WO2004016810A2

WO2004016810A2 - Use of an mrp4 binding substances for the diagnosis and treatment of cancer

Info

Publication number: WO2004016810A2
Application number: PCT/DE2003/002159
Authority: WO
Inventors: Uta Schwidetzky; Christian Pilarsky; Bernd Hinzmann; Thomas Specht; André ROSENTHAL; Rosemarie Lichtner; Edgar Dahl
Original assignee: Metagen Pharmaceuticals Gmbh; Uta Schwidetzky
Priority date: 2002-07-26
Filing date: 2003-06-23
Publication date: 2004-02-26
Also published as: AU2003250766A1; WO2004016810A3; DE10234901A1

Abstract

The invention relates to the uses of Mrp4 for the diagnosis and treatment of prostate and/or bladder and/or ovarian tumours, and for screening for substances for such purposes.

Description

Uses of substances that bind to Mrp4 for the diagnosis and treatment of cancer.

Field of the Invention

The invention relates to new uses of Mrp4 or sequences derived therefrom for screening for substances which bind to it, and to the use of substances which bind to rp4 for the diagnosis and / or treatment of tumor diseases.

Background of the Invention and Prior Art

Mrp4 is part of the ATP binding cassette superfarαil of the transport proteins, which use the energy of the ΑTP hydrolysis for their activity. According to MA Barrand et al., Gen. Pharmacol. 28: 639-645 (1997), Mrp4 is associated with changes in drug accumulation and distribution. In connection with the anti-viral agent PMEA, the anti-cancer agents β-mecaptopurine, 6-thioguanine and estradiol 17-β-D-glucuronide, the transport of these agents through Efflux through the cell membrane by means of rp4 is known (K. Lee et al. , J Natl Cancer Inst 92: 1934-1940 (2000); ZS _. Chen et al., JBC 276 (36): 33747-33754 (2001)). Mrp4 was detected in normal prostate tissue and in stably transfected NIH3T3 cells both in cytoplasm a and in the membrane (K. Lee et al., J Natl Cancer Inst 92: 1934-1940 (2000)). Mrp4 is predominantly expressed in normal prostate tissues and to a lesser extent in various other normal tissues. Expression is also to be observed in various tumor cell lines (P. Borst et al., J Natl Cancer Inst 92: 1295-1302 (2000)).

Resistance to chemotherapy drugs is a major and ongoing problem in the treatment of cancer. Some malignant tumors react poorly to chemotherapy right from the start and are therefore apparently intrinsically resistant. Other tumors initially respond well to chemotherapy, but apparently develop resistance in the course of chemotherapy, which may be due to either a selection process among the malignant cells or a cellular response to the active substance. This widespread phenomenon is called MDR (Multi-Drug Resistance).

Technical problem of the invention

The invention is based on the technical problem of specifying pharmaceutical compositions for diagnosing and / or treating prostate and / or bladder and / or ovarian cancer and means for identifying them. The invention is based in particular on the technical problem of identifying and treating resistant tumors.

Basics of the invention and preferred embodiments.

The invention teaches the use of a nucleic acid coding for Mrp4 and / or a Mrp4 peptide or protein for the detection of prostate and / or bladder and / or ovarian tumors or for the detection of a risk of Disease of prostate and / or bladder and / or ovary tumors, in particular tumors with increased resistance, with a prostate or bladder or ovary tissue sample, for example in vitro, for transcription of Mrp4 RNA or for overexpression of a Mrp4 protein is examined. A nucleic acid coding for Mrp4 or a detector substance that binds to Mrp4 protein or peptide, preferably containing a reporter group, can be used for the detection, whereby binding of said nucleic acid and / or said protein or peptide to the detector substance is detected semi-quantitatively or quantitatively.

The invention further teaches the use of a Mrp4 RNA or a Mrp4 protein or peptide for screening for substances which bind to it, in particular prospective active substances for inhibiting said RNA or said protein or peptide or prospective detector substances, with a prospective substance or a mixture of such prospective substances with said RNA or said protein or peptide is contacted, binding events being ascertained using a binding assay, and a binding prospective substance being selected, if appropriate after deconvolution.

