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1. 发明授权

US5798227A Co-expression of alpha and beta globins 失效
公开(公告)号：US5798227A
公开(公告)日：1998-08-25
申请号：US446105
申请日：1995-05-19
申请人： Stephen J. Hoffman , Douglas L. Looker , Gary L. Stetler , Michael Wagenbach
发明人： Stephen J. Hoffman , Douglas L. Looker , Gary L. Stetler , Michael Wagenbach
IPC分类号： A61K38/16 , A61K38/00 , A61K47/48 , A61P7/08 , C07K20060101 , C07K1/113 , C07K14/805 , C12N15/00 , C12N15/09 , C12N15/12 , C12N15/81 , C12P21/02 , C12R1/19 , C12R1/91 , C12P21/06 , C12N1/20 , C12P21/04
CPC分类号： C07K14/805 , A61K47/48307 , C12N15/81 , A61K38/00 , C07K2319/00 , C07K2319/50 , C07K2319/92
摘要： The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained. We further describe the production of tetrameric human hemoglobin and di-alpha/beta.sub.2 hemoglobin in the yeast Saccharomyces cerevisiae. The synthesis of the protein is directed by a synthetic promotor consisting of two functional parts, an upstream activator sequence (UAS) that confers inducible transcription by galactose from a consensus yeast transcriptional initiation site. The expression construct is designed such that translation is expected to initiate at the same position as the human wild-type genes for .alpha.- and .beta.-globin. Three different types of expression vectors have been used: (1) .alpha.-globin and .beta.-globin contained on two separate plasmids (pGS4688 and pGS4988) in a diploid yeast strain; (2) .alpha.-globin and .beta.-globin each contained on a single plasmid (pGS289 and pGS389) and expressed in either haploid or diploid strains; and (3) di-alpha-globin and beta globin contained on a single plasmid (pGS 3089)and expressed in haploid strains. Finally, we describe the co-expression of alpha and beta globin chains. The chains are folded together and combined intracellularly with heme to form active tetrameric hemoglobin. The hemoglobin may be recovered from the cells' soluble fraction. The invention thus obviates the need to express alpha and beta globin separately, solubilize, renature and purify them, and combine them in vitro with heme to obtain an artificial hemoglobin. By way of comparison, the separately expressed beta globin known in the art is deposited in inclusion bodies. Polycistronic co-expression of alpha and beta globins is particularly preferred.

2. 发明授权

US5744329A DNA encoding fused di-beta globins and production of pseudotetrameric hemoglobin 失效
标题翻译：编码融合的二-β珠蛋白的DNA和伪三聚体血红蛋白的产生
公开(公告)号：US5744329A
公开(公告)日：1998-04-28
申请号：US444942
申请日：1995-05-19
申请人： Stephen J. Hoffman , Douglas L. Looker , Mary S. Rosendahl , Gary L. Stetler , Michael Wagenbach , David C. Anderson , Antony James Mathews , Kiyoshi Nagai
发明人： Stephen J. Hoffman , Douglas L. Looker , Mary S. Rosendahl , Gary L. Stetler , Michael Wagenbach , David C. Anderson , Antony James Mathews , Kiyoshi Nagai
IPC分类号： A61K38/16 , A61K38/00 , A61K47/48 , A61P7/08 , C07K20060101 , C07K1/113 , C07K14/805 , C12N15/00 , C12N15/09 , C12N15/12 , C12N15/81 , C12P21/02 , C12R1/19 , C12R1/91 , C12P21/06 , C07H17/00
CPC分类号： C07K14/805 , A61K47/48307 , C12N15/81 , A61K38/00 , C07K2319/00 , C07K2319/50 , C07K2319/92
摘要： The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.
摘要翻译：在本文中，血红蛋白的α亚基本质上被形成为与β-亚基非共价结合的分离的多肽链，这里以新分子二-α球蛋白的形式提供，单链多肽链通过将两个α亚基连接而定义直接通过肽键或通过柔性氨基酸或肽接头间接。 Di-α珠蛋白可以在体内或体外与β球蛋白和血红素组合以形成血红蛋白。 Di-α球蛋白通过重组DNA技术表达。可以类似地获得二β-珠蛋白。

