DE10113513A1 - Drug combination of kinase inhibitor and neurotropic factor, useful for treating and/or preventing nerve and/or glial cell damage, e.g. associated with neurodegenerative diseases - Google Patents

Abstract

The present invention relates to a product containing at least one kinase inhibitor (A) which has little or no inhibitory activity on Raf and at least one neurotrophic factor (B) and / or at least one substance (C), which in is able to increase and / or cause the cellular formation and / or the release of one or more neurotrophic factors (B) as a combination preparation for use in the prophylaxis and / or therapy of nerve cell and / or glial cell damage.

Description

The present invention relates to a product containing at least a kinase inhibitor (A) that has little or no inhibitory activity on Raf, at least one neurotrophic factor (B) and / or at least a substance (C) which is capable of cellular formation and / or release increase one or more neurotrophic factors (B) and / or effect and which in addition to or instead of the neurotrophic factor (B) the kinase inhibitor (A) can be combined as a combination preparation for use in the prophylaxis and / or therapy of nerve and / or glial cells damage.

The degeneration of nerve cell processes and the death of neuronal ones Cells are an essential characteristic of acute and chronic neuro degenerative diseases. The question of how nerve cells die is in everyone Details not yet sufficiently understood. Likewise, not are the mechanisms for the maintenance of nerve cell process zen, especially of axons.

However, nerve cells are known for their survival, and for outgrowth and maintaining their nerve cell processes a trophic support need.  

It is also known that nerve cells, whether they are sensory or have motor function only in the presence of a number of so-called neurotrophic factors can survive. Withdrawal of this neurotrophic factor ren leads to increased expression of proapopto, at least in cell culture proteins that act on the table, such as, for example, p38, JNK and Fas-L (Le-Niculescu et al Mol Cell Biol 19 (1): 751-763, 1999).

The neurotrophic factors represent members of different families they own the neurotrophins, such as the Brain Derived Neu rotrophic factor (BDNF), neurotrophin-3 (NT-3), NT-4/5, NT-6 (Götz et al Na ture 372: 266-269, 1994) and the Nerve Growth Factor (NGF) (Kaplan and Miller, Curr. Opin. Neurobiol. 10: 381-291, 2000), further the Ciliary Neu rotrophic factor (CNTF), Stöckli et al., Nature 342: 920-923, 1989; J Cell Biol 115: 447-459, 1991), the leukemia inhibitory factor (LIF), the cardiotrophin-1 (CT-1) (Pennica et al., Neuron 17: 63-74, 1996), the insulin-like growth factor (IGF) (Hughes et al., J Neurosci Res 36 (6): 663-671, 1993; Arakawa et al J Neu rosci 10: 3507-3515, 1990), the insulin (Ryu et al. J Neurobiol 39 (4): 536-546, 1999), the hepatocyte growth factor (HGF) (Mama and Klein, Nat. Neurosci. 2: 213-217, 1999), the Gliacell Derived Neurotrophic Factor (GDNF) (Yan et al., Nature 373: 341-344, 1995; Oppenheim et al., Nature 373: 344-346, 1995; Hen derson et al., Science 266: 1062-1064, 1994; Lin et al., Science 260: 1130-1132, 1993; Balok et al., Curr. Opin. Neurobiol. 10: 103-110, 2000), the neurturin (Kotzbauer et al Nature 384: 467-470, 1996), the Artemin (Baloh et al., Neuron 21: 1291-302, 1998), the persephin (Milbrandt et al., Neuron 20: 245-253, 1998), the Midkine (Owada et al., J Neurochem 73 (5): 2084-2092, 1999) and a new one Member of the Transforming Growth Factor family, the GDF-15 (Kriegelstein and Unsicker WO 00/70051)

Such neurotrophic factors act by binding to and activating tyrosine kinase receptors. For example, tie

• - NGF with high affinity for the receptor trkA,
• - BDNF and NT-4/5 at trkB and
• - NT-3 to trkC (Glass and Yancopoulos, Trends Cell Biol 3: 262-268, 1993).
• - CNTF binds to a receptor complex consisting of three subunits, the Re zeptor-a, gp130 and the LIF receptor-b. Binds to this receptor complex also LIF and CT-1.
• - IGF, Insulin, Midkine as well as GDNF bind to their respective specific Receptors.

From these different receptors for neurotrophic factors, the Ak Activation signal via signal-transducing proteins present in the cytoplasm transferred into the cell nucleus. Several signal transmission paths are new here known rones. They include anti-apoptotic activation pathways, such as the PI-3K-AKT signal transmission path and the Ras-Raf- Transmission path. Both activation paths are networked by activated RAS, a protein that plays a key role in the Raf transmission pathway (Yuan and Yankner, Nature 407: 802-809, 2000). Additional Raf-dependent sig Signal transmission paths are the Bag-1-C-Raf signaling path (Wang et al., PNAS USA 93: 7063-7068, 1996) and the Rap-1-B-Raf-cAMP signaling pathway (Grewal et al., J. Biol. Chem. 275: 3722-3728, 2000).

However, they also include proapoptotic activation routes, such as the p38 or JNK transmission paths stimulated by ASK (Sedlacek Drug 59 (3): 435-476, 2000).

Withdrawal of these neurotrophic factors results in nerve cells, at least in the Cell culture, for the increased expression of proapoptotic proteins, such as p38, JNK and Fas-L (Le-Niculescu et al., Mol Cell Biol 19 (1):  751-763, 1999). To a limited extent, the withdrawal of a particular may be new rotrophen factor by adding another neurotrophic factor (Russel et al., J Neurobiol 36 (4): 455-467, 1998). Furthermore is known that the combination of different neurotrophic factors a have additive or synergistic anti-apoptotic effects on nerve cells can (Kobayashi and Matsuoka Neuroreport 11: 2541-2545, 2000).

The pattern of the receptors expressed by the different nerve cells determines the cell specificity of the different neurotrophic factors: how it works NGF neurotrophic on the peripheral sympathetic and on a subpopulation sensitive nerve cells, but has no effect on motor nerves cells. However, BDNF, NT-3 and NT-4 act on motor nerve cells (Griesbeck et al., J Neurosci Res 42: 21-33, 1995; Henderson et al., Nature 363: 266-270, 1993; Sendtner et al Nature 360: 757-758, 1992) as well as CNTF (Sendtner et al., 1990), LIF, CT-1, IGF, insulin, GDNF and neurturin. Darue In addition, BDNF induces neurite growth in cortical, pyramidal Neurons (McAllister et al., Neuron 17: 1057-1064, 1996; Neuron 18: 756-778, 1997) and Purkinje Neurons (Baptista et al., Neuron 12: 243-260, 1994).

