WO2000043505A2 - Protease - Google Patents
Protease Download PDFInfo
- Publication number
- WO2000043505A2 WO2000043505A2 PCT/EP2000/000390 EP0000390W WO0043505A2 WO 2000043505 A2 WO2000043505 A2 WO 2000043505A2 EP 0000390 W EP0000390 W EP 0000390W WO 0043505 A2 WO0043505 A2 WO 0043505A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- protease
- aspartate
- medicament
- diagnostic agent
- proteases
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6478—Aspartic endopeptidases (3.4.23)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
Definitions
- the present invention relates to a protease encoding nucleic acids for the protease and the inhibitors, antibodies, medicaments and diagnostic agents associated therewith.
- the present invention provides a protease with two aspartate residues in a catalytically active structure, wherein a first aspartate residue is in a motif X1GX2GD and a second aspartate residue is in a motif X 3 X 4 DX 5 , wherein Xi, X 2 , X3 and X 5 are independent are selected from each other from Ala, Val, Leu, Met and Ile and X 4 is an aromatic amino acid, and the motifs X ⁇ GX 2 GD and X 3 X 4 DX 5 are located in a transmembrane region.
- proteases are very likely to be involved in the cleavage of the amyloid precursor protein (APP).
- the protease according to the invention represents the hitherto unidentified ⁇ -secretase which is involved in processing the APP to form the amyloid peptides referred to as Aß.
- Preferred proteases of the present invention additionally have a sequence PALX6YX7V, where X 6 and X 7 independently of one another have the same meaning as Xi. However, it is preferred that X 6 and X 7 are leucine or isoleucine.
- the proteases are proteases from mammals, in particular from humans.
- the proteases according to the invention preferably have catalytically active aspartate residues in a region which lies within a transmembrane region.
- Transmembrane regions can be predicted on the basis of different models if the sequence of a protein is known. They are characterized in that in one area there are predominantly hydrophobic amino acids which are flanked by areas in which there are more hydrophilic amino acids.
- An aspartate in a transmembrane region can be detected, for example, by using the "GREASE" program, which is part of the FASTA 2.0 program package. With a window width of 17, this program must be used to calculate a hydrophobicity value of at least 80 for the aspartate.
- the FASTA program package is described in WR Pearson and DJ Lipman, Proc. Natl. Acad. Sci. USA 85 (1988) 2444-2448.
- the GREASE program uses the Kyte / Doolittle algorithm, described in J. Kyte and RF Doolittle, J. Mol. Biol 157: 105-132 (1982).
- proteases of the present invention are referred to as psl 1-5 (human psl 1-5: SEQ ID No. 1 to 4 + 19, murine psl 2-4: SEQ ID No. 5-7, sacc. Cerevisiae psl 3: SEQ ID No. 8, human psl2L SEQ ID No. 18).
- the invention furthermore relates to variants of the proteases according to the invention.
- Variants are proteins which are derived from the proteases according to the invention by one or more mutations, insertions and deletions, in particular by conservative exchanges. special N- or C-terminal shortened or elongated forms.
- the invention also relates to nudeic acids which code for the proteases according to the invention.
- Preferred nucleic acids according to the invention are those with SEQ ID No. 9 - 17 + 20 (human psl 1 - 5: SEQ ID No. 9 - 12 + 20, murine psl 2 - 4: SEQ ID No. 13 - 15, sacc. Cerevisiae psl 3: SEQ ID No. 16, human psl2L SEQ ID No. 17).
- Complementary nudeic acids are also part of the invention.
- the proteases according to the invention are involved in the cleavage of the APP to Aß and are thus indirectly involved in the development of, for example, Alzheimer's disease.
- the invention therefore also relates to inhibitors which inhibit the expression or the activity of the proteases.
- Such inhibitors can be identified in simple procedures.
- Corresponding inhibitors can be identified, for example, by measuring the expression or the activity of the proteases in the presence of potential inhibitors.
- Antibodies directed against the aspartate protesases are particularly suitable for measuring expression and are therefore also part of the invention.
- the aspartate proteases according to the invention are also involved in the cleavage of other transmembrane proteins, in particular the Notch receptor protein and related proteins which play a role in the development of the nervous system.
