MXPA98009698A - Systems of expression by oncogenes or vi - Google Patents

Abstract

The present invention relates to a construction of nucleic acids for the expression of an effector gene, the nucleic acid construct containing a promoter I (component a) that controls the expression of a gel of the transcription factor (component b) also contained in the construction nucleic acids, and containing a promoter II (component c) to which the gene product of the transcription factor gene specifically binds and which controls the expression of an effector gene (component d) also contained in the construction of nucleic acids, characterized in that the activity of the gene product of the transcription factor gene depends on one or more cellular regulatory proteins that specifically bind to this gene product and that affect its activity, to an isolated cell containing the said construct for the preparation of a therapeutic agent and cure for the treatment of diseases, to the use of the aforementioned cell for the same purpose and to a procedure for the preparation of the construction of nucleic acids

Description

Expression systems controlled by oncogenes or viruses The present invention relates to a nucleic acid construct for the expression of an effector gene, the construction of nucleic acids containing a promoter I (component a) that controls the expression of a transcription factor gene (component b) also in the construction of nucleic acids, and containing a promoter II (component c) to which the gene product of the transcription factor gene specifically binds and which controls the expression of an effector gene (component d) also contained in the construction of nucleic acids, characterized in that the activity of the gene product of the transcription factor gene depends on one or more cellular regulatory proteins that specifically bind to this gene product and that affect its activity.

I. Introduction A problem of gene therapy, still insufficiently resolved, is the cell-specific control of the expression of an effector gene, in particular in diseased or otherwise modified cells. The present invention includes a new method of this control. It is based on the recognition (Werness et al., Science 248, 76 (1990)) that in degenerated cells there are regulatory proteins that are modified or diminished so that they no longer bind to their corresponding participating molecules and can interact with them, or they acquire new binding properties with "their corresponding participating molecules or with others." The method according to the invention is further based on the recognition that, for example, the retinoblastoma protein can bind to the activation domains. of the E2F transcription factor and, thereby, can inhibit its activity (Flemington et al., PNAS USA 90, 69-14 (1993)).

Genes for regulatory proteins of this type have already been used for expression systems for the search for inhibitors or stimulators of these regulatory proteins (for example, documents W095 / 19367, W095 / 14777, W097 / 04092). In addition, vector systems have already been disclosed, wherein a first vector expresses a tumor suppressor protein, and a second vector a protein that binds to the tumor suppressor protein and, thereby, inhibits it (W095 / 16771). The two vectors are introduced into a cell. By means of the combination of the two vectors, vectors encoding a tumor suppressor protein can be produced in the cell, without the cell being inhibited in its proliferation by the tumor suppressor protein. Additionally, in WO97 / 12970, expression systems are disclosed, in which the expression of a first gene is controlled by a first promoter, whose function is suppressed in non-tumor cells, and the expression of a second gene, whose Expression product inhibits the expression of the first gene in non-tumor cells, is controlled by a second promoter that is highly regulated in non-tumor cells. The object of the invention is, then, a novel system of simple expression that can only be activated in cells in which regulatory proteins of this type appear in a diminished or modified form. With this, the transcription of an effector gene encoded by the expression system is activated. The expression product of the effector gene has, on its own or in combination with another pharmaceutical active ingredient, a prophylactic or therapeutic effect. - • < II. General description of the invention The expression system corresponding to the invention represents a construction of nucleic acids, whose expression is controlled by oncogenes or viruses through the modification or influence on regulatory proteins caused by the previous ones and which contains, in the case of more simple, the following components: a) at least one activation sequence (promoter unit I) b) at least one gene for a transcription factor, its transcription being controlled by component a) c) at least one other activation sequence (promoter unit II) that controls the expression of component d) through the binding of the transcription factor, encoded by component b) d) at least one effector gene. The arrangement of the different components is reproduced by way of example in Figure 1. According to this invention, two particular embodiments of the nucleic acid construction must be differentiated: a) Embodiment A) This embodiment is characterized by the following properties of the components: Component a) at least one activation sequence (promoter n ° I) Component b) at least one gene for a transcription factor consisting of a fusion protein containing • Component i._) at least one activation domain of a transcription factor • Component b2) at least one binding sequence of a binding protein for a regulatory protein • Component b3) at least one DNA binding domain of a transcription factor Component c) at least one activation sequence (promoter No. II) that is activated by the binding of the transcription factor, encoded by the component b) Component d) at least one effector gene. The arrangement of the various components is reproduced by way of example in FIG. 2. It is a premise for the functionality according to the invention of the expression system that component b2) is included among or in components b_) and b3) so that the binding of the regulatory protein to component b2) inhibits the functionality of the activation domains (component bx) and / or of the DNA binding domains (component b3). This inhibition leads to normal inhibition of expression of the effector gene in normal cells, ie in a normal functional regulatory protein. In a degenerated or infected cell, in which the regulatory protein is modified or complexed, so that it can no longer interact with the corresponding binding protein, either it is no longer present or is only sparingly present, this inhibition is lacking, so that the transcription factor (component b) can freely activate the activation sequence (component c) and, thereby, can initiate the transcription of the effector gene. The transcription of the effector gene is initiated by an activation of the activation sequence [component a)] which results in an expression of the gene for the transcription factor [component b)]. The transcription factor [component b)] binds again to the activation sequence [component c)] that induces an expression of the effector gene [component d)]. In a particular embodiment of this invention, component a) is equal to component c). In this particular embodiment, a slight activation of the activation sequence [promoter I, component a)] leads to an expression of the transcription factor [component b)] that activates both the activation sequence [promoter I, component a) ] as well as the activation sequence [promoter II, component c)] and, thereby, induces the expression of both the effector gene [component d)] and also enhances the expression of the transcription factor [component b)], thereby , again, the expression of the effector gene [component d) J is reinforced. 2) Embodiment B) This embodiment is characterized by the following properties of the components: Component a ') - at least one activator sequence (promoter I), containing • Component a: at least one DNA binding sequence for a regulatory protein and • Component a2): at least one basal promoter, activating component a2 ) the binding of the regulatory protein to the component a Component b ') * at least one gene for a transcription factor acting as a repressor, its expression being induced by component a') Component c ') at least one activation sequence (promoter II) containing • Component c? ): at least one actuation sequence for the induction of the transcription of component d) and • Component c2): at least one DNA sequence for the binding of the repressor (component b '), this binding inhibiting the activation of " transcription of the effector gene located below (component d) Component d) an effector gene The arrangement of the components of embodiment B) is reproduced by way of example in figure 3.

It is a premise for the mode of operation according to the invention of the expression system according to embodiment b) that in the normal cell the binding of a cellular regulatory protein to component a ') of the promoter unit I induces transcription of the repressor gene (component b '), and that the expressed repressor binds to component c2) of promoter unit II and, thereby, inhibits the transcription activation of the structural gene (component d) by the promoter unit II. In a degenerate or infected cell, in which the regulatory protein is modified or complexed, so that it can no longer bind to the DNA binding sequence (component aJ in the unit of prmotor I, or is no longer present or it is only sparingly present, no expression of the gene for the repressor is effected and, thus, no inhibition of the expression of the nucleic acid construction according to the invention In these degenerated or infected cells, in the embodiment B) of the construction of nucleic acids according to the invention transcription of the effector gene (component d) is initiated by the activation of the activation sequence (component c of the promoter unit II.) This expression system can be extended in embodiments A) and B) - by juxtaposing several identical or different sequences for effector genes [components d), d '), d ")] that are linked in each case to each other by the same or different sequences or by activation sequences [components c ") and e")] IRES. In Embodiment A) by the juxtaposition of several identical or different genes for transcription factors [component b)], which in each case are linked together by the same or different IRES sequences or activation sequences [component a)] or [component c)]. In the case of a juxtaposition of genes for different transcription factors, the activation sequences have to be chosen so as to contain nucleotide sequences to which the transcription factor can be linked [component b)]. By the nucleic acid constructs according to the invention, an effector gene [component d)] can be expressed, depending on the choice of the activation sequence [components a) or c_)] in a nonspecific manner, specific to the cell or specific virus or under certain metabolic conditions or also specifically cell cycle. In the case of the effector gene, it is a gene that, in turn, encodes a pharmacologically active substance or an enzyme that separates an inactive precursor from a drug in an active pharmacist. For example, the effector gene can be chosen so that this active substance or this enzyme is expressed as a fusion protein with a ligand, and this ligand binds to the surface of cells, for example endothelial or tumor cells, or leukocytes. . The nucleic acid constructs according to the invention preferably consist of DNA. By the term "nucleic acid constructs" are meant artificial structures based on nucleic acids that can be transcribed in the target cells. Preferably, they are incorporated into a vector, with plasmid vectors or viral vectors being particularly preferred. The nucleic acid construct, optionally incorporated in a vector, is administered to a patient for the prophylaxis or therapy of a disease. The administration must be carried out orally, locally or by injection or infusion. The subject of the present invention are also mammalian cells that contain a nucleic acid construct according to the invention. In a particularly preferred embodiment, the nucleic acid constructs are incorporated into cell lines which, after transfection, can be used as supports for the expression system according to the invention for the expression of the effector gene. Cells of this type can be used for the provision of a therapeutic and curative agent for patients. Alternatively, cells or cell lines such as, for example, tumor cells, of the immune or endothelial system, in which the nucleic acid constructs according to the invention are incorporated, can be administered locally to patients or parenterally, by intravenous, intr-arterial example in a body cavity, in an organ or subcutaneously injected.A preferred use of the nucleic acid construct according to the invention consists, in particular, in the prophylaxis or treatment of a disease, the invention encompassing the in vitro introduction of a nucleic acid construct into a target cell, the non-specific, specific expression of the virus or the target cell, metabolically specific and / or cell cycle-specific of the therapeutic and curative agent in the cell target and local or parenteral administration of the target cell to the patient or local or parenteral administration of the construction of nucleic acids to the patient for the in vivo introduction of a nucleic acid construct into the target cell. The nucleic acid constructs according to the invention do not occur in this form in nature, ie the effector gene for the active substance or for an enzyme or for a ligand-active substance or ligand-enzyme fusion protein is not combined naturally with nucleic acid sequences such as those contained in the nucleic acid construct according to the invention.

