0 0 0 0.1 0.0
121 0 0 0 1 0 0 0 0 0 1 0 1 0 0 1 0 0 0 0.1 0.0121 0 0 0 1 0 0 0 0 0 1 0 1 0 0 1 0 0 0 0.1 0.0
122 0 0 0 1 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0.1 0.0122 0 0 0 1 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0.1 0.0
123 0 0 0 1 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0.1 0.0123 0 0 0 1 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0.1 0.0
124 0 0 0 0 0 0 0 1 1 1 0 1 0 1 1 0 0 0 0.1 0.0124 0 0 0 0 0 0 0 1 1 1 0 1 0 1 1 0 0 0 0.1 0.0
125 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 0.1 0.0125 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 0.1 0.0
126 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0.1 0.0126 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0.1 0.0
127 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0.1 0.0127 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0.1 0.0
128 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0.1 0.0128 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0.1 0.0
129 0 0 0 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0.1 0.0129 0 0 0 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0.1 0.0
130 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0.1 0.0130 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0.1 0.0
131 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0.1 0.0 gesamt 28.7 0 Tabelle 4131 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0.1 0.0 total 28.7 0 Table 4
V1 V2 ZellanZellan¬V1 V2 cell cell cell
Proteincode3 Proteine [1 - 1 3], binär zahl zahl in inProtein code 3 proteins [1 - 1 3], binary number number in
Nr. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1/1000 1/1000No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1/1000 1/1000
1 1 1 1 1 0 0 0 1 1 1 0 0 0 0 1 1 0 0 0.0 0.61 1 1 1 1 0 0 0 1 1 1 0 0 0 0 1 1 0 0 0.0 0.6
2 0 0 1 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0.0 0.62 0 0 1 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0.0 0.6
3 1 1 1 0 0 0 0 1 1 1 0 0 0 0 1 1 0 0 0.0 0.33 1 1 1 0 0 0 0 1 1 1 0 0 0 0 1 1 0 0 0.0 0.3
4 0 0 1 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0.0 0.34 0 0 1 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0.0 0.3
5 0 1 1 1 0 0 0 1 1 1 1 1 0 0 1 1 0 0 0.0 0.25 0 1 1 1 0 0 0 1 1 1 1 1 0 0 1 1 0 0 0.0 0.2
6 0 1 1 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0.0 0.26 0 1 1 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0.0 0.2
7 0 1 0 0 0 0 0 1 1 1 0 0 0 0 0 1 0 0 0.0 0.27 0 1 0 0 0 0 0 1 1 1 0 0 0 0 0 1 0 0 0.0 0.2
8 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.28 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.2
9 0 0 1 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0.0 0.29 0 0 1 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0.0 0.2
10 0 0 0 0 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0.0 0.210 0 0 0 0 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0.0 0.2
11 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0.0 0.211 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0.0 0.2
12 1 1 1 1 0 0 0 1 1 1 1 1 0 0 1 0 0 0 0.0 0.112 1 1 1 1 0 0 0 1 1 1 1 1 0 0 1 0 0 0 0.0 0.1
13 1 1 1 1 0 0 0 0 1 1 0 0 1 1 1 1 0 0 0.0 0.113 1 1 1 1 0 0 0 0 1 1 0 0 1 1 1 1 0 0 0.0 0.1
14 1 1 1 1 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0.0 0.114 1 1 1 1 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0.0 0.1
15 1 1 0 1 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0.0 0.115 1 1 0 1 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0.0 0.1
