MXPA98008157A - Use of benzonaftalenic derivatives for the manufacture of medicines intended for the treatment of diseases of the nervi system - Google Patents

Abstract

The use of the compounds of the general formula (I) for the manufacture of medicaments useful in the treatment of peripheral neuropathies, central neurodegenerative diseases and autoimmune diseases of the nerve system is described

Description

USE OF BENZONAFTALENIC DERIVATIVES FOR THE MANUFACTURE OF MEDICINES INTENDED FOR TREATMENT OF NERVOUS SYSTEM DISEASES Description of the invention The present invention relates to the use of the compounds which correspond to the formula (I) given below, and to their salts for the manufacture of medicaments for the treatment of peripheral neuropathies, central neurodegenerative diseases and autoimmune diseases of the nervous system. these compounds are described, as well as their preparation process, in the European patent EP 0 199 636. In the formula (I): REF. 28488 Ri represents a group -CORß or CH2OH, representing R6 a radical or a radical OR7, wherein R7 represents a hydrogen atom or an alkyl radical having from 1 to 20 carbon atoms, a monohydroxyalkyl radical, or a polyhydroxyalkyl radical, r 'and r "-represents a hydrogen atom, an alkyl radical lower, a mono- or polyhydroxyalkyl radical, an optionally substituted aryl radical or an amino acid or amino sugar residue, or even taken together form a heterocycle, R2 represents a hydrogen atom, an alkyl radical, branched or non-branched, which has from 1 to 15 carbon atoms, an alkoxy radical having from 1 to 4 carbon atoms, or a cycloaliphatic radical, R3 represents a hydrogen atom, a hydroxyl radical, an alkyl radical, branched or not, which has from 1 to 4 carbon atoms, an alkoxy radical having from 1 to 10 carbon atoms, a cycloaliphatic radical substituted or unsubstituted, a thio-cycloaliphatic radical or a radical of the formula -O-Si (CH 3) 2 -Rs, where Rg represents a linear or branched lower alkyl radical, - R and R 5, identical or different, represent an atom of hydrogen, a lower alkyl radical, a hydroxyl radical or a lower acyloxy radical. According to the present invention, among alkyl radicals, branched or non-branched, having 1 to 20 carbon atoms or 1 to 15 carbon atoms, the methyl, ethyl, isopropyl, butyl, tert-butyl radicals can advantageously be mentioned, hexyl, octyl, nonyl, 2-ethylhexyl and dodecyl. Preferably, these radicals have from 1 to 12 carbon atoms. When it is lower, the alkyl radical generally comprises from 1 to 10 carbon atoms. Mention may be made, as the lower alkyl radical, of the radicals "methyl, ethyl, propyl, isopropyl, butyl, tertbutyl and hexyl." By monohydroxyalkyl or polyhydroxyalkyl radical, it is to be understood that a radical having 1 to 6 carbon atoms, and 1 to 5 hydroxyl groups.

Among the monohydroxyalkyl radicals, a radical containing preferably 1 to 3 carbon atoms is prepared, in particular the hydroxymethyl, 2-hydroxyethyl, 2 or 3-hydroxypropyl radicals. Among the polyhydroxyalkyl radicals, a radical having 3 to 6 carbon atoms and 2 to 5 hydroxyl groups, such as 2, 3-dihydroxypropyl, 2,3,4-trihydroxybutyl, 2, 3, 4 radicals is preferred. tetrahydroxypentyl or the pentaerythritol residue. Among the aryl radicals, a phenyl, thiophene or pyridine radical, optionally substituted by at least one halogen atom, a hydroxyl radical, an alkyl radical, a nitro functional group, a methoxy group or an optionally substituted amine functional group, is preferred. An optionally substituted phenyl radical is preferred. By "amino acid residue" is meant primarily a residue that is derived from one of the amino acids such as lysine, glycine, or aspartic acid, and by peptide residue, more particularly is understood a dipeptide or tripeptide residue resulting from the combination of amino acids.