The expression of receptors for neurotrophic factors is not on cells of the Nevensystems limited. For example, osteoblasts trk-A and trk- Form B and thereby NGF and autocrine and / or paracrine BDNF can be stimulated to proliferation (Mogi et al., Life Sci 67 (10): 1197- 1206, 2000). Pheochromocytoma cells (cell line PC12) react to NGF the formation of neurite-like processes, a phenomenon caused by Addition of inhibitors of cell proliferation, such as by Rapamy cin, ciclopirox or flavopiridol can be increased (Parker et al Neu ropharmacology 39 (10): 1913-1919, 2000). More orthotopes as well as ectopes Expression of receptors for neurotrophic factors on cells of different  Tissues as well as on tumor cells have been known for a long time (Sedlacek Drug 59 (3): 435-476, 2000).

A key result of the activation of neuronal and glial cells due to neurotrophic factors is the increased expression of intracellular Pro that protect the cells from controlled cell death (Newmeyer and Green, Neuron 21: 653-655, 1998; Wiese et al., Nature Neurosci. 2: 978-893, 1999). These proteins include the inhibitors of the IAP / ITA family, in particular their IAP-1, IAP-2, x-IAP and survivin. Inhibit IAP / ITA family proteins activation of Procaspase-9 (Deveraux et al., EMBO J. 17: 2215-2223, 1998), which in turn is activated by Cytochrome-C and Apaf 1. Furthermore proteins of the IAP / ITA family inhibit the function of the activated caspases-3.- 6 and -7 and can thereby also the apoptosis caused by these enzymes inhibit (Devereaux et al., Nature 388: 300-304, 1997; Roy et al., EMBO J. 16: 6914-6925, 1997). Transfection of IAP expression plasmids in isolated senso nervous or sympathetic nerve cells can cause cell death caused by withdrawal of Prevent NGF from being triggered. However, the effect of IAPs on the new rite growth much lower than that of NGF and other neurotrophs Factors (Wiese et al., Nature Neurosci 2: 978-893, 1999). Members of the IAP Family can thus convey a survival effect, but are only in small in accordance with the growth and maintenance of nerve processes involved. This indicates that NGF and other neurotrophic factors Sig switch on channels that are different from those of the IAP family.

Akt, a protein of the PI-3K-AKT signaling pathway, has three cellular I somere, of which Akt-3 is particularly expressed in neurons (Datta et al., Ge nes Dev. 13: 2905-2927, 1999). For the Raf signaling pathway in mammals cells, three different Raf proteins play a special role: C-Raf (also called Raf-1), A-Raf and B-Raf. Several of B-Raf exist through un different forms of splicing. In central nervous system cells  B-Raf and C-Raf are predominantly present and active, less so is A-Raf (Morice et al., Eur. J. Neurosci. 11: 1995-2006, 1999, Dugan et al., J Biol Chem 274: 25842-25848, 1999).

B-Raf and C-Raf are not only found in neurons, but also in glial cells (Mikaly et al., Brain Res. 27: 225-238, 1993; Mikaly and Rapp, Acta Histochem. 96: 155-164, 1994).

Although it is relatively clear how neurotrophins and / or membrane depolarization activate signal transduction pathways that ensure the survival of nerve cells, it is largely questionable which mechanisms lead to neurodegenerative diseases to cause dysfunction and neuronal death. Experimental studies (review by Yuan and Yankner, Nature 407: 802-809, 2000) provide clues that nerve cell death can be triggered by, for example

• - the relative activation of proapoptotic factors within the nerves cell, for example due to a lack of phosphorylation and inactivation of proapoptotic proteins (bath, but especially Bax) in the frame the AKT and / or Raf signaling pathways,
• - the lack of activation (e.g. as part of the Raf- Signal transduction path) of transcription factors, which the transcript tion of genes that activate anti-apoptotic proteins (e.g. Bcl-2, be but especially express Bcl-XL),
• - damage to mitochondria with the release of cytochrome C. and the activation of pro-apoptotic enzymes (caspases) for example due to abnormal protein structures or aggregates,  
• - by activating proapoptotic signal cascades (in particular ASK and thus p38 and JNK) by neurotrophins, for example via the Neurotrophin receptor p75 NTR,
• - by oxidative damage, NO overproduction or by enzymatic Malfunctions, e.g. B. by mutations in superoxide dismutase (SOD),
• - through synthesis of pro-apoptotic metabolic products, such as advanced glyco silation end products (AGEPs)
• - by up-regulation and activation of cyclin-dependent kinases (Gio vanni et al., J Biol Chemistry 274/27: 19011-19016, 1999), Park et al. Neuro biol.Aging 21: 771-781, 2000), which are crucial to the Regu cell division involved and / or
• - By pathological protein cleavage on physiological membranes, the z. B. lead to the production of β-amyloid, as well as by specific pathophysio logical processes, which change the folding and structure of Memb Ran proteins from an alpha to a beta structure, such as this. B. can be observed in prion diseases.

In view of these diverse possibilities for the death of nerve cells So far it has been extremely difficult to develop procedures to prevent or treat nerve cells. So far it is not possible the influence of neurotrophic growth factors on nerves to control cells in such a way that only antiapoptotic and not also proa poptotic signal transduction pathways can be activated. For example BDNF the neuronal damage caused by nitrogen monoxide drastically intensify (Ishikawa et al., J Neurochem 75 (2): 494-502, 2000). In clinical Examinations in patients with amyotrophic lateral sclerosis revealed the subcutaneous tissue ne administration of recombinantly produced human CNTF none clear therapeutic effectiveness, but such strong side effects,  that the dose could not be increased (CNTF Treatment Study Group, Neu rology 46 (5): 1244-1249, 1996) and the study had to be ended.

Another research approach attempted using Immunophi lin-binding substances (such as rapamycin, FK506, GPI-1046, Cyclosporin A and analogues of these substances) Damage to nerve cells to prevent or mitigate. The results in cell culture experiments with However, phaeochromocytoma (PC12), neuronal and animal cells were found contradictory (Parker et al., Neuropharmacology 39 (10): 1913-1919, 2000; Costantini and Isacson, Exp. Neurol. 164 (1): 60-70, 2000; Madsen et al., Exp.Neurol. 154 (2): 673-683, 1998; Gold Drug Metab Rev 31 (3): 649-663, 1999; Gold et al., Exp Neurol 147 (2): 269-278, 1997; Steiner et al., PNAS USA, 94 (5): 2019-2024, 1997). Those groups of investigators, what effectiveness in their Models found this effectiveness to be due to inhibition cell division (Parker et al., Neuropharmacology 39 (10): 1913-1919, 2000), a Impairment of the corticoid receptor and Hsp-90 (Gold et al., Drug Metab Rev 31 (3): 649-663, 1999), an inhibition of calcineurin (Marioka et al., Prog Neurobiol 58 (1): 1-30, 1999, an increased expression of the neuronal, "growth associated "factors (GAP 43, Madsen Exp. Neurol 154 (2): 673-683, 1998), a Inhibition of protein phosphatases and / or stimulation of protein kinase C. (Audesirk et al., Brain Res Dev Brain Res 102 (2): 247-260, 1997.