16 1 1 0 0 0 0 0 0 1 1 1 0 0 0 1 0 0 0 0.0 0.116 1 1 0 0 0 0 0 0 1 1 1 0 0 0 1 0 0 0 0.0 0.1
17 1 1 0 0 0 0 0 0 1 1 0 0 1 1 1 1 0 0 0.0 0.117 1 1 0 0 0 0 0 0 1 1 0 0 1 1 1 1 0 0 0.0 0.1
18 1 0 1 1 0 0 1 1 1 1 1 1 0 1 1 1 0 0 0.0 0.118 1 0 1 1 0 0 1 1 1 1 1 1 0 1 1 1 0 0 0.0 0.1
19 1 0 1 1 0 0 1 1 0 1 0 1 0 1 1 1 0 0 0.0 0.119 1 0 1 1 0 0 1 1 0 1 0 1 0 1 1 1 0 0 0.0 0.1
20 1 0 1 1 0 0 1 1 0 1 0 0 0 1 1 0 0 0 0.0 0.120 1 0 1 1 0 0 1 1 0 1 0 0 0 1 1 0 0 0 0.0 0.1
21 1 0 1 1 0 0 0 1 1 1 0 1 0 0 1 0 0 0 0.0 0.121 1 0 1 1 0 0 0 1 1 1 0 1 0 0 1 0 0 0 0.0 0.1
22 1 0 0 1 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0.0 0.122 1 0 0 1 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0.0 0.1
23 1 0 0 1 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0.0 0.123 1 0 0 1 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0.0 0.1
24 1 0 0 0 0 0 0 1 0 0 0 1 0 1 1 0 0 0 0.0 0.124 1 0 0 0 0 0 0 1 0 0 0 1 0 1 1 0 0 0 0.0 0.1
25 1 0 0 0 0 0 0 0 0 1 0 1 0 1 1 1 0 0 0.0 0.125 1 0 0 0 0 0 0 0 0 1 0 1 0 1 1 1 0 0 0.0 0.1
26 1 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0.0 0.126 1 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0.0 0.1
27 0 1 1 1 0 0 0 1 1 0 0 0 0 0 1 0 0 0 0.0 0.127 0 1 1 1 0 0 0 1 1 0 0 0 0 0 1 0 0 0 0.0 0.1
28 0 1 1 1 0 0 0 1 0 1 0 1 0 1 1 0 0 0 0.0 0.128 0 1 1 1 0 0 0 1 0 1 0 1 0 1 1 0 0 0 0.0 0.1
29 0 1 1 1 0 0 0 0 1 1 0 1 0 0 1 0 0 0 0.0 0.129 0 1 1 1 0 0 0 0 1 1 0 1 0 0 1 0 0 0 0.0 0.1
30 0 1 1 0 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.0 0.130 0 1 1 0 0 0 0 1 1 1 1 1 0 1 1 1 0 0 0.0 0.1
31 0 1 1 0 0 0 0 1 1 0 1 1 0 0 1 0 0 0 0.0 0.131 0 1 1 0 0 0 0 1 1 0 1 1 0 0 1 0 0 0 0.0 0.1
32 0 1 1 0 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.0 0.132 0 1 1 0 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.0 0.1
33 0 1 1 0 0 0 0 0 1 1 0 1 0 0 1 1 0 0 0.0 0.133 0 1 1 0 0 0 0 0 1 1 0 1 0 0 1 1 0 0 0.0 0.1
34 0 1 1 0 0 0 0 0 1 1 0 0 1 0 0 1 0 0 0.0 0.134 0 1 1 0 0 0 0 0 1 1 0 0 1 0 0 1 0 0 0.0 0.1
35 0 1 1 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0.0 0.135 0 1 1 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0.0 0.1
36 0 1 0 1 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0.0 0.136 0 1 0 1 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0.0 0.1
37 0 1 0 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0.0 0.137 0 1 0 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0.0 0.1
38 0 1 0 0 0 0 0 1 1 0 1 1 0 0 1 0 0 0 0.0 0.138 0 1 0 0 0 0 0 1 1 0 1 1 0 0 1 0 0 0 0.0 0.1
39 0 1 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0.0 0.139 0 1 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0.0 0.1
1 Proteincode = Binärcode pro Zeile 0 1 0 0 0 0 0 0 1 1 0 1 0 0 0 1 0 0 0.0 0.1 1 protein code = binary code per line 0 1 0 0 0 0 0 0 1 1 0 1 0 0 0 1 0 0 0.0 0.1
0 0 1 1 0 0 1 1 0 1 1 1 0 1 1 0 0 0 0.0 0.10 0 1 1 0 0 1 1 0 1 1 1 0 1 1 0 0 0 0.0 0.1
0 0 1 1 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0.0 0.10 0 1 1 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0.0 0.1
0 0 1 1 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.0 0.10 0 1 1 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.0 0.1
0 0 1 1 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0.0 0.10 0 1 1 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0.0 0.1
0 0 1 0 0 0 1 1 0 1 0 1 0 1 1 1 0 0 0.0 0.10 0 1 0 0 0 1 1 0 1 0 1 0 1 1 1 0 0 0.0 0.1
