ES2534040T3 - Plasmina modificada de forma recombinante - Google Patents
Plasmina modificada de forma recombinante Download PDFInfo
- Publication number
- ES2534040T3 ES2534040T3 ES08857923.0T ES08857923T ES2534040T3 ES 2534040 T3 ES2534040 T3 ES 2534040T3 ES 08857923 T ES08857923 T ES 08857923T ES 2534040 T3 ES2534040 T3 ES 2534040T3
- Authority
- ES
- Spain
- Prior art keywords
- seq
- plasminogen
- domain
- terminal
- tal6003
- 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.)
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Classifications
-
- 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
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6435—Plasmin (3.4.21.7), i.e. fibrinolysin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21007—Plasmin (3.4.21.7), i.e. fibrinolysin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Dispersion Chemistry (AREA)
- Hematology (AREA)
- Diabetes (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Polinucleótido, caracterizado porque comprende una secuencia de nucleótidos que codifica un polipéptido que es por lo menos 95% idéntico a la SEQ ID NO: 2, que tiene a) un dominio kringle N-terminal único homólogo a un dominio kringle del plasminógeno humano nativo, en el que los cuatro últimos residuos de aminoácidos dentro del dominio kringle son V, P, Q y C; y b) un sitio de activación del dominio C-terminal y un dominio de serina proteasa homólogos a los dominios correspondientes en el plasminógeno humano; en los que el polipéptido se une a lisina inmovilizada.
Description
E08857923
30-03-2015
EJEMPLOS
5 La secuencia de aminoácidos para (TLA6003)-plasminógeno se muestra en SEQ ID NO: 2. En un polinucleótido que tiene la secuencia de nucleótidos que codifica (TAL6003)-plasminógeno se optimizaron los codones por la expresión de
E. coli y la estabilidad de ARNm para proporcionar la secuencia de ADN tal como se muestra en SEQ ID NO: 1. Este polinucleótido se clonó en los sitios Ndel y BamHI del vector de expresión de E. coli pET24b(+) (Novagen; Madison, WI) para producir la proteína citosólica.
10 Como se ilustra en la tabla 1, la expresión en bacterias (por ejemplo E. coli) proporciona un polipéptido (TAL6003)-plasminógeno recombinante que tiene la secuencia de aminoácidos tal como se muestra en SEQ ID NO: 2 (es decir, un (TAL6003)-plasminógeno recombinante con una metionina N-terminal (es decir, M1) que precede inmediatamente el residuo de aminoácidos de arginina (es decir, R2) correspondiente a la arginina en la posición 70 (es
15 decir, R70) de la secuencia de aminoácidos de plasminógeno humano nativo mostrado en SEQ ID NO: 4 (véase también, por ejemplo, la Figura 3). Dicho producto recombinante es susceptible a escisión adicional para producir proteínas adicionales que tienen extremos N-terminales diferentes que incluyen una proteína con una lisina N-terminal (es decir, K10) o valina (es decir, V11) correspondiente, respectivamente, a la lisina en la posición 78 (es decir, K78) o la valina en la posición 79 (es decir, V79) de plasminógeno humano nativo.
20
- Tabla 1: Extremos N-terminales de plasmina(plasminógeno) nativo (por ejemplo, basado en SEQ ID NO:4) y (TAL6003)-plasmina(plasminógeno) (por ejemplo, basado en SEQ ID NO: 2, o su variante)
- Plasminógeno nativo que comprende una secuencia lectora de 19 aminoácidos (por ejemplo basado en SEQ ID NO: 4):
- “Lys-plasminógeno” nativo (es decir, escisión de secuencia lectora): (véase SEQ ID NO:5)
- Especies de plasmina nativa posibles basadas en escisión, si es cualquiera, de Lys-Plasminógeno: (véase SEQ ID NO: 14) (véase SEQ ID NO: 13) (véase SEQ ID NO: 12)
- Polipéptidos de (TAL6003)-plasminógeno recombinantes de la presente invención: (por ejemplo, basados en SEQ ID NO: 2)
- Proteínas adicionales basadas en escisión adicional de un (TAL6003)-plasminógeno (por ejemplo, basadas en SEQ ID NO: 2) (véase SEQ ID NO: 11) K10V11YLSEC ….. (SEQ ID NO: 5) V11YLSEC …..
