Coelenterazine analogs

Abstract

Coelenterazine analogs having luminescence properties different from those of known coelenterazine analogs are desired for various luciferases. The invention provides compounds represented by general formula (1) below.

1 - 19 . (canceled) 
     
     
         20 . A method for measuring luminescence, which comprises contacting f-coelenterazine and a luciferase, wherein said luciferase is selected from the group consisting of (a), (b), and (c) below:
 (a) a protein comprising the amino acid sequence of SEQ ID NO: 1;   (b) a protein comprising an amino acid sequence wherein 1 to 8 amino acids are deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO: 1 and having luminescence catalytic activity with luciferin as a substrate; and   (c) a protein comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 1 and having luminescence catalytic activity with luciferin as a substrate,   
     
     
         21 . The Method of  claim 20 , wherein the luciferase comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         22 . A method for assaying the activity of a target promoter comprising:
 providing a polynucleotide encoding a luciferase as a reporter gene and fusing said polynucleotide to said target promoter to construct a vector;   transforming said vector into a host cell; and   contacting said host cell with f-coelenterazine as a luminescence substrate, wherein said luciferase encoded by said polynucleotide is selected from the group consisting of (a), (b), and (c) below:   (a) a protein comprising the amino acid sequence of SEQ ID NO: 1;   (b) a protein comprising an amino acid sequence wherein 1 to 8 amino acids are deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO: 1 and having luminescence catalytic activity with luciferin as a substrate; and   (c) a protein comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ NO: I and having luminescence catalytic activity with luciferin as a substrate.   
     
     
         23 . The method of  claim 22 , wherein said luciferase comprises the amino acid sequence of SEQ ID NO: 1, 
     
     
         24 . The method of  claim 22 , wherein said polynucleotide encoding said luciferase is a polynucleotide selected from the group consisting of (a), (b), (c), and (d) below:
 (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 2;   (b) a polynucleotide comprising a nucleotide sequence in which at least one nucleotide is deleted, substituted, inserted and/or added in the nucleotide sequence of SEQ ID NO: 2 and wherein said polynucleotide encodes a protein having luminescence catalytic activity with luciferin as a substrate;   (c) a polynucleotide comprising a nucleotide sequence having at least 90% identity to the nucleotide sequence of SEQ ID NO: 2 and wherein said polynucleotide encodes a protein having luminescence catalytic activity with luciferin as a substrate; and   (d) a polynucleotide comprising a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 2 and encodes a protein having luminescence catalytic activity with a Iticiferin as a substrate.   
     
     
         25 . The method of  claim 22 , wherein said polynucleotide encoding said luciferase comprises the nucleotide sequence of SEQ ID NO: 2. 
     
     
         26 . A kit comprising:
 (i) f-coelenterazine; and   (ii) a composition selected from the group consisting of a luciferase, a polynucleotide encoding said luciferase, a recombinant vector comprising said polynucleotide encoding said luciferase, and a transformant comprising said polynucleotide encoding said luciferase,   wherein said luciferase is selected from the group consisting of (a), (b), and (c) below:   (a) a protein comprising the amino acid sequence of SEQ ID NO: 1;   (b) a protein comprising an amino acid sequence Wherein 1 to 8 amino acids are deleted, substituted, inserted and/or added in the amino acid sequence of SEQ ID NO: 1 and having luminescence catalytic activity with luciferin as a substrate; and   (c) a protein comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 1 and having luminescence catalytic activity with luciferin as a substrate.   
     
     
         27 . The kit of  claim 26 , wherein said luciferase comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         28 . The kit of  claim 26 , wherein said polynucleotide encoding said luciferase is a polynucleotide selected from the group consisting of (a), (b), (c) and (d) below:
 (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 2;   (b) a polynucleotide comprising a nucleotide sequence in which at least one nucleotide is deleted, substituted, inserted and/or added in the nucleotide sequence of SEQ ID NO: 2 and wherein said polynucleotide encodes a protein having luminescence catalytic activity with luciferin as a substrate;   (c) a polynucleotide comprising a nucleotide sequence having at least 90% identity to the nucleotide sequence of SEQ ID NO: 2 and wherein said polynucleotide encodes a protein having luminescence catalytic activity with luciferin as a substrate; and   (d) a polynucleotide comprising a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of SEQ II) NO: 2 and encodes a protein having luminescence catalytic activity with a luciferin as a substrate.   
     
     
         29 . The kit of  claim 26 , wherein said polynucleotide encoding said luciferase comprises the nucleotide sequence of SEQ II) NO: 2.