WO2006030770A1 - Hla結合性ペプチド、それをコードするdna断片および組み換えベクター - Google Patents
Hla結合性ペプチド、それをコードするdna断片および組み換えベクター Download PDFInfo
- Publication number
- WO2006030770A1 WO2006030770A1 PCT/JP2005/016822 JP2005016822W WO2006030770A1 WO 2006030770 A1 WO2006030770 A1 WO 2006030770A1 JP 2005016822 W JP2005016822 W JP 2005016822W WO 2006030770 A1 WO2006030770 A1 WO 2006030770A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- hla
- binding
- amino acid
- binding peptide
- peptide
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
Definitions
- HLA-binding peptides DNA fragments encoding them and recombinant vectors
- the present invention relates to an HLA-binding peptide, a DNA fragment encoding the peptide, and a recombinant vector.
- CTL cytotoxic T cells
- Patent Document 1 There is a technique described in Patent Document 1 as this kind of technology.
- Patent Document 1 describes that an oligopeptide having a specific amino acid sequence has HLA binding properties.
- Patent Document 1 Disclosure of the Invention of JP-A-8-151396
- HLA-binding peptide described in the above-mentioned document has a binding property to HLA-DQ4. It binds to HLA-A2 molecules (products such as HLA—A * 0201 gene) common in Western countries and HLA—A24 molecules (products such as HLA—A * 2402 gene) common to Japanese species. Whether it is power is unknown.
- An object of the present invention is to provide an HLA-binding peptide that is excellent in binding to a type of HLA molecule.
- an HLA-binding peptide that binds to an HLA-A type molecule comprising one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 60, HLA-binding peptides are also provided that also have lower amino acid residue strength.
- HLA-binding peptide SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 10, 14, 29, 30, 31, 33, 38, 39, 40, 43, 44, 49 and 51 HLA-binding peptides comprising one or more amino acid sequences selected from the group consisting of forces are provided.
- an HLA-binding peptide that binds to an HLA-A type molecule, wherein one or two amino acid residues are missing from the amino acid sequence contained in the HLA-binding peptide.
- an HLA-binding peptide comprising an amino acid sequence which is deleted, substituted or added, and having an amino acid residue strength of 8 or more and 11 or less.
- the structure includes an amino acid sequence in which one or several amino acid residues are deleted, substituted, or added from a specific amino acid sequence having binding properties with an HLA-A molecule. Even if it exists, there exists an effect
- the present invention also provides a DNA fragment comprising a DNA sequence encoding the above-mentioned HLA-binding peptide.
- a recombinant vector comprising a DNA sequence encoding the above HLA-binding peptide is provided.
- an HLA-binding peptide precursor characterized by being converted into the above-mentioned HLA-binding peptide in a mammalian organism.
- an HLA-binding peptide excellent in binding property to an HLA-A type molecule can be obtained.
- FIG. 1 is a schematic diagram for explaining an active learning experiment plan used in an example.
- the amino acid sequence predicted by the hypothesis obtained using the active learning experiment method has an affinity for HLA molecules of 3 or more in terms of log Kd value.
- Peptides containing 8 to 11 amino acid residues were used as HLA-binding peptide candidates. As a result of binding experiments, these peptides were actually
- the HLA-binding peptide according to the present embodiment is an HLA-binding peptide that binds to an HLA-A type molecule, and is one type selected from the group consisting of SEQ ID NOs: 1 to 60 described later. It is an HLA-binding peptide comprising the above amino acid sequence and having an amino acid residue strength of 8 or more and 11 or less.
- HLA-A human immunoglobulin-A
- Westerners such as Germans often have HLA-A2 type.
- sequences are all sequences having 9 amino acid residues contained in a predetermined genomic protein of cancer antigen WT1.
- sequences of SEQ ID NOs: 1 to 30 are sequences having a 9 amino acid residue power contained in a predetermined genomic protein of cancer antigen WT1.
- sequences of SEQ ID NOs: 1 to 30 are sequences with the highest binding properties with the HLA-A24 molecule predicted by the above-described method.
- Sequence numbers 1 to 30 are arranged in the order of excellent binding. That is, SEQ ID NO: 1 is the sequence predicted to have the best binding.
- Each unit of the predicted score for binding to the HLA-A24 molecule of each sequence and the unit of the binding experiment data item is indicated by a log Kd value.
- sequences of SEQ ID NOs: 31 to 60 are sequences having a force of 9 amino acid residues contained in a predetermined genomic protein of cancer antigen WT1.