Finally, the invention teaches the use of a substance which inhibits or binds to Mrp4 for producing a pharmaceutical composition for the treatment of prostate and / or bladder and / or ovary tumors, in particular for the treatment of resistant tumors. The substance can be an antibody which is obtained by immunizing a non-human mammal with a Mrp4 peptide or protein with cDNA coding therefor transfected cells, with tumor cells expressing such a peptide or protein endogenously, or with recombinantly produced Mrp4 peptides or proteins, or a phage display antibody. The substance can also be a mimicry compound of an antibody against a Mrp4 peptide or protein. Finally, the substance can be an aptamer, an antisense RNA, or a ribozyme. The substance can additionally, in particular in the case of a bispecific antibody or a mimicry compound, carry a cytotoxic and / or immunostimulating component. The pharmaceutical composition can be prepared for local application in tissue containing tumor cells.

The invention can be used in a process for

Use diagnosis of a prostate and / or bladder and / or ovary tumor disease, with a detector substance in one embodiment being applied with a reporter group to the tissue to be examined, possibly in vitro after tissue removal, the tissue to be examined then being subjected to a detection method step is which is sensitive to the reporter group, and in the case of detection of a defined minimum value of the reporter group in the tissue the tissue is qualified as, possibly resistant, containing tumor cells, and one

Method for the treatment of a prostate and / or bladder and / or ovary tumor disease, in particular resistant tumors, wherein a pharmaceutical composition according to the invention is administered to a patient in a physiologically effective dose, optionally with, before or after, a different, conventional anti-cancer Active ingredient is presented in the same or different dosage forms. Finally, the invention relates to a pharmaceutical composition comprising at least two components A and B, component A being a substance according to one of claims 4 to 8, component B being a (customary) anti-cancer active ingredient, components A and B being prepared alternatively can be i) as spatially separate components of a multicomponent preparation, intended for simultaneous or successive administration and galenically prepared as identical or different administration forms, or ii) as a combination preparation in the form of a spatially connected and uniform administration form.

The invention is based on the knowledge that Mrp4 is overexpressed in prostate and bladder and / or ovary tumors, ie the expression in said tumor tissues is higher compared to normal cells of the same tissue, and the technical teaching that can be derived therefrom that Mrp4 can be used as a target molecule in the diagnosis and therapy of these diseases, in particular in the case of intrinsic resistance or acquired or acquired resistance. Mrp4 can thus serve as a marker for the identification of tumor cells, in particular resistant tumor cells, in the said tumor tissues. On the other hand, the inhibition of Mrp4 offers the possibility of overcoming resistances or not even allowing them to occur, especially in conjunction with conventional chemotherapy agents. In this respect, the invention can also be used to prevent the development of resistance. Because by preventing the efflux of chemotherapeutic agents given due to Inhibition of Mrp4 ensures the long-term effect of these chemotherapeutic agents in all target cells.

Within the scope of the invention, it may be advisable to take a sample from a tissue which is identified as tumor tissue by other methods, in advance of treatment with a pharmaceutical composition according to the invention, and to examine the tissue sample for expression or overexpression of Mrp4. In this context, it is of particular importance that Mrp4 has substrate specificity, for example to cyclic nucleotides, purine analogs and nucleoside-based antivirals; obtaining the expression pattern for Mrp4 can therefore facilitate the selection of potentially suitable active substances or the exclusion of potentially unsuitable active substances or chemotherapeutic agents. Alternatively, a detector substance according to the invention can be used for diagnosis in vivo to test for Mrp4 phenotype. If an overexpression of Mrp4 compared to normal tissue of the same type is found, the use of the pharmaceutical composition according to the invention is indicated.

It is of independent importance in the context of the invention that the invention can be used particularly advantageously in the case of androgen-sensitive tumors or tumor cells.

In the case of bispecific substances which bind to Mrp4, it is preferred if the substance which binds to Mrp4 additionally carries a cytotoxic and / or immunostimulating component. This ultimately leads to the fact that almost exclusively tumor cells are killed, either by the cytotoxicity or by the attack by the stimulates the immune system, while normal cells are practically completely preserved in the tissue. If a cytotoxic component is used, it will be particularly recommended if the pharmaceutical composition is prepared for local application in tissue containing tumor cells, for example for injection.

The invention further relates to a Mrp4 protein or peptide containing a partial sequence of at least 4 amino acids from SEQ ID 10, in particular from SEQ ID 11, or containing SEQ ID 10, in particular SEQ ID 11. or consisting of such a sequence, but not a protein or peptide according to or encoded by Genbank AccNo NM_005845, AY_081219 and XM_036453. Finally, the invention relates to a nucleic acid coding for such a protein or peptide, in particular a sequence according to SEQ ID 9 or a partial sequence thereof. A protein or peptide according to the invention or a nucleic acid coding therefor can be used in all the contexts described above and below.

