[go: up one dir, main page]

WO1999064574A1 - Procede de preparation de peptides modifies a terminal c - Google Patents

Procede de preparation de peptides modifies a terminal c Download PDF

Info

Publication number
WO1999064574A1
WO1999064574A1 PCT/GB1999/001855 GB9901855W WO9964574A1 WO 1999064574 A1 WO1999064574 A1 WO 1999064574A1 GB 9901855 W GB9901855 W GB 9901855W WO 9964574 A1 WO9964574 A1 WO 9964574A1
Authority
WO
WIPO (PCT)
Prior art keywords
amino acid
peptide
resin
group
molecule
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB1999/001855
Other languages
English (en)
Inventor
Malcolm Douglas Walkinshaw
Paul Taylor
Nicholas John Turner
Sabine Lahja Flitsch
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Edinburgh
Cyclacel Ltd
Original Assignee
University of Edinburgh
Cyclacel Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Edinburgh, Cyclacel Ltd filed Critical University of Edinburgh
Priority to AU42825/99A priority Critical patent/AU4282599A/en
Priority to JP2000553564A priority patent/JP2002517233A/ja
Priority to EP99955499A priority patent/EP1095134A1/fr
Publication of WO1999064574A1 publication Critical patent/WO1999064574A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
    • C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • 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
    • C12N9/90—Isomerases (5.)