- sequences of SEQ ID NOs: 31 to 60 are sequences with the highest binding properties with the HLA-A2 molecule predicted by the method described above. Sequence numbers 31 to 60 are arranged in the order of excellent binding. That is, SEQ ID NO: 31 is the sequence predicted to have the best binding.
- the unit of the predicted score for binding to the HLA-A2 molecule of each sequence and the item of the binding experiment data is V, and the difference is indicated by the log Kd value.
- the binding of the HLA-binding peptide according to the present embodiment to the HLA molecule is -lo In terms of gKd value, it can be 3 or more, particularly preferably 5 or more, and further preferably 5.4 or more.
- the binding property to the HLA molecule is 5 or more in terms of the log Kd value, a peptide having excellent binding property to the HLA molecule can be obtained. It can be suitably used for the development of drugs and the like.
- the binding property to the HLA molecule is 5.4 or more in terms of the log Kd value, a peptide having particularly excellent binding property to the HLA molecule can be obtained. It can be suitably used for the development of effective therapeutic drugs and preventive drugs.
- the HLA-binding peptide according to the present embodiment can be composed of 8 or more and 11 or less amino acid residues.
- cytotoxic T cells are cancer cell-specific cancer antigens (CTL epitopes) that are displayed on HLA class I molecules on the surface of cancer cells and have an amino acid strength of 8 to L1. Recognize specifically, eliminate cancer cells by damaging only cancer cells. And, it is important to make a CTL epitope that also has the amino acid power of 8 ⁇ : L 1 specific to such cancer cells and the like in creating a vaccine for treatment or prevention of cancer and the like.
- the above-mentioned HLA-binding peptide may be a peptide having only amino acid residues, but is not particularly limited.
- it may be an HLA-binding peptide precursor that is modified by modifying a sugar chain or a fatty acid group within a range that does not inhibit the action and effect of the present invention as necessary.
- Such precursors may also be converted into HLA-binding peptides by undergoing changes such as digestion by digestive enzymes in the mammalian body such as the human organs. The same effect as the binding peptide can be obtained with this HLA binding peptide.
- the HLA-binding peptide may be a peptide that binds to human HLA-A24 molecule.
- the HLA-binding peptide may be a peptide that binds to a human HLA-A2 molecule.
- the amino acid sequence contained in the above HLA-binding peptide may be an amino acid sequence derived from a cancer antigen WT1 protein, but is not particularly limited. For example, it may be an amino acid sequence derived from HIV protein or an amino acid sequence derived from cedar pollen protein. It may also contain amino acid sequences derived from other pathogenic or allergenic proteins.
- an HLA-binding peptide that binds to an HLA-A type molecule, wherein one or two amino acid residues of the amino acid sequence contained in the HLA-binding peptide are deleted, substituted, or Provided is an HLA-binding peptide comprising an added amino acid sequence and having an amino acid residue strength of 8 or more and 11 or less.
- the structure includes an amino acid sequence in which one or several amino acid residues are deleted, substituted, or added in a specific amino acid sequence having binding properties with an HLA-A molecule.
- the same effect as the HLA-binding peptide according to Embodiment 1 described above can be obtained.
- amino acid residues in the amino acid sequence predicted by the above-described method and excellent in binding to the HLA-A24 molecule are substituted, deleted, or added. It can be predicted that the same amino acid sequence shows excellent HLA binding properties.
- the amino acid residues to be substituted should be amino acid residues having similar properties to each other, such as hydrophobic amino acid residues.
- the HLA-binding peptides described in Embodiments 1 and 2 can be produced using techniques known to those skilled in the art for V and deviation. For example, it may be artificially synthesized by a solid phase method or a liquid phase method. Moreover, these HLA-binding peptides may be produced by expressing DNA fragments encoding these HLA-binding peptides or recombinant vector. Any of the HLA-binding peptides thus obtained can be identified using methods known to those skilled in the art. For example, the determination can be made using Edman decomposition or mass spectrometry.
- a DNA fragment comprising a DNA sequence encoding the above HLA-binding peptide is provided. Since the DNA fragment according to this embodiment contains a specific DNA sequence, the above HLA-binding peptide can be expressed.
- HLA-binding peptide When the above-mentioned HLA-binding peptide is expressed using the DNA fragment according to the present embodiment, a commercially available artificial fragment may be introduced and expressed in the cell. It can also be expressed using a protein expression kit.
