DE4240798A1 - Neuro-protective agents preventing nerve damage - contain ergot derivs. e.g. clavine lysergic acid amide or 8-alpha-amino:ergoline derivs. - Google Patents

Abstract

Neuroprotective compsns. contain ergot alkaloid derivs. (I). (I) are (a) clavines, esp. nicerogline, lergotril or pergolide; (b) lysergamide derivs., esp. bromocriptine, dihydroergocristine, dihydroergotossine or beta-dehydroergocriptine; or (c) 8alpha-amino-ergolines, esp. lisuride or terguride. The compsns. pref. contain 0.1-100 mg (I) per unit dose and are formulated for oral, sublingual, parenteral, transdermal or nasal admin. USE - The compsns. are useful for preventing death of nerves due to endogenous or exogenous toxic substances; or for neuroprotection in degenerative or ischaemic neurological diseases

Description

The present invention relates to the new therapeuti use of derivatives of clavine (nicergoline, Lergotril, pergolide), the lysergic acid amides (Bromocriptine, dihydroergocristine, dihydroergotosin, β- Dihydroergocriptine) and 8-α-aminoergolines (Lisurid, Tergurid) on neuroprotection in neurologi diseases of degenerative type or ischemic Origin.

All ergot derivatives mentioned have the property involved in dopaminergic, α-adrenergic and serotoninergic Receptors of the central and peripheral nervous system to be able to bind. The most important therapeutic range of use for these medicines Age-related vascular insufficiencies of various kinds Expression, Parkinson's disease, hyperprolac tinemia, depression and headache.

Surprisingly, it was found that said Ergot derivatives the beneficial property of the cere possess brale Neuroprotektion and in particular the caused by endogenous or exogenous toxic substances gently prevent the death of cerebral cells.

A primary culture of rat cerebral neurons is prepared eight days after birth as described by Nico letti et al. (Neurosc, J., 6, 1905, 1986). For this, the cells are cleaved with trypsin, then treated with trypsin and deoxyribonuclease inhibitors, and suspended in Basal Eagle's medium (BMA) containing fetal calf serum (10%), 25 mM KCl, 2 mM glutamine and 50 μg / ml. ml of gentamicin. The suspended cells (10 ⁶ / ml) are streaked onto Petri dishes pretreated with poly-L-lysine (10 μg / ml) and further containing a nutrient medium. After 16-18 hours, cytosine arabinoside (Ara-C) is added to prevent the proliferation of glial and endothelial cells. After 7-9 days of in vitro maturation (DIV), the primary culture consists of a highly homogenous neuronal population composed of cells in a layer of nuclei (<90%) and a small number of GAGAergic neurons (approximately 5%). , as well as a few glial and endothelial cells as an impurity. The cells of the nucleus after one week of ripening reach the defined phenotype after 9 to 11 days. Thereafter, they suffer degradation phenomena, which are mainly induced by the indiscriminate activation of NMDA receptors, which do not adequately counterbalance intracellular energy levels.

The cell vitality is by marking the cells with Fluorescein diacetate and propidinium iodide determined. Fluorescein attacks the esterases of living cells and then emits a yellow-green fluorescence. The Propi dinium iodide permeates the membranes of the cells in the to stood the decomposition, binds to the DNA and ver lends these cells a red color. The vitality will by the percentage with fluorescein mark ter cells and is measured by four determined different cultures. All described Ergot derivatives are used to protect against dying Extremely low concentration cells effective (from 1 nmole to 1 μmol). Figure 1 shows the data which were obtained with bromocriptine.

Recent studies by Manev et al. (J. Pharm. Exp. Therap. 252, 419, 1990) have shown that the breakdown of neurons in cultures is induced by a single "pulse" of glutamic acid, which is accompanied by an increase in basal Ca ²⁺ and 20 to 50 minutes until the glutamine is removed from the incubation solution. Taking into account this fact, the degeneration of the neurons is due to hemostatic destabilization by Ca ²⁺ . This "delayed Ca ²⁺ influx" is detected as follows. The culture is washed three times with a Locke solution containing 154 mM NaCl, 5.6 mM KCl, 5.6 mM glucose, 3.6 mM NaHCO ₃ , 2.3 mM CaCl ₂ and 5 mM HEPES (pH 7.4 ) contains. The Mg ²⁺ ions usually contained in the Locke solution were not used. The glutamic acid is added within 15 minutes at a concentration of 100 μmol. After completion of the "pulse", the cells are washed three times with the Locke solution and incubated at 37 ° C for 40 minutes. Subsequently, the culture is incubated with ⁴⁵ Ca ²⁺ (1 μC / plate) for 10 minutes. The uptake of radioactive Ca ²⁺ is interrupted by washing with a chelating agent (154 mM colin chloride + 1 mM EGTA + 10 mM Tris-HCl, pH 7.2-) maintained at 4 ° C. The intracellular radioactivity is determined after dissolving the cells with 0.5 N NaOH (37 ° C, 30 to 60 minutes).

All Ergot derivatives show the property of being able to protect the cells from the increased "delayed Ca ²⁺ influx" in very low concentrations (from 1 nmole to 1 μmol). Fig. 2 shows the results obtained with lisuride.

The systemic use of 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) effects in humans and similar to Parkinson's disease in monkeys ches syndrome. Low doses of MPTP (1-5 mg / kg) verur In monkeys, a selective loss of nerves  cells in the compact substantia nigra, an irrever sible reduction of dopamine in the stria and a considerable loss of function of the extrapyra mid-motor system. The mouse generates MPTP cerebral damage of considerable magnitude.

