WO2005042461A1 - Compose presentant une affinite avec l'amyloide - Google Patents
Compose presentant une affinite avec l'amyloide Download PDFInfo
- Publication number
- WO2005042461A1 WO2005042461A1 PCT/JP2004/016101 JP2004016101W WO2005042461A1 WO 2005042461 A1 WO2005042461 A1 WO 2005042461A1 JP 2004016101 W JP2004016101 W JP 2004016101W WO 2005042461 A1 WO2005042461 A1 WO 2005042461A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- compound
- amyloid
- formula
- atom
- nmr
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/006—Biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered in the buccal area to detect epithelial cancer cells, dyes used for delineating tissues during surgery
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B23/00—Methine or polymethine dyes, e.g. cyanine dyes
- C09B23/14—Styryl dyes
- C09B23/141—Bis styryl dyes containing two radicals C6H5-CH=CH-
Definitions
- the present invention relates to a novel compound which belongs to the technical field of an in vivo test, and particularly has a affinity for amyloid involved in the onset and progress of Alzheimer's disease and the like, and is useful for the detection thereof.
- the number of patients with Alzheimer's disease is said to be more than 1.5 million in Japan at present, and it is predicted that one out of every 10 elderly people will develop the disease 20 years later.
- the burden is increasing.
- the disease is hereditary, about 10%, and the remaining 90% are sporadic, with everyone at risk of developing the disease. It is characterized by the formation of senile plaques, neurofibrillary tangles, and atrophy of the brain. It is said that these organic changes have been progressing before clinical symptoms appear.
- Non-Patent Document 1 Congo Red has been widely used as a tissue staining agent therefor.
- Congo Red has high affinity for amyloid ⁇ (sometimes abbreviated as ⁇ ) protein, a component of senile plaques, and can be selectively stained.
- ⁇ amyloid ⁇
- in vivo imaging probes for brain amyloid have recently appeared to capture the earliest neuropathological changes, the formation of senile plaques, before birth.
- Congo Red is a fluorescent molecule that has an affinity for the A protein, has a sulfone group in the molecule, and has high hydrophilicity, so it cannot cross the blood-brain barrier and cannot be used in vivo.
- Non-patent document 3 Non-patent document 3
- BSB (trans, trans) _1_bromo_2,5_bis_ (3-hydroxycarbonyl-2-hydroxy) styrylbenzene]
- BSB was intravenously administered to a model mouse of Alzheimer's disease, the mouse was sacrificed, and a brain specimen was stained after death. Good transfer to the brain was confirmed.
- radiolabeling for detection by PET or SPECT is still necessary.
- BSB has also shown excellent staining results as a fluorescent stain for systemic amyloidosis (Y.
- Non-patent document 1 Z.S.Khachaturian, Arch. Neurol., 1985, 42, 1097
- Non-patent document 2 CA Mathis, et al., Bioorg.Med.Chem, Lett., 2002, 12, 295
- Non-patent document 3 MP Kung, et al., J. Mol. Neurosci., 2003, 20, 15
- Non-Patent Document 4 Z.-P. Zhuang, et al "J. Med. Chem., 2003, 46, 237
- Non-Patent Document 5 D.M.Skovronsky et al, Proc.Natl.Acad.Sci., 2000, 97, 7609
- Non-Patent Document 6 Y.Ando et al., Lab.Invest., 2003, in press.
- An object of the present invention is to establish a new technique that enables detection of amyloid such as A / 3 protein in Alzheimer's disease without requiring special equipment. Means for solving the problem
- R and R are each independently a hydroxyl group or a carboxyl group.
- R 123 is a fluorine atom or a bromine atom.
- R is a bromine atom, R must be a fluorine atom
- At least one of the carbon atoms constituting the carboxy group of 12 is carbon-13.
- a preferred specific example of the compound represented by the formula (I) according to the present invention is a styrylbenzyl group represented by the formula (I), wherein R is a hydroxyl group, R is a carboxyl group, and R is a fluorine atom.
- R is a hydroxyl group and R is a carboxy group in the formula (I)
- R is a bromine atom, and a styrylbenne in which the carbon atom at the ⁇ - position and the -position is carbon-13
- Powers of zen compounds include, but are not limited to, these.
