WO2003045375A1

WO2003045375A1 - Novel utilization of cyclopentabenzofurans

Info

Publication number: WO2003045375A1
Application number: PCT/EP2002/012880
Authority: WO
Inventors: Lutz Assmann; Thomas Fahrig; Irene Gerlach; Martin Hendrix
Original assignee: Bayer Aktiengesellschaft
Priority date: 2001-11-29
Filing date: 2002-11-18
Publication date: 2003-06-05
Also published as: DE10158561A1; AU2002356615A1

Abstract

The invention relates to the utilization of cyclopentabenzofurans for the production of medicaments for prophylaxis and/or treatment, especially for neuroprotective treatment of Parkinson's disease.

Description

New use of cyclopentabenzofurans

The invention relates to the use of cyclopentabenzofurans for the manufacture of a medicament for the prophylaxis and / or treatment, in particular for the neuroprotective treatment of Parkinson's disease.

NF-κB (nuclear factor-κB) is a transcription factor that, in its active, DNA-binding form, is composed of dimeric combinations of different members of the NF-B / Rel family of proteins. Five members of this family have been identified so far: NF-κBl (p50 and its precursor protein pl05), NF-κB2 (p52 and its precursor protein plOO), c-Rel, RelA (p65) and RelB (Gosh et al. Ann. Rev. of Immunol. 1998, 16, 225-260). NF-κB is present in unstimulated cells as a cytoplasmic, inactive form by binding to an inhibitory protein (I-κB). After stimulation, I-κB is rapidly phosphorylated by I-κB kinases and proteolytically broken down. This releases NF-κB in its active form and enables its translocation into the cell nucleus. In its capacity as a transcription factor, NF-κB activates or modulates the expression of specific genes in the cell nucleus (Karin, J. Biol. Chem. 1999, 274, 27339-27342).

Parkinson's disease is a chronically progressive, neurodegenerative disease that is characterized by the loss of dopaminergic neurons of the substantia nigra. The massive disturbances of the dopaminergic neurotransmission caused thereby lead to a serious malfunction of the movement-controlling extrapyramidal system. The main characteristics of early signs and symptoms of Parkinson's disease are resting tremors, slowing of movement, muscle stiffness and unstable posture.

The current medications for Parkinson's disease are of a purely symptomatic nature, with substitution therapy with L-dopa being the most frequently used represents a form of therapy. Neither preventive nor restorative therapies are currently available (Mendis et al., Can. J. Neurol. Sei. 1999, 26, 89103).

As in other organ systems, functional NF-κB complexes occur in all cell types of the central nervous system (CNS) (Neill and Kaltschmidt, Trends

Neurosci. 1997, 20, 252-258). In Parkinson's patients, an increased nuclear localization of NF-κB and thus an increased activation of NF-κB-mediated signal transduction in dopaminergic neurons of the substantia nigra have been described (Hunot et al. Proc. Natl. Acad. Sei. USA 1997, 94 , 2642-2647). The functional significance of NF-κB activation in Parkinson's disease is still unclear.

WO 00/07579 and WO 00/08007 describe natural or synthetic rocaglamides as inhibitors for NF-κB-mediated, pathophysiological processes for the treatment of chronic inflammatory diseases such as arteriosclerosis and multiple sclerosis.

The following uses and biological effects of natural and synthetic rocaglamides are also known: pesticides (for example DE-A-199 34 952; Bioorg. Chem. 1999, 18-27.), Fungicidal action (J. Agric. Food Chem. 2000, 48 , 1400-1404), anti-tumor activity [CAPLUS 1999, 49233 (JP-A-

11012279); CAPLUS 1997, 262264 (WO 97/08161)], anti-leukemic effect (US-A-4,539,414).

Surprisingly, it has now been shown that the compounds used according to the invention are for the prophylaxis and / or treatment of Parkinson's

Disease, especially for neuroprotective therapy for Parkinson's disease.

The present invention therefore relates to the use of compounds of the formula (I)

in which

R ¹ and R ³ each independently represent hydrogen, halogen or alkyl,

R ² and R ⁴ each independently represent halogen, alkyl or optionally substituted alkoxy,

R ^{5 represents} hydroxy, alkylamino or the radical -NR ¹² -CHR ¹³ -COOR ¹⁴ , in which

R ^{12 represents} hydrogen or alkyl,

R stands for one of the residues of a natural or synthetic α-amino acid, functional groups being optionally protected, or

R ¹² and R ¹³ together represent - (CH ₂ ) ₃ - and - (CH ₂ ) ₄ -, and

R ^{14 represents} alkyl, benzyl or another C-terminal protective group,

R ^{6 represents} hydrogen or

R ⁵ and R ⁶ together represent oxygen (oxo), R ⁷ and R ⁸ each represent hydrogen,

R ^{9 represents} optionally substituted phenyl or hetaryl,

R ^{10 represents} hydrogen, halogen, alkyl or alkoxy and

R ^{11 represents} hydrogen, halogen or alkyl,

as well as their salts

for the manufacture of a medicament for the prophylaxis and / or treatment of Parkinson's disease.

The compounds used according to the invention can exist in stereoisomeric forms which either behave like image and mirror image (enantiomers) or which do not behave like image and mirror image (diastereomers). The invention relates to both the enantiomers or diastereomers or their respective mixtures. These mixtures of the enantiomers and diastereomers can be separated into the stereoisomerically uniform constituents in a known manner.

The compounds used according to the invention can also be in the form of their salts. In general, salts with organic or inorganic bases or acids may be mentioned here.

Physiologically acceptable salts are preferred in the context of the present invention. Physiologically acceptable salts of the compounds used according to the invention can be salts of the substances according to the invention with mineral acids, carboxylic acids or sulfonic acids. For example, particular preference is given to Salts with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid,

Ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, Acetic acid, propionic acid, lactic acid, tartaric acid, citric acid, fumaric acid, maleic acid or benzoic acid.

Physiologically acceptable salts can also be metal or ammonium salts of the compounds used according to the invention which have a free carboxyl group. For example, particular preference is given to Sodium, potassium, magnesium or calcium salts, as well as ammonium salts, which are derived from ammonia, or organic amines, such as ethylamine, di- or. Triethylamine, di- or triemanolamine, dicyclohexylamine, dimethylaminoethanol, arginine, lysine, ethylenediamine or 2-phenylethylamine.

The substances used in accordance with the invention are generally defined by the formula (I). Preferred substituents or ranges of the radicals listed in the formulas mentioned above and below are explained below.

R and R independently of one another preferably each represent hydrogen, fluorine, chlorine, bromine or CrC ₆ alkyl.

R ² and R ⁴ independently of one another preferably each represent fluorine, chlorine, bromine, dC ₆ -alkyl or C ₁ -C ₄ -alkoxy optionally substituted by fluorine or chlorine.

R ⁵ preferably represents hydroxy, C ₁ -C alkylamino or the rest

-NR ¹² -CHR ¹³ -COOR ¹⁴ .

R preferably represents hydrogen.

R ⁵ and R ⁶ also preferably together represent oxygen (oxo).

R ⁷ and R ⁸ each preferably represent hydrogen. R ⁹ preferably represents phenyl, methylenedioxyphenyl or 5- or 6-membered hetaryl with 1 or 2 heteroatoms from the series nitrogen, oxygen and sulfur, which can each be optionally substituted up to four times by substituents of the group: halogen, C ₁ -C ₆ - Alkyl, hydroxy, C1-C4-alkoxy, Cι-C4-alkylthio, Ci-C ^ alkylcarbonyl, phenyl, phenoxy, hetaryl,

Hetaryloxy, C 1 -C 4 alkyl substituted by fluorine or chlorine, C 1 -C 4 alkoxy substituted by fluorine or chlorine, C 4 -C 4 alkylthio substituted by fluorine or chlorine, C 4 -C 4 alkyl carbonyl substituted by fluorine or chlorine.

