WO1998055149A1 - Utilisation de vecteurs medicamenteux pour produire des medicaments migrants dans les ganglions lymphatiques - Google Patents

Utilisation de vecteurs medicamenteux pour produire des medicaments migrants dans les ganglions lymphatiques Download PDF

Info

Publication number
WO1998055149A1
WO1998055149A1 PCT/JP1998/002373 JP9802373W WO9855149A1 WO 1998055149 A1 WO1998055149 A1 WO 1998055149A1 JP 9802373 W JP9802373 W JP 9802373W WO 9855149 A1 WO9855149 A1 WO 9855149A1
Authority
WO
WIPO (PCT)
Prior art keywords
lewis
monosaccharide
compound
general formula
pullulan
Prior art date
Application number
PCT/JP1998/002373
Other languages
English (en)
Japanese (ja)
Inventor
Kazutoshi Horie
Kazuyoshi Masuda
Masahiro Sakagami
Original Assignee
Shionogi & Co., 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 Shionogi & Co., Ltd. filed Critical Shionogi & Co., Ltd.
Priority to AU74545/98A priority Critical patent/AU7454598A/en
Publication of WO1998055149A1 publication Critical patent/WO1998055149A1/fr

Links

Classifications

    • 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
    • A61K47/56—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof

Definitions

  • the present invention relates to the use of a drug carrier represented by the general formula (I) for producing a lymph node transfer agent, a lymph node transfer agent having a drug compound bound to the drug carrier, and the use of the drug carrier.
  • the present invention relates to a method for producing a lymph node transfer agent.
  • Japanese Patent Application Laid-Open No. 8-857073 discloses a polysaccharide derivative exhibiting organ-directing properties, and in particular, avoids the property of migrating to the liver and inflammatory sites and avoids reticuloendothelial tissues (liver, spleen, etc.). It has been shown to be useful as a drug carrier for antitumor agents and anti-inflammatory agents. Disclosure of the invention
  • lymph node transfer drug a drug that can be selectively transferred to the lymph nodes
  • the present invention relates to the use of a drug carrier represented by the following general formula (I) for producing a lymph node transfer agent.
  • T 1 is —NH—, —NHCO—, —CONH— or one NHCONH—
  • T 2 is — CH 2 CH 2 ( ⁇ CH 2 CH 2 ) m— (m represents an integer of 1 to 10), — (CH 2 ) n— (n represents an integer of 2 to 16) or represented by the following formula (II) Base :
  • F represents a monosaccharide, an N- or 0_acyl derivative of a monosaccharide, an O-alkyl derivative or an ester derivative of a monosaccharide.
  • the oligosaccharide consisting of 2 to 6 monosaccharides or monosaccharide derivatives (provided that the terminal of the monosaccharide and the oligosaccharide is not galactose, mannose, galactosamine or N-acetylgalactosamine)
  • P represents an integer of 0 to 100,000.
  • FIG. 1 is a graph showing the biodistribution of the drug carrier represented by the general formula (I) during primary inflammation in a rat adjuvant arthritis model.
  • FIG. 2 is a graph showing the biodistribution of the drug carrier represented by the general formula (I) during secondary inflammation in a rat adjuvant arthritis model.
  • Figure 3 shows the results of biodistribution experiments for CM pullulan modified with sialyl Lewis X, CM pullulan modified with a sugar chain in which sialic acid and galactose of Lewis X are linked with ⁇ 2-6, and unmodified CM pullulan. It is a craftsman who did.
  • FIG. 5 is an enlarged view of FIG. BEST MODE FOR CARRYING OUT THE INVENTION
  • E is carboxymethyl chitosan (CM chitosan), carboxymethyl pullulan (CM pullulan), carboxymethyl dextran (CM dextran), carboxymethyl mannoglucan (CM mannoglucan), heparin And polysaccharides such as hyaluronic acid and chondroitin sulfate.
  • These polysaccharides may be derivatives, for example, desulfated derivatives (such as de-N-sulfated heparin) and polyalcoholated derivatives (such as polyalcoholized mannoglucan). Further, in addition to these, those converted to have a carboxyl group or an amino group are also included in the polysaccharide derivative.