- the compound of the above general formula (I) can fluorescently stain amyloid. Therefore, according to the present invention, there is provided a fluorescent dye for amyloid comprising the compound represented by the general formula (I). Is done.
- the compound of the above general formula (I) can also be used as an MRI contrast agent having amyloid specificity. Therefore, according to the present invention, the compound represented by the general formula (I) An amyloid-specific MRI contrast agent is provided.
- amyloid used in the context of the present invention is a ⁇ -sheet structure (/ 3 folded structure) that leads to systemic amyloidosis or localized amyloidosis.
- the latter is representative of depositional proteins that also have secondary structural forces, and the latter, which belongs to focal amyloidosis, is due to the protein deposited on the brain of Alzheimer's disease patients as described above.
- the compound of the present invention represented by the formula (I) has a high affinity for amyloid similarly to conventionally known BSB and can fluorescently stain the protein, but is not limited thereto. It also functions as an MRI contrast agent with specificity for myloid.
- FIG. 1 shows a reaction scheme for synthesizing Example compounds (Compounds 1 and 9) of the present invention.
- FIG. 2 shows a 19 F-NMR spectrum (A) of Compound 1 and a 13 C-NMR spectrum (B) of Compound 9.
- FIG. 3 shows the fluorescence spectra of compound 1, compound 9 and BSB.
- FIG. 4 Stained images of temporal cortical sections (fixed with ethanol) of postmortem brain of Alzheimer's disease patient, (A) stained with compound 1, (B) with compound 9, and (C) with BSB. Indicates the case.
- FIG. 5 is a stained image of a frontal cortical slice (fixed with ethanol) of a postmortem brain of an Alzheimer's disease patient, (A) showing a case stained with compound 1 and (B) showing a case stained with compound 9.
- FIG. 6 shows an in vivo 19 F-MR brain image obtained when Compound 1 according to the present invention was administered to amyloid precursor protein transgenic mice.
- FIG. 7 shows a 1 H-MR brain image of a mouse brain which has been subjected to 19 F_MRI measurement and removed and fixed.
- FIG. 1 illustrates a preferred reaction scheme for synthesizing a styrylbenzene compound of the present invention. Briefly, as shown, as shown, OMe or --CO Me (Me is methyl Group) and a phosphonic acid-derived group _P (0) (OEt) (Et is an ethyl group) substituted at both ends with fluorine or bromine and substituted with benzaldehyde (compound indicated by 8 in Fig. 1) P-xylene-hi, hi-
- the compound is reacted with the compound to form a distyrylbenzene structure serving as a skeleton (step d in FIG. 1), and then sequentially demethylated to obtain a compound of the formula (I).
- At least one of the carbon atoms constituting the carboxy group of R and R is carbon 13 ( 13 C).
- R is a fluorine atom
- At least one of the carbon atoms does not have to be carbon 13, i.e., the carbon atoms constituting the carboxyl group at the l-, j3-, ⁇ - and ⁇ -positions and the carboxyl group of R and R are ordinary carbon 12
- the thus-obtained compound of the present invention represented by the formula (I) has an amyloid-specific MRI (magnetic resonance imaging) contrast agent having fluorine and ⁇ or carbon 13 in the molecule and having specificity for amyloid. In this respect, it has features not found in the conventionally known BSB.
- MRI magnetic resonance imaging
- the compounds of the present invention represented by the formula (I) are, similarly to BSB, those having high affinity for amyloid by brain amyloid (protein) and systemic amyloidosis. Miloid can also be fluorescently stained.
- the formula (I) in which R is a fluorine atom is particularly preferred.
- the fluorescence intensity is increased about twice as compared with BSB, in which the fluorescence intensity is high. This is understood because the compound (I) having a fluorine atom in the molecule has no fluorescence quenching due to the heavy atom effect as compared with BSB.
- R is a fluorine atom
- 2,5-Dimethylaniline (10 g, 82 mmol) was suspended in water (30 ml), and concentrated hydrochloric acid (17 ml) was added little by little. After cooling to about 5 ° C, an aqueous solution of sodium nitrite (a solution of 7.0 g dissolved in 10 ml of water; lOlmmol) was added dropwise. The mixture was stirred for 1 hour as it was, and the insoluble material was separated by filtration. To the filtrate was added sodium borofluoride (11.0 g, 10 mmol), and the precipitated diazodium salt was collected by filtration.
- sodium borofluoride (11.0 g, 10 mmol
- Compound 1 expresses a single large peak at ⁇ 118.5 m in 19 F_NMR, and is capable of imaging with 19 F_NMR. It was confirmed that there was.
- Fluorescence spectra were measured for Compound 1 synthesized in Example 1 and Compound 9 synthesized in Example 2, and BSB for comparison.
- the equipment used for the measurement is
- HITACHI F-4500 The measured concentrations were 1 ⁇ M each (in DMSO), and the wavelength of the excitation light was 390 nm.
- Figure 3 shows the measurement results.
- the compounds 1 and 9 of the present invention express strong fluorescence at 511 nm and 521 nm like BSB.
- Compound 1, which has fluorine in the molecule has about twice the fluorescence intensity as compared to BSB.
- Brain amyloid staining Brain amyloid was stained using compound 1 synthesized in Example 1 and compound 9 synthesized in Example 2, and BSB for comparison.
- Non-Patent Document 5 DM Skovronsky et al., Proc. Natl. Acad. ScL, 2000, 97, 7609
- NPF neutral buffer in formalin
- 6- ⁇ paraffin sections were prepared by a conventional method. The sections were immersed in a 0.01% solution of compound 1, compound 9 or BSB in 50% ethanol for 30 minutes, and then immersed in a saturated aqueous solution of lithium carbonate. Washed with 50% ethanol and observed with a fluorescence microscope (V-excited UV light) FIGS.
- FIGS. 4 and 5 show stained images (photographs) observed with a fluorescence microscope.
- FIG. 4 shows a stained image of a temporal cortex section fixed with ethanol
- FIG. 5 shows a stained image of a frontal cortex section (semi-connected slice) fixed with ethanol.
- the images in FIGS. 4 and 5 are actually color images, In order to clearly show the signs, traces of the fluorescent areas are shown below each photographic image. It is understood that the compounds 1 and 9 of the present invention have an affinity for the A / 3 protein constituting the senile plaque and selectively fluorescently stain the protein.
- FIG. 5 shows the same sample stained using Compounds 1 and 9, and the same number in the figure indicates that the sample is derived from the senile plaque in the same place.
- Example 2 shows that the compound of the present invention represented by the formula (I) functions as an amyloid-specific MRI contrast agent.
- FSB compound 1 synthesized in Example 1
- MRI measurement was performed as follows.
- Amyloid precursor protein transgenic mouse (Tg2576.
- FIG. 6 (1) The obtained brain image photograph of 19 F-MR is shown in FIG. 6 (1). Spots presumed to be senile plaques were detected as high signal areas in the 19 F-MR image (white spots in FIG. 6 (1)). Note that FIG. 6 (1) is actually a color image, and the spot-like portions are observed as clear red light-emitting portions.
- Fig. 6 (2) shows the photograph of Fig. 6 (1) by hand to facilitate understanding of Fig. 6 (1).
- FIG. 7 (1) shows a photograph of FIG. 7 (1) traced by hand to facilitate understanding of FIG. 7 (1).
- FIG. 7 shows a photograph of FIG. 7 (1) traced by hand to facilitate understanding of FIG. 7 (1).
- the parts corresponding to FIG. 6 can also be obtained by ⁇ -MRI.
- a T2-weighted image signal is obtained in the area shown in Fig. 6, and it is understood that the spot-like part shown in Fig.
- FIG. 6 is a senile plaque. Furthermore, it has been shown that 19 F-MRI in vivo can detect senile plaques with higher sensitivity than 1 H-MRI of post-mortem brain depending on the part of the brain (for example, the circled part in FIG. 6). Furthermore, frozen brain was MRI measured as described above, 40 zm create a thick cryosections was observed intracerebral distribution of FSB by fluorescence microscopy, 19 F-MRI, as shown in FIG. 6 The abnormal part (spotted part) found in the above was found to have been detected as a result of the accumulation of FSB in lesions that could be morphologically confirmed as senile plaques.
- the compound of the present invention can be used as a fluorescent staining agent for amyloid derived from systemic amyloidosis and local amyloidosis, for example, in histopathological examination of amyloid-related diseases represented by Alzheimer's disease. it can.
- the compound of the present invention also functions as an MRI contrast agent having amyloid specificity, as a simple method using MRI widely used in medical institutions, amyloid correlation such as Alheimer's disease can be used. It can also be used for prenatal diagnosis of disease.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Radiology & Medical Imaging (AREA)
- Oncology (AREA)
- Optics & Photonics (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2005515170A JP3877754B2 (ja) | 2003-10-30 | 2004-10-29 | アミロイド親和性化合物 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2003369797 | 2003-10-30 | ||
JP2003-369797 | 2003-10-30 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2005042461A1 true WO2005042461A1 (fr) | 2005-05-12 |
Family
ID=34543834
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2004/016101 WO2005042461A1 (fr) | 2003-10-30 | 2004-10-29 | Compose presentant une affinite avec l'amyloide |
Country Status (2)
Country | Link |
---|---|
JP (1) | JP3877754B2 (fr) |
WO (1) | WO2005042461A1 (fr) |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007034757A1 (fr) * | 2005-09-20 | 2007-03-29 | Fuence Co., Ltd. | Remède pour des maladies amyloïdes et médicament pour le suivi de celles-ci |
WO2007111179A1 (fr) * | 2006-03-28 | 2007-10-04 | Shiga University Of Medical Science | Agent d'imagerie de diagnostic pour une maladie des neurones incurable |
JP2008231102A (ja) * | 2007-02-23 | 2008-10-02 | Hiroaki Okuno | アミロイドベータ凝集阻害作用を有するフェノール誘導体 |
US8022075B2 (en) | 2005-11-30 | 2011-09-20 | Fujifilm Ri Pharma Co., Ltd. | Diagnostic and remedy for disease caused by amyloid aggregation and/or deposition |
WO2013066181A1 (fr) | 2011-11-04 | 2013-05-10 | Academisch Ziekenhuis Leiden Lumc | Ligands d'imagerie |
JP2014122846A (ja) * | 2012-12-21 | 2014-07-03 | Hamamatsu Photonics Kk | アミロイドの凝集体の定量方法及びその定量装置 |
WO2014109296A1 (fr) | 2013-01-09 | 2014-07-17 | 国立大学法人滋賀医科大学 | Agent de diagnostic irm pour une maladie neurologique réfractaire |
US8956589B2 (en) | 2009-02-27 | 2015-02-17 | Shiga University Of Medical Science | Imaging diagnostic agent and extracorporeal diagnostic agent for incurable neurological diseases |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002016333A2 (fr) * | 2000-08-24 | 2002-02-28 | University Of Pittsburgh | Derives de thioflavine utilises dans un diagnostic antemortem de la maladie d'alzheimer, imagerie in vivo et prevention du depot d'amyloide |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7029655B2 (en) * | 2000-10-04 | 2006-04-18 | California Institute Of Technology | Magnetic resonance imaging agents for in vivo labeling and detection of amyloid deposits |
CA2458961C (fr) * | 2001-04-27 | 2008-07-08 | The General Hospital Corporation | Diagnostic oculaire de la maladie d'alzheimer |
-
2004
- 2004-10-29 WO PCT/JP2004/016101 patent/WO2005042461A1/fr active Application Filing
- 2004-10-29 JP JP2005515170A patent/JP3877754B2/ja active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002016333A2 (fr) * | 2000-08-24 | 2002-02-28 | University Of Pittsburgh | Derives de thioflavine utilises dans un diagnostic antemortem de la maladie d'alzheimer, imagerie in vivo et prevention du depot d'amyloide |
Non-Patent Citations (1)
Title |
---|
LEE C.W. ET AL.: "Isomerization of (Z,Z) to (E,E)1-bromo-2,5-bis-(3-hydroxycarbonyl-4-hydroxy)styrylbenzene in strong base: probes for amyloid plaques in the brain", JOURNAL OF MEDICINAL CHEMISTRY, vol. 44, no. 14, 2001, pages 2270 - 2275, XP002958808 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007034757A1 (fr) * | 2005-09-20 | 2007-03-29 | Fuence Co., Ltd. | Remède pour des maladies amyloïdes et médicament pour le suivi de celles-ci |
US8022075B2 (en) | 2005-11-30 | 2011-09-20 | Fujifilm Ri Pharma Co., Ltd. | Diagnostic and remedy for disease caused by amyloid aggregation and/or deposition |
WO2007111179A1 (fr) * | 2006-03-28 | 2007-10-04 | Shiga University Of Medical Science | Agent d'imagerie de diagnostic pour une maladie des neurones incurable |
JP2008231102A (ja) * | 2007-02-23 | 2008-10-02 | Hiroaki Okuno | アミロイドベータ凝集阻害作用を有するフェノール誘導体 |
US8956589B2 (en) | 2009-02-27 | 2015-02-17 | Shiga University Of Medical Science | Imaging diagnostic agent and extracorporeal diagnostic agent for incurable neurological diseases |
WO2013066181A1 (fr) | 2011-11-04 | 2013-05-10 | Academisch Ziekenhuis Leiden Lumc | Ligands d'imagerie |
JP2014122846A (ja) * | 2012-12-21 | 2014-07-03 | Hamamatsu Photonics Kk | アミロイドの凝集体の定量方法及びその定量装置 |
WO2014109296A1 (fr) | 2013-01-09 | 2014-07-17 | 国立大学法人滋賀医科大学 | Agent de diagnostic irm pour une maladie neurologique réfractaire |
Also Published As
Publication number | Publication date |
---|---|
JP3877754B2 (ja) | 2007-02-07 |
JPWO2005042461A1 (ja) | 2007-05-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5522958B2 (ja) | 多点リン酸化ペプチド(タンパク)認識化合物、それを用いる検出方法 | |
CN107412788B (zh) | 用于对脑内所蓄积的Tau蛋白质进行成像的新的化合物 | |
US20110208064A1 (en) | Curcumin Derivatives for Amyloid-Beta Plaque Imaging | |
EP2023919A2 (fr) | Composés et sondes amyloïdes de ceux-ci pour des utilisations thérapeutiques et en imagerie | |
US20110071301A1 (en) | Abeta-binding small molecules | |
KR20090010987A (ko) | 신규 아밀로이드 친화성 화합물 | |
JP5699286B2 (ja) | 神経難病の画像診断薬及び体外診断薬 | |
KR20100091965A (ko) | 신규 아밀로이드 친화성 화합물 | |
JPWO2009057578A1 (ja) | 新規アミロイド親和性化合物の使用及び製造方法 | |
WO2007148755A1 (fr) | Nouveau composé presentant une affinité pour une substance amyloïde | |
RU2671506C2 (ru) | Имидазо[1,2-а]пиридин-7-амины в качестве средств визуализации | |
JP5048655B2 (ja) | 新規アミロイド親和性化合物 | |
KR20090083414A (ko) | 신규 아밀로이드 친화성 화합물 | |
CN108191789B (zh) | 一种吩噻嗪衍生物、其制备方法和应用 | |
WO2005042461A1 (fr) | Compose presentant une affinite avec l'amyloide | |
CN109180556B (zh) | 一种部花菁类荧光化合物及其制备方法和应用 | |
WO2005016888A1 (fr) | Sonde pour maladies liees a l'accumulation d'amyloide, agent de coloration d'amyloide, remede et agent prophylactique de maladies liees a l'accumulation d'amyloide et sonde de diagnostic et agent de coloration en matiere de changement neurofibrillaire | |
KR20100089858A (ko) | 신규 아밀로이드 친화성 화합물의 사용 및 제조 방법 | |
WO2022045093A1 (fr) | NOUVEAU COMPOSÉ, LIANT POUR AGRÉGAT D'α-SYNUCLÉINE ET UTILISATION ASSOCIÉE | |
CN114656447A (zh) | 一类基于高时空分辨率的近红外荧光、磁共振Aβ双模式成像探针及其制备方法和应用 | |
JP7008299B2 (ja) | 神経難病の画像診断薬及び体外診断薬 | |
KR20100094478A (ko) | 신규 아밀로이드 친화성 화합물의 사용 및 제조 방법 | |
CN108290883B (zh) | 用于tau成像的氮杂环丁烷衍生物 | |
JP2010189359A (ja) | ベンゾチアゾール誘導体含有診断用組成物 | |
WO2023098622A1 (fr) | LIGAND DE LIAISON À PETITE MOLÉCULE D'AGRÉGAT D'α-SYNUCLÉINE, PROCÉDÉ DE PRÉPARATION ET UTILISATION ASSOCIÉS |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 2005515170 Country of ref document: JP |
|
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 KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL 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): BW GH 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 IT LU 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 |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
122 | Ep: pct application non-entry in european phase |