R ¹⁰ preferably represents hydrogen, fluorine, chlorine, d-Ce-alkyl or dC ₆ -alkoxy.

R ¹¹ preferably represents hydrogen, fluorine, chlorine, bromine or d -C ₆ alkyl.

R ¹² preferably represents hydrogen or C ₁ -C 4 alkyl.

R ¹³ preferably represents hydrogen, C ₁ -C 4 -alkyl optionally substituted by amino or hydroxy, or mercaptomethyl, methylthioethyl, carboxymethyl, carboxyethyl, carbamoylmethyl, carbamoylethyl, guanidino-propyl or optionally substituted by amino, nitro, halogen, hydroxyl or

Methoxy substituted phenyl or benzyl or for naphthylmethyl, indolylmethyl, imidazolylmethyl, triazolylmethyl or pyridylmethyl, where functional groups may optionally be protected.

R ¹² and R ¹³ also preferably together represent - (CH ₂ ) ₃ - or - (CH ₂ ) ₄

R ¹⁴ preferably represents Ci-Cβ-alkyl, benzyl or another C-terminal protective group.

R ¹ and R ³ independently of one another particularly preferably each represent hydrogen, fluorine, chlorine, bromine, methyl or ethyl. R ² and R ⁴ independently of one another particularly preferably each represent fluorine, chlorine, bromine, C ₁ -C ₄ -alkyl or methoxy or ethoxy which is optionally substituted by fluorine or chlorine.

R particularly preferably represents hydroxyl, C ₁ -C -alkylamino or the radical -NR ¹² -CHR ¹³ -COOR ¹⁴ .

R ⁶ particularly preferably represents hydrogen.

R ⁵ and R ⁶ also particularly preferably together represent oxygen (oxo).

R ⁷ and R ⁸ each particularly preferably represent hydrogen.

R ⁹ particularly preferably represents phenyl, methylenedioxyphenyl or 5- or

6-membered hetaryl with 1 or 2 heteroatoms from the series nitrogen, oxygen and sulfur, each of which can optionally be substituted up to 3 times by substituents from the group: fluorine, chlorine, bromine, iodine, C1-C4-alkyl, hydroxy, C ₁ -C alkoxy, C ₁ -C 4 alkylthio, C ₂ -C 4 alkylcarbonyl nyl, Phe, phenoxy, furyl, thienyl, pyrrolyl, thiazolyl, pyridyl, furyloxy,

Thienyloxy, pyrrolyloxy, thiazolyloxy, pyridyloxy, C1-C4-alkyl substituted by fluorine or chlorine, C-C3-alkoxy substituted by fluorine or chlorine, Ci-C _ß- alkylthio substituted by fluorine or chlorine, Cι-C4 substituted by fluorine or chlorine alkylcarbonyl.

R ¹⁰ particularly preferably represents hydrogen, fluorine, -CC ₄ -alkyl or CrC ₄ -alkoxy.

R ¹¹ particularly preferably represents hydrogen, fluorine, chlorine, bromine or C 1 -C ₄ -alkyl. R ¹² particularly preferably represents hydrogen or methyl.

R ¹³ particularly preferably represents hydrogen, methyl, isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl, 3-aminopropyl, 4-aminobutyl, carboxymethyl, 2-carboxyethyl , Carbamoylmethyl, 2-carbamoylethyl, 3-guanidinopropyl, phenyl, benzyl, 4-hydroxybenzyl, 4-methoxybenzyl, 2-nitrobenzyl, 3-nitrobenzyl, 4-nitτobenzyl, 2-aminobenzyl, 3-aminobenzyl, 4-aminobenzyl , 3,4-dichlorobenzyl, 4-iodobenzyl, -naphthylmethyl, ß-naphthylmethyl 3-indolylmethyl, 4-imidazolylmethyl, 1,2,3-triazol-l-yl-methyl, 1,2,4-triazol-l -yl-methyl, 2-pyridylmethyl or 4-pyridylmethyl, where functional groups may optionally be protected.

R ¹² and R ¹³ are also particularly preferably together - (CH ₂ ) ₃ - or - (CH ₂ ) ₄ -.

R ¹⁴ particularly preferably represents C ₁ -C ₄ -alkyl, benzyl or another C-terminal protective group.

R ¹ and R ³ very particularly preferably each represent hydrogen, chlorine or

Bromine.

R ² and R ⁴ very particularly preferably each represent methoxy, ethoxy, trifluoromethoxy, difluoromethoxy, chlorodifluoromethoxy, 2-chloro-1,1,2-trifluoroethoxy, 1,1,2-trifluoroethoxy, 1,1,2,2 -Tetrafluoroethoxy, 2,2,2-

Trichloro-1,1-difluoromethoxy or 1,1-difluoroethoxy, especially for methoxy or ethoxy.

R ⁵ very particularly preferably represents hydroxyl, methylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino, sec.-butylamino, tert.-butylamino or the radical -NR ¹² -CHR ¹³ -COOR ¹⁴ . R ⁶ very particularly preferably represents hydrogen.

R ⁵ and R ⁶ also very particularly preferably stand together for oxygen (oxo).

R and R very particularly preferably each represent hydrogen.

R ⁹ very particularly preferably represents phenyl, methylenedioxyphenyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl or pyridyl, which can in each case optionally be mono- or disubstituted by substituents from the group: fluorine, chlorine, bromine, iodine, methyl, ethyl , n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, trifluoromethyl, difluoromethyl, chlorodifluoromethyl, 2-chloro-l, l, 2-trifluoroethyl, 1,1,2 Trifluoroethyl, 1,1,2,2-tetrafluoroethyl, l, l-difluoro-2,2,2-trichloroethyl, 2,2,2-trifluoroethyl,

1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,3-pentafluoropropyl, hydroxy, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec.-butoxy, tert.-butoxy, trifluoromethoxy, difluoromethoxy, chlorodifluoromethoxy, 2-chloro-l, l, 2-trifluoroethoxy, 1,1,2-trifluoroethoxy, 1,1, 2,2-tetrafluoroethoxy, 2,2,2-trichloro-l, l-difluoromethoxy, 1,1-difluoroethoxy,

Methylthio, ethylthio, trifluoromethylthio, 1,1-difluoroethylthio, 2,2,2-trifluoroethylthio, phenyl, phenoxy, acetyl, propionyl, propylcarbonyl, butylcarbonyl or 2-methylpropylcarbonyl.

R ¹⁰ very particularly preferably represents hydrogen, fluorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, n -Butoxy, isobutoxy, sec-butoxy or tert-butoxy.

R ¹¹ very particularly preferably represents hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl. R ¹² very particularly preferably represents hydrogen.

R ¹³ very particularly preferably represents hydrogen, methyl, isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl, 3-aminopropyl, 4-aminobutyl, carboxymethyl, 2-carboxyethyl,

Carbamoylmethyl, 2-carbamoylethyl, 3-guanidinopropyl, phenyl, benzyl, 4-

Hydroxybenzyl, 3-indolylmethyl or 4-imidazolylmethyl.

R ¹⁴ very particularly preferably represents methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl or benzyl.

The general definitions or explanations of residues or explanations listed above or in preferred areas can be combined with one another, that is to say also between the respective areas and preferred areas. They apply accordingly to the end products as well as to the preliminary and intermediate products.

