公开(公告)号：WO1989006226A1

公开(公告)日：1989-07-13

申请号：PCT/US1989000016

申请日：1989-01-03

Applicant: QUEST SYSTEMS, INC. ,  EDWARDS, Brooks ,  BRONSTEIN, Irena, Y.

Inventor： QUEST SYSTEMS, INC.

IPC: C07C41/01

CPC classification number: C07H15/203 ,  C07C41/01 ,  C07C67/29 ,  C07C2603/74 ,  C07D321/00 ,  C07F9/12 ,  C07F9/65512 ,  C07F9/65742 ,  C07H15/26 ,  C12Q1/34 ,  G01N21/76 ,  G01N33/581 ,  G01N33/582 ,  C07C43/215 ,  C07C43/1788 ,  C07C69/157

Abstract: Compounds having formula (I), wherein T is a polycycloalkylidene group (e.g., adamant-2-ylidene); R is a C1-20 alkyl, aralkyl or cycloalkyl group; and Y is a fluorescent chromophore (e.g., m-phenylene), produced by reacting a compound having formula (II) with an R-ylating agent (e.g., R2SO4) in the presence of an alkali metal alkoxide in a polar aprotic solvent. Also, compounds having formula (III) are produced by reacting a compound having formula (IV) with formula (V), wherein X is an electronegative leaving group (e.g., a halogen anion such as chloride ion) in the presence of a Lewis base (e.g., a trialkyl-amine) dissolved in an aprotic organic solvent (e.g., benzene or toluene). Also, compounds having formula (VI) are produced by reacting a compound of formula (VII) with a tetra-O-acylated-O-hexopyranoside halide, then hydrolyzing off the protective acyl groups. The aforementioned compounds and procedures are useful in the synthesis of enzyme-cleavable 1,2-dioxetane ring systems that can serve as members of a binding pair employed, for example, in chemiluminescent immunoassays, DNA probe assays, and direct assays for an enzyme.

Abstract translation: 具有式（I）的化合物，其中T是多亚烷基（例如，金刚烷-2-亚基）; R是C 1-20烷基，芳烷基或环烷基; Y是通过在极性非质子溶剂中在碱金属醇盐存在下使具有式（II）的化合物与R-衍生剂（例如R 2 SO 4）反应制备的荧光发色团（例如间亚苯基）。 此外，具有式（III）的化合物通过在路易斯碱存在下使具有式（IV）的化合物与式（V）反应来制备，其中X是电负离子基团（例如卤素阴离子如氯离子） （例如三烷基胺）溶于非质子有机溶剂（如苯或甲苯）中。 此外，具有式（VI）的化合物通过使式（VII）的化合物与四-O-酰化的O-六吡喃糖苷卤化物反应，然后水解保护性酰基来制备。 上述化合物和方法可用于合成可切割的1,2-二氧杂环丁烷环系统，其可以用作结合对的成员，例如在化学发光免疫测定，DNA探针测定和酶的直接测定中使用。

公开(公告)号：WO1992004341A1

公开(公告)日：1992-03-19

申请号：PCT/US1991006096

申请日：1991-08-30

Applicant: TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  JUO, Rouh-Rong

Inventor： TROPIX, INC.

IPC: C07D321/00

CPC classification number: C12Q1/34 ,  C07C43/1788 ,  C07C43/196 ,  C07C2603/74 ,  C07D321/00 ,  C07F9/12 ,  C07F9/65512

Abstract: Enzymatically cleavable chemiluminescent 1,2-dioxetane compounds capable of producing light energy when decomposed, substantially stable at room temperature before a bond by which an enzymatically cleavable labile substituent thereof is intentionally cleaved, are disclosed. These compounds can be represented by formula (I) wherein X and X each represent, individually, hydrogen, a hydroxyl group, a halo substituent, an unsubstituted lower alkyl group, a hydroxy (lower) alkyl group, a halo (lower) alkyl group, a phenyl group, a halophenyl group, an alkoxyphenyl group, a hydroxyalkoxy group, a cyano group, a carboxyl or substituted carboxyl group or an amide group, with at least one of X and X being other than hydrogen; and R1 and R2 individually or together, represent an organic substituent that does not interfere with the production of light when the dioxetane compound is enzymatically cleaved and that satisfies the valence of the dioxetane compound's 4-carbon atom, with the provisos that if R1 and R2 represent individual substituents the R2 substituent is aromatic, heteroaromatic, or an unsaturated substituent in conjugation with an aromatic ring, and that at least one of R1 and R2 is, or R1 and R2 taken together are, an enzymatically cleavable labile group-substituted fluorescent chromophore group that produces a luminescent substance when the enzymatically cleavable labile substituent thereof is removed by an enzyme. The corresponding dioxetanes which, instead of being substituted at the 5' or 7', or at the 5' and 7' positions, instead contain a 4' methylene group, are also disclosed, as are intermediates for all these 3-substituted adamant-2'-ylidenedioxetanes, and their use as reporter molecules in assays.

Abstract translation: 公开了能够在分解时产生光能的酶促切割的化学发光的1,2-二氧杂环丁烷化合物，在其中有可能切割其可酶切的不稳定取代基的键之间在室温下基本稳定。 这些化合物可以由式（I）表示，其中X和X 1各自表示氢，羟基，卤素取代基，未取代的低级烷基，羟基（低级）烷基，卤代（低级） ）烷基，苯基，卤代苯基，烷氧基苯基，羟基烷氧基，氰基，羧基或取代的羧基或酰胺基，X和X 1中的至少一个不同于氢 ; R1和R2分别或一起表示当二氧杂环丁烷化合物被酶解时不干扰光的产生并且满足二氧杂环丁烷化合物的4-碳原子的化合价的有机取代基，条件是如果R1和R2 代表各个取代基，R 2取代基是芳族的，杂芳族的，或与芳香环共轭的不饱和取代基，R1和R2中的至少一个是或R1和R2一起是可酶切割的不稳定基团取代的荧光发色团 当其酶促切割不稳定取代基被酶除去时产生发光物质的组。 也公开了相对于在5'或7'或5'和7'位置取代含有4'亚甲基的二氧杂环丁烷，以及所有这些3-取代的金刚烷基 - 2'-亚基二氧杂环丁烷，以及它们在测定中作为报道分子的用途。