0 0 1 0 0 0 0 1 1 1 0 1 0 0 1 1 0 0 0.0 0.10 0 1 0 0 0 0 1 1 1 0 1 0 0 1 1 0 0 0.0 0.1
0 0 1 0 0 0 0 1 1 0 1 0 0 0 0 0 0 0 0.0 0.10 0 1 0 0 0 0 1 1 0 1 0 0 0 0 0 0 0 0.0 0.1
0 0 1 0 0 0 0 1 0 1 1 1 0 1 0 0 0 0 0.0 0.10 0 1 0 0 0 0 1 0 1 1 1 0 1 0 0 0 0 0.0 0.1
0 0 1 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0.0 0.10 0 1 0 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0.0 0.1
0 0 1 0 0 0 0 1 0 1 0 1 0 1 0 0 0 0 0.0 0.10 0 1 0 0 0 0 1 0 1 0 1 0 1 0 0 0 0 0.0 0.1
0 0 1 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0.0 0.10 0 1 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0.0 0.1
0 0 1 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0.0 0.10 0 1 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0.0 0.1
0 0 0 1 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0.0 0.10 0 0 1 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0.0 0.1
0 0 0 1 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0.0 0.10 0 0 1 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0.0 0.1
0 0 0 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0.0 0.10 0 0 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0.0 0.1
0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0.0 0.10 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0.0 0.1
0 0 0 0 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.0 0.10 0 0 0 0 0 0 1 1 0 0 1 0 1 1 0 0 0 0.0 0.1
0 0 0 0 0 0 0 1 0 1 1 1 0 1 1 0 0 0 0.0 0.10 0 0 0 0 0 0 1 0 1 1 1 0 1 1 0 0 0 0.0 0.1
0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0.0 0.10 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0.0 0.1
0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0.0 0.1 gesamt 0.0 9.1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0.0 0.1 total 0.0 9.1

Claims

Verwendung eines Aminopeptidasen-InhibitorsANSPRÜCHE:Verwendung mindestens eines Aminopeptidasen-Inhibitors zur Herstellung eines Arzneimittels zur Behandlung von Tumorerkrankungen und/oder Immunkrankheiten, wobei der mindestens eine Aminopeptidasen-Inhibitor eine Blockierung einer Polarisierung von invasiven menschlichen oder tierischen Tumor- und/oder Immunzellen durch Veränderung mindestens eines Oberflächenproteins CD13 als Mitglied eines Proteinnetzwerkes auf der Oberfläche der Tumor- und/oder Immunzellen bewirkt, wobei das Proteinnetzwerk bis zu 30 Oberflächenproteine aus einer Gruppe bestehend aus Use of an aminopeptidase inhibitor Claims: Use of at least one aminopeptidase inhibitor for the manufacture of a medicament for the treatment of tumor diseases and / or immune diseases, wherein the at least one aminopeptidase inhibitor blocks the polarization of invasive human or animal tumor and / or immune cells by altering at least causes a surface protein CD13 as a member of a protein network on the surface of the tumor and / or immune cells, the protein network comprising up to 30 surface proteins from a group
1. CD41. CD4
2. CD8 3. HLA-DR 4. HLA-DQ 5. CD32. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
6. CD26 7. CD38 8. CD45RA 9. CD16 10. CD576. CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
11. CD56 12. CD7 13. CD54 14. CD58 15. CD13811. CD56 12. CD7 13. CD54 14. CD58 15. CD138
16. CD13 17. CD62L 18. CD71 19. CD11 b 20. CD3616. CD13 17. CD62L 18. CD71 19. CD11 b 20. CD36
21. CD29 22. CD49d 23. CD18 24. CD49f 25. CD1921. CD29 22. CD49d 23. CD18 24. CD49f 25. CD19
26. CD2 27. CD20 28. CD10 29. CD44 30. CD8026. CD2 27. CD20 28. CD10 29. CD44 30. CD80
umfaßt.includes.