According to the invention, preference is given to using the compounds of the formula (I) in which there is a combination of the meanings listed above as preferred (preferred).

According to the invention, particular preference is given to using the compounds of the formula (I) in which there is a combination of the meanings listed above as being particularly preferred.

According to the invention, the compounds of

Formula (I) in which there is a combination of the meanings given above as being particularly preferred.

According to a second aspect, compounds of the formula (I) in which R ¹ and R ³ each independently represent hydrogen, halogen or alkyl,

R ^{12 represents} hydrogen or alkyl,

R ^{13 stands} for one of the residues of a natural or synthetic α-amino acid, functional groups being optionally protected, or

R ¹² and R ¹³ together represent - (CH ₂ ) ₃ - and - (CH ₂ ) -, and

R ^{14 represents} alkyl, benzyl or another C-terminal protective group,

R ^{6 represents} hydrogen or

R ⁵ and R ⁶ together represent oxygen (oxo),

R ⁷ and R ⁸ each represent hydrogen,

R ^{9 represents} optionally substituted phenyl or hetaryl,

R ^{10 represents} hydrogen, alkyl or alkoxy and

R ^{π represents} hydrogen, halogen or alkyl. According to the second aspect of the invention, the use of compounds of the formula (I) in which

R ¹ and R ³ each represent hydrogen, fluorine, chlorine, bromine, methyl or ethyl,

R ² and R ⁴ each represent methoxy, ethoxy, trifluoromethoxy, difluoromethoxy, chlorodifluoromethoxy, 2-chloro-1, 1, 2-trifluoroethoxy, 1, 1, 2-trifluoroethoxy, 1,1,2,2-tetrafluoroethoxy, 2 , 2,2-trichloro-l, l-difluoromethoxy or 1,1-difluoroethoxy,

R ^{5 represents} hydroxy, methylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino, sec.-butylamino, tert.-butylamino or the radical -NR ¹² -CHR ¹³ -COOR ¹⁴ ,

R ^{6 represents} hydrogen, or

R ⁵ and R ⁶ together represent oxygen (oxo),

R and R each represent hydrogen,

R ⁹ for optionally single or double by fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, trifluoromethyl, difluoromethyl, chlorodifluoromethyl, 2- Chloro-l, l, 2-trifluoroethyl, 1,1,2-trifluoroethyl, 1,1,2,2-tetrafluoroethyl, l, l-difluoro-2,2,2-trichloroethyl, 2,2, 2-trifluoroethyl, 1,1,2,3,3,3-hexafluoropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,3-pentafluoropropyl, hydroxy, methoxy, ethoxy, n -Propoxy, isopropoxy, n-butoxy, isobutoxy, sec.-butoxy, tert.-butoxy, trifluoromethoxy, difluoromethoxy, chlorodifluoromethoxy, 2-chloro-l, l, 2-trifluoroethoxy, 1,1,2-trifluoroethoxy, 1,1 , 2,2-tetrafluoroethoxy, 2,2,2-trichloro-l, l-difluoromethoxy or 1,1-difluoroethoxy substituted phenyl, methylenedioxyphenyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl or pyridyl,

R ¹⁰ for hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec. Butoxy or tert-butoxy,

R ^{11 represents} hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl,

R ^{12 represents} hydrogen,

R ¹³ for hydrogen, methyl, iso-propyl, iso-butyl, sec.-butyl, hydroxymethyl, 1-

Hydroxyethyl, mercaptomethyl, 2-methylthioethyl, 3-aminopropyl, 4-aminobutyl, carboxymethyl, 2-carboxyethyl, carbamoylmethyl, 2-carbamoyl-ethyl, 3-guanidinopropyl, phenyl, benzyl, 4-hydroxybenzyl, 3-indolyl-methyl or 4-imidazolylmethyl, and

R ^{14 represents} methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, benzyl or another C-terminal protective group.

Saturated or unsaturated hydrocarbon radicals such as alkyl or alkenyl can also be used in connection with heteroatoms, e.g. in alkoxy, where possible, be straight-chain or branched.

Optionally substituted radicals can be mono- or polysubstituted, whereby in the case of multiple substitutions the substituents can be the same or different.

In the context of the invention, hetaryl generally represents an aromatic, optionally benzo-fused 5- to 7-membered, preferably 5- to 6-membered Heterocycle, which can contain up to 3 heteroatoms from the S, N and / or O series. Examples include: indolyl, isoquinolyl, quinolyl, benzorbythiophene, benzo [b] furanyl, pyridyl, thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, imidazolyl, morpholinyl or piperidyl. Furyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, pyridyl and thienyl are preferred.

The protective groups known from peptide chemistry are listed, for example, in TW Greene, PGM Wuts, Protective Groups in Organic Synthesis, 2 ^nd Ed., John Wiley & Sons, New York 1991. Examples of suitable protective groups are C 1 -C ₄ -alkyl and benzyl in particular for hydroxyl or carboxy groups (C-terminal protective groups); Acetyl, trifluoroacetyl, trichloroacetyl, benzoyl, phenylacetyl, tert-butoxycarbonyl (Boc), benzyloxycarbonyl, (Cbz), (9-fluorenyl) methoxycarbonyl (Fmoc) and benzyl, in particular for hydroxyl and amino groups (N-terminal protective groups).

Of these, tert-butyl, benzyl, acetyl, Boc and Cbz are particularly preferred.

The use according to the invention of the preparation examples described in WO 00/08007 is very particularly preferred:

Example 1-1

(1 α, 3α, 3 aα, 8bα) -3, 3 a-dihydro-6,8-dimethoxy-3 a- (4-methoxyphenyl) -3 -phenyl- 1 H -cyclopenta [b] benzofuran-1, 8b (2H) -diol

1H-NMR (CDC1 ₃ ): δ (ppm) 2.38 (ddd, 1 H), 2.51 (s, 1 H), 2.63 (m, 1 H), 3.08 (d, 1

H), 3.46 (dd, 1 H), 3.72 (s, 3 H), 3.84 (s, 3 H), 3.86 (s, 3 H), 4.80 (dt, 1 H), 6.10 (d, 1

H), 6.27 (d, 1 H), 6.69 (d, 2 H), 6.98-7.12 (m, 5 H), 7.20 (d, 2 H).

Example 1-2

(3, 3aα, 8bα) -2,3,3a, 8b-tetrahydro-8b-hydroxy-6,8-dimethoxy-3a- (4-methoxyphenyl) -3-phenyl-cyclopenta [b] benzofuran-1 - on

1H-NMR (CDCI ₃ ): δ (ppm) 2.94-3.10 (m, 2 H), 3.10 (s, 1 H), 3.70 (s, 3 H), 3.83 (s, 3 H), 3.85 (s, 3 H), 3.89 (dd, 1 H), 6.10 (d, 1 H), 6.34 (d, 1 H), 6.68 (d, 2 H), 6.92-6.98 (m, 4 H), 7.08-7.13 ( m, 3 H). Example 1-3

(1 α, 3 ß, 3 aß, 8b ß) -3, 3 a-dihydro-6, 8-dimethoxy-3 a- (4-methoxyphenyl) -3-phenyl-1 H-cyclopenta [b] benzofuran-1 , 8b (2H) diol.