Definitions.

The following sequences are known for Mrp4: Genbank AccNo NM_005845, AY__081219 and XM_036453.

In the context of this description, the term Mrp4 is used for all human isoforms, known or new, based on nucleic acids or amino acids. These terms also include those in the context of these

Description disclosed short sequences derived from the isoforms, for example immunization sequences. Also included are homologs, the Homology is at least 80%, preferably more than 90%, most preferably more than 95%. In the case of the nucleic acid sequences, complementary or allelic variants are also included. Furthermore, sequences are included which only represent partial sequences of the explicitly disclosed sequences, for example one exon or several exons, or sequences complementary thereto, with the proviso that, in the case of the nucleic acids, these partial sequences are of sufficient length for hybridization with a nucleic acid according to the invention, have at least 30 to 50 bases and, in the case of proteins or peptides, bind to a protein- or peptide-specific target molecule with at least the same affinity. Furthermore, all nucleic acids hybridizing with nucleic acids according to the invention are included, namely those which are under stringent conditions (5 ° C. to 25 ° C. below the melting temperature; see additionally JM Sambrook et al., A laboratory manual, Cold Spring Harbor Laboratory Press (1989 ) and EM Southern, J Mol Biol, 98: 503ff (1975)) hybridize. It goes without saying that the invention also includes expression cassettes, ie one or more of the nucleic acid sequences according to the invention with at least one control or regulatory sequence. Such an expression cassette can also comprise a sequence for a known protein, a fusion protein being formed in the course of the translation from a known protein and a protein or peptide according to the invention. Antisense sequences to the above nucleic acid sequences are also included. Finally, RNA and correlating DNA and vice versa are included, as are genomic DNA and correlated cDNA and vice versa and RNAi. In connection with uses according to the invention, the terms of Mrp4 nucleic acids or protein or peptides also include partial sequences in addition to the full lengths of the sequences mentioned (see also the preceding paragraph), with a minimum length of 12 to 30 nucleotides, for example 30 to 90 nucleotides, in the case of nucleic acids and a minimum length of 4 to 10 amino acids, for example 10 to 30 amino acids, in the case of peptides or proteins.

The term treatment also includes prophylaxis.

An inhibitor is a compound or substance which either inhibits the formation of Mrp4 or reduces the activity of Mrp4 formed, based on the Mrp4 activity in the absence of the inhibitor. In this respect, an inhibitor can be a substance that interferes with the cascade of Mrp4. On the other hand, an inhibitor can be a substance that binds with Mrp4 formed, in such a way that further physiological interactions with endogenous substances are at least reduced.

Mimicry molecules are compounds that simulate the variable region, in particular the binding region of an antibody, and bind to a target molecule in the same place as the underlying antibody.

The term antibody includes polyclonal antibodies, monoclonal antibodies, non-human, human and humanized antibodies, anti-idiotypic antibodies and phage display antibodies, but also chimeric antibodies as well specific fragments of the light and / or the heavy chain of the variable region of the underlying antibodies of the above type. The production or production of such antibodies with predetermined immunogens is well known to the average person skilled in the art and need not be explained in more detail. The term antibody also includes bispecific antibodies. Bispecific antibodies combine a defined immune cell activity with a specific tumor cell recognition, whereby tumor cells are killed. A bispecific antibody binds on the one hand to a trigger molecule of the immune effector cell (eg CD3, CD16, CD64) and on the other hand to antigens of the tumor target cell.

The pharmaceutical preparation of a pharmaceutical composition according to the invention can be carried out in a customary manner, both for systemic and for local administration. Counter ions for ionic compounds are, for example, Na ⁺ , K ⁺ , Li ⁺ or cyclohexylammonium. Suitable solid or liquid pharmaceutical preparation forms are, for example, granules, powders, dragees, tablets, (micro) capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions (IV, IP, IM) as well as preparations with a protracted active ingredient -Release, in the production of which conventional auxiliaries such as carriers, explosives, binders, coatings, swelling agents, lubricants or lubricants, flavoring agents, sweeteners and solubilizers are used. Magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talcum, milk protein, gelatin, starch, cellulose and its derivatives, animal and vegetable oils such as cod liver oil, sunflower, peanut or sesame oil may be used as auxiliary substances. Polyethylene glycols and solvents, such as sterile water and mono- or polyhydric alcohols, for example glycerol, are mentioned. A pharmaceutical composition according to the invention can be produced by mixing at least one Mrp4 inhibitor used according to the invention in a defined dose with a pharmaceutically suitable and physiologically compatible carrier and, if appropriate, other suitable active ingredients, additives or auxiliaries with a defined inhibitor dose and preparing the desired dosage form.