Among the aminated sugar residues, we can mention the residues derived from glucosamine, galactosamine and mannosamine. The term "heterocyclic" is preferably understood to mean a piperidino, morpholino, pyrrolidino or piperazino radical optionally substituted in the 4-position by an alkyl radical of 1 to 6 carbon atoms or polyhydroxyalkyl as defined below. Among the alkoxy radicals that have 1 to carbon atoms, the methoxy, ethoxy, isopropoxy, hexyloxy and decyloxy radical can be mentioned in particular. Among the lower alkyloxy radicals are radicals having 1 to 4 carbon atoms, such as, for example, acetyloxy or propionyloxy radicals. By cycloaliphatic radical is meant a cyclic or polycyclic alkane radical containing from 1 to 10 carbon atoms, optionally substituted by one or more halogen atoms, or one or more hydroxyl radicals. The adamantyl or 1-methylcyclohexyl radicals may be mentioned in particular. The preferred thiocycloaliphatic radical is the 1-adamantylthio radical.

Among the halogen atoms, fluorine or chlorine is preferred. The study of the compounds (I) in several tests measuring neurotrophic activity has also been carried out.

Effects of the compounds on the production of NGF by astrocytes in culture The primary cultures of astrocytes are prepared from the cerebral cortex of neonatal rats from 1 to 2 days. After dissection, the cortices are mechanically dissociated by passing through a Blutex nylon (83 mM mesh). Cells are cultured in a DMEM / F12 medium (Gibco, Brl 1: 1) containing 10% fetal calf serum, 2.5 mM L-glutamine, 100 U / ml penicillin and 100 mg / ml streptomycin (a bark for 30 ml) in 12-well Corning boxes at a rate of 1 ml of cell preparation per well. The boxes are placed at 37 ° C in a saturated incubator. with 5% C02, the medium is replaced after three days and the cultures are used on the 7th day of cultivation (80-90% confluence). The treatments with the different compounds are then carried out with 500 ml per well of the medium described above for 40 hours. At the end of the treatment, the NGF, secreted in the medium, is dosed by the immunoenzymatic administration and the proportion or rate of membrane proteins is evaluated by the Bradford technique with the help of coomassie blue.

Dosage of NGF The NGF rates are evaluated by an immunoenzymatic technique. The 96-well plates are treated for two hours at 37 ° C with an anti-NGF monoclonal antibody (0.67 mg / ml) solubilized in 50 mM Na2C03 buffer, pH 9.6. The cast plates are then incubated 1 hour at room temperature with 1% bovine serum albumin solubilized in the same buffer, and washed with a 50 mM Tris-HCl buffer (pH 7.0) containing 200 mM NaCl, 10 mM CaCl 2, 0.1% Triton X-100 and 0.05% NaN3 (Shock Absorber A). The medium of the astrocytes diluted to 9/10 in an extraction buffer (50 mM Tris-HCl, pH 7, 200 mM NaCl, 1% BSA, 0.1% Triton X-100, 4 mg / ml aprotinin, EDTA 2 mM, chloride 0.1 M benzethonium and 0.05% NaN3) and the NGF standards (range of 10 to 320 pg / ml) are then added in duplicate (0.1 ml / wells). The plates are incubated approximately 16 hours at 4 ° C. After washing with buffer A, the plates are incubated for 4 hours at 37 ° C with a second anti-NGF monoclonal antibody coupled to. ß-galactosidase (0.13-0.15 U / ml). After washing with buffer A, the plates are incubated for 2 to 3 hours at 37 ° C with the substrate of β-galactosidase, red-β-galactopyranoside chlorophenol (2 mg / ml), prepared in 10 mM HEPES ( pH 7.0) containing 150 M NaCl, 2 M MgCl 2, 0.1% NaN 3 and 1% BSA. The product of the reaction is measured at 570 nM by a plate reader Bio Kínetics Reader EL 340.

Results The compounds of the invention increase the production of NGF. At the concentration of 100 nM, the more active compounds of the invention increase the production of NGF to more than 70% in relation to the controls.