Preferred effector genes that are incorporated into an expression system according to the invention encode a pharmacologically active substance. These are proteins and glycoproteins, chosen from the group that contains cytokines, growth factors, receptors for cytokines or growth factors, antibodies or fragments of antibodies, antiproliferative or cytostatic action proteins, apoptotic or anti-apoptotic action proteins, tumor antigens , inhibitors of angiogenesis, thrombosis inducing proteins, coagulation inhibitors, fibrinolytic action proteins, blood plasma proteins, complement activating proteins, envelopes of viruses and bacteria, hormones, peptides with activity on blood circulation, neuropeptides, enzymes, mediators, unmodified regulatory proteins that occur naturally and ribozymes or ribonucleotides (antisense) that act as inhibitors on gene expression. Preferably, in the case of the transgene, it is an effector gene that encodes a ribozyme that inactivates the mRNA that encodes a protein chosen from the group that contains proteins for the control of the cell cycle, in particular cyclin A, cyclin B, cyclin DI, cyclin E, E2F1-5, cdc2, cdc25C or DPI, or virus proteins or cytokines or growth factors or their receptors. In another embodiment, the effector gene can encode a ligand-active substance fusion protein, the ligand being an antibody, an antibody fragment, a cytokine, a growth factor, an adhesion molecule or a peptide hormone and the active substance a pharmacologically active substance as described above, or an enzyme. For example, the effector gene can encode a ligand-enzyme fusion protein, wherein the "enzyme dissociates a 'precursor from a drug in a drug and the ligand binds to a surface of the cell, preferably to endothelial cells or cells tumor III. Detailed description of the features of the embodiment A) 1) The component b) 1. 1) Sequence of binding for a regulatory protein [component b2)] Numerous cellular binding proteins for regulatory proteins have been described [Zwicker and Müller, Progress in Cell Cycle Res. 1:91 (1995); Boulikas et al., Int. J. Oncol. 6: 271 (1995); Pawson, Nature 373: 573 (1995); Cotter, Leuk. Lymph. 18: 231 (1995); Hesketh, the Oncogene Facts Book Acad. Press, ISBN 0-12-344550-7 (1995); Miller and Sarver, Nature Med. 3: 389 (1997)]. In the sense of the invention, binding proteins or their binding sequences for regulatory proteins of this type that are only rarely expressed in diseased cells are inhibited in their binding to the binding sequence, due to an excess of the sequence. of binding do not occur in free form or only do so sparingly or are affected or otherwise modified in their function such as, for example, by mutation. Regulatory proteins of this type include, for example, the expressed proteins of tumor suppressor genes. A non-limiting choice of the invention of regulatory proteins of this type and their corresponding binding proteins and their binding sequences is reported in the following examples: Regulatory protein Component b2) _ (cell binding protein with binding sequence for regulated protein- = Ü p53 MDM-2 pRb • transcription factor E2F, -1, -2, -3 • cyclin-D ?; -D2, -D3 or -C • cyclin-A, -E • transcription factor PU.l • transcription factor Elf-l pl30 • transcription factor E2F-5 • cyclin-A, -E Max Myc MAD • Myc VHL elongina.-C, -B Cdk4 pl4, pl5, pld, pl8, p27, p57, p21 MTS-1 (pl6) cdk4 WT-1 • p53 SMAD2 (MADR2: • DPC4 DPC-4 SMAD2 / 3-catenin LEF-1 LEF-1 ß-catenin In a particular embodiment of this invention, component b2) is a binding-protein binding-non-self-binding sequence of the cell for a regulatory protein. A non-cell binding sequence of this type can, for example, be of viral, bacterial or parasitic origin. The use of a non-cell binding sequence of this type makes it possible for the function of component b) in normal cells to be inhibited due to the binding of the respective regulatory protein to component b2). However, the corresponding regulatory protein is extensively bound in infected cells due to the intracellular production of the binding protein containing the binding sequence by the respective infectious agent. Thus, in these cells component b) is free and functional.

In another particular embodiment of this invention, component b2) is an antibody or a part of an antibody with binding sequences (VH and VL) for a regulatory protein. A non-limiting choice of the invention of non-cell binding sequences is gathered in the following examples: Regulatory protein Component b2) (viral binding protein with binding sequence for the regulated protein - raj p53 • IE 84 of CMV (Speir et al., Science 265, 391 (1994) • E1B (55 Kd) of AV (Sarnow et al., Cell 28., 387 (1982); Liu et al., Cold Spring Harbor Symp. On Quantitative Biol. LIX, 215 (1995)) • EBV EBV-5 (Szekeley et al., PNAS USA £ 0, 5455 (1993)) • EBV BHFR1 (Theodorakis et al., Oncogene 12., 1707 (1996)) • E6 HPV-16 or -18 (Dyson et al., Science 243, 934 (1989); Howes et al., Genes Dev. 8., 1300 (1994)) • HBV HBV protein (Wang et al., PNAS USA £ 1, 2230 (1994)) • SV40 T antigen (Lane et al., Nature 278, 261 (1979), Linzer et al., Cell 17, 43 (1979)) pRb • E1A AV (Nevis Science 258, 424 (1992)) • EBV EBNA-2 • EBNA-1 or -5 for EBV • E7 for HPV • T antigen for SV40 pl30 • E1A for AV (Li et al., Genes Dev. 7 2366 (1993)) CBF-1 (RBP-JK) • EBV EBNA-2 (Zimber-Strobl et al., EMBO J. 13, 4973 (1994)) NF-kappa B • HIV Tax (Suzuki et al., Oncogene 9, 3099 (1994)) Lyn-tyrosine kinase • EBV LMP-1 • LMP-2A or EBV bak LMP-2B • E1B (16 Kd) AV (Farrow et al., Nature 374, 731 (1995)) bax • E1B (19 Kd) of AV (Han et al., Genes Dev. 10, 461 (1996)) Regulatory protein Antibodies or fragments of antibodies with binding sequence (VH, V for the regulatory protein p53 • monoclonal antibodies specific for the non-mutated DNA binding domains (Legros et al., Oncogene JL 2071 (1994); 1, 3689 (1994); Hupp et al., Cell 71, 875 (1992); Abarzna et al. ., Cancer Res. 55, 3490 (1995), Bonsing et al., Cytometry 28, 11 (1997), Thomas et al., J. Clin. Path. 50, 143 (1997); Jannot et al., BBRC 230, 242 (1997)) pRb • monoclonal antibodies specific for active pRb (non-phosphorylated) (Hu et al., Mol Cell Biol. 11, 5792 (1991)) In the case of the choice of an antibody, the parts of the antibody that bind to the FVH epitope FVH, in the case of murine origin in humanized form, are preferably used as component b2). The humanization is carried out in the manner described by Winter et al. (Nature 349, 293 (1991) and Hoogenbooms et al (Rev. Tr. Transfus, Hemobiol 36, 19 (1993)) Antibody fragments are prepared according to the state known in the art, for example in the manner described by Winter et al., (Nature 349, 293 (1991), Hoogenbooms et al., Rev. Tr. Transfus, Hemobiol 36., 19 (1993), Girol, Mol.Immunol.28, 1379 ( 1991) or Huston et al., Int. Rev. Immunol., 10. 195 (1993) A detailed description of the preparation of antibodies, antibody fragments and recombinant antibody fragments was performed in the patent application DE 196 49 645.4 The recombinant antibody fragments are prepared directly from existing hybridomas or isolated with the help of "phage display" technology from libraries of urinic or human antibody fragments (Winter et al., Annu., Rev. Immunol. 12, 433 (1994).) These fragments of antibodies are then used in the genetic plane directly to the coupling with components b and b3). For the preparation of recombinant antibody fragments from hybridomas, the genetic information, which codes for the antigen-binding domains (VH, VL) of the antibodies, is obtained by isolating the mRNA, the reverse transcription of the RNA in the CDNA and the subsequent amplification by chain reaction with polymerase and oligonucleotides complementary to the 5 'or 3' ends of the variable fragments. The DNA fragments thus obtained which encode the VH and VL fragments are then cloned into bacterial expression vectors and, in this way, Fv fragments, single chain Fv fragments (scFv) or Fab fragments can be expressed, for example. New fragments of antibodies can be isolated, by means of the "phage display" technology, also directly from antibody libraries (immune libraries, native libraries) of murine or human origin. In the case of the "phage display" of anti-body fragments, the genes of domains that bind antigens are cloned, in the form of fusion genes, with the g3P envelope protein gene of filamentous bacteriophages in the phage genome or in phagemid vectors in the form of scFv fragment genes or in the form of Fab fragment genes. The phages that bind to antigens are selected in plastic containers loaded with antigens (in English panning), in paramagnetic "beads" conjugated with antigens or by binding to cell surfaces. The immune libraries are prepared by PCR amplification of the genes of the variable antibody fragments from B lymphocytes of animals immunized patients. For this, combinations of oligonucleotides that are specific for murine or human immunoglobulins or for families of human immunoglobulin genes are used. With the use of non-immunized donors as a source of the immunoglobulin genes, native libraries can be prepared. Alternatively, genes from the immunoglobulin germ band can be used for the preparation of semi-synthetic antibody repertoires, the region determining the complementarity of the variable fragments being completed by PCR with the aid of degenerate primers. These so-called "single-container" libraries have the advantage, in contrast to immune libraries, that the antibody fragments can be isolated against a plurality of antigens from a single library. The affinity of antibody fragments can be further increased by "phage display" technology, by preparing new libraries of existing antibody fragments by casual mutagenesis, based on codoxy or pre-established by "chain disorder" ("chain shuffling") of individual domains with fragments of native repertoires or with the aid of bacterial mutant strains, and are isolated by reselection under limiting conditions of antibody fragments with improved properties. Additionally, fragments of murine antibodies can be humanized by stepwise exchange of one of the variable domains by a human repertoire and subsequent selection with the original antigen ("guided selection"). In turn, the humanization of murine antibodies is effected by pre-established exchange of the hypervariable regions of human antibodies by the corresponding regions of the original murine antibody. 1. 2) Activation domains [bx component]] and DNA binding domains [component b3)] In the sense of the invention, all genes available from activation domains and DNA binding domains of a transcription factor for component b) can be used. Examples for this, but whose description should not limit the invention, are.- - Activation domains [bx component]] of at least one sequence • of the cDNA for the transactivation domain (TAD) of acidic character of HSV1-VP16 (amino acids 406 a 488, - Triezenberg et al., Genes Developm. 2: 718 (1988); Triezenberg, Curr. Opin. Gen. Developm. 5: 190 (1995) or amino acids 413 to 490; Regier et al., Proc. Nati Acad. Sci. USA 90, 883 (1993)) or • the activation domain of Oct-2 (amino acids 438 to 479; Tanaka et al., Mol. Cell. Biol. 14: 6046 (1994) or amino acids 3 to 154; et al., Nature 374: 657 (1995)) OR • the activation domain of SP1 (amino acids 340 to "485, Courey and Tij'an, Cell 55, 887 (1988)) or • the activation domain of NFY ( amino acids 1 to 233, Li et al., J. Biol., Chem. 267, 8984 (1992) van Hujisduijnen et al., EMBO J. 9, 3119 (1990) Sinha et al., J. Biol. Chem. 92 ., 1624 (1995) Coustry et al., J. Biol. Chem, 270, 468 (1995)) or • the activation domain of ITF2 (amino acids 2 to 452; Seipel et al., EMBO J. 13, 4961 (1992)) or • the activation domain of c-Myc (amino acids 1 to 262; Eilers et al.) Or • the activation domain of CTF (amino acids 399 to 499; Mermod et al., Cell 58/741 (1989); Herr, Nature 374, 657 (1995)) - DNA binding domains [component b3)] of at least one sequence of the cDNA for the DNA binding domain of the Gal4 protein (amino acids 1 to 147; Chasman and Kornberg, Mol. Cell. Biol. 10: 2916 (1990)) or • of the LexA protein (amino acids 1 to 81; Kim et al., Science 255: 203 (1992) or of the complete LexA protein (amino acids 1 to 202; Brent et al., Cell 43: 729 (1985)) or • of the lac repressor protein (lacl) (Brown et al., Cell 49: 603 (1987); Fuerst et al., PNAS USA 86: 2549 (1989)) or, • of the tetracycline repressor protein (tet R) (Gossen et al., PNAS USA 89; 5547 (1992); Dingermann et al., EMBO J. 11: 1487 (1992)) or • of the ZFHD1 protein (Pomerantz et al. al., Science 267: 93 (1995)) It is advantageous in the sense of the invention to incorporate a nuclear localization signal (NLS) at the 3 'end of the DNA binding domain. 2) Activation sequence activatable by component b) of promoter unit II [component c)] - The choice of this activation sequence is oriented according to the choice of the DNA binding domain [component b3)] in the gene for a transcription factor [component b)]. For the examples included in section 1.2 of the DNA binding domains, there are, for example, the following possibilities: 2. 1) Possibility A) an activation sequence with at least one binding sequence [nucleotide sequence: 5'-CGGACAACTGTTGACCG-3 '] (SEQ ID N0: 1) for the Gal4 protein (Chasman and Kornberg, Mol. Cell Biol. 10, 2916 (1990)) and (at the 3 'end of which) the SV40 basal promoter (nucleotides 48 to 5191; Tooze (comp.), DNA Tumor) is incorporated.