- the above DNA fragment can be continuously expressed, for example, by introducing it into a human cell. Therefore, the introduction of a DNA fragment that encodes an HLA-binding peptide into the cell will cause the HLA-binding peptide to be present in the cell continuously, rather than when the HLA-binding peptide itself is introduced into the cell. Can do. In the case where an HLA-binding peptide is used as a vaccine, such a continuous expression is advantageous for enhancing the effectiveness of the vaccine.
- the DNA fragment according to the present embodiment can be produced using a technique known to those skilled in the art. For example, it may be synthesized artificially by a commercially available DNA synthesizer. Alternatively, it may be excised from the HCV genome using restriction enzymes. Alternatively, a primer may be used to amplify the HCV genome by PCR. In addition, any DNA fragment thus obtained can be identified using techniques known to those skilled in the art. For example, It can be identified by a commercially available DNA sequencer.
- a recombinant vector containing a DNA sequence encoding the above HLA-binding peptide can be obtained. Since the recombinant vector according to this embodiment contains a specific DNA sequence, it can express the above HLA-binding peptide.
- this recombinant vector may be introduced into cells and expressed. It can be expressed using a protein expression kit! /.
- the above-described recombinant vector can be continuously expressed by, for example, introduction into human cells. Therefore, the introduction of a recombinant vector that encodes an HLA-binding peptide into the cell may cause the HLA-binding peptide to exist in the cell continuously, rather than the introduction of the HLA-binding peptide itself into the cell. it can . In the case where an HLA-binding peptide is used as a vaccine, such continuous expression is advantageous for enhancing the effectiveness of the vaccine.
- the regulatory region for transcription 'expression such as a promoter region upstream of the DNA sequence encoding the above-mentioned HLA-binding peptide is used as an arbitrary sequence, so that the HLA-binding peptide Can be accurately controlled.
- the above-described recombinant vector can contain any sequence other than the DNA sequence encoding the above-mentioned HLA-binding peptide.
- it may contain the sequence of a marker gene such as a drug resistance gene.
- the recombinant vector according to the present embodiment can be produced using techniques known to those skilled in the art. For example, it can be obtained by cleaving a multicloning site of a commercially available vector such as pBR322 or pUC19 at an arbitrary restriction enzyme site, inserting the DNA fragment into the site, and ligating. In addition, any of the recombinant vectors thus obtained can be identified using techniques known to those skilled in the art.
- the length of the cleaved DNA fragment corresponds to the cleavage map of a commercially available vector such as pBR322 or pUC19, and is confirmed by agarose gel electrophoresis, and the DNA sequence is included in the DNA sequence excised from the cloning site. Can be identified by DNA sequencer etc.
- a predetermined genomic protein of the cancer antigen WT1 (SEQ ID NO:
- HLA-binding peptides containing amino acid sequences derived from HLA-binding peptides other than WT1 may be HLA-binding peptides against pathogens such as HIV viruses, and include amino acid sequences derived from allergen-derived proteins such as cedar pollen It may be a binding peptide.
- HLA-binding peptides are mainly for infectious diseases (influenza, SARS, HIV, HCV, etc.), cancer immunotherapy, allergic diseases (hay fever, rheumatism, atopy, asthma, etc.), autoimmunity Can be suitably used for treatment or prevention of diseases
- Figure 1 shows a schematic diagram of the active learning experiment plan used here.
- the peptide at the selected question point is produced by the synthesis' purification method described later, and the actual binding ability is measured by an experiment described later and added to the accumulated data.
- a supervised learning algorithm of a hidden Markov model was used as the lower learning algorithm, and the initial data power of 223 peptides was started, and 20 to 30 peptides were predicted per experiment. The above procedure was repeated 4 times, and a total of 341 data were obtained.
- binding ability was measured. If the binding ability was high, it was selected as a candidate for an HLA-binding peptide that can be used as a vaccine material.
- the obtained result was input to a learning system including a learning machine that uses a hidden Markov model as a mathematical algorithm, and a rule was created.
- the learning machines sampled different results and created rules. It should be noted that the rules that are expressed differ depending on the learning machine.
- the rules and experimental data obtained are stored as accumulated data as needed.
- Peptides were synthesized manually using Merrifield's solid phase method using Fmoc amino acids. After deprotection, reverse phase HPLC purification using a C18 column was achieved to a purity of 95% or higher. Peptide identification and purity were confirmed by MALDI-TOF mass spectrometry (Voyager DE RP, PerSeptive). Peptide quantification was performed with Micro BC A Atsey (Pierce) using BSA as a standard protein.