1H-NMR (CDC1 ₃ ): δ (ppm) 2.20 (dd, 1 H, J = 13.6, 6.6), 2.36 (s, 1 H), 2.75 (td, 1 H,

J = 13.8, 6.4), 3.29 (s, 1 H), 3.71 (s, 3 H), 3.84 (s, 3 H), 3.91 (s, 3 H), 4.01 (dd, 1 H, J

= 14.0, 6.5), 4.82 (d, 1 H, J = 6.3), 6.15 (d, 1 H, J = 1.9), 6.29 (s, 1 H, J = 1.9), 6.68

(d, 2H, J = 8.9), 6.98-7.19 (m, 7H).

[α] _D = -131.4 (c = 0.4 CHC1 ₃ )

Example 1-4

(lα, 3ß, 3aß, 8bß) -5,7-dibromo-3,3a-dihydro-6,8-dimethoxy-3a- (4-methoxyphenyl) -3-phenyl-1 H-cyclopenta [b] benzofuran-1 , 8b (2H) diol

1 H NMR (CDC1 ₃ ): δ (ppm) 2.23 (dd, 1 H), 2.81 (td, 1 H), 3.46 (s, 1 H), 3.72 (s, 3 H), 3.95 (s, 3 H) ), 4.02 (s, 3 H), 4.84 (d, 1 H), 6.71 (d, 2 H), 7.01-7.19 (m, 7 H). Example 1-5

(1 α, 3 ß, 3 aß, 8b ß) -N- (3, 3 a-dihydro-6, 8-dimethoxy-3 a- (4-methoxyphenyl) -3 -phenylcyclopenta [b] benzofuran-8b (2H) -ol-l-yl) -2-aminoessigsäuremethylester. 1H-ΝMR (CDC1 ₃ ): δ (ppm) 2.19 (ddd, 1 H), 2.62 (td, 1 H), 3.50 (AB, 2 H), 3.69 (s, 3 H), 3.73 (s, 3 H) ), 3.77 (dd, 1 H), 3.83 (s, 3 H), 3.88 (s, 3 H), 4.11 (dd, 1 H), 6.11 (d, 1 H), 6.24 (d, 1 H), 6.64 (d, 2H), 7.04-7.27 (m, 9H).

Example 1-6

(I-6a) (I-6b) trans isomer: (lα, 3ß, 3aα, 8bα) -3,3a-dihydro-6,8-diethoxy-3a- (4-methoxyphenyl) -3- phenyl-lH- cyclopenta [b] benzofuran-1,8b (2H) -diol (I-6b):

1H-NMR (CDC1 ₃ ): δ (ppm) 1.40 (m, 6 H), 2.12 (dt 1 H), 2.23 (dd, 1 H), 2.39 (s, 1 H), 3.81 (s, 3 H) , 3.99 (q, 2 H), 4.04 (q, 2 H), 4.26 (dd, 1 H), 4.56 (d, 1 H), 6.01 (d, 1 H), 6.07 (d, 1 H), 6.90 (d, 2 H), 7.02 (m, 2 H), 7.18 (m, 3 H), 7.42 (d, 2 H), cis isomer: (lα, 3α, 3aα, 8bα) -3,3a-dihydro-6,8-diethoxy-3a- (4-methoxyphenyl) -3-phenyl-1H-cyclopenta (] benzofuran-1, 8b (2H ) diol (I-6a):

1H-NMR (CDC1 ₃ ): δ (ppm) 1.44 (m, 6 H), 2.48 (ddd, 1 H), 2.62 (m, 1 H), 3.47 (m,

1 H), 3.71 (s, 3 H), 4.07 (m, 4 H), 4.80 (t, 1 H), 6.09 (d, 1 H), 6.23 (d, 1 H), 6.70 (d, 2

H), 6.99-7.12 (m, 5 H), 7.19 (d, 2 H).

Example 1-7

(3α, 3aα, 8bα) -8b-hydroxy-6,8-diethoxy-3a- (4-methoxyphenyl) -3-phenyl-2,3,3a, 8b-tetrahydrocyclopenta- [bjbenzofuran-1-one

Example 1-8

(lα, 3ß, 3aß, 8bß) -3,3a-dihydro-6,8-diethoxy-3a- (4-methoxyphenyl) -3-phenyl-lH-cyclopenta [b] benzofuran-1,8b (2H) diol 1H-NMR (CDCI ₃ ): δ (ppm) 1.48 (m, 6 H), 2.11 (dd, 1 H), 2.67 (dt, 1 H), 3.65 (s, 3 H), 3.92 (m, 1 H) ), 3.98 (m, 2 H), 4.08 (m, 1 H), 4.78 (d, 1 H), 6.06 (d, 1 H), 6.19 (d, 1 H), 6.61 (d, 2 H), 6.91-7.07 (m, 7 h).

Examples 1-9 to 1-79

Table 2

t

t

t

I

t

O

*): The diastereomer pairs (3S *; 3R *) were only separated at this stage

Table 2 (continued)

Table 2 (continued)

©

Table 2 (continued)

w

Table 2 (continued)

I to

Table 2 (continued)

Table 2 (continued)

Table 2 (continued)

Table 2 (continued)

Table 2 (continued)

1

Table 2 (continued)

O

Table 2 (continued)

VO

Table 2 (continued)

I

©

Table 2 (continued)

Table 2 (continued)

t

Table 2 (continued)

The compounds of formula (I) used according to the invention are described in WO 00/08007. For the preparation of the compounds of formula (I) used according to the invention, reference is expressly made to the disclosure of WO 00/08007.

Parkinson's disease, idiopathic, is a chronic, progressive neurological disorder that belongs to a broader classification of neurological disorders, known as Parkinsonism. It is clinically defined by the appearance of at least two of the four cardinal symptoms: bradykinesia, resting tremor, muscle stiffness and postural and movement disorders. The idiopathic form of Parkinson's disease is pathologically characterized by the loss of pigmented nerve cells, particularly in the area of the substantia nigra of the brain. Idiopathic Parkinson's disease accounts for approximately 75% of all Parkinsonism diseases. The remaining 25% of cases are called atypical park sonism and include diseases such as multiple system atrophy,

Striatonigral degeneration or vascular parkinsonism.

The present invention also includes pharmaceutical preparations which, in addition to inert, non-toxic, pharmaceutically suitable auxiliaries and excipients, contain one or more compounds of the general formula (I) or which consist of one or more compounds of the formula (I), and processes for the preparation of these preparations.

The compounds of the formula (I) should be present in these preparations in a concentration of 0.1 to 99.5% by weight, preferably 0.5 to 95% by weight, of the total mixture.

In addition to the compounds of formula (I), the pharmaceutical preparations can also contain other active pharmaceutical ingredients. The pharmaceutical preparations listed above can be prepared in a customary manner by known methods, for example using the excipient or excipients.

The new active compounds can be converted in a known manner into the customary formulations, such as tablets, dragées, pills, granules, aerosols, syrups, emulsions, suspensions and solutions, using inert, non-toxic, pharmaceutically suitable excipients or solvents. Here, the therapeutically active compound should in each case be present in a concentration of about 0.5 to 90% by weight of the total mixture, i.e. in amounts sufficient to achieve the dosage range indicated.

The formulations are prepared, for example, by stretching the active ingredients with solvents and / or carriers, optionally using emulsifiers and / or dispersants, e.g. in the case of the use of water as a diluent, organic solvents can optionally be used as auxiliary solvents.

The application is carried out in the usual way, preferably orally, transdermally or parenterally, in particular perlingually or intravenously. It can also be inhaled over

Mouth or nose, for example using a spray, or topically on the skin.

In general, it has proven to be advantageous to administer amounts of approximately 0.001 to 10 mg kg, with oral administration preferably approximately 0.005 to 3 mg / kg body weight in order to achieve effective results.