- proteolytic cleavage inside membranes is also involved in other important processes, e.g. :
- Proteolytic cleavage of the ER stress sensor protein Irel The endoplasmic reticulum has a mechanism that detects the accumulation of unfolded or incorrectly folded proteins and sends a signal to the cell nucleus. This mechanism is called "unfolded protein response" or UPR, and leads to the increased formation of proteins that facilitate folding. In mammals there are two sensor proteins (Irelalpha and Irelbeta) that react to folding defects and are then proteolytically cleaved within the membrane.
- the processes mentioned can be influenced therapeutically by using the protease or its inhibitors according to the invention.
- Zeil lines which do not express any of the proteases or nucleic acids according to the invention and preferably also contain no homologous proteases or nucleic acids are particularly suitable for this. These can be used to test the activity of the proteases or to determine inhibitors according to Example 2, preferably in accordance with the method described in Example 1. Saccharomyces cerevisiae is particularly suitable for this.
- SEQ ID No. 8 and 16 can be used to produce yeast strains by known methods which no longer contain this protein or the nucleic acid. They are therefore preferably suitable as an expression system for characterizing the aspartate proteases according to the invention or for identifying suitable inhibitors.
- yeast cell lines preferably yeast cell lines and the use of the protein with SEQ ID No. 8 as aspartate protease and the nucleic acid with SEQ ID No. 16 for the expression of a protease are therefore also the subject of the invention.
- proteases, nudeic acids, inhibitors and antibodies according to the invention can be contained in medicinal and diagnostic agents. They are particularly suitable for the treatment or diagnosis of diseases which are causally linked to the cleavage of the amyloid precursor protein, in particular Alzheimer's disease.
- the putative ⁇ -secretases are transfected stably or transiently in cos-7 cells, which additionally express SpA4CT (signal peptide fused to ßA4 followed by the APP C-terminus).
- SpA4CT signal peptide fused to ßA4 followed by the APP C-terminus.
- ⁇ -Secretase activity is recognizable in this system by the generation of a 4.6 KDa peptide or by the disappearance of the 11 KDa band of the complete SpA4CT. If the pathologically relevant ⁇ -secretase is present, both fragments should be detectable inside the cell.
- ßA4 which is generated by an endogenous plasma membrane-constant ⁇ -secretase activity, which, however, plays no role in the pathogenesis of Alzheimer's.
- the transfected cells are washed three times with cold DMEM and then harvested on ice with a cell scraper.
- the cells (approx. 5X10 6 cells) are collected by centrifugation and in 1 ml Lysis buffer (150 mM NaCl, 50 mM Tris-HCl pH 7.5, 1% NP-40, 1% Triton-x-100, 2 mM EDTA) lysed.
- the kernels are centrifuged off at 11,000 g.
- the supernatant is subjected to immunoprecipitation.
- 1 ⁇ g of the cell lysate is mixed with 2 ⁇ g / ml WO2 immunoglobulin (anti-ßA4 antibody) and shaken overhead at 4 ° C.
- the Protein-G Sepharose is consecutively two times with the buffers A, B and C (A: 150 mM NaCI, 10 mM Tris-HCl pH 7.5, 0.2% NP-40, 2 mM EDTA; B: 500 mM NaCI, 10 mM Tris-HCI pH 7.5, 0.2% NP-40, 2 mM EDTA; C: 10 mM Tris-HCI pH 7.5), mixed with 20 ⁇ l 3X sample buffer, heated to 95 ° C and the supernatant to a 12% Tris Tricine gel applied. After the gel electrophoretic size fractionation, the proteins are transferred to a PVDF membrane and subsequently detected with an anti-ßA4 antibody.
- the enzyme is coexpressed according to the above instructions in Cos-7 cells with SpA4CT.
- the cells are brought into contact with the substance to be investigated in a suitable manner (in the presence or absence of membrane-permeabilizing agents).
- the intracellularly formed ⁇ A4 is then detected as described above.