Definitions

  • This invention relates to the manufacture of peptides and peptide analogues.
  • c' is a substituted or unsubstituted carbon atom
  • X is a C x to C 4 chain in which each of the carbon atoms may be substituted or unsubstituted; and R 1 and R 2 are groups mimicking the side chains of a natural or unnatural amino acid.
  • the invention provides a method of forming a complex between a protein and a molecule having analogous structure to at least a portion of the natural binding partner of the protein, wherein the molecule consists of or contains a group of formula I .
  • molecules of this type will be as pharmaceuticals capable of affecting the activity of the bound protein, whether by inhibition or potentiation, or as templates for the design of such pharmaceuticals.
  • Other important applications will include the use in assays, as affinity ligands in purification and as diagnostic molecules for the natural binding partners.
  • the carbonyl group and the adjacent methylene and CR 1 R 2 groups mimic the structure of an amino acid (apart from the amino group) .
  • the identity of groups R 1 and R 2 and of any further substituents may vary widely, and may for example be chosen from alkyl, aryl, heterocyclic, alkanol, carboxyl, amino, imino, imido, ether, ester, thiol and thioether groups.
  • R 1 and R 2 may be chosen as follows:
  • R 1 and R 2 Other species may also be chosen as groups R 1 and R 2 , in order to produce a wide range of different amino acid mimicking molecules .
  • ring structure of Formula I imparts a high degree of conformational restriction, which limits the degrees of freedom available to the resultant molecules. This leads to tighter ligand binding and thus increases the chances of identifying active compounds.
  • the preferred number of atoms in the ring lies in the range 4 to 7 : 3-membered rings exhibit excessive strain, and rings with more than 7 members are generally too flexible.
  • a 5- or 6-membered ring is employed.
  • cyclic 1,3-diones especially those having 6-membered rings of the following formula:
  • the invention therefore provides, in another aspect, a method of manufacturing a peptide or peptide analogue comprising the steps of reacting a molecule having a free carboxyl group with a compound of formula I (as defined above) .
  • the molecule having a free carboxyl group may be produced by reacting the terminal hydroxyl group of a resin-bound linker with a substituted chloroformate to produce a resin-bound molecule having an activating group attached via a carbonate linkage. An amino acid is then added, which displaces the activating group to form a carbamate linkage, resulting in a resin-bound molecule terminating in an amino acid residue with a terminal carboxyl group.
  • chloroformate is p-nitrophenyl chloroformate
  • the amino acid is valine.
  • suitable reaction steps for producing the initial resin-bound linker which are known in the art.
  • polypeptide precursor is a key intermediate in the peptide formation process of the invention. It may be used in a number of ways to produce polypeptide molecules in the "reverse" manner, the incorporation of Cyclodiones being only one example.
  • the precursor may be reacted with a further substituted chloroformate to produce a mixed carbonic anhydride (i.e. a molecule having a -CO-O-CO-O- linkage) .
  • a mixed carbonic anhydride i.e. a molecule having a -CO-O-CO-O- linkage
  • the resulting polypeptide may then be directly reacted with a group of Formula I or Formula II, in the presence of a suitable coupling reagent (such as DCC/DMAP) , as illustrated in the following example: - ⁇ : UNK; ⁇
  • a suitable coupling reagent such as DCC/DMAP
  • the terminal amino acid residue may be cyclised by treatment with a coupling reagent to form an oxazolone, which is itself a significant intermediate in the synthesis of peptides and peptide- mimicking molecules.
  • the oxazolone may readily be alkylated at the C-4 position to produce an ot, a- dialkylated amino acid, which is a group known to induce certain elements of secondary structure.
  • the oxazolone may be reacted with a further (free) oxazolone to produce a "R ⁇ gheimer dimer" . Both reactions are illustrated in the following example:
  • the invention provides a method of manufacturing a peptide or polypeptide comprising the initial steps of: (a) forming a resin-bound peptide molecule with a terminal amino acid residue having a free carboxyl group; and (b) performing a cyclising condensation reaction between the carbonyl of the peptide linkage and the free carboxyl group of the terminal amino acid residue to form a terminal oxazolone group.
  • the invention permits the creation of a very substantial range of different peptides and peptide- mimicking molecules . These in turn will provide a large "library” of compounds for potential therapeutic use.
  • polypeptide-mimicking molecules provided by the invention may be seen from the following illustrative example relating to the interaction of Cyclodiones with cyclophilin.
  • Cyclophilins are potential drug targets in a number of different disease areas, including immunosuppressive disorders, parasite infection 1 , rheumatoid arthritis 2 and AIDS 3 ' 4 .
  • the only known cyclophilin-binding drug is cyclosporin A, an immunosuppressant undecapeptide drug used to prevent organ rejection after transplant operations 5,6,7 .
  • Cyclodiones can mimic peptides by forming a crystal complex of the parent dimedone with the protein human cyclophilin A.
  • a protein X-ray structure resolved to a resolution of 0.2 nm clearly shows the presence of dimedone in the cyclophilin A active site.
  • the position of the dimethyl group of the dimedone group mimics the position of a valine side chain adopted by the cyclosporin A inhibitor.
  • hCypA Recombinant human cyclophilin
  • Novartis AG Recombinant human cyclophilin
  • Crystals of the hCypA were grown by vapour diffusion at 17°C by the hanging drop method.
  • the precipitating solution in the well consisted of lOOmM Tris.HCl (pH 8.0), 22% (w/v) PEG 8000, 5% (v/v) DMSO, 0.02% NaN 3 .
  • the initial 8 ⁇ l drop consisted of 50mM Tris.HCl (pH 8.0), 11% (w/v) PEG 8000, 2.5% (v/v) DMSO, 0.02% NaN 3 , 0.4mM hCypA .
  • the dimedone ligand was introduced into the crystal (0.2mm x 0. lmm x 0.025mm) using a stepwise soaking procedure in which the DMSO concentration was gradually reduced and the ligand concentration gradually increased. This procedure was required to prevent crystal damage and also to prevent competition of DMSO binding at the active site.
  • a single crystal of hCypA was soaked in a precipitating solution containing 30mM ethyl-1-piperidine- glyoxylate and a reduced (4%) concentration of DMSO. After one hour the crystal was transferred to a fresh soaking solution containing 30 mM ligand and 3% DMSO. The crystal was transferred a total of 6 times over a period of 6 hours. The final soak was for 2.5 hours in a solution containing no DMSO and 30mM dimedone.
  • Flash freezing of the crystal in liquid nitrogen was carried out after soaking in a cryoprotectant solution consisting of lOOmM Tris.HCl (pH 8.0), 22% (w/v), PEG 8000, 0.02% NaN 3 , and 30mM ligand 30% glycerol .
  • the refined structure of hCypA from the X-ray structure of the hCypA/Cyclosporine A complex was used as a starting model.
  • a rotation and translation search using AMORE was used to position the molecule and gave a good starting model.
  • Subsequent rigid body refinement and refinement of individual atom isotropic temperature factors gave an R factor of 21% for data between 15 and 1.7A.
  • a difference Fourier map calculated at this stage showed clear electron density for the bound ligand.
  • the structure was built using the WITNOTP software.
  • Novel cyclophilin binding ligands are highly likely to be medically useful as inhibitory drugs .
  • the discovery that inhibition of cyclophilin prevents its incorporation into the HIV protein coat indicates that families of inhibitors unrelated to the immunosuppressant cyclosporins may provide potential anti-HIV drugs.
  • the development of species- specific cyclophilin inhibitors may also provide a route to novel anti-parasitic drugs.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Epidemiology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicinal Preparation (AREA)