Verwendung nach Anspruch 1 , dadurch gekennzeichnet, daß der mindestens eine Aminopeptidasen-Inhibitor ein Aminopeptidasen-Inhibitor vom Homophtalimid-Typ und/oder Actinonin und/oder Bestatin und/oder ein Antikörper, insbesondere ein monoklonaler Antikörper, gegen eines der Oberflächenproteine ist.Use according to claim 1, characterized in that that the at least one aminopeptidase inhibitor is an aminopeptidase inhibitor of the homophthalimide type and / or actinonin and / or bestatin and / or an antibody, in particular a monoclonal antibody, against one of the surface proteins.
3. Verwendung nach einem der vorhergehenden Ansprüche, dadurch gekennzeichnet, daß die Immunkrankheiten Autoimmunkrankheiten oder Abstoßungsreaktionen von transplantierten Organen oder Allergien, insbesondere Al- lergien der Atemwege, sind.3. Use according to one of the preceding claims, characterized in that the immune diseases are autoimmune diseases or rejection reactions of transplanted organs or allergies, in particular allergies to the respiratory tract.
4. Verwendung nach einem der vorhergehenden Ansprüche, dadurch gekennzeichnet, daß zur Herstellung des Arzneimittels mindestens ein weiterer Inhibitor eingesetzt wird, der mindestens ein Oberflächenprotein inhibiert, das keine Aminopeptidase ist.4. Use according to one of the preceding claims, characterized in that at least one further inhibitor is used for the production of the medicament which inhibits at least one surface protein which is not an aminopeptidase.
5. Verwendung nach einem der vorhergehenden Ansprüche, dadurch gekennzeichnet, daß mindestens ein Aminopeptidasen-Inhibitor und/oder mindestens ein weiterer Inhibitor eine Veränderung mindestens eines Oberflächenproteins der Tumor- und/oder Immunzellen bewirkt, das die Adhäsion an Endothelzellen und/oder extrazelluläre Strukturen, insbesondere organspezifische Endothelzellen und/oder organspezifische extrazelluläre Strukturen, vermittelt.5. Use according to one of the preceding claims, characterized in that at least one aminopeptidase inhibitor and / or at least one further inhibitor brings about a change in at least one surface protein of the tumor and / or immune cells which results in the adhesion to endothelial cells and / or extracellular structures, in particular organ-specific endothelial cells and / or organ-specific extracellular structures.
6. Verwendung nach einem der vorhergehenden Ansprüche, dadurch gekennzeichnet, daß mindestens ein Aminopeptidasen-Inhibitor und/oder mindestens ein weiterer Inhibitor eine Veränderung der adhäsiven Funktionen von Endothelzellen bewirkt. 6. Use according to one of the preceding claims, characterized in that at least one aminopeptidase inhibitor and / or at least one further inhibitor brings about a change in the adhesive functions of endothelial cells.
7. Verwendung nach einem der vorhergehenden Ansprüche, dadurch gekennzeichnet, daß durch mindestens einen Aminopeptidasen-Inhibitor und/oder durch mindestens einen weiteren Inhibitor die Expression mindestens eines Oberflächenproteins, insbesondere eines Adhäsionsmoleküls, beeinflußbar ist.7. Use according to any one of the preceding claims, characterized in that the expression of at least one surface protein, in particular an adhesion molecule, can be influenced by at least one aminopeptidase inhibitor and / or by at least one further inhibitor.
