The invention discloses fluorescent chelator compounds that are especially useful to monitor and measure cytosolic concentrations of alkali metal cations such as Na⁺, K⁺, and Li⁺. The new compounds are comprised of: (1) crown ethers (that may or may not have substituent groups attached to the core carbons, but will always contain at least one core nitrogen) that are linked via the core nitrogen(s) to at least one (2) fluorophore that contains an additional heteroaromatic liganding center. In the currently preferred dye, SBFI, the core compound is crown ether 1,7-diaza-4,10,13-trioxacyclopentadecane and the heteroaromatic fluorophores are benzofurans that are linked to isophthalate groups. Selectivities for Na⁺ over K⁺ of about 20 are observed, resulting in effective dissociation constants for Na⁺ of about 20 mM against a background of 120 mM K⁺. Increasing [Na⁺] increases the ratio of excitation efficiency at 330-345 nm to that at 370-390 nm with emission collected at 450-550 nm, so that ratio fluorometry and imaging work at the same wavelengths as used with the well-known Ca²⁺ indicator fura-2. If the macrocyclic ring is increased in size to a 1,10-diaza-4,7,13,16-tetraoxacyclooctadecane, the chelators become selective for K⁺ over Na⁺. If the ring is decreased in size, for example to a 1,7-diaza-4,10-dioxacyclododecane, the chelators become selective for Li⁺ over Na⁺.
本发明公开了荧光
螯合剂化合物,特别适用于监测和测量碱
金属阳离子(如 Na⁺、K⁺ 和 Li⁺)的细胞浓度。新化合物由以下部分组成:(1)
冠醚(可能有取代基团,也可能没有,但总是含有至少一个核心氮),通过核心氮与至少一个(2) 含有额外杂芳香族配位中心的荧光团相连。在目前首选的
染料 SBFI 中,核心化合物是
冠醚 1,7-二氮杂-4,10,13-三氧杂
环十五烷,杂芳香族荧光团是与
间苯二甲酸酯基相连的
苯并呋喃。观察到 Na⁺ 对 K⁺ 的选择性约为 20,在 120 mM K⁺ 的背景下,Na⁺ 的有效解离常数约为 20 mM。增加[Na⁺]可提高 330-345 nm 处的激发效率与 370-390 nm 处的激发效率之比,并在 450-550 nm 处收集发射光谱。如果大环的尺寸增加到 1,10-二氮杂-4,7,13,16-四氧杂环
十八烷,
螯合剂对 K⁺ 的选择性就会高于 Na⁺。如果环的尺寸减小,例如减小到 1,7-二氮杂-4,10-二氧杂
环十二烷,
螯合剂对 Li⁺ 的选择性就会大于 Na⁺。