Nevertheless, it may be necessary to deviate from the amounts mentioned, depending on the body weight or the type of route of administration, on the individual's behavior towards the medication, the type of medication

Formulation and the time or interval at which the administration takes place. In some cases it may be sufficient to make do with less than the aforementioned minimum quantity, while in other cases the above upper limit must be exceeded. In the case of application of larger quantities, it may be advisable to distribute them in several individual doses over the day.

The NF-κB inhibitory activity of the compounds according to the invention can be determined in vitro as follows:

1. Inhibition of NF-κB-mediated gene expression of the tumor necrosis factor-α (TNFα) gene in human monocytes

The promoter of the human TNFα gene contains three NF-κB binding sequences, which are designated as kl, k2 and k3. The NF-κB binding sequences are found in the promoter of the TNFα gene at the nucleotide positions kl = -587 to -577, k2 = -210 to -202 and k3 = -98 to -87 and these DNA sequences bind NF-κB specifically ( AE

Goldfield et al., Proc. Natl. Acad. Scri. USA 87, 9769-9773, 1990). Lipopolysaccharide or phorbol ester (such as phorbol myristate acetate) induce NF-κB release (Lenardo, M.J., et al., Cell 58, 227-229, 1989).

The following biological test is therefore carried out to detect the inhibition of NF-κB-mediated TNFα gene expression by the substances described here.

Donor blood mononuclear cells are isolated with Vacutainer CPT ™ tubes (Becton Dickinson and Company, Franklin Lakes, NJ. 07417-1885) according to the manufacturer's instructions. The Vacutainer CPT tubes contain 1.0 ml of phosphate-buffered saline with 120 USP units of sodium heparin over 3.0 g of a polyester gel that covers 2.0 ml of a Ficoll solution. After the donor blood has been centrifuged, the monocytes are taken from a zone above the polyester gel and sown for cell culture with a cell density of 250 × 10 ⁵ cells per well and 96-well microtiter plates. The cells are incubated for 4 to 6 hours in medium RPMI-1640 (Gibco BRL, Life Technologies GmbH, Dieselstrasse 5, 76344 Eggenstein). The culture supernatant is then aspirated off, medium RPMI-1640 is again added and the substances to be tested are usually in concentrations between 0 μM

(Negative control) and 20 μM added. Bacterial lipopolysaccharide (LPS), Sigma-Aldrich Chemie GmbH, Grünwalder Weg 30, 82039 Deisenhofen, order no .: L 4391, in a concentration of 125 ng / ml to stimulate NF-B- mediated TNFα gene expression added. After a further incubation of 18 hours at 37 ° C in a 5% CO ₂ atmosphere, the

Microtiter plate culture supernatant is taken and the TNFα content therein is determined quantitatively using commercially available enzyme-linked immunosorbent assays (ELISA).

TNFα ELISA for concentration determination are e.g. from Sigma-Aldrich

Chemie GmbH, Grünwalder Weg 30, 82039 Deisenhofen under the name Human TNFα ELISA Kit, order no .: CKH-200A. In accordance with the manufacturer's instructions for use, the concentration of the TNFα formed in the culture supernatant of the monocyte culture after LPS stimulation is determined quantitatively with and without inhibitor substance.

The results of the TNFα concentration determination are plotted against each other in an xy-axis diagram. The graph of the y-coordinates (TNFα concentration in the culture supernatant) and the x-coordinates (concentration of the inhibitor used) enables the inhibition of NF-B-mediated TNFα synthesis to be read as a function of the concentration of the inhibitor substance. In this way, the active substance concentration of the inhibitor added which, for example, inhibits TNFα synthesis by 50% can be read off from the diagram. This concentration of active substance, which causes 50% inhibition, is called the effective inhibitor concentration for 50% inhibition (IC ₅₀ ). The following compounds, for example, showed that they are potent inhibitors of NF-κB-mediated TNFα synthesis:

2. Inhibition of NF-κB-mediated gene expression of the human tissue factor gene in monocytes

Tissue factor is a membrane-bound protein that is the primary initiator of the blood coagulation cascade and a key function in cardiovascular diseases such as unstable angina pectoris, acute implications after plaque rupture, vascular occlusions of various etiology, arteriosclerotic processes and other diseases such as septic shock or cancer. The tissue factor gene is particularly induced in monocytes and endothelial cells by NF-κB activation. The promoter of the human tissue factor gene contains an NF-κB binding site which makes a decisive contribution to activating the promoter (P. Oeth et al. Arteriosclerosis, Thrombosis, and Vascular Biology 17, 365-374, 1997).

The following biological test was therefore carried out to further demonstrate the inhibition of NF-κB-mediated gene expression by the inhibitors according to the invention:

The promoter fragment of the human tissue factor gene, which contains the NF-κB binding sequence, was with oligonucleotide primers of the sequence 5'-TCC CTC GAG ATC TCC CAG AGG CAA ACT GCC AGA T-3 '(5' primer of the position -925) and

5'-TCC TCG AGC CAT GGC TAG CAG TTG GGC GGC GAG ATC-3 '(3' primer containing the ATG start codon of the coding sequence of the tissue factor gene) cloned by the polymerase chain reaction (PCR) and fused by Ncol / Xhol cloning with the luciferase start codon in the plasmid pGL3-Basic Vector (Promega Corp. 2800 Woods Hollow Road, Madison, WI 53711-5399 USA). This regulates the expression of the luciferase in the resulting recombinant plasmid by the human tissue factor promoter. This expression construct was subjected to the analysis of DNA sequencing and transfected into the monocyte cell line RAW 264.7 (American Type Culture Collection 12301 Parklane Drive, Rockville, Maryland 20852, USA). The transfection, selection and clone analysis were carried out according to standard methods as described (see Transfection of Mammalian Cells in Culture, LG Davis et al. Basic Methods in

Molecular Biology, Elsevier Sei. Publishing Co., New York 1986). After the selection of RAW clones, which had integrated the expression construct stably in the genome, one of these transfectants, called RAW-A3, was selected for the test of the inhibitors.

Test procedure:

106 RAW-A3 cells are sown in each well in 12-well microtiter plates. In three days, the serum concentration in the RPMI medium is gradually reduced from 10% total calf serum to 0.5% and 0.1% serum content in order to reduce the serum-dependent tissue factor promoter activation to a minimum.

After 24 hours of culture in medium with 0.1% serum, the NF-κB inhibitor is added and then serum is added up to a concentration of 15% in the medium for promoter induction.

After a further 6 hours, the culture supernatant is aspirated and the cells are used to measure the luciferase activity in accordance with the "Luciferase Assay System" (Technical Bulletin from Promega, 2800 Woods Hollow Road, Madison, WI 53711-5399 USA, products E4030 , E1483, El 501) The cell lysate is incubated with the luciferase assay substrate and measured in a luminometer to measure the emitted light, when plotted in an xy-axis diagram with the respective emitted light (specified in relative light units) as The y coordinate and the inhibitor concentrations as x coordinates result in a graph for f (x), in which the half-maximum y value is to be assigned to an inhibitor concentration x which corresponds to the inhibitory concentration IC ₅₀ in this test.

For example, the IC ₅₀ value of the compound according to preparation example 1-1 is 20 μM and 1-5 2 μM. The NF-κB-mediated expression of luciferase by the inhibitors according to the invention with a concentration in the lower micromolar or in the sub-micromolar range indicates the high effectiveness of the inhibition of NF-κB-mediated gene expression.