  • CM chitosan means that about 1 to 200% of D-darcosamine or N-acetyl-D-dalcosamine constituting chitosan is carboxymethylated.
  • CM pullulan means that about 1 to 300%, preferably about 20 to 120% of D-dalcos constituting pullulan is carboxymethylated.
  • CM dextran means that about 1 to 300% of D-glucose constituting dextran is carboxymethylated.
  • CM mannoglucan means that about 1 to 300%, preferably about 20 to 120% of D-mannose constituting mannoglucan is carboxymethylated.
  • the polysaccharides and derivatives thereof in E are one or more monovalent monosaccharides from which the functional groups (for example, amino group, carboxyl group, hydroxyl group, etc.) used for binding to T1 have been removed. (Up to p) or a derivative thereof.
  • the monosaccharide represented by F include monosaccharides other than galactose, mannose, galactosamine, and N-acetylgalactosamine, such as glucose.
  • Hexoses such as coace, fructos, fucose, neuraminic acid, and peronic acid; hexosamines such as glucosamine; and bentoses such as reportose, deoxyribose, arabinose, and xylose.
  • the monosaccharide may be a derivative thereof, and examples thereof include an N- or dimethyl derivative, an O-alkyl derivative, and an ester derivative of sulfuric acid, phosphoric acid, or the like.
  • these derivatives include sialic acid, which is an N- and / or acyl derivative of neuraminic acid (eg, N-acetyl neuraminic acid); and N-amino acid, which is a hexosamine N-acyl derivative Cetyldarcosamine; 6-O-benzoyl-glucose with 6-position of glucose benzoylated; 6-O-carboxymethyl-N-acetyl with 6-hydroxyl group of N-acetyldarcosamine carboxymethylated Darukosamin: 2-position amino group of Darukosamin is Mononobe Njiru reduction has been 2 one N- base Njiru - Darukosamin the like.
  • the oligosaccharide represented by F is a monosaccharide selected from the group consisting of galactose, glucose, fructose, mannose, fucose, neuraminic acid, peruronic acid, galactosamine, dalcosamine, ribose, deoxyribose, arabinose and xylose.
  • it refers to a linear or branched hetero-oligomer or homo-oligomer composed of 2 to 6, preferably 2 to 4 monosaccharide derivatives (for example, the derivatives exemplified above).
  • Such oligosaccharides include, for example, saccharides in which any of the hydroxyl groups of sucrose, Lewis X or Lewis A galactose have been sialylated (eg, sialyl Lewis A, sialyl Lewis X and ⁇ 2-6 sialyl Lewis X (sialic Examples thereof include saccharides in which the 2-position of the acid is ether-linked to the 6-position of galactose), sialyl ⁇ -acetyl lactosamine, lactose, maltose, Lewis X, sulfated Lewis X, and the like.
  • the non-reducing end of the oligosaccharide is not mannose, galactose, galactosamine or peracetyl galactosamine.
  • the above “monosaccharide” and “oligosaccharide” refer to one of the hydroxyl groups (preferably the anomeric hydroxyl group) of the sugar molecule (in the case of oligosaccharide, the sugar molecule binding to T 2 ).
  • Shall mean the removed one.
  • F and T 2 are bonded by an ⁇ - ⁇ -daricoside bond or a ⁇ -glycoside bond.
  • m represents an integer of 1 to 10, and is preferably 2 to 5.
  • N represents an integer of 2 to 16, and preferably 2 to 8.
  • the molecular weight of the drug carrier represented by the general formula (I) is preferably from 50 to 500 K, particularly preferably from 150 to 400 K.
  • the molecular weight is preferably about 190K.
  • E is a CM pull run
  • p is an integer other than 0, and D is 1.
  • T 2 is —CH 2 CH 2 (OCH 2 CH 2 ) 5 — and F is sialyl Lewis X, ⁇ 2-6 sialyl Lewis X or sialyl diacetylacetyltosamine have a molecular weight of Approximately 2401: up to approximately 350 mm is preferred.
  • ⁇ is CM pullulan; p is an integer of 20 or more; F is sialyl Lewis X, ⁇ 2-6 sialyl Lewis X or sialyl diacetyl lactosamine; — C ONH—; and ⁇ 2 is _CH 2 CH 2 (OCH 2 CH 2 ) 5—.
  • the compound represented by the general formula (I) has a property of selectively transferring to lymph nodes.
  • lymph node refers to all lymph nodes present in the body of the animal, and includes, for example, mesenteric lymph node, subknee lymph node, and face lymph node.