- ↓ indica sitios de escisión potencial
El vector de expresión que comprende el ADN que codifica (TAL6003)-plasminógeno se transforma en una variedad de
25 células que incluyen Bl21(DE3)RIL (Stratagene, La Jolla, CA), BL21(DE3) (genotipo: F -ompT hsdSB (rB-mB-) gal dcm (DE3)) (EMB Biosciences, Inc., San Diego, CA), y BLR(DE3) (genotipo: F-ampT hsdSB (rB -mB -) gal dcm (DE3) Δ(srl-recA)306::Tn10(TetR)), y la sobreexpresión de proteína seguida por la inducción por IPTG 1mM (isopropil-beta-D-tiogalactopiranósido) se analizó por SDS-PAGE. Los estimados de expresión fueron, como mínimo, de aproximadamente 250 mg/l de cultivo celular en matraces agitadores.
30 La célula tipo BL21(DE3)RIL se diseña para expresar ARNt de E. coli raros que codifican para Arg, IIe, y Leu. Además, tanto BL21(DE3) como BLR(DE3) son cepas B de E. coli que se clasifican como no patogénicas para humanos y animales basados en la ausencia de factores de virulencia y colonización. Las células BLR(DE3) carecen del gen recA para recombinación de ADN, y no se ha informado de la inducción de fago lambda con estas células. Un banco celular
11
Claims (1)
-
imagen1
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US99114807P | 2007-11-29 | 2007-11-29 | |
| US991148P | 2007-11-29 | ||
| PCT/US2008/084645 WO2009073471A1 (en) | 2007-11-29 | 2008-11-25 | Recombinantly modified plasmin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2534040T3 true ES2534040T3 (es) | 2015-04-16 |
Family
ID=40379999
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES08857923.0T Active ES2534040T3 (es) | 2007-11-29 | 2008-11-25 | Plasmina modificada de forma recombinante |
Country Status (18)
| Country | Link |
|---|---|
| US (3) | US8101394B2 (es) |
| EP (1) | EP2220221B1 (es) |
| JP (1) | JP5539896B2 (es) |
| KR (1) | KR101529743B1 (es) |
| CN (1) | CN101918548B (es) |
| AU (1) | AU2008331545B2 (es) |
| BR (1) | BRPI0819780B1 (es) |
| CA (1) | CA2707266C (es) |
| ES (1) | ES2534040T3 (es) |
| HK (1) | HK1146089A1 (es) |
| HU (1) | HUE024916T2 (es) |
| IL (1) | IL205936A (es) |
| MX (1) | MX2010005947A (es) |
| NZ (2) | NZ585715A (es) |
| PL (1) | PL2220221T3 (es) |
| PT (1) | PT2220221E (es) |
| WO (1) | WO2009073471A1 (es) |
| ZA (1) | ZA201003730B (es) |
Families Citing this family (27)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8034913B2 (en) | 2004-04-22 | 2011-10-11 | Grifols Therapeutics Inc. | Recombinantly modified plasmin |
| NZ585715A (en) | 2007-11-29 | 2011-11-25 | Talecris Biotherapeutics Inc | Recombinantly modified plasmin |
| AU2009256168B2 (en) | 2008-06-04 | 2014-06-05 | Grifols Therapeutics Inc. | Composition, method and kit for preparing plasmin |
| US9206410B2 (en) * | 2009-03-03 | 2015-12-08 | Grifols Therapeutics Inc. | Compositions, methods and kits for preparing plasminogen and plasmin prepared therefrom |