For some substances, indications of a therapy could be found in animal models effect can be found (Parker et al., Neuropharmacology 39 (10): 1913-1919, 2000, Madsen et al., Exp Neurol 154 (2): 673-683, 1998, Gold et al., Exp Neurol 147 (2): 269-278, 1997, Steiner et al PNAS USA 94 (5): 2019-2024, 1997). In contrast to this, after clinical administration of Im munphilin-binding drugs, such as cyclosporin A, rapamycin or FK506 for immunosuppression no neuroprotective effects on the treated patients.  

It could also be shown that inhibitors of cell proliferation, such as for example those that inhibit cyclin-dependent kinases (such as as Flavopiridol, Roscovitin or Olomoucin (Park et al., J Biol Chemistry 271: 21898-21905, 1996; Maas et al., J Neurochem 70 (4): 1401-1410, 1998) or also different inhibitors of cell proliferation (such as chloro phenylthio-CAMP, N-acetylcysteine, Deferoxamine, Ciclopirox or Mimosin, Park et al., 17: 1256-1270, 1997), or selected immunophilin binding substances (such as Rapamycin, Parker et al, Neuropharmacology 39 (10): 1913- 1919, 2000), are capable of pheochromocytoma (PC12) or nerve cells Protect in cell culture from the influence of cell-damaging substances.

Inhibitors of apoptotic proteins, especially caspases (Park et al., J. Neurosccience 18: 830-840, 1998) can indeed the rapid apoptosis of ge Prevent damaged nerve cells, but not the delayed ones Cell death.

Substances used in these experiments to target nerve cells Damage and lead to their apoptosis include the cytostatics Camptothecin, cytosine arabinoside, etoposide, teniposide or mitoxanthrone (Park et al., J Neuroscience 17: 1256-1270, 1997), or the proapoptotic B-amyloid Protein (Giovanni et al., J Biol Chemistry 274/27: 19011-19016, 1999, Park et al., Neurobiol Aging 21, 771-781, 2000). There are indications that the a Poptotic effect of these substances is essentially due to the fact that they are in Nerve cells for an increased expression of cyclin-dependent kinases (CDKs) and proapoptotic signaling proteins such as p38 and JNK (Court ney and Coffey, Eur J Neurosci 11 (3): 1073-1084, 1999).  

As a mechanism for the neuronal protective effect of inhibitors of the Cell proliferation has been postulated to increase the activity of CDKs and / or of signal transduction kinases (pro gently, for example due to substances damaging the nerve cells) can reduce different ways. For example, suppose that the nerve cell protective effect of inhibitors of cell proliferation like flavopiridol for direct inhibition of CDKs (especially cdk4 and cdk6). This inhibition consequently causes an inhibition of the Phosphorylation of pRB and P107 and thus an inhibition of transcription activity of E2F / Dp (Sedlacek, Critic. Rev. Onc./Heam, 37 (2001).

This is contradicted by the fact that for butyrolactone-1 (a relatively highly specific CDK- Inhibitor) no protective effect on nerve cells can be demonstrated could (Maas et al., J Neurochem 70 (4): 1401-1410, 1998) and staurosporins (a broad-acting kinase inhibitor) induced apoptosis in nerve cells (Thomas and Mayle Brain Res 884 (1-2): 163-173, 2000).

On the other hand, cycloheximide appears to inhibit protein synthesis by A poptose of nerve cells after treatment with Ara-C (and the ver increased expression of proapoptotic proteins such as p38 and JNK) to reduce ren.

The clinical application of the previously known antiproliferative and nerves cell cultures under the respective experimental protective conditions Punching is significantly restricted by its toxicity as well due to the risk that these substances also in the presence of neurotrophic Substances can damage nerve cells.  

So some inhibitors of the cyclin dependent kinases were chosen because of their tiproliferative efficacy already tested on tumor patients. In these studies there was strong dose-limiting toxicity, for example for flavopiridol watery diarrhea, and after their successful pharmacological treatment lung, severe hypotension (Senderowicz et al., J Clin Oncol 16: 2986-2999, 1998).

Treatment with kinase inhibitors also carries the risk that in Nerve cells are not just proaptotic kinases (such as those on cyclin-dependent kinases involved in cell division [CDK]) are inhibited, but also anti-apoptotic kinases, for example signal transduction kinases such as B-Raf (Wiese et al., Nature Neurobiol 4: 137-142, 2001) or but those such as PKC, the inhibition of which reduces the Function and axon growth of nerve cells (Audesirk et al., Brain Res Dev Brain Res 102 (2): 247-260, 1997). The inhibition of such, the Ki survival and the function of neurons supporting kinases nose inhibitors, should rather increase the damage to nerve cells. So is of staurosporine (an inhibitor of a number of kinases including from CDKs) is known to increase apoptosis of nerve cells (Thomas and Mayle Brain Res 884 (1-2): 163-173, 2000; Lankiewicz et al., J Biol Chem 275 (22): 17064-17071, 2000; Choi et al., J Neurochem 74 (4): 1621-1626, 2000). Wortmannin, a kinase inhibitor with special activity on the phosphatidyli nositol 3-kinase, inhibits the antiapoptotic activity of IGF-1 (Yamaguchi et al., J Biol Chem Oct 27, 2000). Flavopiridol not only inhibits CDKs, but also receptor-associated kinases such as PKC are also very effective (Sedlacek et al., Int J Oncol 9: 1143-1168, 1996; Sedlacek, Crit Rev Oncol Hematol 2000). Through the Inhibition of such anti-apoptotic kinases could be the administration of Fla vopiridol involve the risk of nerve cell damage. In this To context it must be taken into account that an isolated inhibition of the PKC Can inhibit neurite growth and possibly also become trophic  Disorders in the nerve cell processes occurs, which are the neurodegenerative ones Reinforce changes.

For the therapeutic effect of kinase inhibitors as well as for any other, possibly nerve cell protective substance, is therefore significant in what extent they neurite growth in clinically relevant target cells promotional and / or inhibiting mechanisms can influence whether for example, is able to control the axon growth of motor neurons, sensory Neurons, dopaminergic neurons, Purkinje cells, cholinergic projection new promote or inhibit rons and retinal ganglion cells.