3. Inhibition of NF-κB-mediated gene expression of the tumor necrosis factor-α (TNFα) gene in human monocytes as a function of three different TNFα synthesis stimuli

The NF-κB-mediated induction of TNFα synthesis is independent of the different stimuli used by LPS, opsonized zymosan or phorbol myristate acetate (PMA) for the activation of the TNFα promoter (A. Baldwin, Annual Rev. Immunology 14, 649, 1996). The use of the inhibitors according to the invention should therefore also result in a comparatively strong inhibition of the

TNFα synthesis perform regardless of the type of stimulation of TNFα synthesis.

The tests for this detection of the stimulus-independent inhibition of TNFα synthesis were carried out in the same way as in the example of the test procedure

A is described with the difference that in addition to LPS as a stimulus, cell culture batches with 100 nM PMA or with 100 μg / ml opsonized zymosan were also stimulated. Zymosan and phorbol myristate acetate (PMA) can be obtained from Sigma, Grünwalder Weg 30, 82041 Deisenhofen, Germany under order no. Z4250 and P8139 are available. Zymosan is opsonized in human serum. The NF-κB inhibitors according to the invention inhibit TNFα synthesis in human monocytes in a comparable manner, independently of the stimulus with IC ₅₀ values in the submicromolar range, as was shown in Example A. The IC50 value according to PMA stimulus is 0.10 μM for the connection according to preparation example II and 0.09 μM for connection 1-5.

4. Inhibition of NF-κB Mediated Gene Expression of the Adhesion Protein ELAM-1 on Human Umbilical Cord Endothelial Cells (HUVEC)

The recruitment of leukocytes from the blood circulation into the extravascular space is essential for inflammatory responses and for repairing tissue damage. The process of leukocyte immigration involves several steps in series. The initial interaction between leukocytes and the endothelium of the blood vessels is determined by TNF and II-1 dependent expression of the

Adhesion protein ELAM-1 mediated on the endothelium. It mediates the so-called rolling of the leukocytes along the blood vessel wall. The transcriptional regulation of ELAM-1 expression is dependent both on the nuclear factor-κB (NF-κB) activation and binding to the ELAM-1 promoter and on the “AP-1 binding site [MA. Read et., Biol. Chem. Vol. 272, 2753-2761, (1997)].

The influence of compound 1-3 -R on the expression of ELAM-1 was checked in two different test approaches. The adhesion of human neutrophils to TNF-α stimulated HUVEC cells was measured in a functional approach. The expression of ELAM-1 on the surface of the HUVECs was determined using a fluorescence-labeled ELAM-1-specific monoclonal antibody by means of FACS (cell sorting) analysis.

Carrying out the experiment: ELAM-dependent neutrophil adhesion to endothelial cells The neutrophils were isolated from human blood (100 ml). For this purpose, 3.5 ml of polyprep was placed in a centrifuge beaker and carefully overlaid with 5 ml of blood. After centrifuging at 2 100 min ^{-1 for} 30 minutes, the neutrophil band was sucked off in the middle of the centrifuge cup. After 1: 2 dilutions in Endothelial Cell Basal Medium (EBM) from Clonetics was again

Centrifuged at 1000 min ^{-1 for} 20 minutes and then taken up to a cell concentration of 10 ⁶ cells / ml.

The umbilical cord endothelial cells were grown to confluence in 96-well microtiter plates in EBM + 10% FCS. At the start of the experiment, the medium was exchanged for EBM without FCS and the test substances were then used. After 20 minutes, the HUVECs were stimulated with 10 nM TNF-α. After 4 hours of incubation, 200 μl / well of the neutrophil suspension was exchanged. The neutrophils were previously labeled with a 25 μM BCECP phosphor fluorescent dye solution for 20 minutes. After 30 minutes of incubation, the BCECP neutrophil

The solution was drawn off with excess neutrophils and exchanged for 200 μl / well of 0.5% NaOH solution. The fluorescence of the adhering neutrophils was then measured in the fluorescence photometer.

Both the adhesion of the neutrophils to TNF-α stimulated HUVECs and that

Expression of ELAM-1 on the cell surface could be inhibited by the compound by 50%. The maximum inhibition of cell adhesion was determined to be 50 nM for compound I-3-R. The maximum 50% inhibition of ELAM-1 expression is in good agreement with the simultaneous NF-B and AP-1-dependent regulation of the ELAM-1 promoter.

Carrying out the experiment Quantitative measurement of the TNF-induced expression of ELAM-1 in HUVECs

Umbilical cord endothelial cells (HUVEC) were cultured as described above and in the presence or absence of 10 ng / ml TNF in the presence or absence incubated by test substances for 4 hours. The cells were dissolved out of the microtiter plates by incubation with 5 mM EDTA in PBS, centrifuged for 5 minutes at 1000 min ⁻¹ , and in 100 μl PBS plus 1% bovine serum albumin (BSA, Sigma-Aldrich GmbH, Best No. A 7906) and an anti-ELAM 1 antibody (monoclonal antibody, Becton and Dickinson, Erembodegem, Belgium, order no.

No. 550023, 30 μg / ml) at RT. After 15 minutes the cells were centrifuged again, the supernatant was discarded and the cells were washed in PBS plus 1% BSA. After renewed centrifugation, the cell pellet obtained was placed in PBS plus 1% BSA and a goat anti-mouse antibody (Dianova, Raboisen 5, Hamburg; order no. 115-096-062; 30 μg / ml) for a period of 15 Minutes incubated. After centrifuging and washing again, the cells were taken up in 1 ml of PBS plus 1% BSA and measured in a flow cytometer (from Becton and Dickinson) at 488 nm. This method measures the intensity of the fluorescence as a function of the bound anti-ELAM-1 antibody per cell. 5,000 cells were measured for each value.

HUVECs stimulated with TNF for 4 hours showed significantly stronger fluorescence signals after incubation with anti-ELAM-1 antibodies than cells, on the other hand, which were not incubated with TNF. The compound from preparation example 1-3 -R was able to significantly inhibit this induced expression of ELAM-1 in a concentration range between 0.05 μM and 5 μM, but the expression of ELAM 1 was not completely inhibited at any of the concentrations examined.

5. Inhibition of interleukin-2 synthesis

To test the inhibitory effect of cyclopentabenzene derivatives on the interleukin

2 synthesis, a reporter gene cell line was used which contains the interleukin-2 promoter coupled to the luciferase gene. The promoter contains the DNA sequence from

-480 to +4. The vector used is pGL3; the starting cell line into which the entire construct was stably transfected is SS-1. The culture medium for this cell line was RPMI 1640 (Gibco, Rockille). It also contained: 100 μg / ml streptomycin, 100 U / ml penicillin, 2 mM L-glutamine, 10% heat-inactivated FBS and 800 μg / ml G418 sulfate.

Test procedure

The reporter gene test cell line was seeded in phenol red-free RPMI with the additions of 1 × 10 ⁶ cells per well in 96-well plates and with phorbol 12-myristat 13-acetate (PMA; 5 ng / ml) and ionomycin (10 M ; 0.4 μg / ml) for 24 hours

Incubated 37 ° C in an atmosphere of 5% CO2, 95% air. The test substances were added simultaneously with PMA.

Measurement of luciferase activity:

To generate the luminescence, LucLite ™ solution (Packard, Meriden, CT) was added to a concentration of 100 μl / well and the luminescence was measured in the luminometer (Luminoskan, Labsystems) immediately after the addition.

The compound from Preparation 1-3 -R was able to induce the

6. Inhibition of interleukin-8 release from human endothelial cells

Cyclopentanbenzofurans were investigated for their inhibitory effect on the interleukin-lß (IL-lß) stimulated release of interleukin-8 (IL-8) from human umbilical cord endothelial cells (HUVEC). IL-1β induced IL-8 formation is mediated by an NF-κB dependent signal transduction pathway.

