DE10142175A1 - Use of beta-adrenoceptor agonists, e.g. reproterol, salmeterol or terbutaline, for restoring and/or maintaining function of damaged nerve cells, e.g. for treatment of neurodegenerative diseases - Google Patents

Abstract

Use of beta -adrenoceptor agonists (I) for restoring and/or maintaining the function of partially or completely damaged cells of the central nervous system,

Description

The present invention relates to the use of β-adrenoceptor agonists for Treatment of neurodegenerative diseases.

The human brain is a highly complex organ with more than 100 billion Nerve cells (= neurons) and about 10,000 connections (= synapses) per cell. The The brain is the central organ of the conscious and unconscious processing of the brain Human stimuli, thinking and feeling, the targeted Doing, learning and remembering. One of the most important achievements of the human Brain is information processing in language; also control center of a variety of organ functions as well as breathing, heart rate and temperature control.

There are a variety of diseases that lead to the death of nerve cells and / or one Decreased synapses and thus reduced brain performance. Examples of such clinical pictures are Alzheimer's disease, cerebrovascular dementias, Parkinson's disease, Pick's disease, Huntington's chorus, amyotrophic lateral sclerosis, Lewy Body dementia, stroke and brain trauma, such as contusio and commotio cerebri as well Brain and spinal cord injuries or cross-sectional injuries, spina bifida, and Diseases of the inner ear, for example disorders associated with the appearance of a Tinnitus, such as subacute or chronic tinitus, are associated with sudden hearing loss, disease Menière, and diseases with hearing impairment or Decrease in vision, etc. A clinically established neuroprotective So far there is no therapy for the clinical pictures mentioned. After the appearance of the Symptoms are only treated, but not their causes.

The goal of a causal therapy for brain disorders is the demise of To prevent nerve cells.

There is currently no established therapy with which it is possible to pre-nerve cells Protect or regenerate damage. An essential element in therapy of the diseases mentioned above is to prevent damage processes to nerve cells, increase cerebral blood flow or when a vessel closes again to minimize the risk of damage. However, this form of therapy is If at all, only successful if used quickly after the acute event can be.

The present invention was based on the object of finding drugs which Protect nerve cells from damage and the function of partial or complete can at least partially restore degenerated cells.

Surprisingly, it was found that the activation of astrocytes with Drugs such as β-adrenoceptor agonists, endogenous processes of neuroprotection in Gear is set, which reduces the damage or destruction of nerve cells and in some cases can even be prevented.

The present invention accordingly relates to the use of β-adrenergic Agonists to restore and / or maintain the function of partial or completely damaged cells of the central nervous system and / or other nerve cells.

For the purposes of the present invention, "damaged cell" means that the cell is damaged external influences has been damaged or in the sense of degeneration by in the cell running processes is partially or completely destroyed, with an impairment can go hand in hand with bodily functions. The term "cell damage" includes both the damage to individual cells or cell types and the damage to Strands or pathways of nerve cells.

In addition to the cells of the central nervous system, the nerve cells also include the cells of the spinal cord and all other nerve cells in the body.

β-adrenoceptors respond in particular to adrenergic drugs. Examples of β- adrenergic agonists in the present invention because of their good potency preferably used are formoterol, fenoterol, salbutamol, orciprenaline, Isoetharine, cimaterol, ractopamine, reproterol, salmeterol, terbutaline, their isomers, Acid addition salts, analogs and any mixtures of the foregoing.

Β1-adrenoceptor agonists such as dopamine can also activate astrocytes and thereby achieve the protection of the neurons.

Experiments have shown, for example, that the lipophilic β- Adrenoceptor agonists can permeate into the brain and there the β-adrenoceptors Stimulate astrocytes. The stimulation of these receptors in turn leads to one Activation of the astrocytes and, as a result, an increased release of Growth factors, such as NGF, which nerve cells before a disease-related Can protect damage.