  • the compound represented by the general formula (I) is taken up by medullary and peripheral sinus macrophages of these lymph nodes. Therefore, the compound represented by the general formula (I) can be used as a drug carrier for supplying a drug to lymph nodes.
  • drugs used include immunosuppressants, anticancer drugs for cancer lymph node metastasis, and anti-HIV drugs for HIV growth suppression. In addition, it can be used as an immunoadjuvant for application to vaccines and to diagnostic agents for the purpose of imaging the immune system.
  • the compounds contained in the general formula (I) are all known compounds and can be produced according to the method described in JP-A-8-85703.
  • a drug compound represented by the following general formula (III) is conjugated to a drug carrier represented by the general formula (I) directly or via a spacer, A drug and a method for producing the drug are provided.
  • Z is a pharmaceutical compound
  • Y is a bond or Represents a spacer
  • p + Q l to an integer of 1 to 1,000 (however, when Z is an antitumor agent and an anti-inflammatory agent, P is 0)]
  • Pharmaceutical compounds include, for example, immunosuppressants such as brequinal, methotrexate, FK506, cyclosporin, NL9C (compound PA-48153C described in JP-A-5-310726); Anti-tumor agent for the lymph node metastasis of cancer such as sorbicin, 5-FU, cis-bratin, and axbra; For the suppression of HIV growth such as reverse transcriptase inhibitor, HIV protease inhibitor, and integration inhibitor Anti-HIV agents (eg, AZT, ddI, etc.), but are not limited thereto.
  • immunosuppressants such as brequinal, methotrexate, FK506, cyclosporin, NL9C (compound
  • a spacer represented by Y is a group that facilitates complexing of a drug and a carrier, and releases a pharmaceutical compound at a certain desired rate in lymph nodes. It can also play a role.
  • the spacer is not particularly limited, but as described above, the compound represented by the general formula (I) is incorporated into the medullary macula phage and the hepatic sinus macrophage of the lymph node. For this reason, it is preferable to use a marker that is cleaved by these macrophages.
  • spacers include alkyl chains having 2 to 18 carbon atoms, preferably 2 to 8 carbon atoms, and ethers (eg, hexethylene glycol, ethylene glycol, diethylene glycol, triethylene glycol, etc.).
  • peptides preferably has 2 to 10 amino acids, more preferably 2 to 4 amino acids.
  • the amino acids constituting the peptides may be any of neutral amino acids, basic amino acids, and acidic amino acids, and may be aliphatic amino acids or aromatic amino acids.
  • the peptide sequence preferably includes the following sequences: one Gly—Phe—, one Phe_Gly, and -GlyPhe-GlyGlv-.
  • Solvents that do not participate in the reaction after converting the carboxyl group of the drug having a hydroxyl group into an active ester for example, aqueous sodium hydrogen carbonate, potassium hydrogen carbonate, pH 7.4 phosphate buffer, etc.
  • an active ester for example, aqueous sodium hydrogen carbonate, potassium hydrogen carbonate, pH 7.4 phosphate buffer, etc.
  • Neutral or weakly basic solution in a compound represented by the general formula (I) by reacting with a polysaccharide moiety having an amino group
  • a drug having a leaving group or a drug converted to have a leaving group, and a polysaccharide moiety having an amino group of the compound represented by the general formula (I) are reacted with a solvent that does not participate in the reaction (eg, sodium bicarbonate).
  • a solvent that does not participate in the reaction eg, sodium bicarbonate.
  • Aqueous solution, aqueous potassium bicarbonate solution or a weakly basic solution or
  • a drug having no formyl group or a compound converted to have a formyl group and a polysaccharide moiety having an amino group of the compound represented by formula (I) are reacted with a solvent that does not participate in the reaction (for example, acetic acid aqueous solution, etc.). ) In the presence of sodium cyanoborohydride at room temperature to 80 at room temperature.
