Benzothiazole dioxetanes

1. A 1,2-dioxetane compound capable of producing light energy when decomposed, represented by the formula:

wherein T is:

and wherein R may independently be any branched alkyl or cycloalkyl group which provides stabilization for the dioxetane or where both R groups together form a cycloalkyl or polycycloalkyl moiety spiro bound to the dioxetane ring, wherein each R group or the spiro bound moiety may be unsubstituted or substituted with one or more electron-withdrawing groups or electron donating groups, and wherein R 1 is an aryl group, or an alkyl group of 1-20 carbon atoms, which may be optionally substituted with 1 or more halogen atoms, and wherein Y may be H, or an electron donating or withdrawing group, or an organic linker group which may be attached to (1) an ancillary fluorophore, or (2) any biological moiety, and wherein X may be any protecting group which is removed by non-enzymatic chemical or enzymatic means.

2. The compound of claim 1 , wherein X is a phosphate, both R groups together form a spiroadamantyl group, and R 1 is methyl, and Y is hydrogen.

3. The compound of claim 1 , wherein at least one of R group, and Y bear a substituent which improves the solubility of said dioxetane in an aqueous solution.

4. The compound of claim 3 , wherein each substituent improving solubility is a carboxylic acid moiety, a sulfonic acid moiety or a phosphoric acid moiety.

5. The compound of claim 1 , wherein X is an enzyme removable group.

6. The compound of claim 1 , wherein X is a moiety which is removed by a change in pH or heat.

7. The compound of claim 1 , wherein Y is an alkyl or alkoxy moiety linked to a fluorophore in an energy transfer relationship with said dioxetane.

8. A kit for detecting a first substance in a sample, comprising the 1,2-dioxetane compound of claim 1 .

9. The kit of claim 8 , further comprising an enzyme which, in the presence of said dioxetane, causes said dioxetane to decompose.

10. The kit of claim 9 , further comprising a water soluble enhancing substance which enhances a chemiluminescence emission detectable from the decomposition of said dioxetane.

11. The kit of claim 10 , wherein said enhancing substance is quaternary onium polymer.

12. The kit of claim 10 , wherein said kit further comprises an enhancement additive which improves the enhancement of detected chemiluminescence emission effected by said enhancing substance.

13. The kit of claim 8 , wherein at least one of R and Y bear a substituent which improves the solubility of said dioxetane in aqueous solution.

14. A method for detecting a first substance in a sample, comprising adding the compound of claim 1 to said sample, wherein X is removed by said first substance, incubating said sample and inspecting said sample for the generation of light, wherein light so generated is indicative of the presence, and the amount of light detected is indicative of the amount, of said first substance.

15. The method of claim 14 , wherein said method further comprises adding an enhancement agent to said sample, to enhance the amount of light detected.

16. The method of claim 14 , wherein said light is detected by a CCD camera.

17. The method of claim 15 , wherein said enhancement agent is an onium quaternary polymer.

18. The method of claim 17 , wherein said method further comprises the addition of an enhancement additive, to improve the enhancement of detected chemiluminescent emission effected by said enhancement agent.

19. The method of claim 14 , wherein said first substance is an enzyme.

20. The method of claim 19 , wherein said enzyme is completed with a biological moiety of interest.

21. A 1,2-dioxetane compound capable of producing light energy when decomposed, represented by the formula:

wherein T is:

and wherein Y may be H, or an electron donating or withdrawing group, or an organic linker group which may be attached to (1) an ancillary fluorophore, or (2) any biological moiety, and wherein X may be any protecting group which is removed by non-enzymatic chemical or enzymatic means, where R 2 -R 6 are each independently H, alkyl of 1-20 carbon atoms, or an aryl group, where each of R 2 -R 6 may be unsubstituted or substituted one or more with electron-donating groups or electron-withdrawing groups, wherein R 3 and R 4 may be joined as a spiro-fused cycloalkyl group.

22. The compound of claim 21 , wherein at least one of R 2 -R 6 is substituted with an electron-active group.

23. The compound of claim 21 , wherein X is a phosphate and Y is hydrogen.

24. The compound of claim 21 , wherein at least one of R 2 -R 6 and Y bear a substituent which improves the solubility of said dioxetane in an aqueous solution.

25. The compound of claim 24 , wherein the substituent improving solubility is a carboxylic acid moiety, a sulfonic acid moiety or a phosphoric acid moiety.

26. The compound of claim 21 , wherein X is an enzyme removable group.

27. The compound of claim 21 , wherein X is a moiety which is removed by a change in pH or heat.

28. The compound of claim 21 , wherein Y is an alkyl or alkoxy moiety linked to a fluorophore in an energy transfer relationship with said dioxetane.

29. A kit for detecting a first substance in a sample, comprising the 1,2-dioxetane compound of claim 21 .

30. The kit of claim 29 , further comprising a water soluble enhancing substance which enhances a chemiluminescence emission detectable from the decomposition of said dioxetane.

31. The kit of claim 30 , further comprising an enhancement additive which improves the enhancement of detected chemiluminescence emission effected by said enhancing substance.

32. The kit of claim 29 , wherein Y bears a substituent which improves the solubility of said dioxetane in aqueous solution.

33. A method for detecting a first substance in a sample, comprising adding the compound of claim 21 to said sample, wherein X is removed by the first substance, incubating the sample and inspecting the sample for the generation of light, wherein light so generated is indicative of the presence, and the amount of light detected is indicative of the amount, of the first substance.

34. The method of claim 33 , wherein the first substance is an enzyme.

35. The method of claim 34 , wherein the enzyme is complexed with a biological moiety.