Abstract

Des molécules non peptidiques de formule (I) peuvent être utilisées pour imiter la structure de peptides et pour se lier à des protéines qui se lient naturellement à des ligands de peptides. Dans la formule (I), C' représente un atome de carbone substitué ou non substitué; X représente une chaîne C1 à C4 dans laquelle chaque atome de carbone peut être substitué ou non substitué; et R1 et R2 représentent des groupes qui imitent les chaînes latérales d'un acide aminé naturel ou non naturel. Les molécules peuvent être utilisées pour elles-mêmes ou bien être incorporées dans des molécules plus grosses telles que des polypeptides. Dans les deux cas, elles peuvent être utilisées en tant que substances thérapeutiques pour altérer l'activité de la protéine. De plus, elles peuvent être utilisées pour analyser et titrer les protéines ou bien pour agir en tant que ligands d'affinité dans leur purification. On décrit également des procédés de production de polypeptides comprenant les molécules de l'invention.
PCT/GB1999/001855 1998-06-11 1999-06-11 Procede de preparation de peptides modifies a terminal c Ceased WO1999064574A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU42825/99A AU4282599A (en) 1998-06-11 1999-06-11 Method to prepare c-terminal modified peptides
JP2000553564A JP2002517233A (ja) 1998-06-11 1999-06-11 Ｃ末端修飾ペプチドの製造方法
EP99955499A EP1095134A1 (fr) 1998-06-11 1999-06-11 Procede de preparation de peptides modifies a terminal c

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9812675.8 1998-06-11
GBGB9812675.8A GB9812675D0 (en) 1998-06-11 1998-06-11 Peptides

Publications (1)

Publication Number Publication Date
WO1999064574A1 true WO1999064574A1 (fr) 1999-12-16

Family

ID=10833628

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1999/001855 Ceased WO1999064574A1 (fr) 1998-06-11 1999-06-11 Procede de preparation de peptides modifies a terminal c

Country Status (5)

Country Link
EP (1) EP1095134A1 (fr)
JP (1) JP2002517233A (fr)
AU (1) AU4282599A (fr)
GB (1) GB9812675D0 (fr)
WO (1) WO1999064574A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7223795B2 (en) 2000-12-13 2007-05-29 Cyclacel Limited Modified peptide derivatives and methods of use thereof
EP1958961A2 (fr) 1998-11-13 2008-08-20 Cyclacel Limited Vecteurs de transport
WO2018047062A1 (fr) 2016-09-06 2018-03-15 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Dérivés de pro-insuline
US10195287B2 (en) 2013-12-05 2019-02-05 Chemical & Biopharmaceuticals Laboratories Of Patras S.A. Biologically active insulin derivatives
WO2019069274A1 (fr) 2017-10-04 2019-04-11 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Procédé de préparation d'un peptide de type glucagon
WO2022106951A1 (fr) 2020-11-20 2022-05-27 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Procédé de production d'egf
US11634455B2 (en) 2013-08-29 2023-04-25 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Amino diacids containing peptide modifiers

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994000509A1 (fr) * 1992-06-30 1994-01-06 Legomer Partners, L.P. Matieres derivees d'oxazolone
WO1998008956A2 (fr) * 1996-08-27 1998-03-05 President And Fellows Of Harvard College Utilisation d'une proteine de fixation de macrolide mutee pour prevenir la reaction du greffon contre l'hote

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994000509A1 (fr) * 1992-06-30 1994-01-06 Legomer Partners, L.P. Matieres derivees d'oxazolone
WO1998008956A2 (fr) * 1996-08-27 1998-03-05 President And Fellows Of Harvard College Utilisation d'une proteine de fixation de macrolide mutee pour prevenir la reaction du greffon contre l'hote