Effects of the compounds on the neuritogénes is and the neuritic growth of the dorsal root ganglia The dorsal root ganglia (DRG) are removed from the rat fetuses of 14 days of gestation. Samples are taken. at 3 different levels of the spinal cord (cervical, thoracic and sacral). The DRGs are cultured for 24 hours in a DMEM medium (Gibco, Brl) containing 5% fetal calf serum, 100 U / ml penicillin and 100 μg / ml streptomycin, plus or minus the compound to be studied in a incubator saturated to 5% C02. The medium is then replaced with an identical medium devoid of fetal calf serum containing 1 μm of cytosine arabinoside (SIGMA) to inhibit the proliferation of neuronal cells. The effects of the compounds on neuritogénesis and neuritic growth are evaluated 48 hours after the start of culture using a morphometric technique with the help of a BIOCOM image analysis system.

Results Several compounds of the invention increase the neuritogenesis of dorsal root ganglia explants by more than 100% with respect to controls, at a concentration of 100 nM. The therapeutic potential of the compounds of the invention for the treatment of autoimmune diseases of the nervous system has also been evaluated.

Effects of mouse lymphocyte proliferation compounds that exhibit experimental autoimmune encephalitis (EAE) EAE induction Female SJL / L 'mice are injected subcutaneously, to JO and J7 with the myelin basic protein (MBP, 400 μg / mouse) emulsified in complete Freund's adjuvant. Approximately 20 days after the first immunization, the lymph nodes of the animals that develop the clinical signs of EAE are used for the proliferation test.

Measurement of lymphocyte proliferation The ganglia are removed and dissociated mechanically by passing through a metal screen. The cells are cultured in RPMI medium (Gibco, BRL) containing 10% depleted fetal calf serum, 2.5 rmM L-glutamine, 100 U / ml penicillin, 100 μg / ml streptomycin and 0. 5% of 2-mercaptoethanol at a rate of 250,000 cells per well in 96-well TPP boxes. The layers are placed at 37 ° C in an incubator saturated with 5% C02. The cells are treated on the day of culture with the different retinoic acid analogues, in the absence or in the presence of MBP (25 μg / ml, antigenic stimulation) for 96 hours. Proliferation is evaluated by measuring the incorporation of tritiated thymidine added to the culture medium during the last 16 hours of incubation.

Results The analysis of the results shows that the compounds of the invention inhibit the proliferation of the lymphocytes that come from EAE mice with IC50 ranging from 0.1 to 100 nM.

Effects of the compounds on the production of TNF-α (Factor A of Tumor Necrosis.) By the activated microglial cells.

Primary cultures of microglial cells: The microglial cells are prepared from the bark of neonatal rats for 1 day.

The cells are cultured in a DMEM medium (Gibco, BRL) containing 10% fetal calf serum (inactivated myoclonus), 2.5 mM L-glutamine, 4.5 g / 1 L-glucose and 100 μg / ml gentamicin in 96-well TPP plates. The cells are treated with different retinoic acid analogues and stimulated with 1 μg / m? of lipopolysaccharide (LPS) in order to induce the production of TNFa. After 18 hours of incubation the culture media are collected for the dose of TNFa.

Dosage of TNFa: The rates or proportions of TNFα in supernatants of microglial cells are determined with the help of. a cytotoxicity test on the L929 cell line. The test is revealed thanks to the MTT colorimetric dosage.

Results The compounds of the invention decrease the production of TNFa by activated microglial cells in culture, with effects ranging up to 50% decrease at the concentration of 10 nM. The results show that the compounds of the invention have a neurotrophic activity that can find an application for the treatment of peripheral neuropathies of the traumatic, ischemic, metabolic, infectious, alcoholic, iatrogenic or genetic type, in the treatment of the diseases that affect the motor neurons, such as amyotrophic lateral sclerosis and spinal amyotrophies. These can also be used for the preparation of medicines useful in the treatment of retinopathies, cerebral senility, dementia consecutive to multiple infarcts, vascular dementia, in the treatment of olivo-ponto-cerebellar atrophy and other neurodegenerative diseases, for example Alzheimer's disease, Pick's disease or Huntington's chorea. The compounds of the invention can also find an application in the prevention of neuronal death subsequent to a cerebrovascular accident or spinal or cranial trauma. These can also be checked as active in pathologies of autoimmune origin such as sclerosis in plaques, Guillain-Barré disease and myasthenia gravis. The most active compounds respond to the following formula: COR, in which - R6 represents a radical or an OR7 radical, R representing a hydrogen atom, an alkyl radical having from 1 to 20 carbon atoms, a monohydroxyalkyl radical, or a polyhydroxyalkyl radical, r 'and r "represent a hydrogen atom, a lower alkyl radical, a mono- or polyhydroxyalkyl radical, an optionally substituted aryl radical or an amino acid or amine sugar residue, or even taken together form a heterocycle, R represents a hydrogen atom, a hydroxyl radical, a branched or non-branched alkyl radical, having 1 to 4 carbon atoms or an alkoxy radical of 1 to 4 carbon atoms "6- [3- (1-adamantyl) -4-methoxyphenyl] -2-naphthoic acid, and 6- [3- (1 α-methyl) -4-hydroxy-phenyl] -2-naphthoic, their esters (in particular their methyl ethers) and their amides, are the preferred compounds.

