Substituted thiazoline-dioxetan substrates, process for the prod

Details Substituted thiazoline-dioxetan substrates, process for the prod Substituted thiazoline-dioxetan substrates, process for the prod

1993-11-30

1995-10-03

Gerstl, Robert

Organic compounds -- part of the class 532-570 series

Organic compounds

Heterocyclic carbon compounds containing a hetero ring...

C07D51310

Patent

active

054553571

DESCRIPTION:

BRIEF SUMMARY
This application is a 371 of PCT/EP93/00752 filed on Mar. 29, 1993.
The invention concerns thiazoline-dioxetan substrates, processes for the production and use in enzymatic analytical methods.
The reaction of luciferin (formula I) with luciferase, oxygen and ATP leads to the formation of oxiluciferin. In this reaction, light (maximum wavelength at 562 nm) is emitted as chemiluminescence. In this process the dioxetan of formula II is presumably formed as a high-energy intermediate (F. McCapra, Chem. Commun. 155 (1968)). Numerous chemiluminescing 1,2-dioxetan compounds have been developed on the basis of this postulate. The adamantyl residue has been described for the stabilization of the instable 1,2-dioxetans (EP-A 0 254 051, EP-A 0 352 713, WO 91/03479, WO 90/07511 as well as the publications which they cited). However, none of the known compounds is based on the activated intermediate stage of luciferin according to formula II as the parent substance, a thiazoline derivative. ##STR3##
The object of the present invention was to provide a 1,2-dioxetan based on the activated intermediate stage of thiazolines and preferably of luciferin which is stable and only decomposes when reacted with an activating agent with formation of chemiluminescence. This object is achieved by a compound of the general formula III, ##STR4## in which R is either ##STR5## and R.sub.1 and R.sub.2 are in each case either the same or different and represent hydrogen or a lower alkyl C.sub.1 -C.sub.6 which is straight-chained or branched, X is a cleavable group which can be cleaved by an activating agent and at least one of the groups R.sub.4 or R.sub.5 represents a group stabilizing the dioxetan structure and at most one of the groups R.sub.4 or R.sub.5 represents hydrogen.
Acids, bases, salts, enzymes, inorganic or organic catalysts and electron donors are preferably used as activating agents.
The group --O--X can be located at any position on the phenyl ring. However, if R is a benzthiazole group, the group --O--X is preferably situated at the 5 position. If R is a phenyl group, the group --O--X is preferably situated at the 3 position. This applies analogously for the --OR.sub.3 group from which the --O--X group is formed.
--O--X is preferably the hydroxy salt of an oxy acid, phosphate, aryl- or alkylcarboxyl ester, alkyloxy or arylsilyloxy, sulfate, oxypyranoside.
The composition of the aryl and alkyl residues is not critical. Any person skilled in the art can select suitable residues for X without difficulty. It must merely be soluble and it must be possible to cleave X by activating agents.
R.sub.1 and R.sub.2 are preferably hydrogen, methyl or ethyl groups. If for example phosphate is used as --O--X, then the chemiluminescence reaction can be induced by addition of alkaline phosphatase. If the galactoside is used, the chemiluminescence reaction can be induced by .beta.-galactosidase. If a silyloxy residue is used as --O--X, then the chemiluminescence can be induced by addition of fluoride.
Groups R.sub.4 and/or R.sub.5 are suitable for stabilizing the dioxetan structure that are those which protect the dioxetan group from reactions. This stabilization is preferably effected by steric screening of this group. Adamantanyl, phenyl, cyclohexyl, secondary and tertiary aliphatic alkyl groups (such as e.g. the t-butyl group) in a substituted and unsubstituted form are for example suitable. In this connection R.sub.4 and R.sub.5 can be the same or different. In the case of the adamantyl residue, this is preferably bound in such a way that R.sub.4 and R.sub.5 denote parts of the ring structure and are accordingly bridged (formula IIIa): ##STR6##
The compounds of the general formula III are novel. A process for the synthesis of these compounds has not been previously known.
Accordingly the invention also concerns a process for the production of the compounds of formula III which is characterized in that a compound of formula IV or IVa ##STR7## in which R.sub.3 represents alkyl which is straight-chained or branched with 1-6 C-atoms and pr

REFERENCES:
patent: 4857652 (1989-08-01), Schaap
patent: 4959182 (1990-09-01), Schaap
patent: 4962192 (1990-10-01), Schaap
patent: 4983779 (1991-01-01), Schaap
patent: 5004565 (1991-04-01), Schapp
patent: 5013827 (1991-05-01), Schaap
patent: 5068339 (1991-11-01), Schaap et al.
patent: 5089630 (1992-02-01), Bronstein et al.
White, E. II., et al. (1980) Jour. Amer. Chem. Soc., 102:9, pp. 3199-3208.

Affiliated with

Also associated with

Rating

Substituted thiazoline-dioxetan substrates, process for the prod does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Substituted thiazoline-dioxetan substrates, process for the prod, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Substituted thiazoline-dioxetan substrates, process for the prod will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-1078467