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
B HENKEL ET AL.: "Investigations on solid-phase synthesis in N-to-C direction (inverse synthesis)", LIEBIGS ANNALEN/RECUEIL: ORGANIC AND BIOORGANIC CHEMISTRY - A EUROPEAN JOURNAL., no. 10, 1997, VCH PUBLISHERS, FLORIDA., US, pages 2161 - 2168, XP002116055, ISSN: 0947-3440 *
M DAVIES & M BRADLEY: "Internal resin capture - a self purification method for the synthesis of C-terminally modified peptides", TETRAHEDRON., vol. 55, no. 15, 9 April 1999 (1999-04-09), ELSEVIER SCIENCE PUBLISHERS, AMSTERDAM., NL, pages 4733 - 4746, XP002116057, ISSN: 0040-4020 *
R P SHARMA ET AL.: "A novel method of solid phase peptide synthesis", PEPTIDES. CHEMISTRY, STRUCTURE AND BIOLOGY. PROC. XIIITH AMERICAN PEPTIDE SYMPOSIUM, JUNE 20-25, 1993, 1994, Escom, Leiden, 194, pages 127 - 129, XP002116056 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1958961A2 (fr) 1998-11-13 2008-08-20 Cyclacel Limited Vecteurs de transport
US7223795B2 (en) 2000-12-13 2007-05-29 Cyclacel Limited Modified peptide derivatives and methods of use thereof
US11634455B2 (en) 2013-08-29 2023-04-25 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Amino diacids containing peptide modifiers
US12441760B2 (en) 2013-08-29 2025-10-14 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Amino diacids containing peptide modifiers
US10195287B2 (en) 2013-12-05 2019-02-05 Chemical & Biopharmaceuticals Laboratories Of Patras S.A. Biologically active insulin derivatives
WO2018047062A1 (fr) 2016-09-06 2018-03-15 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Dérivés de pro-insuline
US11230585B2 (en) 2016-09-06 2022-01-25 Chemical & Biopharmaceutical Laboratories Of Patra Proinsulin derivatives
WO2019069274A1 (fr) 2017-10-04 2019-04-11 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Procédé de préparation d'un peptide de type glucagon
WO2022106951A1 (fr) 2020-11-20 2022-05-27 Chemical & Biopharmaceutical Laboratories Of Patras S.A. Procédé de production d'egf

Also Published As

Publication number Publication date
AU4282599A (en) 1999-12-30
GB9812675D0 (en) 1998-08-12
JP2002517233A (ja) 2002-06-18
EP1095134A1 (fr) 2001-05-02

Similar Documents

Publication Publication Date Title
Chow et al. Ligation technologies for the synthesis of cyclic peptides
US5965695A (en) Modified peptide and peptide libraries with protease resistance, derivatives thereof and methods of producing and screening such
EP0535155B1 (fr) Banques de peptides modifies resistant a la protease
JP6254527B2 (ja) 構造化ポリペプチドの特異性のモジュレーション
US5635597A (en) Peptides that bind IL-2 receptors
JP4928263B2 (ja) ペプチド結合サロゲートを組み入れた空間的に規定された大環状分子
CZ407392A3 (en) Bank of oligomers
JPH09501490A (ja) 位相学的に分離された、コードされた固相ライブラリー
Goodman et al. New reagents, reactions, and peptidomimetics for drug design
Freeman et al. Microwave-assisted solid-phase aza-peptide synthesis: aza scan of a PKB/Akt inhibitor using aza-arginine and aza-proline precursors
US5948693A (en) Solid phase synthesis of immunosuppressive agents
Abboud et al. A straightforward methodology to overcome solubility challenges for N-terminal cysteinyl peptide segments used in native chemical ligation
EP1095134A1 (fr) Procede de preparation de peptides modifies a terminal c
EP0448099A2 (fr) Peptides nouveaux
JP2007534619A (ja) Ｃ５ａ受容体拮抗物質
US20030125243A1 (en) Synthesis of cyclic peptides
US6787612B1 (en) Resin derivatization method and uses thereof
Hu et al. Cyclosporin analogs modified in the 3, 7, 8-positions: substituent effects on peptidylprolyl isomerase inhibition and immunosuppressive activity are nonadditive
Cudic et al. Peptidomimetics: Fmoc solid-phase pseudopeptide synthesis
Maeda et al. Solid phase synthesis of α-amino squaric acid-containing peptides
JP3584283B2 (ja) オリゴチロシンを有するペプチド
Nunez SURE PROTEIN FOR PEPTIDE CYCLIZATION
Davies Cyclic, modified and conjugated peptides
Stokes New methods for the synthesis of biologically active cyclosporins
Bourne et al. A convenient method for synthesis of cyclic peptide libraries

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK 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 MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ 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

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1999955499

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWE Wipo information: entry into national phase

Ref document number: 09719383

Country of ref document: US

WWP Wipo information: published in national office

Ref document number: 1999955499

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1999955499

Country of ref document: EP