- C1R-A24 cells were exposed to acidic conditions at pH 3.3 for 30 seconds, and originally bound to HLA-A * 2402 molecules, and were associated with HLA class I molecules in common. Thus, the light chain j8 2m was dissociated and removed. After neutralization, purification
- MHC-pep three-membered aggregate
- JY cells were exposed to acidic conditions at pH 3.8 for 30 seconds to dissociate and remove the endogenous peptide and light chain i8 2m that had been noncovalently associated with the HLA-A * 0201 molecule. After neutralization, a reassociation experiment was conducted.
- the dissociation constant Kd value was calculated by the method published in (Udaka et al., Immunogenetics, 51, 816-828, 2000).
- sequences of SEQ ID NOS: 1 to 30 in Table 1 are sequences having 9 amino acid residue power included in the full-length sequence of a predetermined protein of WT1 registered in GENBANK.
- sequences of SEQ ID NOs: 1 to 30 are sequences with the highest binding to the HLA-A24 molecule predicted by the hypothesis obtained by the experimental design method described in the first embodiment.
- Sequence numbers 1 to 30 are arranged in the order of excellent binding. That is, SEQ ID NO: 1 is the sequence that is predicted to have the best binding.
- the full-length amino acid sequence of a predetermined protein of WT1 is shown in SEQ ID NO: 61.
- Table 1 lists amino acid sequences corresponding to the top 30 scores of prediction results using the above-described prediction program, prediction scores, and binding experiment data corresponding thereto. All the binding experiment data were obtained by artificially synthesizing the peptide sequence of WT1 by the synthesis method described above.
- sequences of SEQ ID NOs: 31 to 60 in Table 2 are sequences having a 9 amino acid residue power included in the full-length sequence of a predetermined protein of WT1 registered in GENBANK.
- sequences of SEQ ID NOs: 31 to 60 are sequences with the highest binding to the HLA-A2 molecule predicted by the hypothesis obtained by the experimental design described in Embodiment 1.
- Sequence numbers 31 to 60 are arranged in the order of excellent binding. That is, SEQ ID NO: 31 is the sequence predicted to have the highest binding.
- the full-length sequence of a given protein of WT1 is SEQ ID NO: 61 (MGSDVRDLNALLPA
- Table 2 lists amino acid sequences corresponding to the top 30 scores of prediction results using the above-described prediction program, prediction scores, and binding experiment data corresponding thereto. All the binding experiment data were obtained by artificially synthesizing the peptide sequence of WT1 by the synthesis method described above.
- amino acid residues of the amino acid sequence having excellent binding properties with the HLA-A molecule are predicted. Even if it is an amino acid sequence formed by substitution, deletion, or addition, it can be said that the same excellent HLA binding property is exhibited.
- the amino acid residues to be substituted are preferably amino acid residues having similar properties to each other, such as hydrophobic amino acid residues.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP05783164A EP1801211A4 (en) | 2004-09-17 | 2005-09-13 | BINDING PEPTIDE HLA MOLECULE, DNA FRAGMENT ENCODING SAME, AND RECOMBINANT VECTOR |
US11/574,963 US20080293916A1 (en) | 2004-09-17 | 2005-09-13 | Hla-Binding Peptide, and Dna Fragment and Recombinant Vector Coding for Said Hla-Binding Peptide |
JP2006535143A JPWO2006030770A1 (ja) | 2004-09-17 | 2005-09-13 | Hla結合性ペプチド、それをコードするdna断片および組み換えベクター |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004-272385 | 2004-09-17 | ||
JP2004272385 | 2004-09-17 | ||
JP2005-046459 | 2005-02-23 | ||
JP2005046459 | 2005-02-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2006030770A1 true WO2006030770A1 (ja) | 2006-03-23 |
Family
ID=36060021
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2005/016822 WO2006030770A1 (ja) | 2004-09-17 | 2005-09-13 | Hla結合性ペプチド、それをコードするdna断片および組み換えベクター |
Country Status (4)
Country | Link |
---|---|
US (1) | US20080293916A1 (ja) |