8. Pharmazeutisches Präparat, das unter einer Verwendung mindestens eines Aminopeptidasen-Inhibitors oder einer Kombination von minde- stens einem Aminopeptidasen-Inhibitor und mindestens einem weiteren8. Pharmaceutical preparation which uses at least one aminopeptidase inhibitor or a combination of at least one aminopeptidase inhibitor and at least one other
Inhibitor nach den Ansprüchen 1 bis 7 herstellbar ist.Inhibitor according to claims 1 to 7 can be produced.
9. Verfahren zur Identifikation mindestens eines Aminopeptidasen- Inhibitors, der eine Blockierung einer Polarisierung von invasiven menschlichen oder tierischen Tumor- und/oder Immunzellen bewirkt, folgende Schritte umfassend:9. A method for identifying at least one aminopeptidase inhibitor which blocks the polarization of invasive human or animal tumor and / or immune cells, comprising the following steps:
a) Detektieren von Oberflächenproteinkombinationen eines Proteinnetzwerkes, die sich auf der Oberfläche der unbehandelten Tu- mor- und/oder Immunzellen befinden, wobei das Proteinnetzwerk bis zu 30 Oberflächenproteine aus einer Gruppe bestehend ausa) Detecting surface protein combinations of a protein network which are located on the surface of the untreated tumor and / or immune cells, the protein network comprising up to 30 surface proteins from a group
1. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD31. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
6. CD26 7. CD38 8. CD45RA 9. CD16 10. CD576. CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
11. CD56 12. CD7 13. CD54 14. CD58 15. CD 13811.CD56 12.CD7 13.CD54 14.CD58 15.CD 138
16. CD13 17. CD62L 18. CD71 19. CD11b 20. CD3616. CD13 17. CD62L 18. CD71 19. CD11b 20. CD36
21. CD29 22. CD49d 23. CD18 24. CD49f 25. CD1921. CD29 22. CD49d 23. CD18 24. CD49f 25. CD19
26. CD2 27. CD20 28. CD10 29. CD44 30. CD8026. CD2 27. CD20 28. CD10 29. CD44 30. CD80
umfaßt; b) Behandeln der oder gleichartiger Tumor- und/oder Immunzellen mit mindestens einem Aminopeptidasen-Inhibitor;comprises; b) treating the or similar tumor and / or immune cells with at least one aminopeptidase inhibitor;
c) Detektieren der Oberflächenproteinkombinationen des Protein- netzwerkes, die sich auf der Oberfläche der behandelten Tumor- und/oder Immunzellen befinden; undc) detecting the surface protein combinations of the protein network which are located on the surface of the treated tumor and / or immune cells; and
d) Vergleichen der in Schritt a) und in Schritt c) detektierten Oberflä- chenproteinkombinationen, wobei der mindestens eine Amino- peptidasen-lnhibitor bei einer Abweichung der in Schritt a) detektierten Oberflächenproteinkombinationen von den in Schritt c) detektierten Oberflächenproteinkombinationen durch mindestens eine Veränderung des Oberflächenproteins CD13 die Blockierung der Polarisierung der Tumor- und/oder Immunzellen bewirkt.d) comparing the surface protein combinations detected in step a) and in step c), the at least one amino peptidase inhibitor if the surface protein combinations detected in step a) differ from the surface protein combinations detected in step c) by at least one change in the Surface protein CD13 causes the polarization of the tumor and / or immune cells to be blocked.
10. Verfahren nach Anspruch 9, dadurch gekennzeichnet, daß das Verfahren nach Schritt d) einen weiteren Schritt umfaßt, in dem der mindestens eine in Schritt d) identifizierte Aminopeptidasen-Inhibitor zu mindestens einer polarisierenden Tumor- und/oder Immunzelle zugegeben wird und die weitere Entwicklung der mindestens einen polarisierenden Tumor- und/oder Immunzelle detektiert wird.10. The method according to claim 9, characterized in that the method after step d) comprises a further step in which the at least one aminopeptidase inhibitor identified in step d) is added to at least one polarizing tumor and / or immune cell and the further Development of the at least one polarizing tumor and / or immune cell is detected.
