US4952707A
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Enzymatically-cleavable chemiluminescent fused polycyclic ring-containing 1,2-dioxetanes
Assignee: Applied Biosystems LLC
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Filing: 1988-06-30
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Granted: 1990-08-28
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Expires: 2008-06-30
Abstract
Abstract Chemiluminescent 1,2-dioxetane compounds are disclosed in which the molecule is stabilized at the 3-position on the dioxetane ring against decomposition prior to the molecule's coming in contact with a labile group-removing substance (e.g., an enzyme that will cleave the labile group to cause the molecule to decompose to form at least one light-emitting fluorophore) and substituted at the 4-position on the dioxetane ring with a fused polycyclic ring-containing fluorophore moiety bearing a labile ring substituent whose point of attachment to the fused polycyclic ring, in relation to this ring's point(s) of attachment to the dioxetane ring, is such that the total number of ring atoms separating these points of attachment, including the ring atoms at the points of attachment, is an odd whole number. These odd pattern substituted compounds decompose to emit light of greater intensity and of a different wavelength than that emitted by the corresponding even pattern substituted isomers. They are useful in detecting the presence or determining the concentration of chemical or biological substances in immunoassays, chemical assays and nucleic acid probe assays, and in chemical/physical probe procedures for studying the microstructures of macromolecules. Two or more of them can also be used in combination, or one or more of them can be used together with other chemiluminescent compounds, in multi-channel assays to detect two or more different analytes simultaneously. Novel intermediates used in the preparation of these odd pattern substituted compounds are also disclosed.
Inventors
- 👤 Brooks Edwards
- 👤 Irena Y. Bronstein
- 👤 Alison A. Laird
- 👤 John C. Voyta
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 12:42
CPC Classifications
C; C07; C07C; C07C43/00; C07C43/02; C07C43/20; C07C43/215; C07C43/225; C07C43/23; C07C49/00; C07C49/76; C07C49/84; C07C69/00; C07D; C07D321/00; C07F; C07F9/00; C07F9/02; C07F9/547; C07F9/655; C07F9/6551; C07F9/65512; C12; C12Q; C12Q1/00; C12Q1/34; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; C07C2603/00; C07C2603/56; C07C2603/58; C07C2603/70; C07C2603/74; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S435/973
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
192 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound having the formula: ##STR9## wherein R 1 is hydrogen, or a bond when R 2 is a substituent bound to the dioxetane ring through a spiro linkage, or an organic substituent that does not interfere with the production of light but satisfies the valence of the dioxetane ring carbon atom to which it is attached; R 2 is a fused polycyclic ring-containing fluorophore moiety bound to the dioxetane ring through a single bond or a spiro linkage and having an enzymatically cleavable labile ring substituent containing a bond which, when cleaved, renders the polycyclic moiety electron-rich to in turn render the dioxetane compound decompasable to emit light, the enzymatically cleavable labile ring substituent's point of attachment to the fused polycyclic moiety, in relation to the fused polycyclic moiety's point of attachment to the dioxetane ring, being such that the total number of ring atoms separating these points of attachment, including the ring atoms at these points of attachment, is an odd whole number; and T is a stabilizing group that prevents the dioxetane compound from decomposing before the enzymatically cleavable labile ring substituent's bond is cleaved.
Claim 2
Dependent Claim
192 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound having the formula: ##STR9## wherein R 1 is hydrogen, or a bond when R 2 is a substituent bound to the dioxetane ring through a spiro linkage, or an organic substituent that does not interfere with the production of light but satisfies the valence of the dioxetane ring carbon atom to which it is attached; R 2 is a fused polycyclic ring-containing fluorophore moiety bound to the dioxetane ring through a single bond or a spiro linkage and having an enzymatically cleavable labile ring substituent containing a bond which, when cleaved, renders the polycyclic moiety electron-rich to in turn render the dioxetane compound decompasable to emit light, the enzymatically cleavable labile ring substituent's point of attachment to the fused polycyclic moiety, in relation to the fused polycyclic moiety's point of attachment to the dioxetane ring, being such that the total number of ring atoms separating these points of attachment, including the ring atoms at these points of attachment, is an odd whole number; and T is a stabilizing group that prevents the dioxetane compound from decomposing before the enzymatically cleavable labile ring substituent's bond is cleaved.
