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11. 发明授权

US5696087A Method and compositions for reducing cholesterol absorption 失效
标题翻译：减少胆固醇吸收的方法和组合物
公开(公告)号：US5696087A
公开(公告)日：1997-12-09
申请号：US347718
申请日：1994-12-01
申请人： Jordan J. N. Tang , Chi-Sun Wang
发明人： Jordan J. N. Tang , Chi-Sun Wang
IPC分类号： A61K38/00 , A61K51/04 , C12N9/20 , A61K38/16 , A61K38/08 , A61K38/10
CPC分类号： A61K51/0493 , C12N9/20 , A61K38/00
摘要： Compositions including all or a portion and chemically or recombinantly synthesized analogues of the carboxy terminal region of bile salt-activated lipase (BAL) are described, which, when orally ingested, compete with native BAL in binding to the intestinal surface and thus reduce the amount of cholesterol taken into the blood stream.
摘要翻译：描述了包括胆汁盐活化脂肪酶（BAL）的羧基末端区域的全部或部分化学或重组合成的类似物的组合物，其在口服摄取时与天然BAL竞争结合肠表面，从而减少量的胆固醇摄入血液。

12. 发明授权

US5821226A BAL C-tail drug delivery molecules 失效
标题翻译： BAL C尾药物递送分子
公开(公告)号：US5821226A
公开(公告)日：1998-10-13
申请号：US482262
申请日：1995-06-07
申请人： Jordan J. N. Tang , Chi-Sun Wang
发明人： Jordan J. N. Tang , Chi-Sun Wang
IPC分类号： A61K38/00 , A61K51/04 , C12N9/20 , A61K38/16 , A61K38/08 , A61K38/10
CPC分类号： C12N9/20 , A61K51/0493 , A61K38/00
摘要： Drug delivery conjugates of including a BAL C-tail peptide including all or a portion of the carboxy terminal region of human bile salt-activated lipase (BAL) conjugated to a biologically active substance are described. The C-tail peptide-drug conjugates, when orally ingested, compete with native BAL in binding to the intestinal surface, and, as a result, permit drug compositions to be delivered specifically to the intestine. Useful C-tail peptides are derivatives of the carboxy terminal region of BAL derived from all or portion of the region containing amino acid residues 539 to 722, and have a mucin-like structure containing at least three of the repeating proline-rich units of eleven amino acid residues each.
摘要翻译：描述了包含与生物活性物质缀合的人胆汁盐活化脂肪酶（BAL）的全部或部分羧基末端区域的BAL C尾肽的药物递送缀合物。当口服摄取时，C尾肽 - 药物偶联物与天然BAL竞争结合肠表面，结果允许药物组合物被特异性递送至肠道。有用的C尾肽是衍生自含有氨基酸残基539至722的全部或部分区域的BAL的羧基末端区的衍生物，并且具有包含至少三个重复的富含脯氨酸的单元的粘蛋白样结构氨基酸残基各自。

13. 发明授权

US5681819A Method and compositions for reducing cholesterol absorption 失效
标题翻译：减少胆固醇吸收的方法和组合物
公开(公告)号：US5681819A
公开(公告)日：1997-10-28
申请号：US479160
申请日：1995-06-07
申请人： Jordan J. N. Tang , Chi-Sun Wang
发明人： Jordan J. N. Tang , Chi-Sun Wang
IPC分类号： A61K38/00 , A61K51/04 , C12N9/20 , A61K38/08 , A61K38/10 , A61K38/16
CPC分类号： C12N9/20 , A61K51/0493 , A61K38/00
摘要： Compositions derived from all or a portion of the carboxy terminal region of human bile salt-activated lipase (BAL) are described, which, when orally ingested, compete with native BAL in binding to the intestinal surface, thus reducing the physiological role of BAL in mediating the transfer of cholesterol into the intestinal cells, and, as a result, reducing the amount of cholesterol absorbed from the intestine into the blood stream. Useful derivatives of the carboxy terminal region of BAL are derived from all or portion of the region containing amino acid residues 539 to 722, and have a mucin-like structure containing at least three of the repeating proline-rich units of eleven amino acid residues each.
摘要翻译：描述了衍生自人胆汁盐活化脂肪酶（BAL）的羧基末端区域的全部或部分的组合物，其在口服摄取时与天然BAL竞争结合肠表面，从而降低BAL在生理学上的作用介导胆固醇转移到肠细胞中，结果减少从肠吸收到胆固醇的胆固醇的量。 BAL的羧基末端区域的有用的衍生物衍生自含有氨基酸残基539至722的所有或部分区域，并且具有含有至少三个重复的富含脯氨酸的单元的粘蛋白样结构，每个氨基酸残基为11个氨基酸残基。

14. 发明授权

US06545127B1 Catalytically active recombinant memapsin and methods of use thereof 失效
标题翻译：催化活性重组膜突触蛋白及其使用方法
公开(公告)号：US06545127B1
公开(公告)日：2003-04-08
申请号：US09604608
申请日：2000-06-27
申请人： Jordan J. N. Tang , Xinli Lin , Gerald Koelsch , Lin Hong
发明人： Jordan J. N. Tang , Xinli Lin , Gerald Koelsch , Lin Hong
IPC分类号： G01N3348
CPC分类号： C07K5/1021 , A61K38/00 , A61K39/00 , C07K1/1136 , C07K5/06026 , C07K5/06043 , C07K5/0806 , C07K2299/00 , C12N9/6421 , C12N9/6478 , Y02A90/26 , Y10S514/879
摘要： Methods for the production of purified, catalytically active, recombinant memapsin 2 have been developed. The substrate and subsite specificity of the catalytically active enzyme have been determined. The substrate and subsite specificity information was used to design substrate analogs of the natural memapsin 2 substrate that can inhibit the function of memapsin 2. The substrate analogs are based on peptide sequences, shown to be related to the natural peptide substrates for memapsin 2. The substrate analogs contain at least one analog of an amide bond which is not capable of being cleaved by memapsin 2. Processes for the synthesis of two substrate analogues including isosteres at the sites of the critical amino acid residues were developed and the substrate analogues, OMR99-1 and OM99-2, were synthesized. OM99-2 is based on an octapeptide Glu-Val-Asn-Leu-Ala-Ala-Glu-Phe (SEQ ID NO:28) with the Leu-Ala peptide bond substituted by a transition-state isostere hydroxyethylene group (FIG. 1). The inhibition constant of OM99-2 is 1.6×10−9M against recombinant pro-memapsin 2. Crystallography of memapsin 2 bound to this inhibitor was used to determine the three dimensional structure of the protein, as well as the importance of the various residues in binding. This information can be used by those skilled in the art to design new inhibitors, using commercially available software programs and techniques familiar to those in organic chemistry and enzymology, to design new inhibitors to memapsin 2, useful in diagnostics and for the treatment and/or prevention of Alzheimer's disease.
摘要翻译：已经开发了用于生产纯化的，催化活性的重组突变蛋白2的方法。已经确定了催化活性酶的底物和亚位点特异性。底物和亚位点特异性信息用于设计可以抑制膜蛋白2功能的天然memapsin 2底物的底物类似物。底物类似物基于肽序列，显示与memapsin 2的天然肽底物相关。底物类似物含有至少一个酰胺键的类似物，该类似物不能被膜蛋白2切割。开发了两个底物类似物合成的关键氨基酸残基位点处的等位基因，底物类似物OMR99- 1和OM99-2。 OM99-2基于由过渡态等电位羟基亚乙基取代的Leu-Ala肽键的八肽Glu-Val-Asn-Leu-Ala-Ala-Glu-Phe（SEQ ID NO：28）（图1 ）。 OM99-2的抑制常数为1.6×10 -9 M，与重组pro-memapsin2结合使用与此抑制剂结合的胶原蛋白2的结晶学，以确定蛋白质的三维结构，以及各种残基在结合中的重要性。本领域技术人员可以使用本信息来设计新的抑制剂，使用商业上可获得的有机化学和酶学方面熟悉的软件程序和技术来设计新的抑制剂2，可用于诊断和治疗和/或预防阿尔茨海默病。

15. 发明授权

US5215907A Thermostable acid protease from sulfolobus acidocaldarius 失效
标题翻译：来自硫酸杆菌的热稳定性酸性蛋白酶
公开(公告)号：US5215907A
公开(公告)日：1993-06-01
申请号：US827892
申请日：1992-01-30
申请人： Jordan J. N. Tang , Xin-Li Lin
发明人： Jordan J. N. Tang , Xin-Li Lin
IPC分类号： C12N9/52 , C12N15/57 , C12N15/74
CPC分类号： C12N9/52 , C12N15/74
摘要： A 46,000 Dalton thermostable, very acidic protease, which has been named thermopsin, was purified to homogeneity from the culture medium of Sulfolobus acidocaldarius by a five-step procedure including column chromatographies on DEAE-Sepharose CL-6B, phenyl-Sepharose CL-4B, Sephadex G-100, MonoQ (FPLC), and gel filtration (HPLC). The enzyme is a single polypeptide chain having proteolytic activity over pH range 0 to 11 at temperatures between 0.degree. C. and 100.degree. C., with maximal activity at approximately pH 2 and 90.degree. C. Antibodies directed against thermopsin have been prepared. Through studies using various aspartic protease inhibitors, thiol and metalloprotease inhibitors, and serine protease inhibitors, it was determined that, although similar to some aspartic proteases, the active site of thermopsin is clearly not identical to that of other aspartic proteases. Thermopsin hydrolyses the following bonds: Leu-Val, Leu-Tyr, Phe-Phe, Phe-Tyr, and Tyr-Thr, indicating that the specificity of thermopsin is similar to that of pepsin for the large hydrophobic residues at both sides of the scissile bond. In addition, thermopsin is resistant to detergent inactivation, the protein retaining proteolytic activity even in the presence of high concentrations of sodium dodecyl sulfate.
摘要翻译：已经命名为Thermopsin的46,000道尔顿热稳定的非常酸性的蛋白酶通过五步法从Sulfolobus acidocaldarius的培养基纯化至均匀，包括在DEAE-Sepharose CL-6B，苯基琼脂糖CL-4B上的柱色谱， Sephadex G-100，MonoQ（FPLC）和凝胶过滤（HPLC）。该酶是在0℃至100℃的温度范围内在pH范围0至11下具有蛋白水解活性的单一多肽链，在约pH和90℃下具有最大活性，已经制备了针对热蛋白的抗体。通过使用各种天冬氨酸蛋白酶抑制剂，硫醇和金属蛋白酶抑制剂和丝氨酸蛋白酶抑制剂的研究，确定尽管与一些天冬氨酸蛋白酶类似，但是热诱导因子的活性位点与其它天冬氨酸蛋白酶的活性位点显然不完全相同。 Thermopsin水解以下键：Leu-Val，Leu-Tyr，Phe-Phe，Phe-Tyr和Tyr-Thr，表明热敏蛋白的特异性与胃蛋白酶的特异性相似，对于两侧的大型疏水残基键。此外，热敏蛋白耐洗涤剂失活，蛋白质即使在高浓度十二烷基硫酸钠存在下也能保持蛋白水解活性。

16. 发明授权

US06303380B1 Construction of retroviral producer cells from adenoviral and retroviral vectors 有权
标题翻译：从腺病毒和逆转录病毒载体构建逆转录病毒生产细胞
公开(公告)号：US06303380B1
公开(公告)日：2001-10-16
申请号：US09301846
申请日：1999-04-29
申请人： Xinli Lin , Jordan J. N. Tang
发明人： Xinli Lin , Jordan J. N. Tang
IPC分类号： C12N510
CPC分类号： C12N15/86 , C12N2710/10344 , C12N2740/13043 , C12N2740/13052
摘要： A combination of adenoviral and retroviral vectors used to construct second generation packaging cells that deliver marker genes to target cells is described. A vector based upon Moloney murine leukemia virus (MLV) was used to deliver marker genes, and an adenovirus-based delivery system was used to deliver MLV structural genes (gagpol and env) to cultured cells. The procedure transformed the cells into new retroviral producer cells, which generate replication-incompetent retroviral particles in the culture supernatant for transferring marker genes to target cells. The titer of the retroviral-containing supernatant generated from the second generation producer cells reached above 105 cfu/ml which is comparable to the MLV-based producer cell lines currently used in human gene therapy trials. The vector and procedures are adaptable for experimental human gene therapy in which the new producer cells are transplanted into patients for continuous gene transfer.
摘要翻译：描述了用于构建将标记基因递送至靶细胞的第二代包装细胞的腺病毒和逆转录病毒载体的组合。使用基于莫洛尼鼠白血病病毒（MLV）的载体递送标记基因，并且使用基于腺病毒的递送系统将MLV结构基因（gagpol和env）递送至培养的细胞。该过程将细胞转化为新的逆转录病毒生产细胞，其在培养上清液中产生复制不足的逆转录病毒颗粒，用于将标记基因转移至靶细胞。从第二代生产细胞产生的含有逆转录病毒的上清液的滴度高于105cfu / ml，这与目前用于人类基因治疗试验的基于MLV的生产细胞系相当。载体和程序适用于实验性人类基因治疗，其中新的生产细胞被移植到患者中用于连续基因转移。