Tumor cells overexpress Mrp4 specifically or differentially if Mrp4 is expressed in at least 10% higher amounts than normal cells of the same tissue.

Cytotoxic components or groups are compounds which directly or indirectly induce apoptosis or lead to necrosis or at least inhibit growth. In addition to being coupled to a substance according to the invention, these can also be used as component B. In addition to radioisotopes (for example 188Re, 213Bi, 99mTc, 90Y, 131J, 177Lu), such groups or compounds can in particular be cytostatic agents which are used in tumor therapy. Examples of these are: alkylating agents (for example mechlorethamine, ifosfamide, chlorambucil, cyclophosphamide, melphalan, alkylsulfonates, busulphan, nitrosoureas, carmustine, lomustine, semustin, triazenes, dacarbazine), antitamabolites (for example folic acid antagonists, methotrexate, fluorimidiloxane, fluorimidine) , Fluorodeoxyuridine, cytarabine, gemcitabine, purine analogues, mercaptopurine), mitosis inhibitors (e.g. vinca alkaloids, voncristine, vinblastine, paclitaxal, docetaxel, protaxel), epipodophyllotoxins (e.g. etoposide, teniposide), antibiotics (e.g. Dactinomycin, daunorubicin, idarubicin, anthracycline, bleomycin, L-asparaginase), platinum complex compounds (e.g. cisplatin), hormones and related compounds (e.g. adrenal cortex steroids, aminogluthetimide, gestagens, estrogens, androgens, anti-estrogens, tamoxifen, tamoxifen). When such a compound is bound to a substance which binds to Mrp4, the coupling takes place in such a way that the affinity for Mrp4 is reduced by not more than 90%, preferably 50%, based on the substance without a cytostatic group, and the cytostatic effect of the group is not reduced by more than 90%, preferably 50%, based on the compound without substance.

An immunostimulating component is usually a protein or an effective component thereof, which stimulates cells of the immune system. Examples include: cytokines such as M-CSF, GM-CSF, G-CSF, interferons such as IFN-alpha, -beta, -gamma, interleukins such as IL-1 to -16 (except -8), human LIF, Che okines such as Rantes, MCAF, MIP-1-alpha, -beta, NAP-1 and IL-8. This component can also be used as a component B.

A reporter group is an atom, molecule or a compound which, in conjunction with an assay placed thereon, enables the detection of the reporter group and the compound or substance thus connected to the reporter group. Examples of reporter groups and associated detection methods are: 32P labeling and intensity measurement using phosphoimager. Many more. Examples are known to the person skilled in the art and do not need to be listed in detail. A substance that binds to Mrp4 can be a substance that binds a Mrp4 protein or a Mrp4 RNA.

Resistance refers to the resistance of malignant cells to chemotherapy drugs. A cell is in the

In the case of resistance that can be acquired, if the time to cell death is extended compared to a reference cell of the same cell type or cell line and the same dosage, or if the proliferation rate of the resistant cell under the same conditions is increased compared to that of the reference cell. In the case of initial resistance, a cell is resistant if the time to cell death is prolonged or the proliferation rate is increased compared to a non-resistant reference cell of different cell types and the same dosage.

Definitions expanded in the context of the above definition in relation to the narrow sense of the word also include the specific terms in the narrow sense of the word.

Examples.

The invention is explained in more detail below on the basis of examples and figures which merely illustrate preferred embodiments. Show it:

1: Chip analysis for the differential expression of Mrp4 in protate tumor tissues from patients, 2: Cancer Profiling Array for the differential expression of Mrp4 in protate and ovarian tumor tissues from patients,

3: Mrp4 TaqMan in prostate tumor and protate abnormal tissues from the same patient,

4: Mrp4 TaqMan in different human tumor cell lines,

5: various hammerhead ribozymes which cut Mrp4 RNA,

6: immunization peptides,

7: a nucleic acid coding for Mrp4 (SEQ ID 9), and

8: a Mrp4 protein or peptide (SEQ ID 10), the labeled partial sequence (SEQ ID 11) being of independent importance.

Example 1:

Electronic Northern were produced in accordance with the literature reference DE 198 11 194 AI, the assembled sequence from ESTs or Contigs given in the sequence listing being used (ok_582). Overexpression in tumor tissue with a significance of> 0.90 was found for prostate and bladder tissue.

Example 2: Overexpression in prostate tumor Paired prostate tumor and normal tissues were removed by laser microdissection from 54 patients. The RNA prepared from it was amplified, labeled with digoxygenin and hybridized on a DNA chip using Affymetrix technology. The results of the analysis of the chip are shown in FIG. 1. It can be seen that in 55% of the prostate tumors (28 out of 54 patients) the expression is increased by at least a factor of 2.

Example 3: Overexpression in prostate and ovarian tumor.

FIG. 2 shows results of the expression of Mrp4 on prostate and bladder tumors and the associated normal tissues, obtained as stated above, from a cancer profiling array. For this, the Mrp4 sequence was labeled with 32P-dCTP using random hexamer priming and hybridized to the cancer profiling array. This contains 225 cDNA pairs, each pair representing tumor and normal tissue from one patient. It can be seen that Mrp4 is overexpressed in several prostate and ovarian tumors.

Example 4: Overexpression in prostate tumor.

Mrp4-specific oligonucleotide primers were generated and used for quantitative PCR (TaqMan), the first strand of cDNA coming from different tissues and cell lines, as indicated in FIG. 3. Mann first recognizes that there is significant overexpression of Mrp4 (4-9-fold) in 5 out of 15 prostate tissue pairs. No expression takes place in other normal tissues of FIG. 3. The same applies to the cell lines examined, with the exception of the androgen-sensitive tumor cell line LNCaP.

A detailed representation of the investigations on cell lines is given in FIG. 4, with the strong expression in the prostate tumor cell line LNCaP first becoming more apparent. However, it can also be seen that expression in non-androgen-sensitive cell lines, such as, for example, PC-3 and DU-145, is comparatively low.

Example 5: Detection of Mrp4 using antibodies

In this example, the labeling of a tumor or its metastases by an anti-Mrp4 antibody in vivo (mouse model) is described. An anti-Mrp4 antibody is labeled with a marker molecule (e.g. radioisotope). In NMRI nude mice, 3 10 ⁶ Mrp4-transfected human cells or tumor cells with high endogenous Mrp4 expression, for example LNCaP cells, are transplanted. After a period of time sufficient to develop metastases to a secondary site in the body, for example 30 days after the transplant, the mice become infected with labeled antibodies. The control animals are treated with an irrelevant antibody. A few hours after the antibody application, the animals are sacrificed and tissue sections are made from all organs. These sections are examined for the presence of labeled anti-Mrp4 antibody.

The anti-Mrp4 antibodies are polyclonal antibodies against human Mrp4 protein conjugated to a carrier protein, raised in rabbits and affinity-purified with the specific immobilized peptides.

Examples of immunization peptides are given in Figure 6 (SEQ ID 1 to 4). Cells transfected with Mrp4 cDNA, or partial sequences thereof, such as, for example, COS cells or NIH3T3 cells, can also be used as immunogens. Tumor cells expressing Mrp4 endogenously are also suitable. Furthermore, recombinantly produced Mrp4 or partial sequences thereof, which are expressed in producer cells such as E. coli or insect cells, can also be used for the immunization.

Example 6: Immunohistochemical detection of tumor cells.

Primary tumors are isolated from the patients with prostate, bladder and / or ovary tumors and prepared as paraffin or frozen sections. These sections are examined with an anti-Mrp4 antibody for the overexpression of Mrp4 in tumor cells. The immunohistological examination with the Mrp4 antibody shows higher expression of Mrp4 in the tumor cells compared to surrounding normal tissue. The investigation is carried out in detail by incubation with the anti-Mrp4 antibody as primary antibody, a biotinylated secondary anti-rabbit antibody and a streptavidin-coupled horseradish peroxidase. The coloring is done with DAB as a chromogenic substrate (brown coloring). Counterstaining is done with hemalaun solution (blue staining). Malignant and non-malignant cells can be distinguished, the malignant cells having a strong staining, ie high Mrp4 content, have, while the non-malignant cells are only moderately stained.

Example 7: RNA inhibitors

FIG. 5 (SEQ ID 5 and 6) shows various hammerhead ribozymes which cut Mrp4 at the points shown and thus inhibit or at least reduce the activity of any translation products. Suitable antisense RNA sequences are, for example, 5'-UCACCUCCUGGUACACGGGCAG-3 '(Seq. ID 7) and 5'-GCAGAUGUUCGCGUCCUGCAGC-3 "(Seq. ID 8). It is also possible to construct primers by means of which RNAi probes are generated to inhibit Mrp4 by gene silencing (RNA interaction).

Claims

Claims:

1. Use of a nucleic acid coding for Mrp4 and / or a Mrp4 peptide or protein for the detection of prostate and / or bladder and / or ovary tumors, or for the detection of a risk of the disease of prostate and / or bladder and / or ovary tumors , in particular of tumors with increased resistance, wherein a prostate or bladder or ovary tissue sample is examined for over-transcription of Mrp4 RNA or for overexpression of a Mrp4 protein.

2. Use according to claim 1, wherein a nucleic acid coding for Mrp4 or a detector substance that binds to Mrp4 protein or peptide, preferably containing a reporter group, is used, binding of said nucleic acid and / or said protein or peptide to the detector substance being detected semi-quantitatively or quantitatively becomes.

3. Use of a Mrp4 RNA or a Mrp4 protein or peptide for screening for substances which bind to it, in particular prospective active substances for inhibiting said RNA or said protein or peptide or prospective detector substances, a prospective substance or a mixture of such prospective substances with said RNA or said protein or peptide is contacted, whereby binding events are determined with a binding assay, and wherein a binding prospective substance, if necessary after deconvolution, is selected.

4. Use of a substance which inhibits or binds to Mrp4 for producing a pharmaceutical composition for the treatment of prostate and / or bladder and / or ovary tumors, in particular for the treatment of resistant tumors.

5. Use according to claim 4, wherein the substance is an antibody which is obtainable by immunizing a non-human mammal with a Mrp4 peptide or protein, or a cDNA coding therefor, or a phage display antibody.

6. Use according to claim 4, wherein the substance is a facial expression compound of an antibody against a Mrp4 peptide or protein.

7. Use according to claim 4, wherein the substance is an aptamer, an antisense RNA, or a ribozyme.

8. Use according to any one of claims 4 to 7, wherein the substance additionally carries a cytotoxic and / or immune stimulating component.

9. Use according to one of claims 4 to 8, wherein the pharmaceutical composition is prepared for local application in tissue containing tumor cells.

10. Procedure for diagnosing a prostate and / or

Bladder and / or ovary tumor disease, in particular for the diagnosis of resistant tumors, wherein a detector substance is applied in one embodiment with a reporter group to the tissue to be examined, the tissue to be examined then being subjected to a detection method step which is sensitive to the reporter group , and in the case of the detection of a defined minimum value of the reporter group in the tissue, the tissue is qualified as containing, possibly resistant, tumor cells.

11. A method for the treatment of a prostate and / or bladder and / or ovarian tumor disease, in particular resistant tumors, wherein a pharmaceutical composition according to one of claims 4 to 9 is administered to a patient in a physiologically effective dose, optionally at the same time, before, or then a different anti-cancer drug is presented in the same or different dosage form.

12. Pharmaceutical composition containing at least two components A and B, wherein component A is a substance according to one of claims 4 to 8, wherein component B is an anti-cancer agent, wherein the Components A and B can alternatively be prepared i) as spatially separate components of a multicomponent preparation, intended for simultaneous or successive administration and galenically prepared as the same or different administration forms, or ii) as a combination preparation in the form of a spatially connected and uniform administration form.

13. A Mrp4 protein or peptide containing a partial sequence of at least 4 amino acids from SEQ ID 10, in particular from SEQ ID 11, or containing SEQ ID 10, in particular SEQ ID 11, or consisting of such a Chen sequence, but not a protein or peptide according to or encoded by Genbank AccNo NM_005845, AY_081219 and XM_036453 and not SEQ ID 1 according to WO0058471.

14. A Mrp4 nucleic acid coding for a protein or peptide according to claim 13, in particular containing a partial sequence of at least 12 nucleotides from SEQ ID 9.