HUVEC P-115 endothelial cells were manufactured by CellSystems (St. Katharinen,

Germany) and cultivated according to the supplier's instructions. 5-7 days Before carrying out the experiment, the cells were distributed on 96-well microtiter plates with a density of 5000 cells per well. 16 hours before the addition of the test substances and stimulus (IL-1ß), the cell culture medium was replaced by 170 μl of fresh medium (EBM2 medium, Cell Systems). Then 20 μl of the test substance was added in the desired concentrations and the IL-8 synthesis and release was induced by adding 10 μl IL-lß (final concentration 10 ng / ml; Biosource GmbH, Ratingen, Germany). After 6 hours, 150 μl of the cell culture supernatant were removed and the IL-8 concentration released was determined using an IL-8 ELISA (Biosource GmbH, Ratingen, Germany). The ELISA was operated according to the manufacturer's instructions.

For example, the compound from Preparation 1-40 was able to inhibit the release of IL-8 by 50% (IC50) at a concentration of 250 to 300 nM.

EXAMPLES

Example 1: MPP ⁺ toxicity in primary rat dopaminergic neurons

MPTP (= l-methyl-4-phenyl-l, 2,3,6-tetrahydropyridine) is a neurotoxin that

Humans and animals that cause the degeneration of dopaminergic neurons in the substantia nigra characteristic of Parkinson's syndrome and the motor symptoms typical of Parkinsonism. In the brain, MPTP is metabolized to MPP ⁺ (l-methyl-4-phenylpyridinium), which is taken up into dopaminergic neurons via the dopamine transporter and exerts its toxic effect there. In cultured dopaminergic neurons, the addition of MPP ^{+ is} similarly toxic.

Dopaminergic neurons were obtained from brains of 15-day-old rat embryos. For this purpose, the ventral parts of the midbrain were dissected out, the tissue was enzymatically dissociated and the cell suspension obtained with a density of 80,000

Cells / well distributed on 96 well microtiter plates. The cells were cultivated for 7 days in cell culture medium consisting of DMEM / Ham's F12, 0.5% BSA, 6 mg / ml glucose, 2 mM L-glutamine and N2 additives. Various concentrations of the test substance (Example 1-41) were then added and the cell death of dopaminergic neurons was induced by adding 1 μM MPP ⁺ . The functionality and survival of dopaminergic neurons was determined after 24 hours by measuring the dopamine uptake. For this purpose, the cell culture medium was replaced by a buffered saline solution (25 mM HEPES, 125 mM NaCl, 4.8 mM KC1, 1.2 mM KH ₂ PO, 1.3 mM CaCl ₂ , 1.2 mM MgCl ₂ , 5.6 mM glucose) and the dopamine intake by adding radiolabeled dopamine started. After 30 minutes of incubation at 37 ° C., the buffer was removed and the cells were washed with cold buffer. Finally, the radioactivity remaining in the cells (= amount of dopamine taken up) was determined.

MPP ⁺ treatment of the cells led to a 35% reduction in dopamine uptake, synonymous with a significant loss of dopaminergic function Neurons. Concentrations of the test substance> 10 nM significantly increased the dopamine uptake of MPP ⁺ -treated dopaminergic neurons (FIG. 1). These results show that the test substance has neuroprotective properties in vitro.

Fig. 1: Primary dopaminergic neurons were treated with MPP ⁺ in the presence and absence of various concentrations of the test substance for 24 hours. The functionality of the dopaminergic neurons was then determined by measuring the dopamine uptake. The dopamine uptake of untreated cells (Ko) was set equal to 100% and all other values were calculated in relation. * p <0.05,

Comparison of substance-treated with MPP ⁺ -treated cells using the Students' T test; SD, standard deviation.

Example 2: MPTP mouse model

The neuroprotective in vivo effect of the compounds according to the invention was demonstrated in a mouse model for Parkinson's syndrome, the so-called MPTP model.

The mice were given 3 mg / kg MPTP i.p. on 3 consecutive days. applied (Bezard et al. Neurosci. Lett. 1997, 234, 47-50). The experimental animals then show a reduced number of dopaminergic neurons in the substantia nigra pars compacta.

The extent of cell damage is quantified by a histological examination on the tenth day after MPTP injection. Dopaminergic neurons are made immunohistochemically visible as cells that contain the enzyme tyrosine hydroxylase, which is essential in dopamine metabolism. The remaining number of dopaminergic neurons is quantified using a computer program (Nelson et al., J. Comp. Neurol. 1996, 369, 361-371). Mice that received Example 1-41 at a dose of 3 mg / kg bid ip for 10 days from the first day of MPTP intoxication had significantly more dopaminergic neurons in the substantia nigra pars compacta than control animals that had only received MPTP (Fig. 2).

Fig. 2: Mice were administered 4 mg / kg MPTP for three days. The test substance (Subst, 3 mg / kg) was administered i.p. twice a day starting with the first MPTP administration over a period of 10 days. injected. The number of dopaminergic neurons in the substantia nigra was then quantified. The number of tyrosine hydroxylase immunoreactive neurons (TH-LR neurons) in control animals (Ko) became the same

100% set and the remaining values calculated in relation to it. * p <0.01 (Fisher's LSD test)

Claims

Patentansprflche

1. Use of compounds of the formula (I)

in which

R ¹ and R ³ each independently represent hydrogen, halogen or alkyl,

R ^{12 represents} hydrogen or alkyl,

R ¹² and R ¹³ together represent - (CH ₂ ) ₃ - and - (CH ₂ ) ₄ -, and R ^{14 represents} alkyl, benzyl or another C-terminal protective group,

R ^{6 represents} hydrogen or

R ⁵ and R ⁶ together represent oxygen (oxo),

R ⁷ and R ⁸ each represent hydrogen,

R ^{9 represents} optionally substituted phenyl or hetaryl,

R ^{10 represents} hydrogen, halogen, alkyl or alkoxy and

R ^{u represents} hydrogen, halogen or alkyl,

and their salts for the manufacture of a medicament prophylaxis and / or treatment of Parkinson's disease.

Use according to claim 1 of compounds of the formula (I),

embedded image in which

R ¹ and R ³ each independently represent hydrogen, fluorine, chlorine, bromine or -Ce alkyl,

R ² and R ⁴ each independently represent fluorine, chlorine, bromine, Ci-Cβ-alkyl or C ₁ -C ₄ alkoxy optionally substituted by fluorine or chlorine,

R ⁵ for hydroxy, CC ₄ - alkylamino or for the radical -NR -CHR ¹

COOR ¹⁴ stands, R »6 represents hydrogen, or

R> 5 and R together represent oxygen (oxo),

R ⁷ and R ⁸ each represent hydrogen,

R ^{9 is} phenyl, methylenedioxyphenyl or 5- or 6-membered

Hetaryl with 1 or 2 heteroatoms from the series nitrogen, oxygen or sulfur, each of which can optionally be up to four times substituted by substituents from the group: halogen, Ci-Cg-alkyl, hydroxy, Ci-C ^ alkoxy, Cj-C4 -Alkylthio, -C-C4-alkylcarbonyl, phenyl, phenoxy, hetaryl, hetaryloxy, substituted by fluorine or chlorine -CC4-alkyl, substituted by fluorine or chlorine

C 1 -C 4 -alkoxy, C 1 -C 4 -alkylthio substituted by fluorine or chlorine, C 1 -C 4 -alkylcarbonyl substituted by fluorine or chlorine,

R ^{10 represents} hydrogen, fluorine, chlorine, d-Cβ-alkyl or d-Ce-alkoxy,

R ^{11 represents} hydrogen, fluorine, chlorine, bromine or -Ce alkyl,

R ^{12 represents} hydrogen or -Gralkyl,

R ¹³ for hydrogen, optionally substituted by amino or hydroxy (CrC ^ alkyl or for mercaptomethyl, methylthioethyl, carboxymethyl, carboxyethyl, carbamoylmethyl, carbamoylethyl, guanidinopropyl or for optionally substituted by amino, nitro, halogen, hydroxy or methoxy tuated phenyl or benzyl or for naphthylmethyl, indolylmethyl, imidazolyethyl, triazolyethyl or pyridylmethyl, where functional groups can optionally be protected, or

R ¹² and R ¹³ together represent - (CH) ₃ - or - (CH ₂ ) ₄ -,

R ¹⁴ for (Ci-C _ö ) - alkyl, benzyl or another C-terminal

Protection group stands,

and their salts.