| KR20120050442A (ko) | 2009-07-10 | 2012-05-18 | 쓰롬보제닉스 엔.브이. | 플라스미노겐 및 플라스민의 변이체 |
| EP2480249B1 (en) | 2009-08-28 | 2015-01-28 | ThromboGenics N.V. | Plasmin for the treatment of filtration failure after trabeculectomy |
| CN102121023B (zh) * | 2010-12-22 | 2012-07-04 | 中山大学 | 突变型人纤溶酶原kringle5及其制备方法及应用 |
| JP6085568B2 (ja) | 2011-01-05 | 2017-02-22 | スロンボジェニックス・ナムローゼ・フェンノートシャップＴｈｒｏｍｂｏＧｅｎｉｃｓ ＮＶ | プラスミノーゲンおよびプラスミンの変異体 |
| AU2012296884B2 (en) | 2011-08-12 | 2015-02-05 | Thrombogenics N.V. | Plasminogen and plasmin variants |
| CN103205410B (zh) * | 2012-01-13 | 2018-03-06 | 蔺莹莹 | 重组微纤维蛋白溶酶及其制备方法和应用 |
| UY35457A (es) | 2013-03-14 | 2014-10-31 | Bayer Healthcare Llc | ANTICUERPOS MONOCLONALES CONTRA ANTITROMBINA ß |
| DE102014112212A1 (de) * | 2014-08-26 | 2016-03-03 | Akesion Gmbh | Rekombinante Fusionsproteine zur Vorbeugung oder Behandlung von Adhäsionen bei Geweben oder Organen |
| FR3035120B1 (fr) * | 2015-04-15 | 2020-02-07 | Arcadophta | Composition de plasminogenase immobilisee, procede de preparation, utilisation et dispositif comprenant une telle composition |
| TWI653981B (zh) | 2015-12-18 | 2019-03-21 | 深圳瑞健生命科學硏究院有限公司 | Use of plasminogen for the preparation of a medicament for preventing or treating a condition associated with diabetic nerve injury |
| US10709771B2 (en) | 2015-12-18 | 2020-07-14 | Talengen International Limited | Method for preventing or treating diabetic retinopathy |
| CA3008686C (en) | 2015-12-18 | 2023-03-14 | Talengen International Limited | Method for preventing and treating diabetic nephropathy |
| CA3008186A1 (en) | 2015-12-18 | 2017-06-22 | Talengen International Limited | Novel method for preventing and treating angiocardiopathy |
| CA3047175A1 (en) | 2016-12-15 | 2018-06-21 | Talengen International Limited | Method for mitigating heart disease |
| TWI644682B (zh) * | 2016-12-15 | 2018-12-21 | 深圳瑞健生命科學硏究院有限公司 | 一種纖溶酶原在製備促進胰島細胞損傷修復的藥物上的用途 |
| CN110191718A (zh) | 2016-12-15 | 2019-08-30 | 泰伦基国际有限公司 | 一种预防和治疗组织器官纤维化的方法 |
| JP7214225B2 (ja) * | 2016-12-15 | 2023-01-30 | タレンゲン インターナショナル リミテッド | 脂肪肝を予防および治療するための方法 |
| CN108210905A (zh) * | 2016-12-15 | 2018-06-29 | 深圳瑞健生命科学研究院有限公司 | 预防和治疗皮肤纤维化的药物及其用途 |
| WO2018108161A1 (zh) | 2016-12-15 | 2018-06-21 | 深圳瑞健生命科学研究院有限公司 | 一种预防和治疗肥胖症的方法和药物 |
| CN111344004A (zh) | 2017-06-19 | 2020-06-26 | 泰伦基国际有限公司 | 一种调控glp-1/glp-1r 的方法和药物 |
| US11140020B1 (en) | 2018-03-01 | 2021-10-05 | Amazon Technologies, Inc. | Availability-enhancing gateways for network traffic in virtualized computing environments |
| WO2020152713A1 (en) * | 2019-01-25 | 2020-07-30 | Council Of Scientific & Industrial Research | Fibrinolytic composition and method of its preparation |
| KR20230005298A (ko) * | 2020-05-11 | 2023-01-09 | 탈렌젠 인터내셔널 리미티드 | 척수성 근위축증의 치료 방법 및 약물 |
Family Cites Families (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4652639A (en) | 1982-05-06 | 1987-03-24 | Amgen | Manufacture and expression of structural genes |