EP (2) | EP1801211A4 (ja) |
JP (1) | JPWO2006030770A1 (ja) |
WO (1) | WO2006030770A1 (ja) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008096831A1 (ja) | 2007-02-07 | 2008-08-14 | The Research Foundation For Microbial Diseases Of Osaka University | 癌の治療剤 |
WO2013089252A1 (ja) | 2011-12-14 | 2013-06-20 | 国立大学法人 高知大学 | ヘルパーt細胞誘導性ポリペプチドの改変 |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2216041A4 (en) * | 2007-11-20 | 2012-10-24 | Nec Corp | METHOD FOR INDUCING CYTOTOXIC LYMPHOCYTE T, CYTOTOXIC LYMPHOCYTE T INDUCER, AND PHARMACEUTICAL COMPOSITION AND VACCINE COMPRISING EACH INDUCER |
US9670529B2 (en) | 2012-02-28 | 2017-06-06 | Population Genetics Technologies Ltd. | Method for attaching a counter sequence to a nucleic acid sample |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11316754A (ja) * | 1998-05-06 | 1999-11-16 | Nec Corp | 実験計画法及び実験計画プログラムを記録した記録媒体 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH08151396A (ja) | 1994-11-28 | 1996-06-11 | Teijin Ltd | Hla結合性オリゴペプチド及びそれを含有する免疫調節剤 |
US20030072767A1 (en) * | 1998-09-30 | 2003-04-17 | Alexander Gaiger | Compositions and methods for WT1 specific immunotherapy |
-
2005
- 2005-09-13 US US11/574,963 patent/US20080293916A1/en not_active Abandoned
- 2005-09-13 JP JP2006535143A patent/JPWO2006030770A1/ja active Pending
- 2005-09-13 EP EP05783164A patent/EP1801211A4/en not_active Withdrawn
- 2005-09-13 WO PCT/JP2005/016822 patent/WO2006030770A1/ja active Application Filing
- 2005-09-13 EP EP20130153899 patent/EP2636679B1/en not_active Not-in-force
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11316754A (ja) * | 1998-05-06 | 1999-11-16 | Nec Corp | 実験計画法及び実験計画プログラムを記録した記録媒体 |
Non-Patent Citations (4)
Title |
---|
GAO L ET AL: "Selective elimination of leukemic CD34(+) progenitor cells by cytotoxic T lymphocytes specific for WT1.", BLOOD., vol. 95, no. 7, 2000, pages 2198 - 2203, XP002337253 * |
MAKITA M ET AL: "Antilung Cancer Effect of WT1-Specific Cytotoxic T Lymphocytes.", CLINICAL CANCER RESEARCH., vol. 8, 2002, pages 2626 - 2631, XP002993869 * |
OHMINAMI H ET AL: "HLA class I-restricted lysis of leukemia cells by a CD8(+) cytotoxic T-lymphocyte clone specific for WT1 peptide.", BLOOD., vol. 95, no. 1, 2000, pages 286 - 293, XP002190642 * |
TSUBOI A. ET AL: "WT1 Peptide-Based Immunotherapy for patients with Lung Cancer: Report of Two Cases.", MICROBIOL. IMMUNOL., vol. 48, no. 3, 2004, pages 175 - 184, XP002391404 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008096831A1 (ja) | 2007-02-07 | 2008-08-14 | The Research Foundation For Microbial Diseases Of Osaka University | 癌の治療剤 |
CN105664147A (zh) * | 2007-02-07 | 2016-06-15 | 日本电气株式会社 | 用于癌症的治疗剂 |
WO2013089252A1 (ja) | 2011-12-14 | 2013-06-20 | 国立大学法人 高知大学 | ヘルパーt細胞誘導性ポリペプチドの改変 |
Also Published As
Publication number | Publication date |
---|---|
EP1801211A1 (en) | 2007-06-27 |
EP2636679A1 (en) | 2013-09-11 |
EP1801211A4 (en) | 2008-06-11 |
JPWO2006030770A1 (ja) | 2008-05-15 |
EP2636679B1 (en) | 2015-05-20 |
US20080293916A1 (en) | 2008-11-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US9353151B2 (en) | HLA-binding peptide, and DNA fragment and recombinant vector coding for said HLA-binding peptide | |
JP5614659B2 (ja) | Hla結合性ペプチド、それをコードするdna断片および組み換えベクター | |
US20150087809A1 (en) | Hla-binding peptides, precursors thereof, dna fragments and recombinant vectors that code for those peptide sequences | |
WO2006030770A1 (ja) | Hla結合性ペプチド、それをコードするdna断片および組み換えベクター | |
JP2013078325A (ja) | Hla結合性ペプチド、その前駆体、それをコードするdna断片および組み換えベクター | |
JP5648927B2 (ja) | Hla結合性ペプチド、それをコードするdna断片および組み換えベクター | |
JP2006230269A (ja) | Hla結合性ペプチド、それをコードするdna断片および組み換えベクター |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU LV MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2006535143 Country of ref document: JP |
|
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: 11574963 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 200580031564.0 Country of ref document: CN |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2005783164 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 2005783164 Country of ref document: EP |