3. Use according to claim 1 of compounds of formula (I),

wherein

R ¹ and R ³ each independently represent hydrogen, fluorine, chlorine, bromine, methyl or ethyl,

R ² and R ⁴ independently of one another each for fluorine, chlorine, bromine, C _\ -

C ₄ alkyl or methoxy or ethoxy which is optionally substituted by fluorine or chlorine,

R ^{5 represents} hydroxy, dC ₄ - alkylamino or the radical -NR ¹² - CHR ¹³ -COOR ¹⁴ ,

R ^{6 represents} hydrogen, or

R ⁵ and R ⁶ together represent oxygen (oxo),

R ⁷ and R ⁸ each represent hydrogen, R ^{9 is} phenyl, methylenedioxyphenyl or 5- or 6-membered

Hetaryl with 1 or 2 heteroatoms from the nitrogen series,

Oxygen and sulfur, each of which may optionally be substituted up to 3 times by substituents on the

Group: fluorine, chlorine, bromine, iodine, C 1 -C ₄ -alkyl, hydroxy, C _r C ₄ -alkoxy, C _r C ₄ -alkylthio, C _r C4-alkylcarbonyl, phenyl, phenoxy, furyl, thienyl, pyrrolyl, thiazolyl, Pyridyl, furyloxy, thienyloxy, pyrrolyloxy, thiazolyloxy, pyridyloxy, C ₁ -C -alkyl substituted by fluorine or chlorine, Cj-C _ß- alkoxy substituted by fluorine or chlorine, Ci-Cß-alkylthio substituted by fluorine or chlorine, by fluorine or Chlorine substituted C _j -C4 alkylcarbonyl,

R ¹⁰ ₄ alkyl or Cι-C ₄ -alkoxy represents hydrogen, fluorine, C ₁ -C

R ^{11 represents} hydrogen, fluorine, chlorine, bromine or C 1 -C ₄ alkyl,

R ^{12 represents} hydrogen or methyl,

R ¹³ for hydrogen, methyl, isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl, 3-aminopropyl, 4-aminobutyl, carboxymethyl, 2-car- boxyethyl, carbamoylmethyl, 2-carbamoylethyl, 3-guanidinopropyl, phenyl, benzyl, 4-hydroxybenzyl, 4-methoxybenzyl, 2-nitrobenzyl, 3-nitrobenzyl, 4-nitrobenzyl, 2-amino-benzyl, 3-aminobenzyl, 4-aminobenzyl, 3,4-dichlorobenzyl, 4-iodobenzyl, α-naphthylmethyl, β-naphthylmethyl 3-indolylmethyl, 4-imidazolylmethyl, 1,2,3-triazol-l-yl-methyl, 1,2,4 - Triazol-1-yl-methyl, 2-pyridylmethyl or 4-pyridylmethyl stands, where functional groups can optionally be protected, or

R ¹² and R ¹³ together represent - (CH ₂ ) ₃ - or - (CH ₂ ) ₄ -, and

R ^{14 represents} C ₁ -C ₄ alkyl, benzyl or another C-terminal protective group,

and their salts.

4. Use according to claim 1 of compounds of formula (I),

wherein

R ² and R ⁴ each represent methoxy, ethoxy, trifluoromethoxy, difluoromethoxy, chlorodifluoromethoxy, 2-chloro-l, l, 2-trifluoroethoxy, 1,1,2-trifluoroethoxy, 1,1,2,2-tetrafluoroethoxy, 2.2 , 2-trichloro-

1,1-difluoromethoxy or 1,1-difluoroethoxy,

R ^{6 represents} hydrogen, or

R ⁵ and R ⁶ together represent oxygen (oxo), R and R each represent hydrogen,

R ⁹ for phenyl, methylenedioxyphenyl, furyl, thienyl, pyrrolyl,

Oxazolyl, isoxazolyl, thiazolyl or pyridyl, each of which may optionally be mono- or disubstituted by substituents from the group fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec.-butyl , tert-butyl, trifluoromethyl, difluoromethyl, chlorodifluoromethyl, 2-chloro-l, l, 2-trifluoroethyl, 1,1,2-trifluoroethyl, 1,1,2,2-tetrafluoroethyl, l, l-difluoro- 2,2,2-trichloroethyl, 2,2,2-

Trifluoroethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3-tetrafluoropropyl, 2,2,3,3,3-pentafluoropropyl, hydroxy, methoxy, ethoxy, n-propoxy, Isopropoxy, n-butoxy, isobutoxy, sec.-butoxy, tert.-butoxy, trifluoromethoxy, difluoromethoxy, chlorodifluoromethoxy, 2-chloro-l, l, 2-trifluoroethoxy, 1,1,2-

Trifluoroethoxy, 1,1,2,2-tetrafluoroethoxy, 2,2,2-trichloro-l, l-difluoromethoxy, 1,1-difluoroethoxy, methylthio, ethylthio, trifluoromethylthio, 1,1-difluoroethylthio, 2,2,2- Trifluoroethylthio, phenyl, phenoxy, furyl, thienyl, pyrrolyl, thiazolyl, pyridyl, furyloxy, thienyloxy, pyrrolyloxy, thiazolyloxy, pyridyloxy, acetyl, propionyl, propylcarbonyl, butylcarbonyl or 2-methylpropylcarbonyl,

R ¹⁰ for hydrogen, fluorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec.-butyl, tert.-butyl, methoxy, ethoxy, n-

Propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy,

R ^{11 represents} hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, R ^{12 represents} hydrogen,

R ¹³ for hydrogen, methyl, isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl, 3-aminopropyl, 4-aminobutyl, carboxymethyl, 2-car- boxyethyl, carbamoylmethyl, 2-carbamoylethyl, 3-guanidino-propyl, phenyl, benzyl, 4-hydroxybenzyl, 3-indolylmethyl or 4-imidazolylmethyl, and

R ¹⁴ for methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec.-

Butyl, tert-butyl or benzyl,

and their salts.

5. Use of compounds according to one of claims 1 to 4 for the manufacture of a medicament for the neuroprotective treatment of Parkinson's disease.

6. Medicament containing a compound according to any one of claims 1 to 4 for the prophylaxis and / or treatment of Parkinson's disease.

7. Medicament containing a compound according to any one of claims 1 to 4 for the neuroprotective treatment of Parkinson's disease.

8. Medicament according to claim 6 or 7, wherein the compound is present in admixture with at least one pharmaceutically acceptable, essentially non-toxic carrier or excipient.