Method of preparing D-luciferin derivatives

• US 5,374,534 A
• Filed: 05/14/1993
• Issued: 12/20/1994
• Est. Priority Date: 07/19/1990
• Status: Expired due to Fees

• Current Assignee: Charm Sciences, Inc.
• Original Assignee: Charm Sciences, Inc.
• Inventors: Charm, Stanley E., Zomer, Eliezer, Saul, Steven
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): Leary, Louise N.
• Application Number:
  US08/062,315
• Time in Patent Office: 585 Days
• Field of Search: 435/8, 435/25, 435/7.72, 435/4, 435/23, 540/469, 548/178, 436/546, 536/24, 530/330
• US Class Current: 435/8
• CPC Class Codes: C07D 277/64 with only hydrocarbon or substituted hydrocarbon radicals attached in position 2
• CPC Class Codes: C12Q 1/66 involving luciferase
• CPC Class Codes: G01N 33/567 utilising isolate of tissue or organ as binding agent
• CPC Class Codes: G01N 33/60 involving radioactive labelled substances

Abstract

A method of preparing a 6-substituted D-luciferin ester suitable for use in the determination of pesticides by a bioluminescence method. The 6-substituted D-luciferin esters, such as 6-D-luciferin acetate, are prepared in a liquid solvent containing N-acetyl imidazole with D-luciferin and recovering as the reaction product 6-D-luciferin acetate.

Claims

1. A method of preparing a 6-substituted D-luciferin ester compound, which method consists essentially of:

a) reacting in a liquid solvent a substituted N-acylated imidazole compound with D-luciferin; and

b) recovering a solution of 6-substituted D-luciferin ester compound.

2. The method of claim 1 wherein the liquid solvent comprises a water solvent mixture.

3. The method of claim 1 wherein the liquid solvent comprises water.

4. The method of claim 1 wherein the substituted N-acylated imidazole compound comprises N-acetylimidazole in an acetone-water solvent.

5. The method of claim 1 which includes monitoring the decrease in bioluminescence of the luciferin as a measure of the completeness of the reaction.

6. The method of claim 1 wherein the substituted group of the N-acylated imidazole compound is selected from the group consisting of:

alkyl, benzyl and phenyl radicals.

7. The method of claim 1 wherein the liquid solvent comprises methanol, and the D-luciferin is a synthetic D-luciferin.

8. The method of claim 1 wherein the N-acylated imidazole compound is N-acetyl imidazole.

9. The method of claim 8 wherein the substituted N-acylated imidazole compound is used in stoichiometric excess.

10. A method of preparing a 6-substituted acetyl D-luciferin ester compound, which method consists essentially of:

a) reacting in a liquid solvent a stoichiometric excess of N-acetyl imidazole with D-luciferin to provide 6-acetyl-D-luciferin ester compound and imidazole; and

b) recovering a liquid solvent solution of the 6-acetyl-D-luciferin ester compound for use in a bioluminescence test method.