Viruses (Cold Spring Harbor New York, New York, Cold Spring Harbor Laboratory) or • the c-fos promoter (Das et al., Nature 374, 657 (1995)) or • the U2 sn RNA promoter or • the promoter of HSV TK (Papavassiliou et al., J. Biol. Chem. 265, 9402 (1990), Park et al., Molec. Endocrinol, 7, 319 (1993)). 2. 2) Possibility B) - an activation sequence with at least one binding sequence [nucleotide sequence: 5 '-TACTGTATGTACATACAGTA-3'] (SEQ ID NO: 2) for the LexA protein [LexA operator; Brent et al., Nature 612, 312 (1984)] and (at which extreme 3 ') is incorporated • the SV40 basal promoter (nucleotides 48 to 5191; Tooze (comp.), DNA Tumor Viruses (Cold Spring Harbor New York, New York, Cold Spring Harbor Laboratory) or other promoter (see possibility TO) . 2. 3) Possibility C) - an activation sequence with at least one binding sequence of the Lac operator (nucleotide sequence: 5 '-GAATTGTGAGC-GCTCACAATTC-3') (SEQ ID NO: 3) for the lac repressor protein I (Fuerst et al., PNAS USA 86, 2549 (1989); Simons et al., PNAS USA 81, 1624 (1984)) and (at whose 3 'end) the SV40 basal promoter is incorporated (nucleotides 48 to 5191; Tooze (comp.), DNA Tumor Viruses (Cold Spring Harbor, New York, NY, Cold Spring Harbor Laboratory) or another promoter (see possibility A). 2. 4) Possibility D) - an activation sequence • with at least one tetracycline operator binding sequence (tet 0) (nucleotide sequence: 5 '-TCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAG-3') (SEQ ID NO: 4) for the tetracycline repressor protein (tet R) and (at the 3 'end of which) the SV40 basal promoter is incorporated (nucleotides 48 to 5191; Tooze (comp)), DNA Tumor Viruses (Cold Spring Harbor, New York, N.Y., Cold Spring Harbor Laboratory) or another promoter (see possibility A). 2. 5) Possibility E) - an activation sequence with at least one binding sequence [nucleotide sequence: 5 '-TAATGATGGCG-3') (SEQ IDNO: 5) for the ZFHD-1 protein (Pomerantz et al., Science 267, "" 93 (1995)) and (in -whose 3 'end) is incorporated • the SV40 basal promoter (nucleotides 48 to 5191; Tooze (comp)), DNA Tumor Viruses (Cold Spring Harbor New York, New York, Cold Spring Harbor Laboratory) or another promoter (see possibility A).

IV) Detailed description of the features of the embodiment B) 1) The activation sequence of the promoter unit I [component a ')] 1. 1) The DNA binding sequence for a regulatory protein [component a._)] It belongs to the DNA binding sequences for transcription factors that are prevented by mutation in their DNA binding capacity or are increased or reduced quantitatively in the cell. Transcription factors and their modifications were represented in the form of a compendium, for example, by Nichols et al., Blood 8.0, 2953 (1992); Crepieux et al., Crit. Rev. Oncogen. 5, 615 (1994); La Thangue, TIBS 1 £, 108 (1994); Lipton, Nature Med. 3, 20 (1997)). For example, they belong to at least one DNA binding sequence for the p53 protein [ATAATTGGGCAAGTCTAGGAA-3; (SEQ ID NO: 6) Kern et al., Science 252, 1708 (1991), Cho et al., Science 265, 346 (1994) or - (G / A) - (G / A) - (G / A) -C- (A / T) - (T / A) -G; Cho et al. , Science 265, 346 (1994)] for the Wt-1 protein (Wang et al., Oncogene 10., 415 (1995); Borel et al., Biochem. 35/37, 12070 (1996)) for the NF-kappa B protein (5'-GGGACTTTCC-3 'nucleotide sequence) (SEQ ID NO: 7); Urban et al., Genes and Developm. 4, 1975 (1990), - Roug et al., Virol. 189; 750 (1992)) or HIV-LTR (Gimble et al., J. Virol. 62., 4104 (1988)) for the E2F / DP-1 complex (at least one 5'-nucleotide sequence -TTTTCCCGCCAAAA (SEQ ID NO: 8); Ó 5 '-TTTTCCCGCCTTTTTT (SEQ ID NO: 9); Ó 5 '-TTTTCCCGCGCTTTTTT) (SEQ ID NO: 10) (Ouellete et al., Oncogene 7, 1075 (1992)) for the Myc / Max protein (at least one 5'-CACGTG-3 'nucleotide sequence) (Walhout et al., Nucí Acids Res. 25., 1493 (1997); Nozaki et al., J. Biochem. 121, 550 (1997)) or 5 '-CATGTG-3' (Físher et al., EMBO J. 12, 5075 (1993)) 1. 2) the basal promoter [component a2)] To him belong, for example: the basal promoter of SV40 (nucleotides 48 to 5191; Tooze (comp.), DNA Tumor Viruses (Cold Spring Harbor New York, New York, Cold Spring Harbor Laboratory) or the c-fos promoter (Das et al., Nature 374, 657 (1995)) or the Us sn RNA promoter or the HSV TK promoter (Papavassiliou et al., J. Biol. Chem. 265, 9402 (1990), Park et al., Mol.? Ndocrin. 319 (1993)) ) The repressor [component b ')] They include, for example, the lac repressor (Brown et al., Cell 49, 603 (1987); Fürst et al., PNAS USA 86, 2549 (1989) or the tetracycline repressor (Gossen et al., PNAS USA 89., 5549 (1992), Dingermann et al., EMBO J. 11, 1487 (1992)) ) The activation sequence [component cx)] influenced by component b ') To it belong, for example, all the activation sequences listed below in section V). 4) the DNA binding sequence for the repressor [component c2)] To it belong, for example: at least one binding sequence of the lac operator (nucleotide sequence: 5 '-GAATTGTGAGCGCTCACAATTC-3') (SEQ ID NO: 3) for the lac I repressor protein (Fürst et al., PNAS USA 86, 2549 (1989), Simons et al., PNAS USA 81, 1624 (1984)) or at least one tetracycline operator binding sequence (tet O) (nucleotide sequence: 5 '-TCGAGTTTAC- CACTCCCTATCAGTGATAGAGAAAAGTGAAAG-3 ') (SEQ ID NO: 4) for the tetracycline repressor protein (tet R).

V) the activation sequence I [component a) in embodiment A) and component c_) in embodiment B)] In the sense of the invention, nucleotide sequences which, after the binding of transcription factors, activate the transcription of a gene located adjacent to the 3 'end have to be used as activation sequences. The choice of the activation sequence is oriented according to the disease to be treated and the target cell to be transduced. Thus, the activation sequence [component a)] can be unlimitedly activated, in a specific manner of the target cell, in certain metabolic conditions, in a specific manner of the cell cycle or in a specific manner of the virus. A detailed description of these promoter sequences was already carried out in the patent applications EP 95930524.4, EP95931933.6, EP95931204.2, EP95931205.9, EP97101507.8, EP97102547.3, DE19639103.2 and DE19651443.6. The promoter sequences to be chosen belong, for example: 1) promoters and promoter sequences that can be activated without limit, such as, for example, the promoter of RNA polymerase III, the promoter of RNA polymerase II, the promoter and enhancer of CMV the SV40 promoter 2) Viral promoter and activator sequences such as, for example, - HBV HCV HSV HPV EBV - HTLV HIV In the case of using the HIV promoter, the entire LTR sequence, including the TAR sequence [positions -453 to -80, Rosen et al., Has to be used as the virus-specific promoter. , Cell 41, 813 (1985)]. 3) Metabolically activatable promoter and enhancer sequences such as, for example, the hypoxia-inducible enhancer. 4) Activators that can be activated in a specific way in the cell cycle. Such promoters are, for example, the promoter of the cdc25C gene, the cyclin A gene, the cdc2 gene, the B-myb gene, the DHFR gene, the E2F-1 gene, the gene cdc25B, or binding sequences for transcription factors that manifest or are activated during cell proliferation. These binding sequences include, for example, binding sequences for c-myc protein. Monomers or multimers of the nucleotide sequence designated Myc E box [5'-GGAAGCAGACCACGTGGTCTGCTTCC-3 ') (SEQ ID NO: 11) must be included in these binding sequences; Blackwood and Eisenmann, Science 251: 1211 (1991)].

