# US 5,707,559 A · Chemiluminescent 1,2-dioxetane compounds > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,707,559 A - Document ID: US5707559A - Status: Granted - Kind: A - Application number: 07/023,649 - Filed: 1987-03-09 - Granted: 1998-01-13 - Earliest claimed: 1986-07-17 - Assignees: Tropix, Inc. - Inventors: Arthur Paul Schaap; Irena Y. Bronstein - Web page: https://patentgrep.com/patent/US5707559A ## CPC - C07D 405/12: linked by a chain containing hetero atoms as chain links - C07C 37/00: Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring - C07C 37/055: the substituted group being bound to oxygen, e.g. ether group - C07C 37/62: by introduction of halogen; by substitution of halogen atoms by other halogen atoms - C07C 51/15: by reaction of organic compounds with carbon dioxide, e.g. Kolbe-Schmitt synthesis - C07D 207/46: with hetero atoms directly attached to the ring nitrogen atom - C07D 311/82: Xanthenes - C07D 321/00: Heterocyclic compounds containing rings having two oxygen atoms as the only ring hetero atoms, not provided for by groups C07D317/00 - C07D319/00 - C07D 407/10: linked by a carbon chain containing aromatic rings - C07D 407/12: linked by a chain containing hetero atoms as chain links - C07D 417/12: linked by a chain containing hetero atoms as chain links - C07D 493/10: Spiro-condensed systems - C07D 495/04: Ortho-condensed systems - C07F 7/1804: {Compounds having Si-O-C linkages (Si-O-acyl linkages C07F7/1896)} - C07F 9/6541: condensed with carbocyclic rings or {carbocyclic} ring systems - C07F 9/65512: {condensed with carbocyclic rings or carbocyclic ring systems} - C07F 9/65522: {condensed with carbocyclic rings or carbocyclic ring systems} - C07F 9/65586: {at least one of the hetero rings does not contain nitrogen as ring hetero atom} - C07F 9/6561: containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings - C07H 15/203: Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems - C07C 2603/18: Fluorenes; Hydrogenated fluorenes - C07C 2603/74: Adamantanes - C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08) - C07C: ACYCLIC OR CARBOCYCLIC COMPOUNDS - C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22) - C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13) ## Abstract Novel light producing 1,2-dioxetanes are described of the formula ##STR1## wherein ArOX is an aryl ring substituted with an X oxy group and A are passive organic groups which allow the 1,2-dioxetane to produce light when triggered by removing X. X is a chemically labile group which is removed by an activating agent. The 1,2-dioxetane compounds can be triggered to produce light at room temperatures. ## Claims ### Claim 1 The method for generating light which comprises: (a) providing a stable 1,2-dioxetane of the formula ##STR46## wherein ArOX represents an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent so that the unstable 1,2-dioxetane compound decomposes to form light and two carbonyl containing compounds of the formula ##STR47## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate 1,2-dioxetane and wherein A are passive organic groups which allow the light to be produced; and (b) decomposing the stable 1,2-dioxetane with the activating agent. ### Claim 2 The method for generating light which comprises: (a) providing a stable 1,2-dioxetane compound of the formula ##STR48## wherein R.sub.1 and R.sub.2 together and R.sub.3 and R.sub.4 together can be joined as spirofused alkylene and aryl rings, wherein at least one of R.sub.1 and R.sub.2 or R.sub.3 and R.sub.4 is an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2 dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes to form light and two carbonyl containing compounds of the formula: ##STR49## wherein those of R.sub.1, R.sub.2, R.sub.3 or R.sub.4 which are unsubstituted by an X-oxy group are carbon containing organic groups which provide stability for the stable 1,2-dioxetane compound and wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate; and (b) decomposing the stable 1,2-dioxetane with the activating agent. ### Claim 3 The method for generating light which comprises: (a) providing a stable dioxetane compound of the formula: ##STR50## wherein R.sub.1 is selected from alkyl, alkoxy, aryloxy dialkylamino, trialkyl or aryl silyloxy and aryl groups including spirofused aryl groups with R.sub.2, wherein R.sub.2 is an aryl group which can include R.sub.1 and is substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes to form light and two carbonyl containing compounds of the formula: ##STR51## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate and wherein R.sub.3 and R.sub.4 are selected from aryl and alkyl groups which can be joined together as spirofused polycyclic alkyl and polycyclic aryl groups; and (b) decomposing the stable 1,2-dioxetane with the activating agent. ### Claim 4 (depends on 1) The method of claim 1 wherein the 1,2-dioxetane compound is decomposed by an enzyme from a biological source which removes X. ### Claim 5 (depends on 4) The method of claim 4 wherein the 1,2-dioxetane has a