All Ergot derivatives show the property of protecting the mouse from the neurotoxic effects of MPTP. The administration of MPTP in the CB57 mouse causes severe degeneration of the nigra-strial dopaminergic gene system. Doses of MPTP (20 mg / kg ip) are given at intervals of one hour each. The animals are killed 19 days after treatment with MPTP. After 19 days of treatment, the levels of strontium dopamine and DOPAG reached levels approximately 50% of those found in the control group. The administration of Ergot derivatives at different dosages half an hour after the first dose of MPTP protects against the degenerative effects of the toxic substances. In animals pretreated with such substances, dopamine and DOPAC levels are approximately 3 to 4 times greater than those treated with MPTP alone. Figures 3 and 4 show the data obtained with pergolide.

Monkeys, Macaca fascicularis, with an average amount weight of 3 kg is injected with a neurotoxic substance MPTP at a dosage of 0.2 mg / kg i.v .; eight animals are kept under ketamine narcosis. A group of four monkeys (control group) is single treated with a physiological solution. The Treatment with MPTP took place in eight monkeys once a day for four consecutive days; four of the monkeys received orally a physiological solution (4 ml / kg)  (MPTP group) and four monkeys were given twice daily (every 12 hours) oral bromocriptine (MPTP + BCP group) in a dose of 2 mg / kg (4 ml / kg) over four days. To At the end of treatment with MPTP, the monkeys of the MPTP group for another 10 days once a day with phy solution (4 ml / kg), monkeys of MPTP + BCP Group once a day for 10 more days with Bromcrip tin at a dose of 2 mg / kg (4 ml / kg). The Control group continued to be oral with physiological Solution (4 ml / kg) treated as before in the same way delt. The behavior of the animals is up to the 14. Days observed; on the 15th day, the animals are treated with keta min, Nembutal or Pentbarbital anesthetized and samples taken from the brain. The Putamen and the Kaudatum are homogenized in 0.32 M sucrose and on Dopa min and homovanillic acid examined.

Bromcriptin is capable of keeping the brain in front of the neuro Protect the toxic effects of MPTP, as the Behavior and neurochemistry show (Table 1).  

Table 1

Effect of bromocriptine on the behavior and neurochemistry of monkeys after administration of neurotoxic MPTP

MPTP is probably neurotoxic due to two mechanisms. The metabolism of dopamine induces the formation of H ₂ O ₂ in monoamine oxidase (MAO), which leads to the aging of dopamine neurons. MPTP releases dopamine when applied, thereby forming a significant amount of peroxide radicals.

The other mechanism is characterized in that an intermediate of metabolism of MPTP MAO acts and forms a neurotoxic derivative.

The ergot derivatives mentioned can be administered orally, sublingually, parenterally or percutaneously applied, the Formulation of the pharmaceutical composition for the intended use selected accordingly becomes.

For neuroprotection the daily dosages vary between 0.1 and 100 mg depending on the weight and the condition of the patient and the ergot Derivative.

The pharmaceutical compositions of the invention conditions are prepared by known methods and contain the usual pharmaceutically acceptable and compatible excipients and carriers and given if in addition other active ingredients to the ge desired to achieve effect and appropriate use.

Examples of these preparations include capsules, pil len, tablets, drops, ampoules for intramuscular or intravenous use and optionally forms with prolonged release of the active substance (sustained-release Forms), etc.

There are also those formulations that are not in one named under this invention, provided they the person skilled in the art are familiar.

To give examples, but not restrictive are three pharmaceutical formulations ge called: capsules, tablets and ampoules.  

Examples capsules

1 capsule contains for example:

Bromocriptine | 10 mg Silica, magnesium stearate, maleic acid, amido, lactose, q.b.ad 160 mg

ampoules

For example, 1 ampoule of lyophilizate contains:

Nicergoline | 4 mg tartaric acid 1.04 mg lactose 30 mg

Solvent ampoule Sodium chloride | 17 mg benzalkonium chloride 100 g Aqua bidestillata, q.b.ad 2 mg

tablet

For example, 1 tablet contains:

Lisuride | 1 mg Amido, lactose, magnesium stearate, microcrystalline cellulose, q.b.ad 50 mg

Claims (9)

1. Pharmaceutical compositions for Neuroprotek tion and to prevent the death of nerves due to endogenous or exogenous toxic substances, characterized in that they contain ergot derivatives as active ingredient or pharmaceutically acceptable salts thereof pharmaceutically. 2. Pharmaceutical compositions according to claim 1, characterized in that the ergot derivatives of the group of clavines, lysergic acid amides and 8-α- Aminoergolines are selected. 3. Pharmaceutical composition according to claim 2, characterized in that the clavine Nicergo lin, lergotril and pergolide. 4. Pharmaceutical composition according to claim 2, characterized in that the lysergic acid amides Bromocriptine, dihydroergocristine, dihydroergotossin and β-dihydroergocriptine. 5. Pharmaceutical composition according to claim 2, characterized in that the 8-α-aminoergolines Lisurid and Tergurid include.   6. Pharmaceutical composition according to one of preceding claims, characterized that the content of the active ingredients 0.1 to 100 mg per unit dose. 7. Pharmaceutical composition according to one of preceding claims to oral, sublingual, parenteral, percutaneous or nasal administration chung. 8. A pharmaceutical composition according to claim 7 in Form of tablets, capsules, pills, drops, Retard-Oralia, nose drops and sprays and ampoule len. 9. Use of clavines, lysergic acid amides and 8- α-aminoergolines for the preparation of medicaments for neuroprotection and prevention of Abster nerves due to endogenous or exo contain toxic substances.