A combination of flavopiridol with low concentrations of NGF results on pheochromocytoma cells (such as the PC 12 cells) to an outgrowth of Neurite-like processes (Parker et al., Neuropharmacoly 39 (10): 1913- 1919, 2000). However, this effect was not specific for flavopiridol as Kina se inhibitor, he could also use other cell proliferation inhibitors ration, such as rapamycin or ciclopirox, are not ever seen but with such immunophilin-binding substances as FK506 or GPI 1046, which are not inhibitors of cell proliferation. Furthermore was this effect is not transferable to nerve cells. So FK506 showed (as well Rapamycin) has a neuroprotective effect on cerebral ischemia in the Rat, but not GPI-1046.

Surprisingly, it has now been found that the treatment of nerve cells or of glial cells with a combination of kinase inhibitors that have no or cause only a slight inhibition of the B-Raf kinase (kinase inhibitor (A)) Substances that themselves have a neurotrophic effect (neurotrophic Factor (B)) and / or with substances that cause formation and / or release  of neurotrophic factors (B) and / or cause (substance C) that Survival rate of nerve cells and that of sensory and as well motor neurons significantly increased, their resistance to nerves cell-damaging substances increase the specific axon growth of these ner vein cells increased, nerve cell function improved and / or death rate of nerve cells after exposure to substances that damage nerve cells decreases.

The protective effect of this combination of kinase inhibitor (A) and / or new rotrophem factor (B) and / or substance (C) on nerve cells is significantly stronger than the effectiveness of the individual substances. Especially in the presence of an en cymatically functional B-Rafs and above all with an additionally elevated one Expression members of the IAP gene family. An increase in expression follows signal paths mediated via PI-3K, AKT and NFkB. So the Kombinati on of kinase inhibitor (A) and neurotrophic factor (B) little or none Effect on nerve cells that do not express B-Raf but C-Raf.

The present invention therefore relates to a product comprising at least one kinase inhibitor (A) which has little or no inhibitory activity on Raf and

• a) at least one neurotrophic factor (B) and / or
• b) at least one substance (C) which is capable of cellular formation and / or the release of one or more neurotrophic factors (B) reinforce and / or effect

as a combination preparation in the form of a mixture or as a single compo nent for simultaneous or different use at under different or the same places in the prophylaxis and / or therapy of Ner venzell and / or glial cell damage.  

Basically, it is not essential for the effect of the product according to the invention irrelevant whether the product is the same as the nerve and / or glial cells or in different places, as individual components or as a mixture, the same in time or at different times, preferably within 6 hours, particularly preferably within 12 hours, in particular within 24 Hours, especially within 48 hours, for example simultaneously with egg ner mixture of kinase inhibitor (A) with neurotrophic factor (B) and / or Substance (C), or first with kinase inhibitor (A) and then with new rotrophem factor (B) and / or substance (C) or first with neurotrophic factor (B) and / or substance (C) and subsequently in contact with kinase inhibitor (A) brought.

Preferably, the pretreatment with kinase inhibitor (A) or the same early treatment with kinase inhibitor (A) and / or neurotrophic factor (B) and / or substance (C) carried out.

Preferred for the effect and the choice of administration is that the first Substance in the nerve cell to be treated is still effective when the second Substance is administered. This is usually the case if the distance between preferential administration of the first substance and the second substance wise up to 6 hours, particularly preferably up to 12 hours, in particular up to to 24 hours, especially up to 48 hours.

The contacting can be done, for example, by adding the substances to the Culture medium of a nerve cell or glial cell culture, or by local or systemic administration of the substances into an organism.

Local administration in the sense of this invention can, for example, in a Wound cavity, on the skin, in a body opening, in a body cavity, as in for example in the chest or abdominal cavity, or in an organ, especially in that  Brain, spinal canal, joint, connective or muscle weave. Systemic administration can be oral, rectal or in the Circulation, for example intravenously or intraarterially.

The effectiveness of the method according to the invention can be seen from the Demonstrate the function of nerve cells.

The function of nerve cells, for example, stimulation conduction and all involved biochemical and / or electrochemical processes the. Furthermore, the function of nerve cells encompasses this morphologically tangible growth of neurites or axons and of dendrites. In particular the term function of nerve cells encompasses the survival of nerve cells.

The death of nerve cells can be caused, for example, by in vitro Test procedures with which apoptosis can be detected are observed the. Such test methods are, for example, "tunnel assay" (Gavrielli et al., J. Cell Biol., 119: 493-501, 1992; Gold et al., Lab. Invest. 71: 219-225, 1994), Chromatin fragmentation (Götz et al., Hum. Mol. Genet. 9: 2479-2489, 2000), Enumeration of surviving and dying nerve cells (Arakawa et al., J. Neurosci. 10, 3507-3515, 1990), the use of test substances for quantification detection of cell death in cell culture (Uliasz and Hewett, J. Neurosci. Methods 100, 157-163, 2000), quantification of the expression of cell death-associated genes in Nerve cells such as cyclins (Timsit et al., Eur. J. Neurosci. 11, 263-278, 1999) and determination of neuronal cell death after addition of Aß (Iwasaki et al., Mol. Psych. 1, 65-71, 1996) or after induction of oxidative stress (Manev et al., Exp. Neurol. 133, 198-206, 1995).  

Cells of the nervous system in the sense of the present invention are glial cells or neuronal cells, for example sensory and sympathetic neurons Cells, motor neuronal cells, cholinergic neurons of the basal forebrain, dopaminergic nerve cells of the midbrain (substantia nigra), granule cells and Purkinje cells of the cerebellum and hippocampus, retinal ganglion cells and photoreceptors as well as neural stem cells.

In a preferred embodiment, the kinase inhibitors (A) are included little or no inhibitory activity on B-Raf kinase inhibitory derivatives of 4H-1-benzopyran. Examples of such derivatives are in the patents EP 0137193 and EP 366061 in detail and the structural The relationship between these derivatives has been described by Sedlacek et al., Int J Oncol 9: 1143-1168, 1996). The example in these patents derivatives of flavopiridol and the flavopiridol itself (Sedla cek et al., Int J Oncol 9: 1143-1168, 1996) are expressly the subject of these Invention.

In a further embodiment, the kinase inhibitors (A) are flavopiridol derivatives, represented by Kim et al., J Med Chem 43 (22): 4126-4134, 2000, esp special thio-flavopiridol and oxy-flavopiridol. Furthermore, are examples of Kinase inhibitors (A) the 2- (2-amino-) shown by Ebendal (WO 00/50030) 3-methoxyphenyl) -4-oxo-4H- (1) benzopyran.