phosphate group as the X-oxy group and the enzyme is alkaline phosphatase. ### Claim 6 (depends on 4) The method of claim 4 wherein the 1,2-dioxetane has an acetoxy group as the OX group and the enzyme is acetylcholinesterase. ### Claim 7 (depends on 4) The method of claim 4 wherein the 1,2-dioxetane has an acetoxy group as the OX group and wherein the enzyme is arylesterase. ### Claim 8 (depends on 8) The method of claim 8 wherein the method is performed as part of an immunoassay. ### Claim 9 (depends on 2) The method of claim 2 wherein R.sub.3 and R.sub.4 are combined together to form a polycyclic polyalkylene group containing 6 to 30 carbon atoms. ### Claim 10 (depends on 9) The method of claim 9 wherein R.sub.2 is selected from naphthyl and phenyl groups containing the X-oxy group and wherein R.sub.1 is a methoxy group. ### Claim 11 (depends on 10) The method of claim 10 wherein the X-oxy group is selected from hydroxyl, trialkyl or aryl silyloxy, inorganic oxyacid salt, oxygen-pyranoside, aryl carboxyl esters and alkylcarboxyl esters and wherein X is removed by the activating agent. ### Claim 12 (depends on 1) The method of claim 1 wherein the X-oxy group is a phosphate group and wherein the phosphate group is removed with a phosphatase as the activating agent. ### Claim 13 (depends on 11) The method of claim 11 wherein the X-oxy group is selected from alkyl and aryl carboxyl ester groups and wherein the carboxyl group is removed with an esterase. ### Claim 14 (depends on 2) The method of claim 2 wherein R.sub.3 and R.sub.4 are combined together to form a polycyclic polyalkylene group containing 6 to 30 carbon atoms and wherein R.sub.1 and R.sub.2 are combined together as a spirofused aryl group which is substituted with the X-oxy group. ### Claim 15 (depends on 14) The method of claim 14 wherein the spirofused aryl group is a 9H-xantheny-9-yl group. ### Claim 16 (depends on 15) The method of claim 15 wherein the spirofused aryl group is a 9H-fluoren-9-yl group. ### Claim 17 (depends on 15) The method of claim 15 wherein the 9H-xanthen-9-yl is substituted with an alkyl carboxyl ester group as the OX group and wherein the alkyl carboxyl group is removed by an esterase. ### Claim 18 (depends on 15) The method of claim 15 wherein the 9H-xanthenyl-9-yl group is substituted with a PO.sub.4 group as the OX group and wherein a PO.sub.3 group is removed from the PO.sub.4 group by a phosphatase. ### Claim 19 (depends on 2) The method of claim 2 wherein the 1,2-dioxetane compound is decomposed by an enzyme from a biological source which removes X. ### Claim 20 (depends on 19) The method of claim 19 wherein the 1,2-dioxetane has a phosphate group as the X-oxy group and the enzyme is alkaline phosphatase. ### Claim 21 (depends on 19) The method of claim 19 wherein the 1,2-dioxetane has an acetoxy group as the X-oxy group and the enzyme is acetylcholinesterase. ### Claim 22 (depends on 13) The method of claim 13 wherein the 1,2-dioxetane has an acetoxy group as the X-oxy group and wherein the enzyme is arylesterase. ### Claim 23 (depends on 2) The method of claim 2 wherein the method is performed as part of an immunoassay. ### Claim 24 A method for generating light in a chemiluminescent assay using an enzyme from a biological source which comprises: (a) providing a compound of the formula: ##STR52## wherein R.sub.1 and R.sub.2 taken together are a spirofused polycyclic aryl group having an aryl ring substituted with an X-oxy group wherein X is removeable by enzymatic triggering ahd R.sub.3 and R.sub.4 taken together are an adamantyl group; and (b) triggering the compound to generate light using an enzyme from a biological source. ### Claim 25 (depends on 24) The method of claim 24 wherein the spirofused polycyclic aryl group is xanthenyl with an aryl ring substituted with the X-oxy group. ### Claim 26 (depends on 25) The method of claim 25 wherein the X-oxy group is a phosphate. ### Claim 27 A method for generating light in a chemiluminescent assay using an enzyme from a biological source which comprises: (a) providing a compound of the formula: ##STR53## wherein R.sub.1 is methoxy, R.sub.2 is an aryl group substituted with an X-oxy group wherein X is removable by enzymatic triggering and R.sub.3 and R.sub.4 taken together are an adamantyl group; and (b) triggering the compound to generate light using an enzyme from a biological source. ### Claim 28 In an immunoassay method wherein there is an optically detectable reaction, the improvement wherein said optically detectable reaction includes the reaction, with an enzyme, of a dioxetane having the formula ##STR54## where T is an adamantyl group; Y is naphthyl; X is methoxy; and Z is an enzyme-cleavable group, so that said enzyme cleaves said enzyme-clearable group from said dioxetane to form a negatively charged substituent bonded to said dioxetane, said negatively charged substituent causing said dioxetane to decompose to form a luminescent substance comprising said group Y of said dioxetane. ### Claim 29 (depends on 28) The method of claim 28 wherein Z is acetoxy.