) Promoters activable by tetracycline such as, for example, the tetracycline operator in combination with a corresponding repressor. I 6) Chimeric promoters A chimeric promoter represents the combination of a promoter sequence located above, specific to the cell, activable via the metabolic pathway or specifically of the virus with a promoter module located below that contains the CDE nucleotide sequence. -CHR or E2FBS-CHR to which suppressor proteins bind, which, thereby, can inhibit activation of the activator sequence located higher in the G0 and Gx phases of the cell cycle (PCT / GB94 / 17366; Lucibello et al., EMBO J. 14, 12 (1994)). 7) Cell-specific activatable promoters A preferably preferentially promoters or promoter sequences based on promoters or gene enhancers which, preferably formed for proteins, encode selected cells. For example, in the sense of the invention, promoters for the following proteins should preferably be used in the following cells: 7. 1 Activated promoter and activator sequences in endothelial cells - * • endothelial-1 glucose transporter, brain specific endoglyne receptor-1 VEGF (flt-1) VEGF receptor-2 (flk-1, KDR) - tie-1 or tie-2 B61 receptor (Eck receptor) B61 endothelin, especially endothelin B or endothelin-1 endothelin receptors, in particular endothelin B receptor mannose-6-phosphate receptors von Willebrand factor IL-la, IL- IL-1 receptor - vascular cell adhesion molecule (VCAM-1) synthetic activator sequences As an alternative to specific promoters of natural endothelial cells, synthetic activator sequences consisting of oligomerized binding sites can also be used for transcripts that are preferentially or selectively active in endothelial cells. An example of this is the transcription factor GATA-2, whose binding site in the endothelin-1 gene is 5'-TTATCT-3 '[Lee et al., Biol. Chem. 266, 16188 (1991), Dormann et al., J.

Biol. Chem. 267, 1279 (1992) and Wilson et al., Mol. Cell Biol. , 4854 (1990)]. 7. 2 Promoters or activator sequences, activated in cells in the vicinity of activated VEGF endothelial cells. The gene regulatory sequences for the VEGF gene are the 5 'flanking region, the 3' flanking region, the c-Src gene or the v-gene. Src - * - steroid hormone receptors and their promoter elements (Truss and Beato, Endocr .. Rev. 14, 459 (1993)), in particular the promoter of the mouse breast tumor virus. 7. 3 Promoters or activator sequences, activated in muscle cells, in particular smooth muscle cells tropomyosin a-actin c-myosin-receptor for PDGF receptor for FGF MRF-4 phosphofructokinase A phosphoglyceratomutase - troponin C myogenin receptors for endothelin A desmin VEGF The regulatory sequences of genes for the gene of VEGF have already been collected in the section "Promoters activated in cells in vicinity of activated endothelial cells" (see above) "artificial" promoters Factors of the family helix-loop-helix (HLH) (MyoD, Myf-5, myogenin, MRF4) are described as specific transcription factors of the muscles. In addition, to the specific transcription factors of the muscles belongs the zinc finger protein GATA-4. The HLH proteins as well as GATA-4 show a specific transcription of the muscles not only with promoters of muscle-specific genes, but also in a heterologous context, as well * with artificial promoters. Artificial promoters of this type are, for example, multiple copies of the binding site (DNA) for proteins HLH specific to muscles such as box E (Myo D) (for example, 4x AGCAGGTGTTGGGAGGC) or multiple copies of the DNA binding site for GATA-4 of the a-myosin heavy chain gene (e.g. 5 '-GGCCGATGGGCAGATAGAGGGGGCCGATGGGC-AGATAGAGG3') (SEQ ID NO: 12) Promoters and activator sequences, activated in glia cells They include, in particular, the regulatory sequences of genes or elements based on genes that encode, for example, the following proteins: - the cell-specific protein periaxin Schwann glutaminsintetase protected it from glia-specific cells (glial fibrillary acidic protein = GFAP) - SlOOb IL-6 glia cell protein (CNTF) receptors for 5-HT TNFa 'IL-10 - receptors I and II growth factor-like VEGF insulin The gene regulatory sequences for the gene VEGF have already been collected previously.

Promoters and activator sequences, activated in hematopoietic cells Regulatory sequences of genes of this type belong to the profiotor sequences for genes of a cytokine or its receptor, which are expressed in hematopoietic cells or in neighboring cells such as, for example, the stroma. These include promoter sequences for, for example, the following cytokines and their receptors: primitive cell factor receptor primitive cell factor IL-Ia IL-1 receptor IL-3-receptor (subunit a) receptor of IL-3 (subunit ß) IL-6 receptor of IL-6 GM-CSF - receptor of GM-CSF (chain or;) regulatory factor of interferon 1 (IRF-l) The promoter of IRF-1 is activated of equal way by IL-6 than by IFN? or IFN / 3 erythropoietin - erythropoietin receptor Promoters and activator sequences, activated in lymphocytes and / or macrophages These include, for example, the promoter and activator sequences of the genes for cytokines, cytokine receptors and adhesion molecules and receptors for the Fc fragment of antibodies. To them belong, for example: - IL-1 receptor IL-IL-1 / β IL-2 receptor IL-2 - "IL-3 receptor IL-3 (C subunit) IL-3 receptor (subunit ß) IL-4 receptor for IL-4 - IL-5 IL-6 receptor for IL-6 interferon regulatory factor 1 (IRF-1) (The IRF-1 promoter is activated in the same way by IL-6 as for IFN? Or IFN3) - promoter that responds to IFN? IL-7 IL-8 IL-10 IL-11 - IFN? GM-CSF GM-CSF receptor (chain) IL-13 LIF - colony stimulation factor receptor of macrophages (M-CSF) type I and II macrophage scavengers MAC-1 (leukocyte functional antigen) LFA-la (leukocyte functional antigen) - pl50.95 (leukocyte functional antigen) Activator and promoter sequences, activated in synovial cells They belong to the promoter sequences for matrix metalloproteinases (MMP), for example for: MMP-1 (interstitial collagenase) MMP-3 (stromelysin / transin) They also belong to promoter sequences for tissue inhibitors. metalloproteinases (TIMP), for example TIMP-1 TIMP-2 TIMP-3 Promoters and activator sequences, activated in leukemia cells They include, for example, promoters for c-myc-HSP-70 bcl-1 / cyclin D-1 bcl-2 IL-6 IL-10-TNFa, TNF0 HOX-11 BCR-Abl E2A-PBX-1 PML- RARA (retinoic acid receptor of promyelocytic leukemia) -. c-myc c-myc proteins bind and activate multimers of the nucleotide sequence (5 '-GGAAGCCAGAC-CAGCTGGTCTGCTTCC-3') (SEQ ID NO: 11) designated Myc E box Activators or activated activator sequences in tumor cells As the promoter or activator sequence, a gene regulatory nucleotide sequence is provided, with which transcription factors formed or active in tumor cells interact. In the sense of this invention, sequences or regulatory elements of genes based on genes encoding proteins formed particularly in cancer cells or sarcoma cells belong to the promoters or preferred activator sequences. Thus, in the case of small cell bronchial carcinomas, the promoter of the N-CAM protein is preferably used, in the case of ovarian carcinomas, the promoter of the "hepatitis growth factor" or "L" receptor. plastin, and in the case of pancreatic carcinomas, the promoter of L-plastin or polymorphic epithelial mucin (PEM).

SAW. The effector gene [component d)] In the sense of the invention, the effector genes [component d)] encode an active substance for the prophylaxis and / or therapy of a disease. Genes' effectors and promoter sequences are to be chosen in relation to the type of disease therapy and taking into account the target cell to be transduced. For example, in the case of the following diseases, the following combinations of promoter sequences and effector genes have to be chosen (a detailed description has already been made in patent applications EP95930524.4, EP95931933.6, EP95931204.2, EP95931205 .9, EP97101507.8, DE 19617851.7, DE19639103.2 and DE19651443, 6, to which reference is made). 1) Tumor therapy 1. 1) Target cells.- - proliferating endothelial cells or - stromal cells neighboring the endothelial cell and muscle cells or - tumor cells or leukemia cells 1. 2) Promoters: '' - specific to the endothelial cell and specific to the cell cycle or - non-specific to the cell or specific to the muscle cell and specific to the cell cycle or 1 - specific for the tumor cell (solid tumors, leukemia) and cell cycle specific ) Effector genes for inhibitors of cell proliferation, for example for - the retinoblastoma protein (pRb = pllO) or the related pl07 and pl30 proteins The retinoblastoma protein (pRb / pllO) and the related pl07 and pl30 proteins are inactivated by phosphorylation. Preferably, genes of these cell cycle inhibitors having mutations for the sites of inactivation of the expressed proteins without affecting their function should be used. Examples of these mutations were described for pllO. Analogously, the DNA sequence for the pl07 protein or the pl30 protein is mutated. - the p53 protein The p53 protein is inactivated in the cell by binding to special proteins such as, for example, MDM2 or by oligomerization of p53 through the dephosphorylated C-terminal serine. Preferably, therefore, a DNA sequence is used for a p53 protein that is shortened at the C-terminus in serine 392. - p21 (WAF-1) - the p66 protein - other cdk inhibitors - the GADD45 protein - the protein bak "" - a -Union protein for a regulatory protein (see II.1.) ) Effector genes for coagulation-inducing factors and inhibitors of angiogenesis, for example: - plasminogen activator inhibitor-1 (PAI-1) - PAI-2-PAI-3 - angiostatin - interferons (IFNa, IFN / 3 or IFN?) - platelet factor 4 - TIMP-l - TIMP-2 - TIMP-3 - leukemia inhibitory factor (LIF) - tissue factor (TF) and its fragments' with coagulant activity Effector genes for cytostatic and cytotoxic proteins, for example para-perforin '- granzyme - IL-2-IL-4-IL-12 - interferons such as, for example, IFN-a, IFN3 or IFN? - TNF such as TNFa or TNFß - oncostatin M - sphingomyelinase - magainin and maganinin derivatives Effector genes for cytostatic or cytotoxic antibodies and for fusion proteins between antibody fragments that bind antigens with cytostatic, cytotoxic or inflammatory proteins or enzymes. Cytostatic or cytotoxic antibodies belong to those directed against membrane structures of endothelial cells such as described, for example, by Burrows et al. (Pharmac.Ther.64, 155 (1994)), Hughes et al., (Cancer Res. 49, 6214 (1989)) and Maruyama et al., (PNAS USA 87, 5744 (1990)). In particular, they belong to antibodies against VEGF receptors. - Furthermore, they include cytostatic or cytotoxic antibodies directed against membrane structures in tumor cells. Antibodies of this type were represented synoptically, for example, by Sedlacek et al., Contrib. to Oncol. 32., Karger Publishing House, Munich (1988) and Contrib. to Oncol. 43., Karger Publishing House, Munich (1992). Other examples represent antibodies against Lewis sialyl, - against peptides in tumors, which are recognized by T cells; against proteins expressed by oncogenes, - against gangliosides such as GD3, GD2, GM2, 9-0-acetyl GD3, fucosyl GM1, - contratantigens of blood groups and their precursors, - against antigens in the polymorphic epithelial mucin; against antigens in heat shock proteins In addition, they belong to them antibodies directed against membrane structures of leukemia cells. A large number of monoclonal antibodies of this type have already been described for diagnostic and therapeutic procedures (compilations in Kristensen, Danish Medical Bulletin 41, 52 (1994 Schranz, Hungarian Therapy 8, 3 (1990 Drexler et al., Leuk. Res. 10., 279 (1986 Naeim, Dis. Markers 7., 1 (1989); Stickney et al., Curr .. Opin. Oncol. 4, 847 (1992); Drexler et al., Blut 57, 327 (1988); Freedman et al., Cancer Invest. 9. 69 (1991)). Depending on the type of leukemia, they are suitable, as ligands, for example monoclonal antibodies or their antibody fragments that bind to antigens directed against the following membrane antigens: AML CD13 CD15 CD33 10 Sialosyl-Le membrane antigen cells B-CLL CD5 CDlc CD23 15 idiotypes and isotypes of membrane immunoglobulins T-CLL CD33 20 M38 receptors for IL-2 T cell receptors ALL CALLA 25 CD19 non-Hodgkin's lymphoma The humanization of murine antibodies, the preparation and optimization of genes for Fab and recombinant Fv fragments are carried out in a manner corresponding to the technique known to the person skilled in the art. The fusion of recombinant Fv fragments with genes for proteins or cytostatic enzymes, cyto¬ Toxic or inflammatory reactions are carried out in the same way, in a manner corresponding to the state of the art known to the person skilled in the art. 7) Effector genes for ligand fusion proteins that bind to target cells with cytostatic and cytotoxic proteins. The ligands include all substances that bind to membrane structures or membrane receptors on endothelial cells. For example, they contain cytokines such as, for example, IL-1 or growth factors or their fragments or partial sequences which, expressed in receptors, are bound by endothelial cells such as, for example, PDGF, bFGF , VEGF, TGF. In addition, they belong to adhesion molecules that bind to activated and / or proliferating endothelial cells. They include, for example, SLex, LFA-1, MAC-1, LECAM-1, VLA-4 or vitronectin. They also belong to substances that bind to structures of the membrane or receptors of the membrane of tumor cells or of leukemia. For example, they include, for example, hormones or growth factors or their fragments or part sequences that bind to receptors expressed by leukemia cells or tumor cells. Growth factors of this type have already been described (compilations in Cross et al., Cell 64, 271 (1991), Aulitzky et al., Drugs 48, 667 (1994), Moore, Clin. Cancer Res. 1, 3 (1995), Van Kooten et al., Leuk, Lymph., 12, 27 (1993).) The fusion of the genes of these ligands that bind to the target cell with proteins or cytostatic, cytotoxic or inflammatory enzymes is effected by corresponding to the state of the art with methods known to the person skilled in the art.