3. 发明授权

US5545727A DNA encoding fused di-alpha globins and production of pseudotetrameric hemoglobin 失效
标题翻译：编码融合二-α球蛋白的DNA和伪三聚体血红蛋白的产生
公开(公告)号：US5545727A
公开(公告)日：1996-08-13
申请号：US789179
申请日：1991-11-08
申请人： Stephen J. Hoffman , Douglas L. Looker , Kiyoshi Nagai
发明人： Stephen J. Hoffman , Douglas L. Looker , Kiyoshi Nagai
IPC分类号： A61K38/16 , A61K38/00 , A61K47/48 , A61P7/08 , C07K20060101 , C07K1/113 , C07K14/805 , C12N15/00 , C12N15/09 , C12N15/12 , C12N15/81 , C12P21/02 , C12R1/19 , C12R1/91
CPC分类号： C07K14/805 , A61K47/48307 , C12N15/81 , A61K38/00 , C07K2319/00 , C07K2319/50 , C07K2319/92
摘要： The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.DNA encoding alpha globin fusion proteins is provided.
摘要翻译：在本文中，血红蛋白的α亚基本质上被形成为与β-亚基非共价结合的分离的多肽链，这里以新分子二-α球蛋白的形式提供，单链多肽链通过将两个α亚基连接而定义直接通过肽键或通过柔性氨基酸或肽接头间接。 Di-α珠蛋白可以在体内或体外与β球蛋白和血红素组合以形成血红蛋白。 Di-α球蛋白通过重组DNA技术表达。可以类似地获得二β-珠蛋白。提供了编码α球蛋白融合蛋白的DNA。

4. 发明授权

US07211560B2 Reduced side-effect hemoglobin compositions 有权
标题翻译：降低副作用血红蛋白组成
公开(公告)号：US07211560B2
公开(公告)日：2007-05-01
申请号：US10747580
申请日：2003-12-29
申请人： Douglas L. Looker , Izydor Z. Apostol , Eric A. Brucker , Michael P. Doyle , David L. Foster , Christopher B. Glascock , James C. Hartman , Geoffrey F. Lee , Douglas D. Lemon , Edwin G. Moore , Jane P. Richards , Michael R. Schick , Stephen P. Trimble , David Pereira , Ton-That Hai , Kenneth E. Burhop
发明人： Douglas L. Looker , Izydor Z. Apostol , Eric A. Brucker , Michael P. Doyle , David L. Foster , Christopher B. Glascock , James C. Hartman , Geoffrey F. Lee , Douglas D. Lemon , Edwin G. Moore , Jane P. Richards , Michael R. Schick , Stephen P. Trimble , David Pereira , Ton-That Hai , Kenneth E. Burhop
IPC分类号： A61K35/14
CPC分类号： C07K14/805 , A61K38/00
摘要： The invention relates to novel hemoglobin compositions, particularly novel recombinant mutant hemoglobin compositions, which eliminate or substantially reduce 1) the creation of heart lesions, 2) gastrointestinal discomfort, 3) pressor effects, and 4) endotoxin hypersensitivity associated with the administration of extracellular hemoglobin compositions in various therapeutic applications. Applications described include treatments for anemia, head injury, hemorrhage or hypovolemia, ischemia, cachexia, sickle cell crisis and stroke; enhancing cancer treatments; stimulating hematopoiesis; improving repair of physically damaged tissues; alleviating cardiogenic shock; and shock resuscitation.
摘要翻译：本发明涉及新型血红蛋白组合物，特别是新型重组突变体血红蛋白组合物，其消除或显着减少1）心脏病变的产生，2）胃肠道不适，3）加压作用，和4）与施用细胞外血红蛋白相关的内毒素超敏反应各种治疗应用中的组合物。描述的应用包括贫血，头部损伤，出血或血容量不足，局部缺血，恶病质，镰状细胞危象和中风的治疗。加强癌症治疗; 刺激造血; 改善身体损伤组织的修复; 减轻心源性休克; 和休克复苏。

