Assays using chemiluminescent, enzymatically cleavable substituted 1,2-dioxetanes and kits therefor

• Main Author: BRONSTEIN; IRENA Y
• Format: Patent
• Language: English
• Published: 25.02.1997
• Edition: 6
• Subjects: ACYCLIC OR CARBOCYCLIC COMPOUNDS; ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAININGELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN,SULFUR, SELENIUM OR TELLURIUM; BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; DERIVATIVES THEREOF; ENZYMOLOGY; HETEROCYCLIC COMPOUNDS; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC CHEMISTRY; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; SUGARS; TESTING; VINEGAR; WINE

Dioxetane compounds reactable with an enzyme to release optically detectable eneregy are disclosed. These compounds have the formula: wherein T is a cycloalkyl group or a fused polycyclo-alkylidene group bonded to the dioxetane ring through a spiro linkage; Y is a fluorescent chromophore capable of absorbing energy to form an excited energy state from which it emits optically detectable energy to return to its original energy state; X is hydrogen or an alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl or cycloheteroalkyl group, or an enzyme-cleavable group containing a bond cleavable by an enzyme to yield an electron-rich moiety bonded to the dioxetane ring, Z is hydrogen, hydroxyl or an enzyme-cleavable group containing a bond cleavable by an enzyme to yield an electron-rich moiety bonded to the dioxetane ring, at least one of X and Z being an enzyme-cleavable group, and T also includes a substituent which enhances the solubility of the dioxetane in aqueous solution, or a substituent which facilitates bonding of the dioxetane to a membrane, film, bead, polymer or polymerizable group, or a substituent which enhances the kinetics of the dioxetane enzyme degradation.