In a preferred embodiment, the kinase inhibitors (A)

• - 7,12-dihydro-indolo (3,2-d) (1) benzazepin-6 (5H) -one (NSC 664704 Saus ville et al., Pharmacol. Ther. 82 (2-3): 285-292, 1999),
• - Kinase inhibitors, such as 7OH staurosporins and their kinase inhibiting derivatives, butyrolactones, Roscovitine, Purvalanol A, Emodin, Anilinoquin azolines (PD-168393 and PD 169414) and phenylaminopyrimidines  (STI 571, CGP 78850, CP 358774, CP59326 and CGP 60474), Trioylimidazole (1-779450), as a whole by Meijer et al., Parmacol. Ther. 82 (2-3): 297-284, 1999 and by Sedlacek et al., Int J Oncol 9: 1143-1168, 1996 or by Sedlacek, Drug 59 (3): 435-476, 2000),
• Paullone (Zaharevitz et al., Cancer Res. 59, 2566-2569, 1999),
• - SB203580 [4- (4-fluorophenyl) -2- (4-methylsulfinylphenyl) -5- (4-pyridyl) 1H- imidazole, (Ishikawa et al., J Neurochem 75 (2): 494-502, 2000, Harada and Sugimoto, Jpn J Pharmacol 79 (3) 369-378, 1999)],
• 1,4-diamino-2,3-dicyano-1,4-bis (2aminophenylthio) butadiene [U0126, (Favata et al., J BIOL Chem 273 (29): 18623-18632, 1998; DeSilva et al., J Immunol 160: 4175-4181, 1998)],
• - 2-2'-amino-3'-methoxyphenyl (-oxanaphtalen-4-one) [PD098059, (Dudley et al., PNAS USA 92: 7686-7686, 1995)],
• - 2- (2-chloro-4-iodo-phenylamino) -N-cyclopropylmethoxy-3,4-difluoro-benzamide [PD184352, (Sebolt-Leopold et al., Nature Med 5: 810-816, 1999)],
• Bis-ethylthiomethyl, [CEP-1347 (KT7515) (Maroney et al., J Neurochem 73 (5): 1901-1912, 1999)], analog of indobarbazole [(K252a) (Saporito et al., J Pharmacol Exr Ther 288: 421-427, 1999)]
• Tetrapyrrolic macrocycles such as 5, 10, 15, 20 Tetraarylporphyrin and 5,10,15-triarylcorrol (Aviezer et al., WO 00/27379) and or
• - Pyrimidone derivatives, which inhibit the tau protein kinase and by Ando et al., WO 00/18758 have been presented in detail.

For the protection and / or therapy of nerve cell damage, the kinase inhibitor (A) is preferably administered in a single dose, particularly preferably in a multiple dose, which are less than the maximum tolerable dose (MTD) of the respective substance for humans. A dose is preferably chosen which is half the MTD. For example, for the infusion of flavopiridol, the MTD on the tumor patient is 50 mg / m ² / dx ³ (Sen derowicz et al., J Clin Oncol 16 (9): 2986-2999, 1998). For use in humans, flavopiridol should therefore preferably be administered in a dose of 0.1-50 mg / m ² , particularly preferably 5-30 mg / m ² , in particular 25 mg / m ² . The daily dose can be administered once a day or divided into several portions over the day at approximately the same time intervals.

The kinase inhibitor (A) is preferably administered either locally or systemically, for example only on one day or for several days a day or every second or third day for several weeks until the therapeuti effect becomes visible. A treatment of two to 25 is preferred Weeks in combination with the neurotrophic factor (B) and / or with Substance (C) selected.

In a preferred embodiment, neurotrophic factors (B) are exemplary

• The neurotrophins BDNF and variants of BDNF (Barde et al., US 5438121, Bailey et al., WO 91/03568), NT-3, NT-4/5, NT-6 or variants the NT and NGF, such as the NNT-1 (Chang, US 5,741,772),
• - CNTF and variants of the CNTF (DiMarco and Laufer WO 98/01149),
• - LIF, CT-1, IGF-1, insulin, midkine, HGF, GDNF, artemin, persephin, GDF 15 or neurturin.

These neurotrophic factors (B) are according to the state of the art preferably produced using recombinant DNA technology. virtue the neurotrophic factors (B) are administered locally or systemically, in  a dose which is below the MTD, particularly preferably about half the MTD is. For example, CNTF lies after subcutaneous administration MTD at about 30 mg / kg body weight every other day for 9 months. Preferably, for example, CNTF or CT-1 or CLC / NNT-1 in one Dose of 0.1-30 mg / kg, particularly preferably 2-25 mg / kg, further be preferably administered from 5-20 mg / kg, in particular from 15 mg / kg. The disposition The daily dose for the neurotrophic factor (B) is determined once or in Portions at roughly equal intervals throughout the day. The new rotrophic factor (B) can be used once or daily or every other, third, four th, fifth, or sixth day or once a week. The Similar to the kinase inhibitor (A), the duration of treatment depends on the clinical effect. A treatment period of 2 weeks is preferred up to 25 weeks in combination with a kinase inhibitor (A).

Preferably, the neurotrophic factor (B), for example in the form of a right protein produced in combination, by means of suitable drug pumping systems in the subarachnoid space (as described by Dittrich et al., Exp Neurol 141: 225-239, 1996) into an organ or tissue, such as the brain or into the muscles, or systemically, for example into the blood vessel system enough.

In a further embodiment, the neurotrophic factor (B) is not as Protein but as a nucleotide sequence which is responsible for the neurotrophic factor coded. Such nucleotide sequences are known to those skilled in the art Methods either in plasmids or in viral vectors, for example adenovi rale vectors (e.g., described by Gravel et al., Nature Medicine 3: 765-770, 1997; Cayouette et al. J Neurosci 18: 9282-9293, 1998) or retroviral vectors or vectors derived from other viruses (e.g. AAV, or HIV), for example lentiviruses, in particular of those viruses which have a Tropism for nerve cells (e.g. HPV) or muscle cells inserted.  

Plasmids or viral vectors are used with suitable tools, for example Protease inhibitors, detergents, buffers, and / or transfection reagents, such as z. B. mixed cationic lipids or cationic polymers and the organ must complement the neurotrophic proteins or instead of the neurotrophic ones Proteins administered.