| US4631211A (en) | 1985-03-25 | 1986-12-23 | Scripps Clinic & Research Foundation | Means for sequential solid phase organic synthesis and methods using the same |
| US4774087A (en) | 1987-08-14 | 1988-09-27 | Northwestern University | Micro-size fibrinolytic plasmin |
| US7253264B1 (en) | 1990-06-28 | 2007-08-07 | Sanofi-Arentideutschland GmbH | Immunoglobulin fusion proteins, their production and use |
| US6312893B1 (en) | 1996-01-23 | 2001-11-06 | Qiagen Genomics, Inc. | Methods and compositions for determining the sequence of nucleic acid molecules |
| KR100482917B1 (ko) | 1996-01-23 | 2005-11-11 | 퀴아진 지노믹스, 인코포레이티드 | 핵산분자의서열을결정하는방법및이를위한조성물 |
| US6613508B1 (en) | 1996-01-23 | 2003-09-02 | Qiagen Genomics, Inc. | Methods and compositions for analyzing nucleic acid molecules utilizing sizing techniques |
| JPH10158300A (ja) | 1996-11-28 | 1998-06-16 | Suzuki Motor Corp | 血管新生抑制効果を有するタンパク質とその製造方法及びアンギオスタチンの製造方法 |
| NZ501774A (en) | 1997-07-22 | 2002-02-01 | Qiagen Genomics Inc | Computer method and system for correlating known characteristics of biomolecules to molecular tags with unique molecular weights that are associated with the biomolecule |
| JP2002520367A (ja) | 1998-07-14 | 2002-07-09 | ブリストル−マイヤーズ スクイブ カンパニー | アンギオスタチンのリジン結合フラグメント |
| EP1117437B1 (en) | 1998-09-29 | 2006-12-27 | Leuven Research & Development vzw | Use of compounds that reduce alpha2-antiplasmin in vivo for the preparation of a composition for the treatment of ischemic stroke |
| US6969515B2 (en) | 1999-11-13 | 2005-11-29 | Talecris Biotherapeutics, Inc. | Method of thrombolysis by local delivery of reversibly inactivated acidified plasmin |
| US6964764B2 (en) | 1999-11-13 | 2005-11-15 | Talecris Biotherapeutics, Inc. | Method of thrombolysis by local delivery of reversibly inactivated acidified plasmin |
| US6355243B1 (en) | 1999-11-13 | 2002-03-12 | Bayer Corporation | Method of thrombolysis by local delivery of active plasmin |
| US6664112B2 (en) | 2000-06-02 | 2003-12-16 | Blue Heron Biotechnology, Inc. | Methods for improving the sequence fidelity of synthetic double-stranded oligonucleotides |
| WO2002050290A1 (en) * | 2000-12-21 | 2002-06-27 | Thromb-X Nv | A yeast expression vector and a method of making a recombinant protein by expression in a yeast cell |
| AU2002357249A1 (en) | 2001-12-13 | 2003-07-09 | Blue Heron Biotechnology, Inc. | Methods for removal of double-stranded oligonucleotides containing sequence errors using mismatch recognition proteins |
| DE10390418D2 (de) | 2002-02-06 | 2005-01-13 | N Zyme Biotec Gmbh | Verfahren zur Herstellung von rekombinanten Proteinen in Mikroorganismen |
| US7776026B2 (en) | 2002-02-06 | 2010-08-17 | Nuvue Technologies, Inc. | Method for vitreous liquefaction |
| GB0228409D0 (en) * | 2002-12-06 | 2003-01-08 | Thromb X Nv | Pharmacological vitreolysis |