The β-adrenoceptor agonists are used in the amounts customary for these drugs applied, in particular in an amount of 0.01 to 100 mg / day, preferred Quantity ranges can also depend on the respective β-adrenoceptor agonist. With Substances like formoterol, fenoterol and salmeterol will be a particularly good one get neuroprotective effect when in an amount of 0.01 to 5 mg / day be administered. Terbutaline is preferably used in an amount of 1.0 to 30 mg / day, Salbutamol in an amount of 1.0 to 50 mg / day, and orciprenaline and reproterol in one Amount applied from 1.0 to 100 mg / day.

Β1-adrenoceptor agonists such as dobutamine can also activate astrocytes and thereby protect the neurons.

The β-adrenoceptor agonists used according to the invention and possibly other conventional ones Drugs that do not negatively influence or support the therapy and usual ones Ingredients can be in the usual pharmaceutical dosage forms, especially as a solution, suspension, emulsion, tablets, suppositories, etc. Also the use in special formulations such as liposomes, nanosomes, slow-release pellets etc. is possible. You can in the usual way, for example orally, parenterally, intravenously, inhalatively, rectally, intraventricular, intraarterial, intraperitoneal and / or intramuscular or as an implant be administered. The mode of administration is preferably selected such that the affected cells as quickly as possible from the invention Drug can be achieved.

The β-adrenoceptor agonists used according to the invention are particularly suitable for Manufacture of medicines for the treatment of neurodegenerative diseases. Examples of such diseases are Alzheimer's disease, cerebrovascular dementias, Parkinson's disease, Pick's disease, Huntington's chorus, amyotrophic lateral sclerosis, Lewy Body dementia, stroke and / or brain trauma such as contusio and commotio cerebri as well as brain and spinal cord injuries or cross-sectional injuries, spina bifida, and diseases of the inner ear, for example diseases associated with the occurrence tinnitus, such as subacute or chronic tinitus, sudden hearing loss, disease Menière, and diseases with hearing impairment or Reduction in eyesight, etc.

In a further embodiment of the present invention, the invention compounds used for prevention of the diseases mentioned above used.

In a further embodiment of the present invention, the invention compounds used as an additive (s) for culture media to promote growth and / or differentiation and / or protection of mammalian and human cells used.

Examples 1) Activation of astrocytes

Primary cultures of astrocytes were obtained from the cerebral cortex tissue of newborn Fischer 344 rats within 24 hours after birth. The brains were dissected from the skull cap under sterile conditions, the bark tissue (cortex) was isolated and the cells were dissociated by a close-meshed wire mesh. The cells were placed in cell culture flasks and cultured in serum-containing DMEM solution (containing fetal calf serum and a penicillin-streptomycin mixture) until the cells confluent. Oligodendrocytes and microglia were removed by washing with cold buffer solution. The confluent astrocytes were then removed from the bottom of the culture flasks with a trypsin solution and sown at a density of 20,000 cells / cm ² in petri dishes on cover slips and cultured in serum-containing medium until the cells re-confluent. Two days after confluence, the medium was changed with serum-free medium and after 24 hours another medium change, also with serum-free medium. Twenty-four hours after the second medium change, salmeterol was added. Six hours after the treatment, the astrocytes were photographed in 200-fold microscopic magnification to document the morphological changes ( Fig. 1). The figure shows the astrocytes 6 hours after the start of treatment. The changes from the polygonal, flat and little light-refracting cells in the controls compared to the activated, star-shaped and light-refracting astrocytes in the groups treated with salmeterol can be clearly seen.

2) Neuroprotective effect of salmeterol in vitro

Primary mixed cultures from the hippocampus of the rat were set up as described in Example 2 and, after 14 days in culture, subjected to a medium change to serum-free medium. Twenty-four hours after changing the medium, the β-adrenoceptor agonist salmeterol was added and the cells were incubated for 4 hours. Sister cultures of the hippocampal cells treated in this way received only vehicle, or salmeterol alone. After 4 hours, the medium was changed and the cells were incubated for 1 hour with L-glutamate (1 mM) in serum-free medium. The medium was then changed again with serum-free medium in order to remove the glutamate from the cultures. Salmeterol was added fresh each time the medium was changed and was thus present in the medium during the glutamate treatment and up to 18 hours afterwards. Eighteen hours after the damage to glutamate, the cells were incubated with a trypan blue solution, fixed and the damaged, blue-stained neurons were quantified under a 200-fold microscopic magnification ( Fig. 2a). The values given are mean values and standard deviation from 5-6 cultures per group. * p <0.05; ** p <0.01; and *** p <0.001 compared to the control treated with glutamate (analysis of variance, Scheffé test).