In a preferred embodiment, substances (C) are all substances which stimulate the intracellular formation and / or the release of neurotrophic factors. These substances include, for example

• - Cytokines and Interleukins (IL), in particular also those that have Macropha stimulate genes, lymphocytes, nerve cells and / or glial cells as in for example IL-1, IL-2, IL-3, IL-4, IL-6, TNF-alpha, interferon (IFN) -alpha, beta, and gamma, GM-CSF, M-CSF, (Hughes et al., Brain Res Mol Brain Res 53: 138-151, 1998; Laurenzi et al., Eur J Biochem 223: 733-744, 1994),
• Melanocortin and analogues thereof, for example Org 2766 (Dyer et al., Peptides 16: 515-522, 1995),
• Toluene (Gotohda et al., Horm Metab Res 32: 301-305, 2000),
• D1 / D2 dopamine agonists, for example apomorphins (Ohta et al., BBRC 272: 18-22, 2000),
• Beta-adrenergic agonists, for example isoproteronol (Barouch et al., J Neuroimmunol 103: 112-121, 2000),
• Neurotransmitters, for example substance P (Barouch et al., J Neuroimmunol 103: 112-121, 2000),
• Catechol and catechol derivatives, for example 4-methylcatechol (Nitta et al., J Pharmacol Exp Ther 291: 1276-1283, 1999),
• Diterpenoids, for example scabronin-A and scabronin-B (Obara et al., Eur J Pharmacol 370: 79-84, 1999),  
• Forskolin and forskolin derivatives, for example NKH 477 (Morinobu et al., J Neurochem 72: 2198-2205, 1999),
• - Agonists for cholinergic receptors, for example nicotine, carbachol and Pilocarpine (French et al., Brain Res Mol Brain Res 67: 124-136, 1999),
• - Dihydro-caffeic acid amide derivatives, (Fukazawa et al., WO 92/02490),
• - Optical isomers of 2-methyl-spiro (1,3-oxathiolan-5,3) -quinuclidine, (Fisher et al., EP 303391),
• Quinoline acid (Rocamora et al., Brain Res Mol Brain Res 26: 89-98, 1994),
• Agonists for the glutamate receptor, for example quisqualate, kainate, Alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) or N- methyl-D-aspartate (NMDA) (Bessho et al., Brain Res Mol Brain Res 18: 201- 208, 1993),
• Agonists for AMPA, for example described by Henrich-Noack et al., DE 195 13 630,
• - 2,6-diphenyl substituted pyridine derivatives, described for example by Hanabusa et al., WO 94/27604,
• Ribavirins and ribavirin derivatives, for example 1- (beta-D-ribofuranosyl) 1H-1,2,4-triazole (Gage et al., WO 00/30656),
• Retinoids, for example retinoic acid (Wion et al., BBRC 149: 510-514, 1987)
• Biguanide derivatives, for example metformin,
• - Sulfonyl-urea derivatives, for example glibenclamide, glimepiride, glic lacid, glibornuride, gliquidon, glisoxide pioglitazone, rosiglitazone maelate and / or repaglinide
• Tolbutamide and / or
• - Aldose reductase inhibitors, for example described by Longo and Mizisin, WO 98/42324,

Substance (C) is preferably administered in a dose which is below the Maximum tolerable dose (MTD), for example for humans. Beson more preferably a dose in the range of 5% -90% of the MTD, furthermore preferably chosen from 10% -60%, in particular from 50% of the MTD.

One or more different kinases are preferred in each case. Inhibitor (s) (A) and neurotrophic factor (s) (B) and / or substance (s) (C) administered.

The product containing the kinase inhibitor (A) and the neurotrophic factor (B) and / or the substance (C) in the event of functional disorders administered by cells of the central or peripheral nervous system, for example wise in cerebral ischemia (stroke), amyeotrophic lateral sclerosis (ALS), Alzheimer's disease, nerve lesions, multiple sclerosis, Par's disease kinson, diabetic neuropathy, spinal muscular atrophy and / or prione diseases such as B. Creutzfeld-Jakob disease.

A preferred embodiment is a method for producing an invent Product according to the invention, in which at least one kinase inhibitor (A) at least one neurotrophic factor (B) and / or at least one substance (C) is appropriately combined.

Another embodiment is the use of an Er according to the invention Certificate for the prophylaxis and / or therapy of nerve and / or glial cells damage.

A preferred embodiment is the use of an inventive Product in which at least one kinase inhibitor (A) with at least one  neurotrophic factor (B) and / or at least one substance (C) with a nerve cell len and / or glial cells are brought into contact.

Another embodiment is a method by which an inventive Product is brought into contact with nerve cells and / or glial cells in vitro.

example 1 Isolation and cultivation of neurons

Parents b-raf (+/-) heterozygous or c-raf (+/-) heterozygous were rejected crosses (Wojnowski et al., Mech. Dev. 76: 141-149, 1998; Nature Genet. 16: 293- 297, 1997). Of embryos aged 12.5 days that are homozygous for b-raf (- / -) or c-raf (- / -) were spinal motor neurons using the Panningtech nik (Metzger et al., J. Neurosci. 18: 1735-1742, 1998) using a monoclonal rat anti p75 antibody (Chemicon, Hofheim, Germany) isolated. For this purpose, the ventrolateral parts of the lumbar spinal cord were measured mechanically crushed, in hepespuffer solution (containing 10 µM 2- Mercaptoethanol) transferred and incubated with trypsin (0.05%, 10 min). The one Cell suspension in the supernatant was loaded with the anti p75 antibody layered culture dish and transferred at room temperature for 30 min. incubated.

The individual culture dishes were then washed, followed by the adhering cells from the culture plate containing 0.8% saline 35 mM KCl and 1 µM 2-mercaptoethanol detached.

The cells obtained in this way were sown at a density of 2000 cells / cm ² in culture plates (Greiner, Nuertingen, Germany), which had been coated with polyornithine and laminin before. The cells were kept at 37 ° C. in neurobasal medium (Life Technologies, supplemented with B27 supplement, 10% horse serum, 500 μM glutamax and 50 μg / ml apotransferrin) and in a 5% CO ₂ atmosphere. 50% of the cell culture medium were kept on Day 1 and replaced every other day thereafter.

The analysis of the mRNA for IAP-1, IAP-2, x-IAP and t-IAP (survivin) was carried out using RT-PCR. RNA was isolated using a Trizol reagent (Life Technologies, Karlsruhe) and 10 ng total RNA was used for an RT-PCR reaction. The primer sequences for amplifying IAP-1, IAP-2, x-IAP and t-IAP (survivin) were as follows:
IAP-1f: 5'-TACTACATAGGACCTGGAGA-3 ',
IAP-1r: 5'-CCCACCATCACAGCAAAA-3 ', annealing 55 ° C;
IAP-2f: 5'-GGAGAAGAAAATGCTGACCC-3 ',
IAP-2r: 5'-GCTTGTAAGGGTATCTGTGT-3 'annealing 55 ° C;
x-IAPf: 5'-TGCAAGAGCTGGATTTTATG-3 ',
x-IAPr: 5'-CCCGATCTGGCAGCTGTACC-3 'annealing 55 ° C;
tIAP (survivin) f: 5'-CCA GAT CTG GCA GCT GTA CC-3 ',
tIAP (survivin) r: 5'-GCC AGC TGC TCA ATT GAC TG-3 ', annealing 64 ° C.