Claim 3
Dependent Claim
21 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 1 in which the odd whole number is 5 or greater.
Claim 4
Dependent Claim
39 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 2 in which the fused polycyclic moiety is the residue of a fused polycyclic aromatic hydrocarbon ring fluorophoric compound containing from 9 to about 30 ring carbon atoms, inclusive.
Claim 5
Dependent Claim
28 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 3 in which the residue of a fused polycyclic aromatic hydrocarbon ring fluorophoric compound is a naphthalene residue.
Claim 6
Dependent Claim
28 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 3 in which the residue of a fused polycyclic aromatic hydrocarbon ring fluorophoric compound is an anthracene residue.
Claim 7
Dependent Claim
41 words
An enzymatically cleavable chemiulminescent 1,2-dioxetane compound as recited in claim 2 in which the fused polycyclic moiety is the residue of a less than fully aromatic fused polycyclic hydrocarbon ring fluorophoric compound containing from 10 to about 30 carbon atoms, inclusive.
Claim 8
Dependent Claim
31 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 6 in which the residue of a less than fully aromatic fused polycyclic hydrocarbon ring fluorophoric compound is a fluorene residue.
Claim 9
Dependent Claim
37 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 2 in which the fused polycyclic moiety is the residue of a fused polycyclic heterocyclic ring fluorophoric compound containing from 9 to about 30 ring atoms, inclusive.
Claim 10
Dependent Claim
27 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in claim 8 in which the residue of a fused polycyclic heterocyclic ring fluorophoric compound is a quinoline residue.
Claim 11
Dependent Claim
41 words
An enzymatically cleavable chemiluminescent 1,2-dioxetane compound as recited in any one of claims 3-9, inclusive, in which R 1 is a methoxy group, R 2 is bound to the dioxetane ring through a single bond and T is an adamant-2-ylidene group.
Claim 12
Dependent Claim
23 words
An enzymatically cleavable 1,2-dioxetane compound as recited in claim 10 in which the enzymatically cleavable labile ring substituent is a phosphate ester group.
Claim 13
Dependent Claim
22 words
An enzymatically cleavable 1,2-dioxetane compound as recited in claim 10 in which the enzymatically cleavable labile ring substituent is a galactoside group.
Claim 14
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(6"-phosphoryloxy)naphth-1'-yl-1,2-dioxetane salt.
Claim 15
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(6"-phosphoryloxy)naphth-1'-yl-1,2-dioxetane salt.
Claim 16
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(7"-phosphoryloxy)naphth-2'-yl-1,2-dioxetane salt.
Claim 17
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(7"-phosphoryloxy)naphth-2'-yl-1,2-dioxetane salt.
Claim 18
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(8"-phosphoryloxy)naphth-1'-yl-1,2-dioxetane salt.
Claim 19
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(8"-phosphoryloxy)naphth-1'-yl-1,2-dioxetane salt.
Claim 20
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(5"-phosphoryloxy)naphth-2'-yl-1,2-dioxetane salt.
Claim 21
Dependent Claim
3 words
A 3-(2'-spiroadamantane)-4-methoxy-4-(5"-phosphoryloxy)naphth-2'-yl-1,2-dioxetane salt.
Claim 22
Dependent Claim
15 words
The disodium salt of a 1,2-dioxetane compound as in any one of claims 13-16, inclusive.
Claim 23
Dependent Claim
1 words
3-(2,-spiroadamantane)-4-methoxy-4-(5"-β-galactosyloxy)naphth-2'-yl-1,2-dioxetane.
Claim 24
Dependent Claim
1 words
3-(2,-spiroadamantane)-4-methoxy-4-(5"-β-galactosyloxy)naphth-2'-yl-1,2-dioxetane.
References Cited (Backward Citations)
EP0254051A2
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-08-27 | 1994-02-26 | 1994-08-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-08-27 | 1998-02-26 | 1998-08-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-08-27 | 2002-02-26 | 2002-08-28 | $7700.00 | Window Closed / Lapsed |