17. 发明授权

US5200183A Recombinant bile salt activated lipases 失效
标题翻译：重组胆汁盐活化脂肪酶
公开(公告)号：US5200183A
公开(公告)日：1993-04-06
申请号：US537426
申请日：1990-06-12
申请人： Jordan J. N. Tang , Chi-Sun Wang
发明人： Jordan J. N. Tang , Chi-Sun Wang
IPC分类号： A23C9/152 , A23C9/12 , A23C9/20 , A23L1/305 , A61K38/46 , A61P1/14 , C07H21/04 , C07K14/47 , C12N9/18 , C12N9/20 , C12N15/09 , C12N15/55 , C12N15/85 , C12R1/19
CPC分类号： A23C9/20 , A23C9/1216 , A23L33/17 , C12N15/8509 , C12N9/18 , C12Y301/01003 , A01K2207/15 , A01K2217/00 , A01K2227/10 , A01K2267/01
摘要： The complete structure of human milk BAL cDNA is disclosed. The nucleotide sequences of the cDNA inserts of two clones overlap and together contain 2951 base pairs of BAL cDNA which codes for an open reading frame of 742 amino acid residues between initiation and termination codons. There is a putative signal sequence of 20 residues which is followed by a 61-amino-terminal sequence of BAL. The cDNA sequence also contains a 678-base 5'-untranslated sequence, a 97-base 3'-untranslated region, and a 14-base poly(A) tail. The deduced BAL protein structure contains in the carboxyl-terminal region fourteen repeating unis of 11 amino acids each. The repeating units have the basic structure of Pro-Val-Pro-Pro-Thr-Gly-Asp-Ser-Gly-Ala-Pro-, with only minor substitutions. The cDNA is useful for expression of protein, study of structure, function and the effect of modification or deletion or addition of amino acids, including entire repeating units, and as probes for studies involving BAL or related lipases, including rat pancreatic lysophospholipase, cholinesterase, and acetylcholinesterase.
摘要翻译：公开了人乳BAL cDNA的完整结构。两个克隆的cDNA插入片段的核苷酸序列重叠并一起含有2951个碱基对的BAL cDNA，其编码起始和终止密码子之间742个氨基酸残基的开放阅读框。存在20个残基的推定信号序列，其后是BAL的61个氨基末端序列。 cDNA序列还含有678碱基的5'-非翻译序列，97碱基3'非翻译区和14碱基聚（A）尾。推测的BAL蛋白结构在羧基末端区域中含有十四个重复单元，每个具有11个氨基酸。重复单元具有Pro-Val-Pro-Pro-Thr-Gly-Asp-Ser-Gly-Ala-Pro-的基本结构，只有很少的取代。该cDNA可用于表达蛋白质，研究结构，功能以及修饰或缺失或添加氨基酸（包括整个重复单元）的作用，以及作为涉及BAL或相关脂肪酶的研究的探针，包括大鼠胰腺溶血磷脂酶，胆碱酯酶，和乙酰胆碱酯酶。