A part of the β-actin mRNA was used as a control for the integrity of the RNA amplified following primers: β-actinf: 5'-GTGGGCCGCCCTAGGCACCAG- 3 ', β-actinr: 5'-CTCTTTAATGTCACGCACGATTTC-3', annealing 64 ° C. The RT-PCR was performed according to the manufacturer's protocol with random Hexamer primers performed. The PCR amplification was carried out as follows leads: 94 ° C, 30 sec, indicated annealing temperatures, 1 min. 72 ° C, 1 min. IAP-1 and t-IAP (survivin) were for 33 and 35 cycles, IAP-2 and x-IAP for 28 and 30 cycles and β-actin for 26 and 28 cycles. RT-PCR on RNA from E12.5- Brains from b-raf and c-raf +/- mating showed a significant decrease  by an average of 60% and 55% for IAP-1 for b-raf and c-raf - / - Emb ryons compared to the wild-type control, of 52% for IAP-2 at b-raf - / - and 46% for x-IAP in b-raf - / - embryos compared to the wild-type control. The Expression differences of IAP-2 and x-IAP in c-raf - / -, as well as t-IAP in b-raf - / - and c-raf - / - were not statistically significantly different in comparison for wild type control.

Embryos 12.5 days old that were homozygous for b-raf (- / -) or c-raf (- / -), sensory neurons were further isolated. For this purpose dorsal root ganglia isolated, in PBS and with trypsin (0.05% in hepespuffer) for 30 min. incubated. Trypsin digestion was accomplished by adding L15- Medium containing 10% horse serum was stopped and subsequently the cells in Culture plates plated for 3-4 hours. Cells in the supernatant were centrifuged giert (10 min. 400 g) and the cell sediment just like for spinal moto neurons described held in the neurobasal medium.

Example 2 Isolation and cultivation of neural stem cells

Neural stem cells were isolated from the brain of normal, b-raf (- / -) or c-raf (- / -) deficient mouse embryos. The area of the forebrain was removed under a dissecting microscope, and in more developed embryos the area of the hippocampus and the periventricular zone. These brain areas were then transferred to 200 μl HBSS (Hanks balanced salt solution (HBSS, Life Technologies (Karlsruhe), incubated with 0.1% trypsin (final concentration in HBSS) for 10 min at 37 ° C., the reaction with 0, 1% trypsin inhibitor (trypsin inhibitor from egg yolk sack (Sigma, Deisenhofen), stock solution: I% in HBSS / 25 mM HEPES) final concentration in HBSS was stopped, then the cells were triturated 10 times with a 200 µl pipette and placed in medium [ (Neurobasal medium (Life Technologies), B27 Supplement (Life Technologies Stock 50x, EK 1x) Glutamax II (Life Technologies Stock 100x, EK 1x), basicFGF (20 ng / ml), EGF (20 ng / ml) 1] in transferred to a volume of 5 ml, the dissociated cells were cultivated in Sarstedt dishes (50 ml) (incubator, 37 ° C., 5% CO ₂ moisture-saturated atmosphere), the medium was changed every two days and the cells grew as embroid bodies and Do not attach, so the cells were transferred to a Falcon tube to change the medium d Centrifuged at 400 g for 5 min. The supernatant was aspirated and the cell sediment was triturated and taken up in fresh medium. After 3 passages at the latest, large embroid bodies were formed, which could be trypsinized (see above) and in low cell density (max. 10,000 cells / plate) on 10 cm dishes (Sarstedt). Individual cells were then picked and first expanded into 96-well plates, later into 24-well and 12-well plates. These single cell clones of neural stem cells could then be examined for their differentiation capacity and then used in test procedures.

To get reproducible results, the cells were also called lines established and frozen and stored for later experiments.

The freezing of the neural stem cells was carried out according to the standard protocol, ie after centrifugation, the cells were taken up in medium with 10% DMSO and first cooled to -86 C at 1 ° C./min (in MrFrosti) and then in liquid N ₂ at -186 ° C to be stored.

Example 3 Effect of neurotrophic factors in combination with a kinase inhibitor on neurons

The following are added:

• - to motor neurons as neurotrophic factors GDNF, BDNF and CNTF (each 0.1, 0.3, 0.5, or 1 ng / ml) and
  to sensory neurons NGF (0.1; 0.3; 0.5; or 1 ng / ml), either individually or in combination with the kinase inhibitor flavopiridol (concentration of 0.1; 0.3; 0, 5 µM; structure, molecular weight of flavopiridol see Sedlacek et al., Int J Oncol 9: 1143-1168, 1996; Sedlacek, Critic Rev Oncol Hematol 2000)
• - or flavopiridol in combination with pilocarpine (concentration of 0.01; 0.1 and 0.3 µM).

In another control group, only flavopiridol becomes the neuronal cells added in each case in the stated concentrations.

In control cultures of normal embryonic neurons survive after 3 days ne addition of the respective neurotrophic growth factors only approx. 10-25% of the Cells, depending on the concentration with the addition of the respective growth factors however, about 30-70% of the originally seeded neuronal cells.

The addition of flavopiridol causes a slight increase in survival (Depending on the concentration, 20-40% of the cells survive). The biggest increase The survival rate can be when combining growth factors, flavopiri dol and pilocarpine or when combining flavopiridol with pilocarpine will be seen (depending on the concentration, between 40-90% of the cells survive). The combinations of neurotrophic growth factor (neurotrophic factor (B)), flavopiridol (kinase inhibitor (A)) and pilocarpine (substance (C)) or Fla vopiridol (kinase inhibitor (A)) and pilocarpine (substance (C)) have their strongest synergistic effect at low concentrations of the growth factors or  at low concentrations of flavopiridol (kinase inhibitor (A)) and high Concentrations of the other substance.

During neural cell cultures of c-raf (- / -), c-raf (+/-) or b-raf (+/-) defi In this regard, embryos do not differ from normal cultures Have embryos, no effect of neurotrophic factors alone on the Survival of motor neurons or sensory neurons from b-raf (- / -) defi specific embryos are detected. With b-raf (- / -) deficient neurons the survival rate after 3 days of cell culture with or without neurotrophic Factors (B) for values that are less than 3% of the seeded neurons. The Combination of neurotrophic growth factors with flavopiridol resulted here with only a slight increase in survival.

This experiment can prove that the product according to the invention with Ver administration of a kinase inhibitor (A), the Raf, in particular B-Raf not or only slightly inhibited, in combination with a neurotrophic factor (B) survival of neuronal cells and promotes them from damaging influences protects.

Example 4 Effect of the method on b-raf (- / -) cells in which b-raf was transfected

As further evidence of the dependence of the effectiveness of the invention Products made from an enzymatically active B-Raf protein become sensory neuronal b-raf (- / -) cells with a plasmid (pCDNA3) that is open reading frame of the B-Raf gene contains (Wojnowski et al. Mech. Dev. 91: 97-104 (2000) or with a LacZ expression plasmid with that of Wiese et al., (Nature Neu rosci. 2: 987-983, 1999) and the survival of the method so neurons transfected depending on the addition of the kinase inhibitor  (A) (flavopiridol) and the neurotrophic factor (B) or the combination of Kinase inhibitor (A) and neurotrophic factor (B) (in concentrations as above and the combination of both substances with pilocarpine (substance (C)) determined in the cell culture.