11. Verfahren nach einem der Ansprüche 9 oder 10, dadurch gekennzeichnet, daß das Verfahren nach Schritt d) einen weiteren Schritt umfaßt, in dem die Bindung der unbehandelten Tumor- und/oder Immunzellen an organspezifische Endothelzellen und/oder an organspezifische extrazelluläre Strukturen detektiert wird, in dem die Bindung der mit dem minde- stens einen in Schritt d) identifizierten Aminopeptidasen-Inhibitor behandelten Tumor- und/oder Immunzellen an die organspezifischen Endo- thelzellen und/oder die organspezifischen extrazellulären Strukturen detektiert wird und in dem die detektierten Bindungen verglichen werden.11. The method according to any one of claims 9 or 10, characterized in that the method after step d) comprises a further step in which the binding of the untreated tumor and / or immune cells to organ-specific endothelial cells and / or to organ-specific extracellular structures is detected , in which the binding of the tumor and / or immune cells treated with the at least one aminopeptidase inhibitor identified in step d) to the organ-specific endo- cell cells and / or the organ-specific extracellular structures is detected and in which the detected bonds are compared.
12. Verfahren zur Identifikation mindestens eines Inhibitors, der in Kombination mit mindestens einem Aminopeptidasen-Inhibitor eine Blockierung einer Polarisierung von invasiven menschlichen oder tierischen Tumor- und/oder Immunzellen bewirkt, folgende Schritte umfassend:12. A method for identifying at least one inhibitor which, in combination with at least one aminopeptidase inhibitor, blocks polarization of invasive human or animal tumor and / or immune cells, comprising the following steps:
a) Detektieren von Oberflächenproteinkombinationen eines Proteinnetzwerkes, die sich auf der Oberfläche der unbehandelten Tumor- und/oder Immunzellen befinden, wobei das Proteinnetzwerk bis zu 30 Oberflächenproteine aus einer Gruppe bestehend ausa) Detecting surface protein combinations of a protein network which are located on the surface of the untreated tumor and / or immune cells, the protein network comprising up to 30 surface proteins from a group
1. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD31. CD4 2. CD8 3. HLA-DR 4. HLA-DQ 5. CD3
6. CD26 7. CD38 8. CD45RA 9. CD16 10. CD576. CD26 7. CD38 8. CD45RA 9. CD16 10. CD57
11. CD56 12. CD7 13. CD54 14. CD58 15. CD13811. CD56 12. CD7 13. CD54 14. CD58 15. CD138
16. CD1316. CD13
17. CD62L17. CD62L
18. CD7118. CD71
19. CD11b19. CD11b
20. CD3620. CD36
21. CD2921. CD29
22. CD49d22. CD49d
23. CD1823. CD18
24. CD49f24. CD49f
25. CD1925. CD19
26. CD2 27. CD20 28. CD10 29. CD44 30. CD8026. CD2 27. CD20 28. CD10 29. CD44 30. CD80
umfaßt;comprises;
b) Behandeln der oder gleichartiger Tumor- und/oder Immunzellen mit mindestens einem potentiellen Inhibitor, der nicht gegen eine Aminopeptidase gerichtet ist;b) treating the or similar tumor and / or immune cells with at least one potential inhibitor which is not directed against an aminopeptidase;
c) Detektieren der Oberflächenproteinkombinationen des Proteinnetzwerkes, die sich auf der Oberfläche der behandelten Tumor- und/oder Immunzellen befinden; undc) detecting the surface protein combinations of the protein network which are located on the surface of the treated tumor and / or immune cells; and
d) Vergleichen der in Schritt a) und in Schritt c) detektierten Oberflä- chenproteinkombinationen, wobei der mindestens eine Inhibitor bei einer Abweichung der in Schritt a) detektierten Oberflächen- proteinkombinationen von den in Schritt c) detektierten Oberfiä- chenproteinkombinationen durch mindestens eine Veränderung eines Oberflächenproteins zur Blockierung der Polarisierung der Tumor- und/oder Immunzellen geeignet ist.d) comparing the surface protein combinations detected in step a) and in step c), the at least one inhibitor in the event of a deviation of the surface protein combinations detected in step a) from the surface protein combinations detected in step c) by at least one change in a surface protein is suitable for blocking the polarization of the tumor and / or immune cells.