5. 发明授权

US06274331B1 Method of determining a functional linker for fusing globin subunts 失效
标题翻译：确定用于融合珠蛋白的功能性连接体的方法
公开(公告)号：US06274331B1
公开(公告)日：2001-08-14
申请号：US08444915
申请日：1995-05-19
申请人： Douglas L. Looker , Gary L. Stetler
发明人： Douglas L. Looker , Gary L. Stetler
IPC分类号： G01N3353
CPC分类号： C07K14/805 , A61K38/00 , A61K47/6445 , C07K2319/00 , C07K2319/50 , C07K2319/92 , C12N15/81
摘要： A functional linker for a polypeptide in which two alpha or beta globin-like domains are genetically fused is determined by screening a library of genetically fused polypeptides, in which the linker region is varied, for the ability to participate in the formation of hemoglobin-like protein, as measured by the protein's response to carbon monoxide. In a preferred embodiment, cells expressing the protein turn red as a result of carbon monoxide pressure.
摘要翻译：通过筛选遗传融合多肽的文库，测定其中两个α或β球蛋白样结构域经遗传融合的功能性连接体，其中连接区不同，参与血红蛋白样形成的能力蛋白质，通过蛋白质对一氧化碳的反应测量。在优选的实施方案中，由于一氧化碳压力，表达蛋白质的细胞变红。

6. 发明授权

US06242417B1 Stabilized compositions containing hemoglobin 失效
标题翻译：含有血红蛋白的稳定组合物
公开(公告)号：US06242417B1
公开(公告)日：2001-06-05
申请号：US08417644
申请日：1995-04-05
申请人： Michael J. Gerber , Douglas L. Looker , Bruce A. Kerwin
发明人： Michael J. Gerber , Douglas L. Looker , Bruce A. Kerwin
IPC分类号： A61K3816
CPC分类号： C12N5/0641 , A61K38/42 , C12N2500/24 , C12N2501/125 , C12N2501/14 , C12N2501/23 , Y10T436/105831 , Y10T436/106664 , A61K2300/00
摘要： The present invention relates to a hemoglobin compositions stabilized against the formation of aggregates. The present invention further relates to methods of making such hemoglobin compositions.
摘要翻译：本发明涉及针对聚集体形成而稳定的血红蛋白组合物。本发明还涉及制备这种血红蛋白组合物的方法。

7. 发明授权

US5844089A Genetically fused globin-like polypeptides having hemoglobin-like activity 失效
标题翻译：具有血红蛋白样活性的遗传融合的球蛋白样多肽
公开(公告)号：US5844089A
公开(公告)日：1998-12-01
申请号：US450733
申请日：1995-05-25
申请人： Stephen J. Hoffman , Douglas L. Looker , Mary S. Rosendahl , Gary L. Stetler , Michael Wagenbach , David C. Anderson , Antony James Mathews , Kiyoshi Nagai
发明人： Stephen J. Hoffman , Douglas L. Looker , Mary S. Rosendahl , Gary L. Stetler , Michael Wagenbach , David C. Anderson , Antony James Mathews , Kiyoshi Nagai
IPC分类号： A61K38/16 , A61K38/00 , A61K47/48 , A61P7/08 , C07K20060101 , C07K1/113 , C07K14/805 , C12N15/00 , C12N15/09 , C12N15/12 , C12N15/81 , C12P21/02 , C12R1/19 , C12R1/91
CPC分类号： C07K14/805 , A61K47/48307 , C12N15/81 , A61K38/00 , C07K2319/00 , C07K2319/50 , C07K2319/92
摘要： The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.
摘要翻译：在本文中，血红蛋白的α亚基本质上被形成为与β-亚基非共价结合的分离的多肽链，这里以新分子二-α球蛋白的形式提供，单链多肽链通过将两个α亚基连接而定义直接通过肽键或通过柔性氨基酸或肽接头间接。 Di-α珠蛋白可以在体内或体外与β球蛋白和血红素组合以形成血红蛋白。 Di-α球蛋白通过重组DNA技术表达。可以类似地获得二β-珠蛋白。