3) Neuroprotective effect of salmeterol in vivo

A focal cerebral ischemia of the mouse was produced by ligating the cerebral artery. Male NMRI mice (26-31 g, 10-12 animals per group) were used for the experiments. The animals were anesthetized by an intraperitoneal injection of tribromoethanol (600 mg / kg). The surgical field was then opened through a 2 cm incision between the left eye and ear, the temporalis muscle was removed by a thermocauter and the bone was removed with a fine bur to expose the artery of the media. This artery and its two distal branches were permanently occluded. During the preparation, the body temperature of the mouse was measured and kept constant at 37 +/- 1 ° C by an infrared heat lamp. After the preparation, the animals were left for an additional two hours at an ambient temperature of 30 ° C. To determine the area of the infarct, the mice were anesthetized again with tribromoethanol 48 hours after occlusion of the cerebral artery and perfused with a 1.5% neutral red solution (0.5 ml intraperitoneally). As a result, the perfused brain tissue turned red and the infarcted area remained bright. The isolated brains were fixed with 4% formaldehyde buffer (pH 7.4) for at least 24 hours and then the non-stained area on the brain surface (infarct area) was computer-supported (NIH image software) measured. The salmeterol dissolved in 0.9% NaCl was injected interperitoneally 5 hours before the operation. The animals in the control group received only 0.9% NaCl solution ( Fig. 2b). The values are mean values and standard deviation of 15-16 animals per group. * p <0.05 compared to the control (analysis of variance, Duncan's test)

Claims (9)

1. Use of β-adrenoceptor agonists for recovery and / or Maintenance of the function of partially or completely damaged cells of the Central nervous system and / or other nerve cells. 2. Use according to claim 1, characterized in that the β-adrenergic Agonists are selected from formoterol, fenoterol, salbutamol, orciprenaline, Isoetharine, cimaterol, ractopamine, reproterol, salmeterol, terbutaline, their isomers, Acid addition salts, analogs and any mixtures of the foregoing. 3. Use according to claim 1 or 2, characterized in that astrocytes and / or endogenous protective mechanisms are activated or stimulated. 4. Use according to one of claims 1 to 3, characterized in that the β- Adrenoceptor agonists in an amount of 0.01 to 100 mg / day. 5. Use according to claim 4, characterized in that substances such as Formoterol, fenoterol and salmeterol in an amount of 0.01 to 5 mg / day, terbutaline in an amount of 1.0 to 30 mg / day, salbutamol in an amount of 1.0 to 50 mg / day and orciprenaline and reproterol in an amount of 1.0 to 100 mg / day become. 6. Use according to one of claims 1 to 5, characterized in that β1- Adrenoceptor agonists such as dobutamine can be used. 7. Use according to one of claims 1 to 6, characterized in that the neurodegenerative diseases are selected from Alzheimer's disease, cerebrovascular dementia, Parkinson's disease, Pick's disease, Huntington's chorea, Amyotrophic lateral sclerosis, Lewy body dementia, stroke and / or Brain trauma such as contusio and commotio cerebri as well as brain and Spinal cord injuries or cross-sectional injuries, spina bifida, and Diseases of the inner ear, for example disorders associated with the appearance of a Tinnitus, such as subacute or chronic tinitus, are associated with sudden hearing loss, disease Menière, and diseases with hearing impairment or Reduction in eyesight, etc. 8. Use according to one of claims 1 to 6, characterized in that the Compounds for the prevention of neurodegenerative diseases are used. 9. Use according to one of claims 1 to 6 as an additive for culture media Promote growth and / or differentiation and / or protection of Mammalian and human cells.