The compounds can be used in association with one or more compounds of the following classes: anti-inflammatory steroids and non-steroids, antivirals (in particular antiherpes), immunosuppressants and immunomodulators. The compounds can be presented under all pharmaceutical forms adapted for enteral, parenteral or local administration, in association with the appropriate excipients, for example in the form of tablets, dragees, pills, capsules, suppositories, patches, solutions or suspensions for drinking or injectables. Daily administration may vary from 0.01 to 100 mg of active ingredient.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present invention.

Having described the invention as above, the content of the following is claimed as property:

Claims (4)

1. The use of the compounds that respond to the formula (I) and its salts, wherein - Ri represents a group -CORe or CH20H, represented Rs a radical or a radical OR ?, wherein R7 represents a hydrogen atom 'or an alkyl radical having from 1 to 20 carbon atoms, a monohydroxyalkyl radical, or a polyhydroxyalkyl radical, r' and r "represent a hydrogen atom, a radical lower alkyl, a mono- or polyhydroxyalkyl radical, an optionally substituted aryl radical or a residue of amino acid or amino sugar, or even taken together form a "heterocycle, R2 represents a hydrogen atom, an alkyl radical, branched or not, having 1 to 15 carbon atoms, an alkoxy radical having 1 to 4 atoms of carbon, or a cycloaliphatic radical, R3 represents a hydrogen atom, a hydroxyl radical, an alkyl radical, branched or unbranched, having 1 to 4 carbon atoms, an alkoxy radical having 1 to 10 carbon atoms, a cycloaliphatic radical substituted or unsubstituted, a thio-cycloaliphatic radical or a radical of the formula -O-Si (CH 3) 2 -Rs, where R 8 represents a linear or branched lower alkyl radical, - R 4 and R 5, which are identical or different, represent an atom of hydrogen, a lower alkyl radical, a hydroxyl radical or a lower acyloxy radical for the manufacture of medicaments useful in the treatment of peripheral neuropathies, of central neurodegenerative diseases and of autoimmune diseases of l -nervous system

2. The use of compounds that respond to the formula in which - R6 represents a radical or an OR7 radical, R7 representing a hydrogen atom, an alkyl radical having from 1 to 20 carbon atoms, a monohydroxyalkyl radical, or a polyhydroxyalkyl radical, r 'and r "represent a hydrogen atom, a lower alkyl radical, a mono- or polyhydroxyalkyl radical, an optionally substituted aryl radical or an amino acid or amine sugar residue, or even taken together form a heterocycle, R represents a hydrogen atom, a hydroxyl radical, a branched or non-branched alkyl radical, having 1 to 4 carbon atoms or an alkoxy radical of 1 to 4 carbon atoms, for the manufacture of medicaments useful in the treatment of neuropathies peripheral, central neurodegenerative diseases and autoimmune diseases of the nervous system.

3. The use of 6- [3- (1-adamantyl) -4-methoxyphenyl] -2-naphthoic acid, and its esters and / or its amides for the manufacture of medicaments useful in the treatment of peripheral neuropathies, central neurodegenerative diseases and autoimmune diseases of the nervous system. *

4. The use of 6- [3- (1-adamantyl) -4-hydroxyphenyl] -2-naphthoic acid, "of its esters and / or its amides for the manufacture of medicaments useful in the treatment of peripheral neuropathies, central neurodegenerative diseases and autoimmune diseases of the nervous system.