Effector genes for inflammation inducers, for example for IL-1 - IL-2 RANTES (MCP-2) monocyte activating and chemotactic factor (MCAF) IL-8 - macrophage inflammatory protein -1 (MIP-la, -β) neutrophil activating protein -2 (NAP-2) IL-3 IL-5 - human leukemia inhibitory factor (LIF) IL-7 IL-11 IL-13 GM-CSF-G-CSF M-CSF cobra venom factor (CVF) or partial sequences of FVC that functionally correspond to the human complement factor C3b, that is to say that they can bind to the factor of complement B and that, after separation by the factor D, represent a C3 convex the human complement factor C3 or its partial sequence C3b dissociation products of the human complement factor C3 that are functionally and structurally similar to CVF bacterial proteins that activate the complement and trigger inflammations such as, for example, Salmonella typhi murium proteins, "grouping" factors of Staphylococcus aureus, modulins, in particular of gram-negative bacteria, "main outer membrane protein "Legionellae or Haemophilus influenzae type B 'or Klebsiella or M molecules of group G streptococci.

Effector genes for enzymes for activating precursors for cytostatic agents, for example for enzymes which cleave inactive previous substances (prodrugs) into active cytostatics (drugs). Substances of this type and the corresponding prodrugs and respective drugs have already been described compilatively by Deonarain et al. (Br. J. Cancer 70, 786 (1994)), Mullen, Pharmac. Ther. 63., 199 (1994)) and Harris et al. (Gene Ther., 170 (1994)). For example, the DNA sequence of one of the following enzymes should be used: thymidine kinases of the Herpes simplex virus thymidine kinase virus Varizella zoster bacterial nitroreductase / bacterial 3-glucuronidase, vegetable β-glucuronidase of Sécale cereale / human 3-glucuronidase human carboxypeptidase (CB), for example CB-A of the barley cell, CB-B of the pancreas or bacterial carboxypeptidase - / 3-lactamase bacterial cytosine-deaminase bacterial catalase or human peroxidase phosphatase, in particular human alkaline phosphatase, acid phosphatase of the prostate human or phosphatase type 5-oxidase, in particular human lysyloxidase or Human acid peroxidase D-aminooxidase, in particular human glutathione-peroxidase, human eosinophil peroxidase or human lymphoid ganglion peroxidase-galactosidase of autoimmune diseases and inflammations 1) Target cells: - proliferating endothelial cells or macrophages and / or lymphocytes or synovial cells 2) Promoters: - specific for endothelial cells and cell cycle specific or macrophage specific and / or lymphocyte and / or cell cycle specific or specific synovial and / or cell cycle specific cells 3) Effector genes for allergy therapy, for example for IFN / 3 - IFN? •• IL-10 antibodies or antibody fragments against IL-4 soluble IL-4 receptors - IL-12 TGF / 3 effector genes to prevent rejection of transplanted organs, for example for IL-10 TGF / 3 - soluble IL-1 receptors soluble IL-2 receptor antagonists IL-1 receptors soluble IL-6 receptors immunosuppressive antibodies or their fragments containing VH and VL or their VH and VL fragments bound through a linker. Immunosuppressive antibodies are, for example, antibodies specific for the T cell receptor or its CD3 complex, against CD4 or CD8 and, in addition, against the IL-2 receptor, the IL-1 receptor or the IL-1 receptor. 4 or against adhesion molecules CD2, LFA-1, CD28 or CD40 Effector genes for the therapy of autoimmune diseases mediated by antibodies ", for example for TGF3 IFNa IFN3 - IFN? IL-12 soluble IL-4 receptors soluble IL-6 receptors immunosuppressive antibodies or their fragments containing VH and VL Effector genes for the therapy of cell-mediated autoimmune diseases, for example for IL-6 - IL-9 IL-10 IL-13 TNFa or TNF / 3 an immunosuppressant antibody or its fragments containing VH and VL Effector genes for inhibitors of cell proliferation, proteins and cytostatic or cytotoxic enzymes for the activation of cytostatic agent precursors Examples of genes that encode proteins of this type have already been pointed out in the section "effector genes for tumor therapy". In the same way as described therein, effector genes encoding antibody fusion proteins or recombinant Fab or Fv fragments of these antibodies or other ligands specific for the target cell and the cytokines, growth factors can be used in the sense of the invention. , receptors, proteins and cytostatic or cytotoxic enzymes indicated above.

Effector genes for arthritis therapy In the sense of the invention, effector genes are chosen, whose expressed protein inhibits directly or indirectly inflammation, for example in the joint, and / or promotes the reconstitution of extracellular matrix (cartilage, connective tissue) in the joint. They belong, for example, IL-1 receptor antagonist (IL-1-RA); IL-1-RA inhibits IL-1 binding, soluble IL-1 receptor soluble IL-1 receptor binds and inactivates IL-1 IL-6 IL-6 increases the secretion of TIMP and superoxide and reduces Secretion of IL-1 and TNFa by synovial cells and chondrocytes - soluble TNF receptor soluble TNF receptor binds and inactivates TNF. IL-4 IL-4 inhibits the formation and secretion of IL-1, TNFa and MMP IL-10 inhibits the formation and secretion of IL-1, TNFa and MMP and increases the secretion of TIMP insulin-like growth factor (IGF-1) IGF-1 stimulates the synthesis of extracellular matrix. TGFβ, especially TGF / 31 and TGF32 TGF3 stimulates the synthesis of extracellular matrix superoxide dismutase TIMP, especially TIMP-1, TIMP-2 or TIMP-3 ia of the deficient formation of blood cells ) Target cells: proliferating immature cells of the hematopoietic system stromal cells neighboring the hematopoietic cells Promoters: specific for hematopoietic and / or cell cycle-specific cells - nonspecific for cells and specific for the cell cycle 3.3) Effector genes for the therapy of anemia, for example for erythropoietin 3. 4) Effector genes for the therapy of leukopenia, for example for G-CSF GM-CSF M-CSF 3. 5) Effector genes for the therapy of thrombocytopenia, for example for IL-3 leukemia inhibitory factor (LIF) - IL-11 thrombopoietin rape of nervous system injuries 4. 1) Target cells: glia cells or proliferating endothelial cells 4. 2) Promoters: - glia cell-specific and cell cycle-specific or specific for endothelial cells and cell cycle-specific or nonspecific and cell-cycle specific 4. 3) Effector genes for neuronal growth factors, for example FGF t nerve growth factor (NGF) - neurotropic factor derived from the brain (BDNF) neurotrophin-3 (NT-3) neurotrophin-4 (NT-4) ciliary neurotrophic factor (CNTF) 4. 4) Effector genes for enzymes, for example for tyrosine hydroxylase dopadecarboxylase 4. 5) Effector genes for cytokines and their inhibitors that inhibit or neutralize the neurotoxic effect of TNFa !, for example for TGF / 3 TNF receptors soluble TNF receptors that neutralize TNFa - IL-10 IL-10 inhibits the formation of IFN ?, TNFa, IL-2 and IL-4 receptors of soluble IL-1 receptor I of IL-1 - receptor II of IL-1 soluble IL-1 receptors neutralize the activity of IL-1 receptor antagonist IL-1 soluble IL-6 receptors ) Therapy of disorders of the blood coagulation system and blood circulation . 1) Target cells: Endothelial cells or proliferating endothelial cells or somatic cells in the vicinity of endothelial cells and smooth muscle cells or macrophages. . 2) Nonspecific promoters of the cell and specific cell cycle or specific endothelial cells, smooth muscle cells or macrophages and specific cell cycle Structural genes for the inhibition of coagulation or for the promotion of fibrinolysis, for example para-activator of tissue plasminogen (tPA) urokinase-type plasminogen activator (uPA) hybrids of tPA and uPA protein C-hirudin serine inhibitors proteinase (serpin) such as, for example, inhibitor of C-1S, antitrypsin or antithrombin III inhibitor of the tissue factor pathway (TFPI) Effector genes for the promotion of coagulation, for example for F VIII F IX - von Willebrand factor F XIII PAI - 1 PAI-2 tissue factor and fragments thereof Effector genes for angiogenesis factors, for example for VEGF FGF Effector genes for lowering blood pressure, for example for kallikrein "nitric oxide synthase" of endothelial cells . 7) Effector genes for the inhibition of the proliferation of smooth muscle cells after endothelial layer injury, for example for an antiproliferative, cytostatic or cytotoxic protein or an enzyme for the separation of cytostatic agent precursors in cytostatic agents as indicated before (in the tumor section) or - a fusion protein of one of these active substances with a ligand, for example an antibody or fragments of antibodies specific for muscle cells . 8) Effector genes for other proteins of the blood plasma, for example for albumin inactivator of Cl cholinesterase serum - transferrin 1-antitrypsin 6) Vaccinations 6. 1) Target cells: •• muscle cells or macrophages and / or lymphocytes endothelial cells 6. 2) Promoters: non-specific and specific to the cell cycle or specific to the target cell and specific to the cell cycle 6. 3) Effector genes for the prophylaxis of infectious diseases The possibilities of preparing effective vaccines by conventional means are limited. Therefore, the DNA vaccine technology was developed. These DNA vaccines pose, however, questions regarding the power of activity. According to this invention, it is necessary to have a higher activity of DNA vaccines. As an active substance, a DNA of a protein formed by the infectious pathogenic agent must be chosen., by triggering an immune reaction, ie by binding antibodies and / or by cytotoxic T lymphocytes, leads to neutralization and / or extermination of the pathogenic agent. The so-called neutralization antigens of this type are already used as vaccination antigens (see a compendium in Ellis, Adv. Exp. Med. Biol. 327, 263 (1992)). In the sense of the invention, DNA encoding neutralizing antigens of the following pathogens is preferred: influenza A * virus "HIV rabies virus HSV (Herpes simplex virus) RSV (respiratory syncytial virus) parainfluenza virus - rotavirus V21V (Varicella zoster virus) CMV (cytomegalovirus) measles virus HPV (human papillomavirus) HBV (hepatitis B virus) - HCV (hepatitis C virus) HDV (hepatitis D virus) HEV (hepatitis E virus) ) HAV (hepatitis A virus) Vibrio cholerae antigen - Borrelia burgdorferi Helicobacter pylori malaria antigen Active substances of this type within the meaning of the invention belong, however, also to the DNA of an anti-cancer antibody. idiotype or its fragments that bind antigens, whose antigen binding structures (the "determining regions of complementarity") represent copies of the structure of the protein or carbohydrate of l Neutralization antigen of the infectious pathogen. Anti-idiotype antibodies of this type can replace in particular carbohydrate antigens in the case of bacterial infectious pathogens. Anti-idiotype antibodies of this type and their dissociation products have already been described by way of compendium by Hawkins et al. (J. Immunother: '14, 273 (1993)) and Westerink and Apicella (Springer Seminars in Immunopathol 15, 227 (1993)).