  • the compound 1-4 was purified by column chromatography on silica gel (30 g) (form: methanol-water 10: 3: 1 (the lower layer was used as the solvent)). (60 mg, 50%) was obtained as a colorless oil.
  • the reaction mixture was added to 99.5% ethanol (1600 ml) to precipitate a crude product, and the precipitate was added to 95% ethanol (560 ml) and acetone (560 ml). ) And dimethyl ether (560 ml) in that order, and then dried under reduced pressure.
  • purified water was purified using a dialysis membrane having a molecular weight exclusion limit of 1200 to 140 000 (manufactured by Sekto Tranovoa). (100 ml) was used as the external solution for two times of dialysis (15 hours at room temperature), and the dialysis solution was freeze-dried under reduced pressure to obtain NL9C-HEG-CM pullulan (compound 2 , 2.80 g, ds: 0.04).
  • Ds is the proton at the anomeric position of CM pullulan (5.30 to 5.90 ppm) and the proton derived from the methyl group of NL9C (1.00 to 0.70 p) in iH-NMR. pm) was calculated from the ratio of the integral values.
  • Example 2 A study on the distribution of the drug carrier represented by the general formula (I) in the body (1)
  • CM pullulan No. 1 (CM pullulan), No. 2 (S LN—CM pullulan) and No. 3 (SL e X—CM) shown in Table 1 below as drug carriers represented by the general formula (I). (Pullulan).
  • 3 H was recovered as H 20 using an automatic sample combustion device (Packard 306) in the case of foot, and decolorization (H 2 0 2 ) soluene-350), and then performed with a liquid scintillation counter.
  • Packard 306 automatic sample combustion device
  • decolorization H 2 0 2 soluene-350
  • the cells were rinsed with 0.1 M sodium phosphate buffer (pH 7.2) containing 7% sucrose (20 minutes x 2 times), and ethanol series (50% ⁇ 70% ⁇ 80 ⁇ 90% ⁇ A dehydration operation of 95% ⁇ 100% ⁇ 100%) was performed for 20 minutes each, followed by pre-soaking and embedding with an embedding agent Technovit (trade name, purchased from Heraeus Kulzer GmbH).
  • the resulting block was cut into 1.5 sections, mounted on a slide glass, coated with an autoradiography emulsion (NR-M2 (Konica) or NTB-3 (Kodak)), and placed in a dark box containing silica gel. Exposure (4 ° C). The exposure time was set by performing development over time (about 2 weeks).
  • each compound was not distributed to bone marrow and liver but was distributed in large amounts to lymph nodes, reflecting the results of the above-mentioned biodistribution experiment (1).
  • the major uptake cells were medullary sinus macrophages, and it was thought that marginal sinus macrophages also took up a little. However, it was not incorporated at all into follicular macrophages.
  • a 3H-labeled sample was administered to a 7-week-old female Lewis rat at a dose of lmg / kg into the tail vein, 24 hours later, blood was collected, blood was removed, and each organ was extracted. After the measurement of radioactivity, decolorize organs about O. lg (H 2 0 2) ⁇ dissolved (Soluene-350), liquid scintillation one Performed by Yuichi Shiyonkaun.
  • NL9C-C8-CM pullulan which is an immunosuppressant synthesized in Example 1
  • the evaluation was conducted from the viewpoint of application to organ transplantation using an experimental skin transplantation system in which a strong rejection was observed after bone marrow.
  • NL9C, NL9C-C8-CM pullulan was dissolved in 5% Tween80Zsaline or 0.65% CremophorEL (trade name) / saline, and immediately after transplantation and 2, 4, 7, 9, and 1 after transplantation. On day 1, a tail vein was performed.
  • ds 0.05 (indicating that 5 out of 100 residues of glucose constituting pullulan are modified with sialyl Lewis X) of compound No. 3
  • the lymph node-migrating drug carrier used in the present invention is an immunosuppressant for suppressing autoimmune diseases or rejection during organ transplantation, an anticancer agent for treating lymph node metastasis of cancer, and suppressing HIV proliferation. It is useful for drug delivery systems such as targeted anti-HIV drugs. It can also be applied to vaccines as an immunoadjuvant and to diagnostic agents for lymph node imaging.