- a reduction in the amount of ⁇ A4 formed suggests that the substance is effective as a ⁇ -secretase inhibitor.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Catalysts (AREA)
- Peptides Or Proteins (AREA)
Abstract
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP00903587A EP1144603A3 (fr) | 1999-01-22 | 2000-01-19 | Protease |
AU25421/00A AU2542100A (en) | 1999-01-22 | 2000-01-19 | Protease |
CA002360585A CA2360585A1 (fr) | 1999-01-22 | 2000-01-19 | Protease |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19902550.9 | 1999-01-22 | ||
DE19902550A DE19902550A1 (de) | 1999-01-22 | 1999-01-22 | Aspartatprotease |
DE19925946 | 1999-06-08 | ||
DE19925946.1 | 1999-06-08 | ||
DE19929115.2 | 1999-06-24 | ||
DE19929115 | 1999-06-24 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2000043505A2 true WO2000043505A2 (fr) | 2000-07-27 |
WO2000043505A3 WO2000043505A3 (fr) | 2001-11-29 |
Family
ID=27218935
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2000/000390 WO2000043505A2 (fr) | 1999-01-22 | 2000-01-19 | Protease |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP1144603A3 (fr) |
AU (1) | AU2542100A (fr) |
CA (1) | CA2360585A1 (fr) |
WO (1) | WO2000043505A2 (fr) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003040684A2 (fr) * | 2001-11-07 | 2003-05-15 | Medical Research Council | Dosage |
US6887677B1 (en) | 1999-07-12 | 2005-05-03 | Trustees Of Dartmouth College | Compounds and methods for identifying compounds which inhibit a new class of aspartyl proteases |
US8609087B2 (en) | 1999-07-12 | 2013-12-17 | Trustees Of Dartmouth College | Compounds and methods for identifying compounds which inhibit a new class of aspartyl proteases |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998033916A2 (fr) * | 1997-01-31 | 1998-08-06 | Genetics Institute, Inc. | Proteines secretees et polynucleotides codant celles-ci |
WO1998040404A2 (fr) * | 1997-03-11 | 1998-09-17 | Genetics Institute, Inc. | Proteines secretees et polynucleotides codant ces dernieres |
WO1999003990A1 (fr) * | 1997-07-16 | 1999-01-28 | Human Genome Sciences, Inc. | Serie de 64 proteines humaines secretees |
WO1999033873A1 (fr) * | 1997-12-26 | 1999-07-08 | Ono Pharmaceutical Co. Ltd. | NOUVEAUX POLYPEPTIDES, ADNc CODANT CES POLYPEPTIDES ET UTILISATION DE CEUX-CI |
WO1999043693A1 (fr) * | 1998-02-26 | 1999-09-02 | Human Genome Sciences, Inc. | Proteines secretees par l'homme |
WO2000017222A1 (fr) * | 1998-09-23 | 2000-03-30 | Human Genome Sciences, Inc. | 31 proteines humaines secretees |
-
2000
- 2000-01-19 CA CA002360585A patent/CA2360585A1/fr not_active Abandoned
- 2000-01-19 EP EP00903587A patent/EP1144603A3/fr not_active Withdrawn
- 2000-01-19 AU AU25421/00A patent/AU2542100A/en not_active Abandoned
- 2000-01-19 WO PCT/EP2000/000390 patent/WO2000043505A2/fr not_active Application Discontinuation
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998033916A2 (fr) * | 1997-01-31 | 1998-08-06 | Genetics Institute, Inc. | Proteines secretees et polynucleotides codant celles-ci |
WO1998040404A2 (fr) * | 1997-03-11 | 1998-09-17 | Genetics Institute, Inc. | Proteines secretees et polynucleotides codant ces dernieres |
WO1999003990A1 (fr) * | 1997-07-16 | 1999-01-28 | Human Genome Sciences, Inc. | Serie de 64 proteines humaines secretees |
WO1999033873A1 (fr) * | 1997-12-26 | 1999-07-08 | Ono Pharmaceutical Co. Ltd. | NOUVEAUX POLYPEPTIDES, ADNc CODANT CES POLYPEPTIDES ET UTILISATION DE CEUX-CI |
WO1999043693A1 (fr) * | 1998-02-26 | 1999-09-02 | Human Genome Sciences, Inc. | Proteines secretees par l'homme |
WO2000017222A1 (fr) * | 1998-09-23 | 2000-03-30 | Human Genome Sciences, Inc. | 31 proteines humaines secretees |
Non-Patent Citations (2)
Title |
---|
DATABASE GENEMBL [Online] 1. August 1998 (1998-08-01) LAMERDIN ET AL: "FOS39554_1" XP002141740 * |