Effector genes for "vaccines against tumors" - They belong to antigens based on tumor cells. Antigens of this type were presented as a compendium, for example, by Sedlacek et al., Contrib. to Oncol. 32., Karger Publishing House, Munich (1988) and Contrib. to Oncol. 43., Karger Publishing House, Munich (1992). Other examples are represented by genes for the following protein antigens or for the variable region (VL, VH) of anti-idiotype antibodies corresponding to the following non-protein antigens: - gangliosides: sialyl Lewis peptides on tumors that are recognized by T cells expressed by oncogenes - antigens of blood groups and their antigen precursors on tumor-associated mucin antigens on heat shock proteins The therapy of chronic infectious diseases .1) Target cell: lymphocyte liver cell and / or macrophage - endothelial cell epithelial cell .2) Promoters: specific to the virus or specific to the "cell" and cell cycle specific 3) Effector genes, for example for a protein that has cystotic, apoptotic or cytotoxic effects - an enzyme that dissociates in the active substance a precursor of an antiviral or cytotoxic substance. 7. 4) Effector genes for antiviral proteins, cytokines and growth factors with antiviral activity. They include, for example, IFNcü, IFNjg, IFN ?, TNF ?, TNFa, IL-1 or TGF3 antibodies of a specificity that inactivate the respective virus or its fragments containing VH and VL or its fragments VH and VL linked through a linker, prepared as already described. Antibodies against virus antigen are, for example: anti-HBV anti-HCV anti-HSV anti-HPV anti-HIV 20 anti-EBV anti-HTLV anti-virus Coxsackie anti-virus Hantaan a protein that binds Rev. These proteins are 25 bind to Rev RNA and inhibit the post-transcriptional steps dependent on Rev expression of retrovirus genes. Examples of Rev binding proteins are: RBP9-27 30 - RBP1-8U - ', RBP1-8D pseudogenes of RBP1-8 ribozymes which digest the mRNA of genes for cell cycle control proteins or virus mRNA. Catalytic ribozymes for HIV were described by way of compendium, for example, by Christoffersen et al., J. Med. Chem. 38, 2033 (1995).

) Effector genes for antibacterial proteins Antibacterial proteins include, for example, antibodies that neutralize bacterial toxins or opsonize bacteria. For example, they belong to antibodies against meningococci C or B E. coli Borrelia Pseudomonas Helicobacter pylori Staphylococcus aureus VII. Combination of the same or different effector genes The object of the invention is, furthermore, a self-reinforcing and possibly controllable expression system by pharmacological route, in which a combination of the DNA sequences of two identical effector genes or of two different effector genes [components c) and e 'is present. )]. For the expression of the two DNA sequences, another promoter sequence or, preferably, the cDNA of an "internal ribosome entry site" (IRES) is interposed between the two effector genes as the regulatory element. An IRES enables the expression of two DNA sequences linked together through an IRES. IRES's of this type were described, for example, by Montford and Smith (TIG 11, -179 (1995), Kaufman et al., Nucí Acids Res. 19, 4485 (1991), Morgan et al., Nucí. Acids Res. 20, 1293 (1992), Dirks et al., Gene 128, 247 (1993), Pelle-tier and Sonenberg, Nature 334, 320 (1988) and Sugitomo et al., BioTechn., 12, 694 (1994). ). Thus, for example, the cDNA of the IRES sequence of the poliovirus (position = 140 a = 630 of the 5'UTR).

Preferably, in the sense of the invention, effector genes must be linked through other promoter sequences or an IRES sequence, which have an additive effect. In the sense of the invention, they are preferred combinations of effector genes, for example for 1) tumor therapy equal or different proteins, cytostatic, apoptotic, cytotoxic or inflammatory or enzymes equal or different for the dissociation of the precursor of a cytostatic agent 2) the therapy of autoimmune diseases - cytokines or different receptors with synergistic effect for the inhibition of the cellular and / or humoral immune reaction or TIMP's different or equal 3) the therapy of the deficient formation of blood cells different hierarchically consecutive cytokines such as, for example, IL-1, IL-3, IL-6 or GM-CSF and erythropoietin, G-CSF or thrombopoie 4) nerve cell injury therapy a neuronal growth factor and a cytokine or inhibitor of a cytokine ) the therapy of disorders of the blood coagulation system and blood circulation an antithrombotic agent and a fibrinolytic agent (for example tPA or uPA) or a cytostatic, apoptotic or cytotoxic protein and an antithrombotic agent or a fibrinolytic agent several different blood coagulation factors, acting synergistically, for example F VIII and FvW or F VIII and F IX 6) Vaccinations an antigen and an immunostimulatory cytokine such as, for example, IL-la, IL-1/3, IL-2, GM-CSF, IL-3 receptor or IL-4 different antigens of an infectious pathogen. cioso or of different infectious pathogens or different antigens of a tumor type or of different types of tumors 7) Viral infectious disease therapy an antiviral protein and a cytostatic protein, apoptotic or cytotoxic antibodies against different surface antigens of a virus or of several viruses 8) Therapy of infectious bacterial diseases antibodies against different superficial antigens and / or toxins of a germ VIII. Incorporation of signal sequences and transmembrane domains 1) Reinforcement of the translation To reinforce the translation, the sequence "of nucleotides GCCACC or GCCGCC can be introduced at the 3 'end of the promoter sequence and, directly at the 5' end of the initiation signal (ATG), the signal or transmembrane sequence (Kozak, J. Cell Biol. 108, 299 (1989)). 2) Secretion Facilitation To facilitate the secretion of the expression product of the effector gene, the homologous signal sequence optionally contained in the DNA sequence of the effector gene can be replaced by a heterologous signal sequence that enhances extracellular secretion. Thus, for example, the signal sequences for immunoglobulin (DNA position = 63 a = 107; Riechmann et al., Nature 332, 323 (1988)) or the signal sequence for CEA (DNA position) can be included. = 33 a = 134, - Schrewe et al., Mol.Cell Biol. 10, 2738 (1990), Berling et al., Cancer Res. 50, 6534 (? 990)) or the signal sequence of the virus glycoprotein human respiratory syncytial (amino acid cDNA = 38 a = 50 or 48 to 65; Lichtenstein et al., J. Gen. Virol. 77, 109 (1996)).

Anchoring of the active substance For the anchoring of the active substance in the cell membrane of the transduced cell forming the active substance, a sequence for a transmembrane domain can be introduced alternatively or additionally to the signal sequence. Thus, for example, the transmembrane sequence of the human macrophage colony stimulating factor (DNA position = 1485 a = 1554; Cosman et al., Behring Inst. Mitt. 83., 15 (1988)) or the sequence of DNA for the signal and transmembrane region of the human respiratory syncytial virus (RSV) glycoprotein G (amino acids 1 to 63. or its partial sequences, amino-ci &amps 38 to 63; Vijayá 'et al., Mol Cell Biol. 8, 1709 (1988), Lichtenstein et al., J. Gen. Virol. 77, 109 (1996)) or the DNA sequence for the signaling and transmembrane region of the neuraminidase virus of the influenza (amino acids 7 to 35 or partial sequence, amino acids 7 to 27; Brown et al., J. Virol. £ 2, 3824 (1988)) can be introduced between the promoter sequence and the sequence of the effector gene.

However, for the anchoring of the active substance in the cell membrane of the transduced cells forming the active substance, the nucleotide sequence for a glycophospholipid anchor can also be introduced. The introduction of a glycophospholipid anchor is carried out at the 3 'end of the nucleotide sequence for the effector gene and can be carried out, additionally, for the introduction of a signal sequence. Glycophospholipid anchors are described, for example, for CEA, for N-CAM and for other membrane proteins such as, for example, Thy-1 (see compendium by Ferguson et al., Ann.Rev. Biochem. ., 285 (1988)).