Landscapes

  • Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne l'emploi de vecteurs médicamenteux pour produire des médicaments migrants dans les ganglions lymphatiques, ces vecteurs étant représentés par la formule générale (I) E-(T1-T2-F)p dans laquelle E représente un polysaccharide, tel que le CM chitosane, le CM pullulane ou le CM dextrane, ou bien un dérivé de celui-ci; T1 représente -NH-, -NHCO-, -CONH- ou -NHCONH-; T2 représente -CH¿2?CH2(OCH2CH2)m-, -(CH2)n-, etc.,; F représente un monosaccharide éventuellement N- ou O-acylé, O-alkylé ou bien estérifié, ou encore un oligosaccharide constitué de 2 à 6 molécules du monosaccharide ou d'un de ses dérivés; et p est un entier compris entre 0 et 1000.
PCT/JP1998/002373 1997-06-03 1998-05-29 Utilisation de vecteurs medicamenteux pour produire des medicaments migrants dans les ganglions lymphatiques WO1998055149A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU74545/98A AU7454598A (en) 1997-06-03 1998-05-29 Use of drug carriers for producing lymphnode migrating drugs

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP9/145011 1997-06-03
JP14501197 1997-06-03

Publications (1)

Publication Number Publication Date
WO1998055149A1 true WO1998055149A1 (fr) 1998-12-10

Family

ID=15375389

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1998/002373 WO1998055149A1 (fr) 1997-06-03 1998-05-29 Utilisation de vecteurs medicamenteux pour produire des medicaments migrants dans les ganglions lymphatiques

Country Status (2)

Country Link
AU (1) AU7454598A (fr)
WO (1) WO1998055149A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001081103A (ja) * 1999-09-13 2001-03-27 Denki Kagaku Kogyo Kk ヒアルロン酸結合薬剤
WO2009049378A1 (fr) * 2007-10-17 2009-04-23 Qbiotics Limited Dérivés de lactone

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0539306A (ja) * 1990-12-14 1993-02-19 D D S Kenkyusho:Kk ヒアルロン酸およびコンドロイチン誘導体
JPH0680705A (ja) * 1992-09-02 1994-03-22 D D S Kenkyusho:Kk ヘパリン誘導体
JPH0885703A (ja) * 1994-09-16 1996-04-02 D D S Kenkyusho:Kk 臓器移行性を有する多糖誘導体および薬物担体

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0539306A (ja) * 1990-12-14 1993-02-19 D D S Kenkyusho:Kk ヒアルロン酸およびコンドロイチン誘導体
JPH0680705A (ja) * 1992-09-02 1994-03-22 D D S Kenkyusho:Kk ヘパリン誘導体
JPH0885703A (ja) * 1994-09-16 1996-04-02 D D S Kenkyusho:Kk 臓器移行性を有する多糖誘導体および薬物担体

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001081103A (ja) * 1999-09-13 2001-03-27 Denki Kagaku Kogyo Kk ヒアルロン酸結合薬剤
WO2009049378A1 (fr) * 2007-10-17 2009-04-23 Qbiotics Limited Dérivés de lactone

Also Published As

Publication number Publication date
AU7454598A (en) 1998-12-21

Similar Documents

Publication Publication Date Title
USRE46190E1 (en) High-molecular weight conjugate of steroids
US5763413A (en) Lewis-associated compound, process for producing the same, and anti-inflammatory
NL9201440A (nl) Triantennaire clusterglycosiden, hun bereiding en toepassing.
AU6488099A (en) Dds compounds and method for assaying the same
JP3048198B2 (ja) 硫酸化グリコサミノグリカノイド誘導体
EA005133B1 (ru) Полисахариды с антитромботической активностью, содержащие по меньшей мере одну ковалентную связь с биотином или производным биотина
KR20090031597A (ko) 콤브레타스타틴류의 고분자 결합체
US8492352B2 (en) Polysaccharides with antithrombotic activity, including a covalent bond and an amino chain
HU215152B (hu) Eljárás pentaszacharid egységet tartalmazó szénhidrátszármazékok és ezeket tartalmazó gyógyszerkészítmények előállítására
EP0128097A1 (fr) Nouveaux dérivés immunostimulants, leur préparation et leur application comme médicament
JP2002537263A (ja) 化学物質
JP3001381B2 (ja) 臓器移行性を有する多糖誘導体および薬物担体
Sashiwa et al. Chemical modification of chitosan 8: preparation of chitosan–dendrimer hybrids via short spacer
WO1998055149A1 (fr) Utilisation de vecteurs medicamenteux pour produire des medicaments migrants dans les ganglions lymphatiques
KR970005346B1 (ko) 사카라이드 유도체
JPH11514992A (ja) 変性エーテルグリセログリコ脂質
EP2611854A1 (fr) Dendrimères à terminaison saccharide à visée anti-inflammatoire
JP2589435B2 (ja) 糖修飾ポリリジン誘導体
Tomaszewska et al. Glucosamine-and galactosamine-based monosaccharides with highly fluorinated motifs
CA2771055C (fr) Oligosaccharides n-sulfates activateurs des recepteurs des fgfs, leur preparation et leur application en therapeutique
JPH07507319A (ja) 免疫抑制性および寛容原性修飾ルイス↑ｃおよびＬａｃＮＡｃ化合物
JPH01221387A (ja) 新規ジサッカライド誘導体及びその塩
Kosma Chemical synthesis of core structures
EP3981777A1 (fr) Composés et polymères antiviraux
CN110467646A (zh) 双核苷酸前体药物

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH GM GW HU ID IL IS JP KE KG 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 ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD 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 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
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA