KR910018545A - Ngf/bdnf 계통 신경영양성 단백질의 생물학적 활성형 재조합 멤버의 제조방법 - Google Patents
Ngf/bdnf 계통 신경영양성 단백질의 생물학적 활성형 재조합 멤버의 제조방법 Download PDFInfo
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Abstract
내용 없음
Description
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제 1도는 인체 DNF핵산 및 추정 아미노산 배열을 나타낸다. 성숙(가공)형 BDNF의 추정 아미노산 배열은 굵은 문자로 표시하였다. 제2도는 백터 pT5T내의 대장균 내에서의 인체성숙(가공)형 NGF의 발현을 나타낸다.
상세한 사항은 실시예2의 원문에 제시된다,. 제3도는 진핵세포에서 생성된 성숙형 인체NGF의 수식 및 재중첩시의 생물학적 활성의 상실 및 재취득을 나타낸다.
Claims (39)
- 제1도는 제시된 핵산배열 또는 그 유도체 및 변이체를 포함하는, 인체 뇌유도 신경영양성 인자(BDNF)를 코드화하는 유전자.
- 제1항에 있어서, 위치 196에 G대신에 A로 치환된, 인체 뇌유도 신경영양성 인자(BDNF)를 코드화하는 유전자.
- 제1항에 있어서, 위치 668에 G대신에 A로 치횐된, 인체 뇌유도 신경영양성 인자(BDNF)를 코드화하는 유전자.
- 제1도에 제세된 아미노산 배열 및 본질적으로 상동성인 단백질을 포함하는, 성숙형 인체 뇌유도 신경영양성 인자(BDNF).
- 제4항에 있어서, 아미노산 66에 발린 (valine)대신에 메티오닌(methionine)으로 치환된, 인체 뇌유도 신경 영양성 인자(BDNF).
- 제4항에 있어서, 아미노산 223에 아르기닌(arginine)대신에 라이신(lysine)으로 치환된, 인체 뇌유도 신경 영양성 인자(BDNF).
- 제1도에 제시된 아미노산 배열 및 본질적으로 상동성인 단백질을 포함하는 인체 뇌유도 신경영양성 인자(BDNF)전구체.
- 제7항에 있어서, 아미노산 66에 발린 대신에 메티오닌으로 치환되, 인체 뇌유도 신경영양성 인자(BDNF)전구체.
- 제7항에 있어서, 아미노산 233에 아르기닌 대신에 라이신으로 치환된, 인체 뇌유도 신경영양성 인자(BDNF)전구체.
- 하기의 단계를 포함하는, 박테리아 세포 발현시스템에서 성숙된 생물학적 활성형의 인체 신경영양성 단백질의 제조 방법; 상기 단백질을 코드화하는 유전자를 백터 속으로 삽입; 생물학적으로 불활성형의 상기 단백질을 형성하기 위하여 상기 유전자를 발현; 및 상기 생물학적 불활성형을 재중첩(refolding)및 재생(renaturing).
- 제10항에 있어서, 상기 박테리아 세포가 대장균의 균주(strain)인 방법.
- 제10항에 있어서, 상기 인체 신경영양성 단백질이 NGF인 방법.
- 제10항에 있어서, 상기 인체 신경영양성 단백질이 BDNF인 방법.
- 제10항에 있어서, 상기 인체 신경영양성 단백질이 NGF-3인 방법.
- 제 6도에 제시된 핵산배열 또는 그의 유도체 및 변이체를 포함하는, 인체 성숙형 NGF-3를 코드화하는 유전자.
- 제7도에 제시된 아미노산 배열 및 본질적으로 상동성인 단백질을 포함하는, 인체 성숙형 NGF-3.
- 제7도에 제시된 아미노산 배열 및 본질적으로 상동성인 단백질을 포함하는, 인체 NGF-3전구체.
- 단백질이 본질적으로 완전한 생물학적 활성을 얻는 방법으로서, 하기 단계를 포함하는, 신경영양성 단백질이 재중첩 및 재생방법; 본질적으로 상기 모든 신경영양성 단백질이 에너지적으로 가장 안정한 형태를 취하는 다양한 3차원 구조 및 디술피드 결합(disulfide bond)양식을, 상기 신경영양성 단백질이 갖도록 하는 반응배지를 제조; 및 상기 반응배지로부터 상기 신경영양성 단백질을 유리.
- 제18항에 있어서, 상기 반응배지가 아래의 사항을 포함하는 단계에 의하여 제조되는, 신경영양성 단백질이 재중첩 및 재생방법; 상기 신경영영성 단백질 용액의 모든 분자간 및 분자내 디술피드 결합의 분쇄 및 유리 티올(free-thiol)의 형성; 혼합된 디술피드 결합을 형성하기 위하여, 디술피드 함유 화합물로 상기 유리 티올울 산화; 및 티올 함유 화합물 존재하에 상기 용액을 희석.
- 제19항에 있어서, 상기 반응배지가 혐기성(anaerobic)조건하에서 유지되는, 신경영양성 단백질의 재중첩 및 재생방법.
- 제19항에 있어서, 상기 반응배지가 글리콜 함유 시약을 함유하는, 신경영양성 단백질의 재중첩 및 재생방법.
- 제21항에 있어서, 상기 글리콜 함유 시약이 폴리에틸렌 글리콜린, 신경영양성 단백질의 재중첩 및 재생방법.
- 제19항에 있어서, 상기 분쇄작용이 상기 용액에 변성제 및 환원제를 첨가함으로써 이루어지는, 신경영양성 단백질의 재중첩 및 재생방법.
- 제23항에 있어서, 상기 변성체가 구아니딘 염화수소(guanidine hydrochloide)또는 요소 (urea)인, 신경영양성 단백질의 재중첩 및 재생방법.
- 제19항에 있어서, 상기 디술피드 함유 화합물이 산화된 글리타티온(giutathion), 시스틴(cystine)또는 시스타민(cystamine)인 신경영양성 단백질의 재중첩 및 재생방법.
- 제19항에 있어서, 상기 티올 함유 화합물이 시스테인(cysteine)또는 디티오테레이롤(dithiothreitol)인, 신경영양성 단백질의 재중첩 및 재생방법.
- 허용할 수 있는 약리학적 캐리어(carrier)에, 제12항의 방법에 의하여 제조된 생물학적 활성형의 NGF를 포함하는, 제약 조성물.
- 허용할 수 있는 약리학적 캐리어 내의 생물학적 활성형의 인체 BDNF를 포함하는, 제약 조성물.
- 제28항에 있어서, 상기 BDNF가 제13항의 방법에 의하여 제조되는 제약 조성물.
- 허용할 수 있는 약리학적 캐리어 내의 생물학적 활성형의, NGF-3를 포함하는 제약 조성물.
- 제30항에 있어서, 상기 NGF-3가 제14항의 방법에 의하여 제조되는, 제약 조성물.
- 단백질이 생물학적 활성ㅇ르 얻는 방법으로서, 하기 단계를 포함하는, 대장균에서 재조합적으로 발현된 인체 성숙형NGF의 중첩(folding)방법; 8몰(M)의 요소를 함유하며 pH가 약 3.0인 20mM의 구연산 나트륨에, 상기 대장균 발현 NGF를 약 0.6mg/ml의 농도로 용해; 8M의 요소를 함유하며 pH가 약 8.5인 1M트리스(tris)용액을 첨가하여, 상기 용액의 pH를 상승; 디티오테레이톨을 약 5~15mM의 농도로 첨가하여, 상기 NGF를 환원; 산화된 글루타티온 또는 시스틴을 약 15~50mM의 농도로 첨가하여, 상기 NGF를 산화; 약 3.2~4.2M의 요소를 함유하며 pH가 약 8.0인 100mM Na2HPO4용액으로, 상기 NGF용액을 약 9배로 희석; 글루타티온 또는 시스틴의농도에 비하여 약 2~3배의 시스테인을 첨가하여, 상기 NGF의 디술피드 상호교환을 촉매 작용화;및 상기 반응 혼합물로부터 상기 NGF를 유리.
- 단백질이 생물학적 활성을 얻는 방법으로서, 하기 단계를 포함하는, 박테리아 발현시스템에서 재조합적으로 발현된 인체 성숙형 NGF의 중첩(folding)방법; 상기 박테리아 발현NGF를 함유하는 용액에 디티오테레이톨 또는 β-메르캅토에탄올(β-mercaptoethanol)을 첨가; 상기 용액에 산화된 글루타티온 또는 시스타민을 첨가; 글리골 함유 시약을 함유하는 완충약으로 상기 용액을 희석; 최종 재중첩 혼합물 생성하기 위하여 시스테인 또는 2-메르캅토에틸아민을 첨가; 상기 최종 재중첩 혼합물을 탈기(脫氣);및 상기 최종 재중첩 혼합물로부터 상기 NGF를 유리.
- 신경질환을 치료하기 위하여, 생물학적 활성형 인체 BDNF를 포함하는 제약 조성물을 이용.
- 알즈하이머(Alzheimer)질병을 치료하기 위하여, 생물학적 활성형 인체 BDNF를 포함하는 제약 조성물을 이용.
- 신경질환을 치료하기 위하여, 생물학적 활성형 인체 NGF-3를 포함하는 제약 조성물을 이용.
- 알즈하이머 질병을 치료하기 위하여, 생물학적 활성형 인체 NGF-3를 포함하는 제약 조성물을 이용.
- 신경질환을 치료하기 위하여, 생물학적 활성형의 박테리아 발현 인체 NGF를 포함하는 제약 조성물을 이용.
- 알즈하이머 질병을 치료하기 위하여, 생물학적 활성형의 박테리아 발현 인체 NGF를 포함하는 제약 조성물을 이용.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US50544190A | 1990-04-06 | 1990-04-06 | |
| US07/505,441 | 1990-04-06 | ||
| US54775090A | 1990-07-02 | 1990-07-02 | |
| US07/594,126 US5235043A (en) | 1990-04-06 | 1990-10-09 | Production of biologically active, recombinant members of the ngf/bdnf family of neurotrophic proteins |
| US07/594126 | 1990-10-09 | ||
| US07/547750 | 1990-10-09 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR910018545A true KR910018545A (ko) | 1991-11-30 |
Family
ID=27414291
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1019910005514A KR910018545A (ko) | 1990-04-06 | 1991-04-06 | Ngf/bdnf 계통 신경영양성 단백질의 생물학적 활성형 재조합 멤버의 제조방법 |
Country Status (17)
| Country | Link |
|---|---|
| US (1) | US5235043A (ko) |
| EP (1) | EP0450386B1 (ko) |
| JP (1) | JP2521851B2 (ko) |
| KR (1) | KR910018545A (ko) |
| AT (1) | ATE183232T1 (ko) |
| AU (1) | AU651339B2 (ko) |
| CA (1) | CA2038669C (ko) |
| DE (1) | DE69131514T2 (ko) |
| DK (1) | DK0450386T3 (ko) |
| ES (1) | ES2140379T3 (ko) |
| FI (1) | FI108146B (ko) |
| GR (1) | GR3031632T3 (ko) |
| IE (1) | IE910827A1 (ko) |
| IL (1) | IL97602A (ko) |
| NO (1) | NO308308B1 (ko) |
| NZ (1) | NZ237544A (ko) |
| TW (1) | TW206258B (ko) |
Families Citing this family (53)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5229500A (en) * | 1989-08-30 | 1993-07-20 | Regeneron Pharmaceuticals, Inc. | Brain derived neurotrophic factor |
| US5180820A (en) * | 1989-08-30 | 1993-01-19 | Barde Yves Alain | Brain-derived neurotrophic factor |
| US5606031A (en) * | 1990-04-06 | 1997-02-25 | Lile; Jack | Production and purification of biologically active recombinant neurotrophic protein in bacteria |
| US5986070A (en) * | 1990-04-06 | 1999-11-16 | Amgen Inc. | Production of biologically active NGF proteins |
| ES2119779T3 (es) * | 1990-09-05 | 1998-10-16 | Southern Cross Biotech Pty Ltd | Solubilizacion de proteinas en formas activas. |
| US5530109A (en) * | 1991-04-10 | 1996-06-25 | Ludwig Institute For Cancer Research | DNA encoding glial mitogenic factors |
| NZ243084A (en) * | 1991-06-12 | 1995-08-28 | Regeneron Pharma | Production and recovery of recombinant neurotrophins, genes for a modified lamb signal sequence, fusion genes and plasmids |
| US5389529A (en) * | 1991-06-12 | 1995-02-14 | Regeneron Pharmaceuticals, Inc. | Modified lamβ signal sequence and processes for producing recombinant neurotrophins |
| DE4139000A1 (de) * | 1991-11-27 | 1993-06-03 | Boehringer Mannheim Gmbh | Verfahren zur gentechnologischen herstellung von biologisch aktivem ss-ngf |
| US5488099A (en) * | 1992-03-06 | 1996-01-30 | Persson, Deceased; Hakan B. | Multifunctional chimeric neurotrophic factors |
| US5316921A (en) * | 1992-05-18 | 1994-05-31 | Genentech, Inc. | Single-chain hepatocyte growth factor variants |
| CA2118012A1 (en) * | 1992-05-18 | 1993-11-25 | Paul J. Godowski | Hepatocyte growth factor variants |
| US5328837A (en) * | 1992-05-18 | 1994-07-12 | Genentech, Inc. | Hepatocyte growth factor protease domain variants |
| DE69330809T2 (de) * | 1992-06-08 | 2002-05-16 | Takeda Chemical Industries Ltd | Therapeutisches Agens für Neutropenie |
| US5364645A (en) * | 1992-10-30 | 1994-11-15 | The Regents Of The University Of California | Method of controlling microorganisms by pulsed ultraviolet laser radiation |
| EP0632008B1 (en) * | 1993-06-01 | 1998-02-04 | Ono Pharmaceutical Co., Ltd. | Pentanoic acid derivatives |
| EP0724598B1 (en) * | 1993-10-18 | 1999-01-13 | Regeneron Pharmaceuticals, Inc. | Growth factor heterodimers |
| US5470719A (en) * | 1994-03-18 | 1995-11-28 | Meng; Shi-Yuan | Modified OmpA signal sequence for enhanced secretion of polypeptides |
| US6743628B1 (en) | 1995-08-28 | 2004-06-01 | Washington University | Method of cell culture using neurturin |
| US5739307A (en) * | 1995-08-28 | 1998-04-14 | Washington University | Polynucleotide encoding neurturin neurotrophic factor |
| US5607711A (en) * | 1995-11-01 | 1997-03-04 | The Regents Of The University Of California | Method of controlling insects and mites with pulsed ultraviolet light |
| US5859311A (en) * | 1995-11-27 | 1999-01-12 | University Of Kentucky Research Foundation | Transgenic mice which overexpress neurotrophin-3 (NT-3) and methods of use |
| US6222022B1 (en) | 1996-03-14 | 2001-04-24 | Washington University | Persephin and related growth factors |
| DE69732350T2 (de) | 1996-03-14 | 2005-12-22 | Washington University | Persephin und verwandte wachstumsfaktoren |
| US7015316B1 (en) * | 1996-03-14 | 2006-03-21 | Washington University | Polynucleotides encoding human persephin and related growth factors |
| CA2257122C (en) * | 1996-06-11 | 2004-04-06 | Boehringer Mannheim Gmbh | Method of activating denatured protein |
| US5910435A (en) * | 1996-07-25 | 1999-06-08 | Wisconsin Alumni Research Foundation | Method of folding proteins with synthetic dithiol catalysts |
| BR9713055A (pt) | 1996-11-15 | 2000-04-04 | Genentech Inc | Processo para isolar uma neurotrofina humana recombinante, composição de neurotrofina e processo para purificar uma neurotrofina |
| US6042579A (en) * | 1997-04-30 | 2000-03-28 | Medtronic, Inc. | Techniques for treating neurodegenerative disorders by infusion of nerve growth factors into the brain |
| EP1012184B1 (en) | 1997-07-14 | 2007-10-10 | Bolder Biotechnology, Inc. | Derivatives of growth hormone and related proteins |
| US7495087B2 (en) | 1997-07-14 | 2009-02-24 | Bolder Biotechnology, Inc. | Cysteine muteins in the C-D loop of human interleukin-11 |
| EP0994188B1 (de) * | 1998-10-09 | 2004-01-07 | Scil proteins GmbH | Verfahren zur Gewinnung von aktivem Beta-NGF |
| ATE406907T1 (de) * | 1998-10-28 | 2008-09-15 | Cornell Res Foundation Inc | Methoden zur regulierung der angiogenese und vaskuläre integrität mittels trk rezeptor liganden bdnf, nt-3 und nt-4 |
| US6329136B1 (en) | 1998-10-30 | 2001-12-11 | The Regents Of The University Of California | Method for laser inactivation of infectious agents |
| BR122013003013B8 (pt) * | 1999-01-14 | 2021-07-06 | Bolder Biotechnology Inc | proteína isolada monopeguilada do hormônio do crescimento e método para sua obtenção |
| CN1379819A (zh) * | 1999-06-28 | 2002-11-13 | 俄克拉荷马州医学研究基金会 | 具有催化活性的重组memapsin蛋白酶及其应用方法 |
| US20020049303A1 (en) * | 1999-06-28 | 2002-04-25 | Tang Jordan J. N. | Catalytically active recombinant memapsin and methods of use thereof |
| AU6899800A (en) | 1999-08-11 | 2001-03-05 | General Hospital Corporation, The | Bdnf polymorphism and association with bipolar disorder |
| US7223406B2 (en) | 2000-07-21 | 2007-05-29 | The Regents Of The University Of California | Methods and compositions for preventing and treating male erectile dysfunction and female sexual arousal disorder |
| WO2002007757A2 (en) * | 2000-07-21 | 2002-01-31 | Lue Tom F | Prevention and treatment of sexual arousal disorders |
| US20040121947A1 (en) * | 2000-12-28 | 2004-06-24 | Oklahoma Medical Research Foundation | Compounds which inhibit beta-secretase activity and methods of use thereof |
| US20030232365A1 (en) * | 2001-02-15 | 2003-12-18 | Whitehead Institute For Biomedical Research | BDNF polymorphisms and association with bipolar disorder |
| EP1233075A3 (en) * | 2001-02-15 | 2003-01-08 | Whitehead Institute For Biomedical Research | BDNF polymorphism and association with bipolar disorder |
| US20060234944A1 (en) * | 2001-10-23 | 2006-10-19 | Oklahoma Medical Reseach Foundation | Beta-secretase inhibitors and methods of use |
| CA2464736A1 (en) * | 2001-10-23 | 2003-05-15 | Oklahoma Medical Research Foundation | Beta-secretase inhibitors and methods of use |
| US20030219696A1 (en) * | 2002-05-23 | 2003-11-27 | Moreland Gerald W. | Method and apparatus for preventing backflow in dental saliva evacuators |
| JP2005305107A (ja) * | 2004-03-25 | 2005-11-04 | Sei Matsuoka | 価値的情報値の測定装置およびこれを使用した測定方法 |
| WO2007021886A2 (en) * | 2005-08-10 | 2007-02-22 | Oklahoma Medical Research Foundation | Truncated memapsin 2 for use for treating alzheimer's disease |
| WO2008002572A2 (en) * | 2006-06-27 | 2008-01-03 | The Government Of The United States Of America As Represented By The Secretary Of Health And Human Services | Method for measuring mature brain derived neurotrophic factor |
| IT1394596B1 (it) * | 2008-09-30 | 2012-07-05 | Fond Parco Tecnologico Padano | Produzione di ngf in pianta. |
| EP2594295A1 (en) | 2011-11-16 | 2013-05-22 | Servicio Andaluz De Salud | Nerve implants based on a compacted biomaterial containing cells |
| GB201804641D0 (en) * | 2018-03-22 | 2018-05-09 | Inivata Ltd | Methods of sequencing nucleic acids and error correction of sequence reads |
| IT202000032423A1 (it) | 2020-12-24 | 2022-06-24 | Univ Degli Studi Di Trieste | Metodo per la produzione di forme processate proteoliticamente di fattori trofici o fattori di crescita |
Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4511502A (en) * | 1982-12-22 | 1985-04-16 | Genentech, Inc. | Purification and activity assurance of precipitated heterologous proteins |
| US4599197A (en) * | 1982-12-22 | 1986-07-08 | Genentech, Inc. | Purification and activity assurance of precipitated heterologous proteins |
| US4512922A (en) * | 1982-12-22 | 1985-04-23 | Genentech, Inc. | Purification and activity assurance of precipitated heterologous proteins |
| US4511503A (en) * | 1982-12-22 | 1985-04-16 | Genentech, Inc. | Purification and activity assurance of precipitated heterologous proteins |
| DK161152C (da) * | 1983-03-03 | 1991-11-11 | Genentech Inc | Polypeptid med egenskaber som human beta-nervevaekstfaktor og fremgangsmaade til fremstilling deraf, dna-isolat omfattende en sekvens som koder for polypeptidet, replicerbar udtrykkelsesvektor for dna-sekvensen, rekombinant vaertscelle transformeret med vektoren, farmaceutisk praeparat indeholdende polypeptidet og fremg. der omfatter anvendelsen af polypeptidet til fremst. af et farmaceutisk praeparat |
