CHEMILUMINESCENT FUSED POLYCYCLIC RING-CONTAINING 1,2-DIOXETANES AND ASSAYS IN WHICH THEY ARE USED

摘要

Chemiluminescent 1,2-dioxetane compounds are disclosed inwhich the molecule is stabilized at the 3-position on thedioxetane ring against decomposition prior to the molecule'scoming in contact with a labile group-removing substance (e. g.,an enzyme that will cleave the labile group to cause the moleculeto decompose to form at least one light-emitting fluorophore) andsubstituted at the 4-position on the dioxetane ring with a fusedpolycyclic ring-containing fluorophore moiety bearing a labilering substituent whose point of attachment to the fusedpolycyclic ring, in relation to this ring's point(s) ofattachment to the dioxetane ring, is such that the total numberof ring atoms separating these points of attachment, includingthe ring atoms at the points of attachment, is an odd wholenumber. These odd pattern substituted compounds decompose toemit light of greater intensity and of a different wavelengththan that emitted by the corresponding even pattern substitutedisomers. They are useful in detecting the presence ordetermining the concentration of chemical or biologicalsubstances in immunoassays, chemical assays and nucleic acidprobe assays, and in chemical/physical probe procedures forstudying the microstructures of macromolecules. Two or more ofthem can also be used in combination, or one or more of them canbe used together with other chemiluminescent compounds, inmulti-channel assays to detect two or more different analytessimultaneously. Novel intermediates used in the preparation ofthese odd pattern substituted compounds are also disclosed.