8. 发明授权

US5929031A Storage stable hemoglobin solutions 失效
标题翻译：储存稳定型血红蛋白溶液
公开(公告)号：US5929031A
公开(公告)日：1999-07-27
申请号：US433896
申请日：1995-05-02
申请人： Bruce A. Kerwin , Douglas L. Looker
发明人： Bruce A. Kerwin , Douglas L. Looker
IPC分类号： A61K38/16 , A61P7/00 , C07K14/805 , A61K35/14 , C07K1/00
CPC分类号： C07K14/805
摘要： The present invention relates to storage stable hemoglobin solutions which contain partially deoxygenated and surprisingly low amounts of reducing agents. Methods for preparing such storage stable hemoglobin solutions are also provided as well as a systems for storing the solutions.
摘要翻译：本发明涉及含有部分脱氧和令人惊奇的少量还原剂的储存稳定型血红蛋白溶液。还提供了制备这种储存稳定血红蛋白溶液的方法以及用于储存溶液的系统。

9. 发明授权

US5914391A Stabilized compositions containing hemoglobin 失效
标题翻译：含有血红蛋白的稳定组合物
公开(公告)号：US5914391A
公开(公告)日：1999-06-22
申请号：US913036
申请日：1997-11-17
申请人： Michael J. Gerber , Douglas L. Looker , Bruce A. Kerwin
发明人： Michael J. Gerber , Douglas L. Looker , Bruce A. Kerwin
IPC分类号： A61K38/42 , C12N5/078 , A61K35/14 , A61K38/06
CPC分类号： C12N5/0641 , A61K38/42 , C12N2500/24 , C12N2501/125 , C12N2501/14 , C12N2501/23 , Y10T436/105831 , Y10T436/106664
摘要： The present invention relates to hemoglobin compositions stabilized against the formation of aggregates. Such compositions contain at least a surfactant, said surfactant not being an adduct of a polymer and an anionic ligand. The present invention further relates to methods of making such hemoglobin compositions.
摘要翻译： PCT No.PCT / US95 / 10232 Sec。 371日期：1997年11月17日 102（e）日期1997年11月17日PCT 1995年8月10日PCT PCT。 WO96 / 27388 PCT公开日期1996年9月12日本发明涉及抗凝集物形成稳定的血红蛋白组合物。这种组合物至少含有表面活性剂，所述表面活性剂不是聚合物和阴离子配体的加合物。本发明还涉及制备这种血红蛋白组合物的方法。

10. 发明授权

US06828125B1 DNA encoding fused di-alpha globins and use thereof 失效
标题翻译：编码融合二-α球蛋白的DNA及其用途
公开(公告)号：US06828125B1
公开(公告)日：2004-12-07
申请号：US08453485
申请日：1995-05-30
申请人： Stephen J. Hoffman , Douglas L. Looker , Kiyoshi Nagai
发明人： Stephen J. Hoffman , Douglas L. Looker , Kiyoshi Nagai
IPC分类号： C12P2106
CPC分类号： C07K14/805 , A61K38/00 , A61K47/6445 , C07K2319/00 , C12N15/81
摘要： The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.
摘要翻译：在本文中，血红蛋白的α亚基本质上被形成为与β-亚基非共价结合的分离的多肽链，这里以新分子二-α球蛋白的形式提供，单链多肽链通过将两个α亚基连接而定义直接通过肽键或通过柔性氨基酸或肽接头间接。 Di-α珠蛋白可以在体内或体外与β球蛋白和血红素组合以形成血红蛋白。 Di-α球蛋白通过重组DNA技术表达。可以类似地获得二β-珠蛋白。

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