Another possibility of anchoring active substances to the cell membrane corresponding to the present invention is the use of a DNA sequence for a ligand-active substance fusion protein. The ligand specificity of this fusion protein is directed against a structure of the membrane on the cell membrane of the target cell chosen. Ligands that bind to the surface of cells include, for example, antibodies or fragments of antibodies directed against structures on the surface of, for example, endothelial cells. In particular, they "" belong antibodies against the VEGF receptors or against quinine or muscle cell receptors such as antibodies against actin or antibodies against angiotensin II receptors or antibodies against growth factor receptors such as, for example, against EGF receptors or against PDGF receptors or against FGF receptors or antibodies against endothelin AA receptors the ligands also belong to antibodies or their fragments which are directed against tumor-specific or tumor-associated antigens on the tumor cell membrane. Antibodies of this type have already been described. The murine monoclonal antibodies are preferably used in a humanized form. Fab and recombinant Fv fragments and their fusion products are prepared, as already described, with the technology known to the person skilled in the art. The ligands also belong to all active substances such as, for example, cytokines or adhesion molecules, growth factors or their fragments or their partial sequences, mediators or peptide hormones that bind to membrane structures or receptors of the membrane on the respective chosen cell. For example, they belong to - ligands for endothelial cells such as IL-1, PDGF, bFGF, VEGF, TGG3 or quinine and quinine derivatives or analogues. In addition, adhesion molecules belong to them. Adhesion molecules of this type such as, for example, SLex, LFA-1, MAC-1, LeCAM-1, VLA-4 or vitronectin and derivatives or analogs of vitronectin have already been described for endothelial cells (compendiums in Augustin-Voss et al. al., J. Cell Biol. 119, 483? (1992), Pauli et al., Cancer Metast., Rev. 9, 175 (1990); Honn et al. "., Cancer Metast., Rev. 11, 353 (1992). ); Varner et al., Cell Adh. Commun. 3, 367 (1995)). The invention is described in more detail in the following examples.

IX. Examples for the more detailed description of the object of the invention 1) Preparation of an expression system controlled by oncogenes The expression system controlled by oncogenes according to the invention consists of the following sequences of different nucleotides juxtaposed located lower: Component a) • the promoter of the cdc25C gene (nucleic acids -290 to +121; Zwicker et al., EMBO J. 14, 4514 (1995), Zwicker et al., Nucí Acids Res. 22, 3822 (1995)) Component b) • the SV40 nuclear localization signal (NLS) (SV40 long T, amino acids 126 to 132; PKKKRKV (SEQ ID NO: 13); Dingwail et al., TIBS 16, 478 (1991)) • the transactivation domain (TAD) acid of VP16 of HSV-1 (amino acids 406 to 488; Triezenberg et al., Genes Developm., 2, 718 (1988); Triezenberg, Curr. Opin. Gene Developm., 5, 190 (1995 )) • the RB binding sequence of the E2F-1 protein (amino acids 409 to 426 (LDYHFGLEEGEGIRDLFD) (SEQ ID NO: 14); Flemington et al., PNAS USA 90, 6914 (1993); Helin et al., Cell 70, 337 (1992)) • the cDNA for the DNA binding domains of the Gal4 protein (amino acids 1 to 147; Chasman and Kornberg, Mol. Cell Biol. 10, 2916 (1990)) Component c) • 10 times the binding sequence for the DNA binding sequence of Gal4 with the sequence of nucleotides 5 '-CGGACAATGTTGACCG-3' (SEQ ID NO: 1 ) (Chasman and Kornberg, Mol.Cell Biol. 10, 2916 (1990)) • the basal promoter of SV40 (nucleic acids 48 to 5191; Toóse (comp.) DNA Tumor Viruses; Cold Spring Harbor, New York, New York, Cold Spring Harbor Laboratory) Component d) • the GCCACC sequence (Kodak, J. Cell Biol. 108., 229 (1989)) • the cDNA for the immunoglobulin signal peptide (nucleotide sequence 63 to 107; Riechmann et al., Nature 332, 323 (1988)) • the cDNA of 3-glucuronidase (nucleotide sequence 93 to 1982, Oshima et al, PNAS (USA 84, 685 (1987)) The linkage of the various components of the construction is carried out at Through appropriate restriction sites that are driven together by PCR amplification to the ends of the different elements, the binding is carried out with the help of enzymes and DNA-ligases specific for the restriction sites, known to the person skilled in the art. These enzymes are commercially available.The nucleotide construct, thus prepared, is incorporated by cloning into the plasmid vector pXP2 (Nordeen, BioTechniques 454 (1988)) which is used directly or in colloidal dispersion systems. for an application In vivo treatment With the plasmid described, 3T3 fibroblasts maintained in culture (positive RB) and osteosarcoma cells (SAOS-2, negative RB) are transfected with the method known to the person skilled in the art (Lucibello et al., EMBO J. 132 (1995)) and the amount of 3-glucuronidase produced by the fibroblasts or osteosarcoma cells is measured with the help of 4-methylumbelliferyl- / 3-glucuronide as a substrate. To examine the capacity of the cell cycle, osteosarcoma cells are synchronized for 48 hours in GQ / G! By extraction of methionine.The DNA content of the cells is determined after staining with Hoechst 33258 in a classifier of fluorescence activation cells (Lucibello et al., EMBO J. 132 (1995)). The following results are achieved: In transfected fibroblasts (positive RB) no increase in 3-glucuronidase can be determined in comparison with non-transfected fibroblasts Transfected osteosarcoma cells (RB negative) clearly express more / 3-glucuronidase than non-transfected osteosarcoma cells Proliferating osteosarcoma cells (DNA> 2S; S = single chromosome sequence) clearly segregate more ß-- glucuronidase than osteosarcoma cells synchronized in Go / G, (DNA = 2S) .Therefore, the described expression system leads to a RB-dependent expression of the gene structural ß-glucuronidase that can be regulated depending on the choice of the promoter sequence, for example in a cell cycle dependent manner. 2) Preparation of a virus-controlled expression system The virus-controlled expression system according to the invention consists of the following juxtaposed different nucleotide sequences located below: Component a) • the promoter of the cdc25C gene (nucleic acids -290 to +121; Zwicker et al., EMBO J 14, 4514 (1995), Zwicker et al., Nucí Acids Res. 23., 3822 (1995)) Component b) • the SV40 nuclear localization signal (NLS) (SV40 long T, amino acids 126 to 132, PKKKRKV (SEQ ID NO: 13), Dingwall et al., TIBS 16, 478 (1991)) • "the TR & amp; Activation domain (TAD) HSV-1 VP16 acid (amino acids 406 to 488; Triezenberg et al., Genes Developm., 2, 718 (1988); Triezenberg, Curr. Opin., Gene Developm. 1995)) • the E6 protein of the HPV-18 virus (nucleotide sequence 100 to 578, Roggenbuck et al., J. Virol. 65, 5068 (1991)) • the cDNA for the DNA binding domains of the Gal4 protein (amino acids 1 to 147; Chasman and Kornberg, Mol.Cell Biol. 10, 2916 (1990)) Component c) "10 times the binding sequence for the DNA binding sequence of Gal4 with the nucleotide sequence 5 '-CGGACAATGTTGACCG -3 '(SEQ ID NO: 1) (Chasman and Kornberg, Mol Cell Biol. 10, 2916 (1990)) • the SV40 basal promoter (nucleic acids 48 to 5191); Toóse (comp.) DNA Tumor Viruses; Cold Spring Harbor, New York, New York, Cold Spring Harbor Laboratory) Component d) • the GCCACC sequence (Kodak, J. Cell Biol. 108, 229 (1989)) • the cDNA for the immunoglobulin signal peptide (sequence of nucleotides 63 to 107; Riechmann et al., Nature 332, 323 (1988)) • cDNA of 3-glucuronidase (nucleotide sequence 93 to 1982; Oshima et al, PNAS USA 84., 685 (1987)) The link The different components of the construction are carried out through suitable restriction sites that are driven together through the PCR amplification to the ends of the different elements. The binding is carried out with the help of enzymes and DNA-ligases specific for the restriction sites, known to the person skilled in the art. These enzymes can be purchased commercially. The "construction of nucleotides" thus prepared is incorporated by cloning into the plasmid vector pUC18 / l9 which is used directly or in colloidal dispersion systems for in vivo application With the described plasmid, human fibroblasts maintained in culture (Wi-38, E6 / E7 negative) and neck carcinoma cells (HeLa, HPV-18 E6 / E7 positives) with the method known to those skilled in the art (Lucibello et al., EMBO J. 132 (1995)) and the amount of β-glucuronidase produced by these cells is measured with the help of 4-methylumbelliferyl- / 3-glucuronide as substrate.To examine the specificity of the cell cycle, the HeLa cells are synchronized over 48 hours in G0 / G1 by extraction of methionine .. The DNA content of the cells is determined after staining with Hoechst 33258 in a classifier of fluorescence activation cells (Lucibello et al., EMBO J. 132 (1995 )).: The following are achieved nte results: In transfected fibroblasts no increase in -glucuronidase can be determined in comparison with untransfected fibroblasts. Transfected HeLa cells clearly express more ß -glucuronidase than non-transfected HeLa cells. Proliferating HeLa cells (DNA> 2S; S = single chromosome sequence) clearly secrete more / 3-glucuronidase than HeLa cells synchronized in GQ / G- ^ (DNA = 2S). Accordingly, the described expression system leads to a specific expression of the virus (HPV18) of the structural / 3-glucuronidase gene that can be regulated as a function of the choice of the promoter sequence, for example in a cell-cycle dependent manner. An active substance according to Examples 1 and 2 allows, after local application, for example in the place of the tumor or after intracranial or subarachnoid administration, or of systemic administration, preferably intravenous or intraarterial, which predominately, if not in an "exclusive way, only those cells that have a mutated oncogene or a viral infection secrete / 3-glucuronidase." This / 3-glucuronidase dissociates a doxorubicin -? - glucuronide now injected and well compatible (Jacquesy et al., EP 0 511 917 Al) in cytostatic doxorubicin, which inhibits the proliferation of endothelial cells and acts cytostatically on cells as well as on neighboring tumor cells, thereby inhibiting tumor development.

Legends of the figures Figure 1: Type and arrangement of the general components of a nucleic acid construct according to the invention.

Figure 2: Schematic representation of the arrangement of the general components of a nucleic acid construct according to the invention according to embodiment A.

Figure 3: Schematic representation of the arrangement of the general components of a nucleic acid construct according to the invention according to embodiment B.