DATABASE GENEMBL [Online] 8. August 1996 (1996-08-08) HILLIER ET AL.: "zc51b02.r1 Soares senescentr fibroblasts NbHSF Homo sapiens cDNA clone IMAGE:325803" XP002157596 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6887677B1 (en) | 1999-07-12 | 2005-05-03 | Trustees Of Dartmouth College | Compounds and methods for identifying compounds which inhibit a new class of aspartyl proteases |
US8609087B2 (en) | 1999-07-12 | 2013-12-17 | Trustees Of Dartmouth College | Compounds and methods for identifying compounds which inhibit a new class of aspartyl proteases |
WO2003040684A2 (fr) * | 2001-11-07 | 2003-05-15 | Medical Research Council | Dosage |
WO2003040684A3 (fr) * | 2001-11-07 | 2003-07-03 | Medical Res Council | Dosage |
Also Published As
Publication number | Publication date |
---|---|
WO2000043505A3 (fr) | 2001-11-29 |
EP1144603A3 (fr) | 2002-02-06 |
EP1144603A2 (fr) | 2001-10-17 |
AU2542100A (en) | 2000-08-07 |
CA2360585A1 (fr) | 2000-07-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE69737754T2 (de) | Substanzen für die präsymptomatische erkennung und zielgerichtete therapie der alzheimer-krankheit beim menschen. | |
EP0679187B2 (fr) | Procede permettant d'obtenir des ectodomaines natifs, oligomeriques, glycosyles de proteines virales membranaires, leur utilisation, notamment comme vaccin contre le vih | |
DE60309888T2 (de) | Apolipoprotein l-i zur behandlung von trypanosomen-erkrankungen | |
EP0276723A2 (fr) | Précurseur protéinique de polypeptide-APC, ADN le codant et utilisation diagnostique de cet ADN et de cette protéine | |
EP1399476B1 (fr) | Utilisation de fragments solubles de cytokeratine-1 en diagnostic | |
Yamada et al. | Cathepsin B generates the most common form of amyloid A (76 residues) as a degradation product from serum amyloid A | |
Gupta et al. | Identification of a novel hydroxyproline-rich glycoprotein as the major allergen in Parthenium pollen | |
EP1161524B1 (fr) | CELLULES COEXPRIMANT UNE PROTEINE PRECURSEUR AMYLOIDE ET UNE $g(a)-SECRETASE, ET LEURS UTILISATIONS DANS DES METHODES D'ESSAI ET A DES FINS DE DIAGNOSTIC | |
EP1144603A3 (fr) | Protease | |
DE102008014880A1 (de) | Antientzündliches Polypeptid | |
DE19902550A1 (de) | Aspartatprotease | |
EP1959013B1 (fr) | Peptide inhibiteur de virus humain (VIRIP) et son utilisation | |
DE60126602T2 (de) | Neue collectine | |
DE60123074T2 (de) | Modulation von gamma-secretase aktivität | |
DE19920514A1 (de) | Methoden zur Auffindung von Proteasen, die spezifisch membrangebundene Substrate spalten | |
EP3149485B1 (fr) | Procédé de diagnostic d'une maladie médiée par la voie alterne du système du complément ou d'un risque pour celle-ci | |
Kasik et al. | A novel complementary deoxyribonucleic acid is abundantly and specifically expressed in the uterus during pregnancy | |
WO1999010480A2 (fr) | Nouvelles calpaines specifiques au tissu, leur production et leur utilisation | |
AT407048B (de) | Rekombinantes hauptallergen des pollens von artemisia vulgaris (beifuss) | |
DE69933690T2 (de) | Apoptosis induzierender faktor | |
EP3881862A1 (fr) | Polypeptide destinés au traitement des maladies du rein glomérulaires et analyse de la progression et pronostic des syndromes dépendants | |
DE69932815T2 (de) | Cyclophilin als Allergen identifiziert | |
WO2002066513A2 (fr) | Fragments humains de lekti circulants hf7072, hf7638 et hf14448 ainsi que leur utilisation | |
CA2139552C (fr) | Utilisation des proteines vegetales receptrices de l'autoincompatibilite (slg et srk), pour le traitement des allergies | |
DE19534763C1 (de) | FMR1-verwandtes Protein |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2000903587 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2360585 Country of ref document: CA Ref country code: CA Ref document number: 2360585 Kind code of ref document: A Format of ref document f/p: F |
|
WWP | Wipo information: published in national office |
Ref document number: 2000903587 Country of ref document: EP |
|
AK | Designated states |
Kind code of ref document: A3 Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A3 Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 2000903587 Country of ref document: EP |