| US8034913B2 (en) * | 2004-04-22 | 2011-10-11 | Grifols Therapeutics Inc. | Recombinantly modified plasmin |
| WO2007047874A2 (en) * | 2005-10-20 | 2007-04-26 | Talecris Biotherapeutics, Inc. | Recombinantly modified plasmin |
| NZ585715A (en) | 2007-11-29 | 2011-11-25 | Talecris Biotherapeutics Inc | Recombinantly modified plasmin |
-
2008
- 2008-11-25 NZ NZ585715A patent/NZ585715A/en unknown
- 2008-11-25 HU HUE08857923A patent/HUE024916T2/en unknown
- 2008-11-25 AU AU2008331545A patent/AU2008331545B2/en active Active
- 2008-11-25 KR KR1020107014243A patent/KR101529743B1/ko active IP Right Grant
- 2008-11-25 WO PCT/US2008/084645 patent/WO2009073471A1/en active Application Filing
- 2008-11-25 MX MX2010005947A patent/MX2010005947A/es active IP Right Grant
- 2008-11-25 CN CN2008801232704A patent/CN101918548B/zh active Active
- 2008-11-25 US US12/744,376 patent/US8101394B2/en active Active
- 2008-11-25 ES ES08857923.0T patent/ES2534040T3/es active Active
- 2008-11-25 JP JP2010536117A patent/JP5539896B2/ja active Active
- 2008-11-25 BR BRPI0819780-6A patent/BRPI0819780B1/pt active IP Right Grant
- 2008-11-25 CA CA2707266A patent/CA2707266C/en active Active
- 2008-11-25 NZ NZ593837A patent/NZ593837A/xx unknown
- 2008-11-25 EP EP08857923.0A patent/EP2220221B1/en active Active
- 2008-11-25 PL PL08857923T patent/PL2220221T3/pl unknown
- 2008-11-25 PT PT88579230T patent/PT2220221E/pt unknown
-
2010
- 2010-05-24 IL IL205936A patent/IL205936A/en active IP Right Grant
- 2010-05-25 ZA ZA2010/03730A patent/ZA201003730B/en unknown
-
2011
- 2011-01-12 HK HK11100248.7A patent/HK1146089A1/xx unknown
- 2011-11-22 US US13/302,322 patent/US8182808B2/en active Active
-
2012
- 2012-05-03 US US13/463,437 patent/US8512980B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| US8101394B2 (en) | 2012-01-24 |
| US20120093799A1 (en) | 2012-04-19 |
| BRPI0819780B1 (pt) | 2023-01-24 |
| CN101918548A (zh) | 2010-12-15 |
| PL2220221T3 (pl) | 2015-07-31 |
| ZA201003730B (en) | 2012-11-28 |
| KR20100102135A (ko) | 2010-09-20 |
| IL205936A (en) | 2015-04-30 |
| HK1146089A1 (en) | 2011-05-13 |
| JP5539896B2 (ja) | 2014-07-02 |
| IL205936A0 (en) | 2010-11-30 |
| AU2008331545B2 (en) | 2014-01-16 |
| CN101918548B (zh) | 2013-10-16 |
| KR101529743B1 (ko) | 2015-06-17 |
| MX2010005947A (es) | 2010-09-10 |
| JP2011505140A (ja) | 2011-02-24 |
| CA2707266C (en) | 2013-12-31 |
| US8182808B2 (en) | 2012-05-22 |
| BRPI0819780A2 (pt) | 2015-08-11 |
| EP2220221B1 (en) | 2014-12-31 |
| HUE024916T2 (en) | 2016-02-29 |
| WO2009073471A1 (en) | 2009-06-11 |
| PT2220221E (pt) | 2015-04-16 |
| EP2220221A1 (en) | 2010-08-25 |
| US20110003332A1 (en) | 2011-01-06 |
| US20120276611A1 (en) | 2012-11-01 |
| NZ593837A (en) | 2013-01-25 |
| AU2008331545A1 (en) | 2009-06-11 |
| US8512980B2 (en) | 2013-08-20 |
| NZ585715A (en) | 2011-11-25 |
| CA2707266A1 (en) | 2009-06-11 |
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