13. Verfahren nach Anspruch 12, dadurch gekennzeichnet, daß die oder die gleichartigen Tumor- und/oder Immunzellen in Schritt b) auch mit mindestens einem Aminopeptidasen-Inhibitor behandelt werden, wobei die Kombination von dem mindestens einen Inhibitor und dem mindestens einen Aminopeptidasen-Inhibitor bei einer Abweichung der in Schritt a) detektierten Oberflächenproteinkombinationen von den in Schritt c) detektierten Oberflächenproteinkombinationen durch minde- stens eine Veränderung eines Oberflächenproteins CD 13 die Blockierung der Polarisierung der Tumor- und/oder Immunzellen bewirkt.13. The method according to claim 12, characterized in that the or the same tumor and / or immune cells in step b) are also treated with at least one aminopeptidase inhibitor, the combination of the at least one inhibitor and the at least one aminopeptidase inhibitor if the surface protein combinations detected in step a) deviate from the surface protein combinations detected in step c) by at least one change in a surface protein CD 13, the polarization of the tumor and / or immune cells is blocked.
14. Verfahren nach einem der Ansprüche 12 oder 13, dadurch gekennzeichnet, daß das Verfahren nach Schritt d) einen weiteren Schritt umfaßt, in dem der mindestens eine in Schritt d) identifizierte Inhibitor oder eine Kombination aus dem mindestens einen in Schritt d) identifizierten Inhibitor und mindestens einem Aminopeptidasen-Inhibitor zu mindestens einer polarisierenden Tumor- und/oder Immunzelle zugegeben wird und die weitere Entwicklung der mindestens einen polarisierenden Tumor- und/oder Immunzelle detektiert wird.14. The method according to any one of claims 12 or 13, characterized in that the method after step d) comprises a further step in which the at least one inhibitor identified in step d) or a combination of the at least one inhibitor identified in step d) and at least one aminopeptidase inhibitor is added to at least one polarizing tumor and / or immune cell and the further development of the at least one polarizing tumor and / or immune cell is detected.
15. Verfahren nach einem der Ansprüche 12 bis 14, dadurch gekennzeichnet, daß das Verfahren nach Schritt d) einen weiteren Schritt umfaßt, in dem die Bindung der unbehandelten Tumor- und/oder Immunzellen an organspezifische Endothelzellen und/oder an organspezifische extrazellu- läre Strukturen detektiert wird, in dem die Bindung der mit dem mindestens einen in Schritt d) identifizierten Inhibitor oder mit einer Kombination aus dem mindestens einen in Schritt d) identifizierten Inhibitor und mindestens einem Aminopeptidasen-Inhibitor behandelten Tumor- und/oder Immunzellen an die organspezifischen Endothelzellen und/oder die organspezifischen extrazellulären Strukturen detektiert wird und in dem die detektierten Bindungen verglichen werden. 15. The method according to any one of claims 12 to 14, characterized in that the method after step d) comprises a further step in which the binding of the untreated tumor and / or immune cells to organ-specific endothelial cells and / or to organ-specific extracellu- Lär structures is detected by binding the tumor and / or immune cells treated with the at least one inhibitor identified in step d) or with a combination of the at least one inhibitor identified in step d) and at least one aminopeptidase inhibitor to the organ-specific Endothelial cells and / or the organ-specific extracellular structures is detected and in which the detected bonds are compared.
EP01911521A 2000-01-24 2001-01-24 Utilization of an aminopeptidase inhibitor Ceased EP1210109A2 (en)

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