Claims (32)

1. CLAIMS 1. - Construction of nucleic acids for the expression of an effector gene, the nucleic acid construct containing a promoter I (component a) that controls the expression of a gene of the transcription factor (component b) also contained in the construction of nucleic acids, and containing a promoter II (component c) to which the gene product of the gene specifically binds? transcription factor and that controls the expression of an effector gene (component d) also contained in the construction of nucleic acids, characterized in that the activity of the gene product of the transcription factor gene depends on one or more cellular regulatory proteins that specifically bind to this gene product and that affect its activity.
2. 2. Nucleic acid construction according to claim 1, characterized in that - Component a): is an activation sequence for the transcription of component b), - Component b): it is a transcription factor that contains h) an activation domain b2) a binding sequence for a cellular regulatory protein b3) a DNA binding domain; Component c): it is an activation sequence that is activated by the binding of the product "of expression of component b) and activates the transcription of component d), and Component d): it is an effector gene 3.- Nucleic acid construction according to claim 1, characterized in that components a, b and c are made in the form of components a ', b' and e ': Component a'): an activation sequence for the transcription of component b ') containing the sequence DNA binding for a cellular regulatory protein a2) a basal promoter Component b '): a transcription factor representing a repressor protein and inhibits component c') Component c '): an activation sequence containing cx) a sequence of activation for the transcription of component d) c2) a DNA sequence that binds the repressor protein [component b ')] and, thereby, inhibits the activation of transcription of the component d) Compounds d) an effector gene 4. Nucleic acid construction according to claim 2, characterized in that component a) is equal to component c). 5. Nucleic acid construction according to claim 2 or 3, characterized in that component a) or component c_) represents a non-specific, cell-specific, metabolically specific, virus-specific and / or specific promoter sequence of the virus. of the cell cycle. 6. Nucleic acid construction according to claim 5, characterized in that component a) or component cx) is selected from the group containing activated promoters in endothelial cells, peritoneal cells, pleural cells, epithelial cells of the skin, of the lung of the gastrointestinal tract, the kidneys and the ureters, in muscle cells, in connective tissue cells, in hematopoietic cells, in macrophages, in lymphocytes, in leukemia cells, in tumor cells or in glia cells or promoter sequences of viruses such as HBV, HCV, HSV, HPV, EBV, HTLV, CMV or HIV promoter or enhancer sequences activated by hypoxia or specific cell cycle activation sequences of the genes for cdc25C, cyclin A, cdc2, E2F-1, B -myb and DHFR binding sequences for transcription factors that manifest as a function of cell proliferation or activated such as monomers or multimers of the Myc E box. 7.- Nucleic acid construction according to one or more of claims 2 to 6, characterized in that the activation domain [(component bx)] of component b) is chosen from the group containing the activation domains of the transcription factors Oct-2, Spl, NFY, ITF-2, VP-16, c-Myc and CTF. 8. - Nucleic acid construction according to one or more of claims 2 to 7, characterized in that the binding sequence [(component b2) of component b)] for a cellular regulatory protein is a cellular binding protein or a part of this binding protein. 9. - Nucleic acid constructions according to claim 8, characterized in that the cell binding protein or a part of this binding protein binds to a cellular regulatory protein chosen from a group containing p53, pRb, p30, Max, MAD, VHL, cdk-4, MTS-1 (pl6 ), WT-1, SMAD-2, DPC-4. 10. Nucleic acid construction according to claim 9, characterized in that the component b ') is chosen from a group of cellular protein proteins containing E2F-1, -2, -3, -4, -5, cyclin-D1 (-D2, -D3 or -C, cyclin-A, -E, Myc, transcription factor PU.lo Elf-1, elongin- B, -C, pl4, pl5, pl6, pl8, p21, p27, p53, Myc, cdk-4, DPC-4 and SMAD-2 11.- Nucleic acid construction according to claim 7, characterized in that the sequence of binding [component b2) of component b)] for a cellular regulatory protein is a viral binding protein or a part of this binding protein. 12. Nucleic acid construction according to claim 11, characterized in that the viral binding protein or a part of this binding protein binds to a cellular regulatory protein chosen from a group containing p53, pRb (pllO), NFKB, pl30, CBF-1, Lyn-tyrosine kinase, bak and bax. 13. Nucleic acid construction according to claim 12, characterized in that the component b2): is chosen from a group of viral binding proteins containing IE 84 of CMV, E1B (55 Kd) of AV, EBNA-5 of EBV, BHFR of EBV, E6 of HPV-16 or -18, protein x of HBV, T antigen of SV40, E1A of AV,? BNA-2 of EBV, EBNA-1 of EBV, E7 of HPV, Tax of HIV, LMP-1 from EBV, LMP-2A or LMP-2B from EBV, E1B (16 Kd) from AV, E1B (10 kD) from AV. 14. Nucleic acid construction according to claim 7, characterized in that the binding sequence [component b2) of component b)] for a cellular regulatory protein is an antibody or a part of this antibody. 15. - Nucleic acid construction according to one of the preceding claims, characterized in that the component c) contains at least one DNA sequence for the binding of component b) and, by this binding, activates the expression of component d). 16. Nucleic acid construction according to claim 15, wherein the DNA sequence is chosen from a group containing the binding sequence (5 '-CGGACAACTGTTGACCCG-3', SEQ ID NO.-1) for the "protein Gal4, the binding sequence (5 '-TACTGTATGTA-CATACAGTA-3', SEQ ID NO: 2) for the LexA protein, the binding sequence (5 '-GAATTGTGAGCGCGCACAATTC-3', SEQ ID NO: 3) for the protein lac repressor I, the binding sequence (5 '- TCGAGTTTACCACTCCCTATCAGTGATAGAGAAAAGTGAAAG-3', SEQ ID NO: 4) for the tetracycline repressor protein and the binding sequence (5 '-TAATGATGGGCG-3', SEQ ID NO: 5 ) for the ZFHD-1 protein 17. - Nucleic acid construction according to one or more of claims 11 to 14, characterized in that the DNA binding sequence for a cellular regulatory protein [ax component]] is a DNA binding sequence. DNA chosen from the group that contains p53 protein, W4-1, NF-kappa B, E2F / DP or Myc / Max. nucleic acids according to claim 17, characterized in that the basal promoter [component a2)] is selected from the group containing the SV40 promoters, c-fos, sn U2 RNA and HSV-TK. 19. Nucleic acid construction according to claim 18, characterized in that the repressor [component b ')] is chosen from the group containing the lac repressor gene or the tetracycline repressor gene. 20. Nucleic acid construction according to claim 19, characterized in that the DNA binding sequence for the repressor [component c ')] contains at least one lac operator binding sequence or at least one tetracycline operator binding sequence. . 21.- Nucleic acid construction according to one or more of claims 1 to 20, characterized in that in the case of the effector gene (component d) it is a gene that codes for an active substance that is chosen from the group containing cytokines, chemokines , growth factors, receptors for cytokines, chemokines or growth factors, proteins with antiproliferative or cytostatic or apoptotic action, antibodies, fragments of antibodies, inhibitors of angiogenesis, peptide hormones, coagulation factors, coagulation inhibitors, proteins fibrinolytics, peptides or proteins that act on blood circulation, blood plasma proteins and antigens of infectious pathogens or of cells or tumors, determining the chosen antigen an immune reaction. 22. - Nucleic acid construction according to one or more of claims 1 to 20, characterized in that in the case of the effector gene it is a gene that codes for an enzyme that dissociates a precursor of a drug in a drug. 23. - Nucleic acid construction according to one or more of claims 1 to 20, characterized in that in the case of the effector gene it is a gene that encodes a ligand-active substance fusion protein or a ligand-enzyme fusion protein, The ligand is chosen from a group containing chytokines, growth factors, antibodies, antibody fragments, peptide hormones, mediators and cell adhesion molecules. 24. Nucleic acid construction according to claim 2, characterized in that components a), b), c) and d) are characterized as follows: Component a) • the promoter of the cdc25C gene (nucleic acids -290 to +121) Component b) • the nuclear localization signal (NLS) of (SV40 (SV40 long T, amino acids 126 to 132; PKKKRKV (SEQ ID NO: 13) • the transactivation domain (TAD) of VP16 acid of HSV-l (amino acids 406 to 488) • the RB binding sequence of the E2F-1 protein (amino acids 409 to 426 (LDYHFGLEEGEGIRDLFD) ( SEQ ID NO: 14) • the cDNA for DNA binding domains of the Gal4 protein (amino acids 1 to 147) - Component c) • '• 10 times the binding sequence for the DNA binding sequence of Gal4 with the sequence of nucleotides 5 '-CGGACAATGTTGACCG-3' (SEQ ID N0: 1) • the SV40 basal promoter (nucleic acids 48 to 5191) Component d) • the GCCACC sequence • the cDNA for the immunoglobulin signal peptide (nucleotide sequence 63 to 107) • the cDNA of the / 3-glucuronidase (nucleotide sequence 93 to 1982). 25. Nucleic acid construction according to claim 2, characterized in that components a), b), c) and d) are characterized as follows: Component a) • the promoter of the cdc25C gene (nucleic acids -290 to +121) Component b) • the SV40 nuclear localization signal (NLS) (SV40 long T, amino acids 126 to 132, PKKKRKV (S? Q ID NO: 13) • the HSV-1 VP16 acid transactivation domain (TAD) ( amino acids 406 to 488) • the E6 protein of the HPV-18 virus (nucleotide sequence 100 to 578) «the cDNA for the DNA binding domains of the Gal4 protein (amino acids 1 to 147) Component c) • 10 times the sequence of binding for the Gal4 DNA binding sequence with the 5'-nucleotide sequence -CGGACAATGTTGACCG-3 '(SEQ ID NO: l) • the SV40 basal promoter (nucleic acids 48 to 5191) Component d) • the GCCACC sequence • "the cDNA for the immunoglobulin signal peptide (nucleotide sequence 63 to 107) • e c / 3-glucuronidase cDNA (nucleotide sequence 93 to 1982) 26.- Nucleic acid construction according to one of the preceding claims, characterized in that it is DNA in the case of the nucleic acid. 27. - Nucleic acid construction according to one of the preceding claims, characterized in that the construction of nucleic acids is incorporated in a vector. 28. Nucleic acid construction according to claim 27, characterized in that it is a vector of the plasmid. 29. Nucleic acid construction according to claim 27, characterized in that it is a viral vector. 30.- Construction of nucleic acids according to rune or several of claims 1 to 29, characterized in that it is administered externally, perorally, intravesically, nasally, intrabronchially or in the gastrointestinal tract, or is injected into an organ, into a cavity of the body, in the musculature, subcutaneously or in the blood circulation, for the prophylaxis or therapy of a disease. 31.- Isolated cell, characterized in that it contains a nucleic acid construction according to one of claims 1 - 29. 32.- Use of a nucleic acid construct according to one or more of claims 1 to 29 or a cell according to claim 31 for the preparation of a therapeutic and curative agent for the treatment of a disease chosen from the group contains infections, tumors, leukemias, autoimmune diseases, allergies, arthritis, inflammations, organ rejections, transplantation reactions against host, diseases of blood coagulation, circulatory diseases, anemia, hormonal diseases and injuries of the central nervous system. 33".- Procedure for the preparation of nucleic acid constructions according to one or more of claims 1 to 30, wherein the different elements are linked together in a staggered manner 34.- Use of a cell according to the claim 31, for the preparation of a therapeutic and curative agent for the prophylaxis and therapy of diseases according to claim 32, characterized in that at least one cell is administered externally, intravesically, nasally, intrabronchially, orally or in the gastrointestinal tract or is injected into an organ, in a cavity the body, the musculature, subcutaneously or in the bloodstream.