公开(公告)号：EP0691401A1

公开(公告)日：1996-01-10

申请号：EP95106995.4

申请日：1988-03-25

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Hartman, Jacob R. ,  Beck, Yaffa ,  Nimrod, Abraham

IPC分类号： C12N9/02 ,  A61K38/44

CPC分类号： C12N9/0089 ,  A61K38/446

摘要： There are disclosed a human manganese superoxide dismutase analog in which at least one of the human manganese superoxide dismutase polypeptides differs from naturally-occurring human manganese superoxide dismutase polypeptide by the absence of lysine at the N-terminus, and a human manganese superoxide dismutase analog in which at least one of the human manganese superoxide dismutase polypeptides differs from naturally-occurring human manganese superoxide dismutase polypeptide by the absence of both lysine and histidine at the N-terminus, pharmaceutical compositions comprising same in an amount effective to alleviate the undesirable symptoms associated with the generation of oxygen free radicals and a pharmaceutically acceptable carrier, as well as their use for the production of a pharmaceutical composition for alleviating the symtoms in a subject suffering from a disorder selected from synovial inflammation, arthritis and lung fibrosis.

摘要翻译： 有圆盘游离缺失人锰超氧化物歧化酶模拟其中人锰超氧化物歧化酶多肽的至少一个从天然存在的人锰超氧化物歧化酶多肽由在N末端缺少赖氨酸的不同，和人锰超氧化物歧化酶模拟在 人锰超氧化物歧化酶多肽，其中至少一个从天然存在的人锰超氧化物歧化酶多肽通过在N-末端没有两个赖氨酸和组氨酸的不同。药物组合物在用量能有效地减轻与相关联的不期望的症状包括相同的 产生氧自由基和药学上可接受的载体，以及其用于制备药物组合物以用于从滑液炎症，关节炎和肺纤维化的病症的受试者减轻symtoms使用。

公开(公告)号：EP0483113B1

公开(公告)日：1999-09-22

申请号：EP92101250.6

申请日：1985-08-26

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Hartman, Jacob R. ,  Kanner, Dov ,  Bartfeld, Daniel

IPC分类号： C12N15/53 ,  C12N9/02

CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

公开(公告)号：EP0173280A1

公开(公告)日：1986-03-05

申请号：EP85110683.1

申请日：1985-08-26

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Aviv, Haim ,  Bartfeld, Daniel ,  Gorecki, Marian ,  Hartman, Jacob R. ,  Kanner, Dov ,  Levanon, Avigdor ,  Locker-Giladi, Hilla ,  Oppenheim, Amos B.

IPC分类号： C12N15/73 ,  C12P21/02 ,  C12N15/67 ,  C12P19/34

CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

摘要： An improved vector which upon introduction into a suitable host containing the thermolabile repressor C 1 renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P L O L from lambda bacteriophage; the N utilization site; a first restriction enzyme site which follows thereafter; a ribosomal binding site; an ATG Initiation codon or DNA which is converted Into an ATG initiation codon upon insertion of the desired gene into the vector, a second restriction enzme site for inserting the gene in phase with the ATG initiation codon; an origin of replication and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present In the host or a fragment designated ci 434 which includes the gene for the cl 343 repressor protein and its associated promoter and operator. The distance between the 3' end of P L O L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs.
Vectors of the invention may also include a T,T 2 rRNA transcription termination sequence located 3' of the second restriction enzyme site. The inventive vectors may have origin of replications capable of autonomous production in the host of at least 400 constitutive copies. Plasmids have been constructed from the vectors and used to produce bovine, chicken, porcine and human growth hormones. human apolipoprotein E and human superoxide dismutase and ana- loges thereof In host cells.
Such superoxide dismutase or analogs thereof may be used to catalyze the reduction of superoxide radicais, reduce reperfusion injury, prolong the survival time of isolated organs and prevent neurological InJury.
Enzymatically active eucaryotic superoxide dismutase or an analog thereof may also be produced by an improved method of this invention which employs a production medium having a concentration of Cu ++ greater than about about 2 ppm.
. The invention also concerns improved methods of ro- covering purified enzymatically active eucaryotic superoxide dismutase or analogs thereof.

摘要翻译： （1）向做了引入合适的细菌宿主细胞contg。 不耐热阻遏C（I）使宿主细胞能，当温度。 被升高到使阻遏物失活 - 影响由该基因编码插入以产生多肽（Ⅰ）载体中的目的基因的表达的，电平是新当其包含双链DNA分子包括在5“到 3“顺序。 （A）的DNA序列contg。 从拉姆达噬菌体的启动子和操纵P（L）O（L）; （B）一个氮利用的网站结合抗终止子N蛋白; （C）的第一限制性内切酶位点允许置换的DNA序列contg的。 核糖体结合位点后，继; （D）的DNA序列contg。 用于制备能够结合在宿主细胞中的核糖体的所需基因的mRNA核糖体结合位点; （E）ATG起始密码子或转化成所希望的基因向载体的插入密码子的DNA序列; 和（f）的第二限制性内切酶位点用于将基因导入与ATG起始密码子相所需载体; 和（g）的DNA序列contg。 从能够在宿主细胞中自主复制的细菌质粒的复制起点; 和选择，DNA序列contg。 与禅意当矢量存在于宿主细胞中，或DNA序列contg可选择或可识别的表型性状相关的基因。 片段C（I）434，包括基因的C（I）434阻遏蛋白和其相关联的启动子和操纵基因序列。

公开(公告)号：EP0173280B1

公开(公告)日：1992-08-12

申请号：EP85110683.1

申请日：1985-08-26

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Aviv, Haim ,  Bartfeld, Daniel ,  Gorecki, Marian ,  Hartman, Jacob R. ,  Kanner, Dov ,  Levanon, Avigdor ,  Locker-Giladi, Hilla ,  Oppenheim, Amos B.

IPC分类号： C12N15/73 ,  C12P21/02 ,  C12N15/67 ,  C12P19/34

CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

公开(公告)号：EP0284105A2

公开(公告)日：1988-09-28

申请号：EP88104880.5

申请日：1988-03-25

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Hartman, Jacob R. ,  Beck, Yaffa ,  Nimrod, Abraham

IPC分类号： A61K38/44

CPC分类号： C12N9/0089 ,  A61K38/446

摘要： A double-stranded cDNA molecule which includes DNA encoding human manganese superoxide dismutase has been created. The sequence of one strand of a double-­stranded DNA molecule which encodes human manganese superoxide dismutase has been discovered. Such mole­cules may be introduced in prokaryotic, e.g., bacteri­al, or eucaryotic, e.g., yeast or mammalian, cells and the resulting cells cultured or grown under suitable conditions so as to produce human manganese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants which may then be recovered. By this invention, human man­ganese superoxide dismutase gene fragments from various plasmids may be ligated to yield a complete genomic human manganese superoxide dismutase gene fragment. Human manganese superoxide dismutase, analogs, or mu­tants may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, pro­long the survival time of isolated organs, or treat inflammations.
The invention also concerns a method of producing enzy­matically active human manganese superoxide dismutase compounds in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase compound by maintaining the bac­ terial cell under suitable conditions and in a suit­able production medium. The production medium is sup­plemented with an amount of Mn⁺⁺ so that the concentra­tion of Mn⁺⁺ in the medium is greater than about 2 ppm. Genomic manganese superoxide dismutase DNA should also be capable of expression in eucaryotic cells under suitable conditions.
This invention also concerns a method of recovering purified enzymatically active manganese superoxide dismutase, analogs or mutants from bacterial cells. It should also be possible to recover manganese SOD from genomic manganese superoxide dismutase DNA expressed in eucaryotic cells using similar methods.
The invention further concerns uses of the human manga­nese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants, various new analogs, compositions of the compounds and methods of treating various disorders using these compositions.

摘要翻译： 双链cDNA分子其中包含编码DNA的人锰超氧化物歧化酶已创建。 双链DNA分子的一个海滩的编码人类锰超氧化物歧化酶的序列已被发现。 幅分子可在原核，例如细菌，或真核，例如，酵母或哺乳动物，细胞和培养的或在合适的条件下生长，以产生人锰超氧化物歧化酶，人锰超氧化物歧化酶的类似物或人类锰超氧化物得到的细胞被引入 氧化物歧化酶的突变体则可以回收。 通过本发明，来自各种质粒的人锰超氧化物歧化酶基因的片段可被连接，以产生全基因组人类锰超氧化物歧化酶基因片段。 人锰超氧化物歧化酶，类似物或突变体可用于催化超氧化物自由基的减少，减少再灌注损伤，延长分离的器官的存活时间，或治疗炎症。 本发明因此涉及制造在其中包含与能够表达由保持在合适的条件下，BAC TERIAL细胞，并在合适的生产培养基编码超氧化物歧化酶化合物的DNA序列的细菌细胞酶促活性的人锰超氧化物歧化酶的化合物的方法。 生产培养基在Mn数量<+> <+>所以没有在介质中的Mn <+> <+>的浓度大于约2 ppm的更大的补充有。 因此基因组锰超氧化物歧化酶DNA shoulderstand能够在合适的条件下的真核细胞中的表达。 所以本发明涉及的从细菌细胞回收纯化的酶促活性锰超氧化物歧化酶，类似物或突变体的方法。 所以应当能够在使用类似的方法的真核细胞中过表达的基因组的锰超氧化物歧化酶的DNA回收锰超氧化物歧化酶。 本发明还涉及利用人锰超氧化物歧化酶的，人的锰超氧化物歧化酶的类似物或人类锰超氧化物歧化酶的突变体，各种新的类似物，所述化合物的组合物以及使用这些治疗各种疾病的组合物的方法。

公开(公告)号：EP0284105B1

公开(公告)日：1995-11-15

申请号：EP88104880.5

申请日：1988-03-25

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Hartman, Jacob R. ,  Beck, Yaffa ,  Nimrod, Abraham

IPC分类号： A61K38/44

CPC分类号： C12N9/0089 ,  A61K38/446

公开(公告)号：EP0483113A3

公开(公告)日：1992-05-13

申请号：EP92101250.6

申请日：1985-08-26

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Aviv, Haim ,  Bartfeld, Daniel ,  Gorecki, Marian ,  Hartman, Jacob R. ,  Kanner, Dov ,  Levanon, Avigdor ,  Locker-Giladi, Hilla ,  Oppenheim, Amos B.

IPC分类号： C12N15/53 ,  C12N9/02

CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

摘要： An improved vector which upon introduction into a suitable host containing the thermolabile repressor C I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P L O L from lambda bacteriophage; the N utilization site; a first restriction enzyme site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzme site for inserting the gene in phase with the ATG initiation codon; an origin of replication and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host or a fragment designated cI⁴³⁴ which includes the gene for the cI⁴³⁴ repressor protein and its associated promoter and operator. The distance between the 3' end of P L O L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Vectors of the invention may also include a T₁T₂ rRNA transcription termination sequence located 3' of the second restriction enzyme site. The inventive vectors may have origin of replications capable of autonomous production in the host of at least 400 constitutive copies. Plasmids have been constructed from the vectors and used to produce bovine, chicken, porcine and human growth hormones, human apolipoprotein E and human superoxide dismutase and analogs thereof in host cells. Such superoxide dismutase or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and prevent neurological injury. Enzymatically active eucaryotic superoxide dismutase or an analog thereof may also be produced by an improved method of this invention which employs a production medium having a concentration of Cu⁺⁺ greater than about about 2ppm. The invention also concerns improved methods of recovering purified enzymatically active eucaryotic superoxide dismutase or analogs thereof.

公开(公告)号：EP0483113A2

公开(公告)日：1992-04-29

申请号：EP92101250.6

申请日：1985-08-26

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Aviv, Haim ,  Bartfeld, Daniel ,  Gorecki, Marian ,  Hartman, Jacob R. ,  Kanner, Dov ,  Levanon, Avigdor ,  Locker-Giladi, Hilla ,  Oppenheim, Amos B.

IPC分类号： C12N15/53 ,  C12N9/02

CPC分类号： C12N9/0089 ,  A23K20/168 ,  A61K38/446 ,  C07K14/61 ,  C07K14/775 ,  C12N15/69 ,  C12N15/73

摘要： An improved vector which upon introduction into a suitable host containing the thermolabile repressor C I renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P L O L from lambda bacteriophage; the N utilization site; a first restriction enzyme site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzme site for inserting the gene in phase with the ATG initiation codon; an origin of replication and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host or a fragment designated cI⁴³⁴ which includes the gene for the cI⁴³⁴ repressor protein and its associated promoter and operator. The distance between the 3' end of P L O L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs.
Vectors of the invention may also include a T₁T₂ rRNA transcription termination sequence located 3' of the second restriction enzyme site. The inventive vectors may have origin of replications capable of autonomous production in the host of at least 400 constitutive copies.
Plasmids have been constructed from the vectors and used to produce bovine, chicken, porcine and human growth hormones, human apolipoprotein E and human superoxide dismutase and analogs thereof in host cells.
Such superoxide dismutase or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and prevent neurological injury.
Enzymatically active eucaryotic superoxide dismutase or an analog thereof may also be produced by an improved method of this invention which employs a production medium having a concentration of Cu⁺⁺ greater than about about 2ppm.
The invention also concerns improved methods of recovering purified enzymatically active eucaryotic superoxide dismutase or analogs thereof.

摘要翻译： 一种改进的载体，其在引入含有不耐热阻遏物CI的合适的宿主中时，使宿主能够实现所需基因的表达。 载体是双链DNA分子，其以5'至3'的顺序包括以下：来自λ噬菌体的启动子和操纵子PLOL; N利用场所; 此后第一个限制酶位点; 核糖体结合位点; ATG起始密码子或DNA，将所需基因插入载体后转化为ATG起始密码子; 用于将基因与ATG起始密码子相插入的第二限制性酶切位点; 复制起点和当载体存在于宿主中时表现出的可选择或可识别的表型性状相关的基因或称为cI4 <3>的片段，其包含cI4基因的基因， 3> 4阻抑蛋白及其相关启动子和操纵子。 PLOL的3'端与N使用位点的5'端之间的距离小于约80个碱基对。 N利用位点的3'末端与核糖体结合位点的5'末端之间的距离小于约300个碱基对。 本发明的载体还可以包含位于第二限制酶位点3'的T1T2 rRNA转录终止序列。 本发明的载体可以具有能够在宿主中自主生产至少400种组成型拷贝的重复起源。 已经从载体构建质粒，并用于在宿主细胞中产生牛，鸡，猪和人生长激素，人载脂蛋白E和人超氧化物歧化酶及其类似物。 这种超氧化物歧化酶或其类似物可用于催化超氧自由基的还原，减少再灌注损伤，延长分离的器官的存活时间并预防神经损伤。 酶促活性的真核超氧化物歧化酶或其类似物也可以通过本发明的改进方法生产，其使用浓度大于约2ppm的Cu 2+的生产培养基。 本发明还涉及回收纯化的酶促活性的真核超氧化物歧化酶或其类似物的改进方法。

公开(公告)号：EP0284105A3

公开(公告)日：1989-01-25

申请号：EP88104880.5

申请日：1988-03-25

申请人： BIO-TECHNOLOGY GENERAL CORPORATION

发明人： Hartman, Jacob R. ,  Beck, Yaffa ,  Nimrod, Abraham

IPC分类号： A61K38/44

CPC分类号： C12N9/0089 ,  A61K38/446

摘要： A double-stranded cDNA molecule which includes DNA encoding human manganese superoxide dismutase has been created. The sequence of one strand of a double-­stranded DNA molecule which encodes human manganese superoxide dismutase has been discovered. Such mole­cules may be introduced in prokaryotic, e.g., bacteri­al, or eucaryotic, e.g., yeast or mammalian, cells and the resulting cells cultured or grown under suitable conditions so as to produce human manganese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants which may then be recovered. By this invention, human man­ganese superoxide dismutase gene fragments from various plasmids may be ligated to yield a complete genomic human manganese superoxide dismutase gene fragment. Human manganese superoxide dismutase, analogs, or mu­tants may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, pro­long the survival time of isolated organs, or treat inflammations. The invention also concerns a method of producing enzy­matically active human manganese superoxide dismutase compounds in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase compound by maintaining the bac­ terial cell under suitable conditions and in a suit­able production medium. The production medium is sup­plemented with an amount of Mn⁺⁺ so that the concentra­tion of Mn⁺⁺ in the medium is greater than about 2 ppm. Genomic manganese superoxide dismutase DNA should also be capable of expression in eucaryotic cells under suitable conditions. This invention also concerns a method of recovering purified enzymatically active manganese superoxide dismutase, analogs or mutants from bacterial cells. It should also be possible to recover manganese SOD from genomic manganese superoxide dismutase DNA expressed in eucaryotic cells using similar methods. The invention further concerns uses of the human manga­nese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants, various new analogs, compositions of the compounds and methods of treating various disorders using these compositions.