| US4766205A (en) * | 1985-11-13 | 1988-08-23 | Beatrice Companies, Inc. | Method for isolation of recombinant polypeptides in biologically active forms |
| US4929700A (en) * | 1987-04-16 | 1990-05-29 | Cetus Corporation | Production of purified, biologically active, bacterially produced recombinant human CSF-1 |
| IT1219874B (it) * | 1988-03-18 | 1990-05-24 | Fidia Farmaceutici | Utilizzazione del fattore di crescita nervoso umano e sue composizioni farmaceutiche |
| US6709837B1 (en) * | 1989-03-10 | 2004-03-23 | Takeda Chemical Industries, Inc. | Polypeptide and production thereof |
| JPH05161493A (ja) * | 1989-08-30 | 1993-06-29 | Max Planck Ges Foerderung Wissenschaft Ev | ニューロトロフィン―３ |
| IL95511A (en) * | 1989-08-30 | 2000-10-31 | Max Planck Gesellschaft | Neurotrophin-3 a novel neurotrophic factor related to nerve growth and brain derived neurotrophic factor |
| US5229500A (en) * | 1989-08-30 | 1993-07-20 | Regeneron Pharmaceuticals, Inc. | Brain derived neurotrophic factor |
| US5180820A (en) * | 1989-08-30 | 1993-01-19 | Barde Yves Alain | Brain-derived neurotrophic factor |
-
1990
- 1990-10-09 US US07/594,126 patent/US5235043A/en not_active Expired - Lifetime
-
1991
- 1991-03-12 IE IE082791A patent/IE910827A1/en not_active IP Right Cessation
- 1991-03-18 DE DE69131514T patent/DE69131514T2/de not_active Expired - Lifetime
- 1991-03-18 AT AT91104171T patent/ATE183232T1/de not_active IP Right Cessation
- 1991-03-18 DK DK91104171T patent/DK0450386T3/da active
- 1991-03-18 EP EP91104171A patent/EP0450386B1/en not_active Expired - Lifetime
- 1991-03-18 ES ES91104171T patent/ES2140379T3/es not_active Expired - Lifetime
- 1991-03-19 IL IL9760291A patent/IL97602A/xx not_active IP Right Cessation
- 1991-03-20 CA CA002038669A patent/CA2038669C/en not_active Expired - Lifetime
- 1991-03-22 NZ NZ237544A patent/NZ237544A/xx unknown
- 1991-04-03 NO NO911303A patent/NO308308B1/no not_active IP Right Cessation
- 1991-04-05 FI FI911643A patent/FI108146B/fi active
- 1991-04-05 AU AU74130/91A patent/AU651339B2/en not_active Ceased
- 1991-04-06 KR KR1019910005514A patent/KR910018545A/ko not_active Application Discontinuation
- 1991-04-06 JP JP3073389A patent/JP2521851B2/ja not_active Expired - Lifetime
- 1991-04-08 TW TW080102624A patent/TW206258B/zh not_active IP Right Cessation
-
1999
- 1999-10-21 GR GR990402673T patent/GR3031632T3/el unknown
Also Published As
| Publication number | Publication date |
|---|---|
| NO308308B1 (no) | 2000-08-28 |
| EP0450386A3 (en) | 1992-09-09 |
| US5235043A (en) | 1993-08-10 |
| IL97602A (en) | 2005-08-31 |
| CA2038669A1 (en) | 1991-10-07 |
| AU651339B2 (en) | 1994-07-21 |
| NZ237544A (en) | 1993-07-27 |
| EP0450386B1 (en) | 1999-08-11 |
| GR3031632T3 (en) | 2000-01-31 |
| NO911303D0 (no) | 1991-04-03 |
| AU7413091A (en) | 1991-11-14 |
| DK0450386T3 (da) | 2000-03-06 |
| TW206258B (ko) | 1993-05-21 |
| ATE183232T1 (de) | 1999-08-15 |
| NO911303L (no) | 1991-10-07 |
| FI108146B (fi) | 2001-11-30 |
| CA2038669C (en) | 2004-06-29 |
| IE910827A1 (en) | 1991-10-09 |
| EP0450386A2 (en) | 1991-10-09 |
| FI911643A (fi) | 1991-10-07 |
| DE69131514T2 (de) | 2000-05-04 |
| IL97602A0 (en) | 1992-06-21 |
| FI911643A0 (fi) | 1991-04-05 |
| ES2140379T3 (es) | 2000-03-01 |
| JP2521851B2 (ja) | 1996-08-07 |
| JPH06319549A (ja) | 1994-11-22 |
| DE69131514D1 (de) | 1999-09-16 |
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