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    • 6. 发明申请
    • ASSAYS AND METHODS FOR CELL PROLIFERATION-TARGETED TREATMENT THERAPIES
    • 细胞增殖指标治疗方法的测定和方法
    • US20160299149A1
    • 2016-10-13
    • US15074993
    • 2016-03-18
    • Otto Phanstiel, IVDeborah AltomareLaurence von Kalm
    • Otto Phanstiel, IVDeborah AltomareLaurence von Kalm
    • G01N33/574A61K31/198A61K31/404C12Q1/68A61K31/137
    • G01N33/57496A61K31/404C12Q1/6886C12Q2600/106C12Q2600/158C12Q2600/16C12Y306/03G01N33/57492G01N2333/4703
    • Assays are described which measure polyamine transport activity and markers associated with polyamine transport and metabolism. These data are then used for the selection of treatments using therapies which target polyamine transport and polyamine metabolism. The assay includes a substrate to which a protein-containing cell sample can bind, a solution comprising a first antibody specific against ATP13A3, wherein the first antibody is configured to bind to ATP13A3 protein on the substrate, a solution comprising a second antibody specific against the first antibody, the second antibody comprising an enzyme linked thereto, wherein the second antibody is configured to bind to the first antibody. The assay further includes a substrate specific to the enzyme, wherein upon combining the substrate and the enzyme, the amount of enzyme in the solution can be identified, wherein said amount of enzyme identified is proportional to the amount of ATP13A3 protein in the sample, wherein said ATP13A3 protein is indicative of polyamine transport in the cells of the sample. In addition, primary antibodies covalently attached to respective fluorophores can be used to directly measure the relative expression levels of these biomarkers in histological samples. In addition, several biomarkers are described which allow for therapy selection based upon the expression and relative ratios of specific proteins associated with polyamine transport (c-myc, ATP13A3 Cav-2, and Cav-1 as well as c-Raf).
    • 描述了测定多胺转运活性和与多胺转运和代谢相关的标记的测定。 然后将这些数据用于使用靶向多胺转运和多胺代谢的疗法的治疗选择。 所述测定法包括含有蛋白质的细胞样品可以结合的底物,包含针对ATP13A3特异性的第一抗体的溶液,其中所述第一抗体配置为与所述底物上的ATP13A3蛋白结合,所述溶液包含针对所述底物的第二抗体 第一抗体,所述第二抗体包含与其连接的酶,其中所述第二抗体被配置为结合所述第一抗体。 该测定还包括对酶特异的底物,其中在组合底物和酶时,可以鉴定溶液中的酶的量,其中所鉴定的酶的量与样品中ATP13A3蛋白的量成比例,其中 所述ATP13A3蛋白表明样品细胞中的多胺转运。 此外,共价连接到相应荧光团的一抗可用于直接测量组织学样品中这些生物标志物的相对表达水平。 此外,描述了几种基于与多胺转运(c-myc,ATP13A3Cav-2和Cav-1以及c-Raf)相关的特异性蛋白质的表达和相对比例进行治疗选择的生物标志物。
    • 7. 发明申请
    • FLUORESCENT CYTOTOXIC COMPOUNDS SPECIFIC FOR THE CELLULAR POLYAMINE TRANSPORT SYSTEM
    • 细胞聚氨酯运输系统的荧光细胞毒素化合物
    • US20130337494A1
    • 2013-12-19
    • US13835708
    • 2013-03-15
    • Otto Phanstiel, IV
    • Otto Phanstiel, IV
    • G01N21/64
    • G01N21/6486C07C255/58C07C2603/24G01N33/5008G01N33/533G01N33/574G01N33/582
    • Cyano-substituted anthracene containing polyamines were synthesized and shown to be efficient polyamine transporter ligands. Moreover, these compounds (3 and 4) had improved fluorescence properties over previously known anthryl-polyamine conjugates, which facilitated their intracellular trafficking by confocal microscopy and fluorescence methods. These cytotoxic fluorescent agents may find use as molecular probes which traffic into cells via the polyamine transport system and may also be viable anticancer drugs which are readily quantified in human tissues due to their excellent fluorescence properties: (excitation: λ405 nm) and emission (420 nm) occurs in the visible light range. The ability to excite and emit in the visible range provides an advantage to these probes as these wavelengths are considered non-toxic to human cells (versus ultraviolet mediated excitations, λ
    • 合成含氰基取代的含多胺的多胺,显示为有效的多胺转运体配体。 此外,这些化合物(3和4)具有比先前已知的蒽基 - 聚胺共轭物更好的荧光性能,这促进了它们通过共焦显微镜和荧光方法的细胞内运输。 这些细胞毒性荧光剂可以用作通过多胺转运系统进入细胞的分子探针,并且还可以是由人体组织中容易定量的活的抗癌药物,因为它们具有优异的荧光性质:(激发:λ405nm)和发射(420 nm)出现在可见光范围内。 在可见光范围内激发和发射的能力为这些探针提供了优势,因为这些波长被认为对人体细胞无毒(相对于紫外线介导的激发,λ<400nm),可见光激光器的购买和操作成本较低 UV激光源。