While non-transfected and transfected with the LacZ expression plasmid b-raf (- / -) deficient neurons in cell culture with and without a neurotrophic factor (B) or with and without kinase inhibitor (A), flavopiridol, or the combination of kinase inhibitor (A) and neurotrophic factor (B) to a considerable extent die, b-raf (- / -) deficient neurons with the b-raf gene turn out to be contained plasmid were transfected as survivable. Immunfluoreszen Examinations on surviving b-raf (- / -) deficient neuronal cells that had been transfected with a plasmid containing the b-raf gene with an antibody specific for the B-Raf protein (Sithanandam et al., On cogene 5: 1775-1780, 1990) that these cells contained the B-Raf protein. The Treatment of these transfected cells with the combination of kinase inhibitor (A) and neurotrophic factor (B) leads to an equally high survival rate as the treatment of normal neuronal cells with the same combination of kinase inhibitor (A) and neurotrophic factor (B) and / or substance (C).

These studies show that the effectiveness of the Er testimony is dependent on an enzymatically functional B-Rafund that treatment with the product according to the invention, containing at least a kinase inhibitor (A) which does not or only slightly inhibits B-Raf, and min at least one neurotrophic factor (B) and / or at least one substance (C)), promotes the survival of nerve and / or glial cells and consequently promotes them protects against harmful influences.

Claims (16)

1. Product containing at least one kinase inhibitor (A) which has little or no inhibitory activity on Raf, and

• a) at least one neurotrophic factor (B) and / or
• b) at least one substance (C) which is capable of enhancing and / or causing the cellular formation and / or the release of one or more neurotrophic factors (B)

as a combination preparation in the form of a mixture or as individual components for simultaneous or temporally different application at different or the same locations in the prophylaxis and / or therapy of nerve cell and / or glial cell damage. 2. Product according to claim 1, characterized in that at least one Kinase inhibitor (A) selected from a group of kinase Inhibitors comprising 4H-1-benzopyran-4 derivatives are included. 3. Product according to claim 2, characterized in that the kinase Inhibitor (A) a 2- (2-amino-3-methoxyphenyl) -4-oxo-4H- (1) benzopyran, Is flavopiridol and / or derivatives of flavopiridol. 4. Product according to claim 2 and 3, characterized in that the Kina se inhibitor (A) is a thio-flavopiridol and / or oxy-flavopiridol.   5. Product according to claim 1, characterized in that the kinase Inhibitor (A) a 7,12-dihydroindolo (3,2-d) (1) benzazepin-6 (5H) -one, 7OH-staurosporine and / or its kinase inhibiting derivatives, Butyro lactone, roscovitine, Purvalanol A, emodine, anilinoquin azoline, phenyla mino-pyrimidine, trioylimidazole, Paullon, tetrapyrrolic macrocycles, 4- (4-fluorophenyl) -2- (4-methylsulfinylphenyl) -5- (4-pyridyl) 1H-imidazole, 1,4-diamino-2,3-dicyano-1,4-bis (zaminophenylthio) butadiene, 2-2'- amino-3'-methoxyphenyl (-oxanaphtalen-4-one), 2- (2-chloro-4 iodine phenylamino) -N-cyclopropylmethoxy-3,4-difluoro-benzamide, bis is ethylthiomethyl (analog of indocarbazole) and / or pyrimidone derivative. 6. Product according to claim 5, characterized in that the kinase Inhibitor (A) a 5,10,15,20-tetraarylporphyrin and / or 5,10,15- is triarylcorrol. 7. Product according to claim 1, characterized in that the neurotrophic Factor (B) is a neurotrophic protein, especially a neurotrophin and / or is a variant of the neurotrophic protein. 8. Product according to claim 7, characterized in that the neurotrophic Factor (B) a BDNF, NT-3, NT 4/5, NT-6 and / or NGF, CNTF, LIF, CT-1, IGF, Insulin, Midkine, HGF, GDNF, GDF-15 and / or Neurturin is. 9. Product according to claim 1, characterized in that the neurotrophic Factor (B) represents a nucleotide sequence coding for a neurotrophic Protein.   10. Product according to claim 9, characterized in that the nucleotide sequence is integrated into a plasmid or into a viral vector. 11. Product according to one of claims 1 to 10, characterized in that that the substance (C) is a cytokine, interleukin and / or interferon, toluene, D1 / D2 dopamine agonist, beta-adrenergic agonist, neurotransmitter, Ribavirin and / or ribavirin derivative, catechol and / or catechol Derivative, diterpenoid, aldose reductase inhibitor, forskolin, agonist of cholinergic receptor, melanocortin, quinolinic acid, 2-methyl-spiro (1,3- oxathiolan-5,3) -quinuclidine, agonist for the glutamate receptor, AMPA- Agonist, 2,6-diphenyl substituted pyridine derivative, dihydro-caffeic acid amide derivative, biguanide derivative, sulfonylurea derivative, tolbutamide and / or retinoid. 12. Product according to claim 11, characterized in that the substance (C) an IL-1, IL-2, IL-3, IL-4, IL-6, TNF-alpha, GM-CSF, M-CSF, IFN- alpha, IFN-beta; IFN-gamma, apomorphine, isoproterenol, substance P, 1- (beta-D-ribofuranosyl) -1H-1,2,4-triazole, 4-methylcatechol, scabronine A and B, NKH 477, nicotine, carbachol and pilocarpine, Org2766, optical I somere of 2-methyl-spiro (1,3-oxathiolan-5,3) -quinuclidine, quisqualate, Kainate, Alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and Nmethyl-D-aspartate (NMDA), metformin, glibenclamide, glimepiride, Gliclazide, glibornide, gliquidon, glisoxepid, pioglitazone, Is rosiglitazone maleate and / or retinoic acid. 13. A method for producing a product according to one of claims 1 to 8, characterized in that at least one kinase inhibitor (A) with at least one neurotrophic factor (B) and / or at least one Substance (C) is combined in a suitable manner.   14. Use of a product according to one of claims 1 to 8 for Prophylaxis and / or therapy of nerve cell and / or glial cell damage. 15. Use of a product according to one of claims 1 to 8, because characterized in that at least one kinase inhibitor (A) with min at least one neurotrophic factor (B) and / or at least one sub punch (C) is brought into contact with nerve cells and / or glial cells the. 16. The method, characterized in that a product according to one of the Claims 1 to 8 in vitro with nerve cells and / or glial cells in Kon be brought to the beat.