18. 发明授权

US5173403A Thermostable acid protease from sulfolobus acidocaldarius and gene 失效
标题翻译：来自硫酸多糖和基因的耐热酸性蛋白酶
公开(公告)号：US5173403A
公开(公告)日：1992-12-22
申请号：US467745
申请日：1990-01-19
申请人： Jordan J. N. Tang , Xin-Li Lin
发明人： Jordan J. N. Tang , Xin-Li Lin
IPC分类号： C12N9/52 , C12N15/09 , C12N15/57 , C12N15/74 , C12R1/01
CPC分类号： C12N9/52 , C12N15/74
摘要： A thermostable, very acidic protease, which has been named thermopsin, was purified to homogeneity from the culture medium of Sulfolobus acidocaldarius by a five-step procedure including column chromatographies on DEAE-Sepharose CL-6B, phenyl-Sepharose CL-4B, Sephadex G-100, MonoQ (FPLC), and gel filtration (HPLC). The enzyme is a single polypeptide chain having proteolytic activity over pH range 0 to 11 at temperatures between 0.degree. and 100.degree. C., with maximal activity at approximately pH 2 and 90.degree. C. Antibodies directed against thermopsin have been prepared. Through studies using various aspartic protease inhibitors, thiol and metalloprotease inhibitors, and serine protease inhibitors, it was determined that, although similar to some aspartic proteases, the active site of thermopsin is clearly not identical to that of other aspartic proteases. Thermopsin hydrolyzes the following bonds: Leu-Val, Leu-Tyr, Phe-Phe, Phe-Tyr, and Tyr-Thr, indicating that the specificity of thermopsin is similar to that of pepsin for the large hydrophobic residues at both sides of the scissile bond. In addition, thermospin is resistant to detergent inactivation, the protein retaining proteolytic activity even in the presence of high concentrations of sodium dodecyl sulfate.
摘要翻译：将已被命名为热敏蛋白的热稳定的非常酸性的蛋白酶通过五步法从含有硫酸硫杆菌的培养基中纯化至均匀，包括在DEAE-Sepharose CL-6B，苯基琼脂糖CL-4B，Sephadex G -100，MonoQ（FPLC）和凝胶过滤（HPLC）。该酶是在0至100℃的温度范围内在pH范围0至11下具有蛋白水解活性的单一多肽链，其在约pH 2和90℃下具有最大活性。已经制备了针对嗜热蛋白的抗体。通过使用各种天冬氨酸蛋白酶抑制剂，硫醇和金属蛋白酶抑制剂和丝氨酸蛋白酶抑制剂的研究，确定尽管与一些天冬氨酸蛋白酶类似，但是热诱导因子的活性位点与其它天冬氨酸蛋白酶的活性位点显然不完全相同。 Thermopsin水解下列键：Leu-Val，Leu-Tyr，Phe-Phe，Phe-Tyr和Tyr-Thr，表明热敏蛋白的特异性与胃蛋白酶的特异性相似，对于两侧的大型疏水残基键。此外，嗜热丝蛋白对洗涤剂失活具有抗性，蛋白质即使在高浓度十二烷基硫酸钠存在下也能保持蛋白水解活性。

19. 发明授权

US4944944A Dietary compositions and methods using bile salt-activated lipase 失效
标题翻译：膳食组合物和使用胆汁盐激活脂肪酶的方法
公开(公告)号：US4944944A
公开(公告)日：1990-07-31
申请号：US122410
申请日：1987-11-19
申请人： Jordan J. N. Tang , Chi-Sun Wang
发明人： Jordan J. N. Tang , Chi-Sun Wang
IPC分类号： A23C9/15 , A23C9/12 , A23C9/152 , A23C9/20 , A23L1/03 , A23L1/305 , A61K35/20 , A61K38/46 , A61P1/00 , A61P3/06 , C12N9/20
CPC分类号： C12Y301/01003 , A23C9/1216 , A23C9/20 , A23L29/06 , A23L33/17 , Y10S426/801
摘要： Dietary compositions, especially cow's milk-based infant formulas, are fortified with bile salt-activated lipase. Methods are provided for feeding newborn and premature infants which include administration of bile salt-activated lipase to increase fat digestion and therefore growth rate. Similarly, a method is provided to treat subjects for inadequate pancreatic enzyme production by administration of bile salt-activated lipase in conjunction with ingestion of fats.

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