Coelenterazine analogs

1. A compound represented by general formula (1) below:

wherein in said general formula (1), R 1 is hydrogen, a substituted or unsubstituted aryl a substituted or unsubstituted arylalkyl, a substituted or unsubstituted arylalkenyl, an alkyl which may optionally be substituted with an alicyclic group, an alkenyl which may optionally be substituted with an alicyclic group, an alicyclic group, or a heterocyclic group.

2. The compound of claim 1 , wherein in said general formula (1), R 1 is phenyl, p-hydroxyphenyl, benzyl, α-hydroxybenzyl, p-hydroxybenzyl, phenylethyl, phenylvinyl, cyclopentyl, cyclopentylmethyl, cyclohexyl, cyclohexylmethyl, cyclohexylethyl, methyl, ethyl, propyl, 2-methylpropyl, 2-methylpropenyl, adamantylmethyl or thiophen-2-yl.

3. The compound of claim 2 , wherein in said general formula (1), R 1 is benzyl, α-hydroxybenzyl or p-hydroxybenzyl.

4. The compound of claim 3 , wherein in said general formula (1), R 1 is benzyl.

5. A method for producing a compound represented by general formula (1) below:

wherein in said general formula (1), R 1 is hydrogen, a substituted or unsubstituted aryl, a substituted or unsubstituted arylalkyl, a substituted or unsubstituted arylalkenyl, an alkyl which may optionally be substituted with an alicyclic group, an alkenyl which may optionally be substituted with an alicyclic group, an alicyclic group, or a heterocyclic group; said method comprises reacting a compound represented by general formula (2) below:

wherein in said general formula (2), R 1 is hydrogen, a substituted or unsubstituted aryl a substituted or unsubstituted arylalkyl, a substituted or unsubstituted arylalkenyl, an alkyl which may optionally be substituted with an alicyclic group, an alkenyl which may optionally be substituted with an alicyclic group, an alicyclic group, or a heterocyclic group; with a compound represented by general formula (3) below:

thereby producing said compound represented by general formula (1).

6. The method of claim 5 , wherein in said general formula (1), R 1 is phenyl, p-hydroxyphenyl, benzyl, α-hydroxybenzyl, p-hydroxybenzyl, phenylethyl, phenylvinyl, cyclopentyl, cyclopentylmethyl, cyclohexyl, cyclohexylmethyl, cyclohexylethyl, methyl, ethyl, propyl, 2-methylpropyl, 2-methylpropenyl, adamantylmethyl or thiophen-2-yl.

7. The method of claim 6 , wherein in said general formula (1), R 1 is benzyl, α-hydroxybenzyl or p-hydroxybenzyl.

8. The compound of claim 7 , wherein in said general formula (1) R 1 is benzyl.

9. A kit comprising:

(i) the compound of claim 1 ; and,

(ii) a composition selected from the group consisting of a luciferase, a polynucleotide encoding the said luciferase, a recombinant vector comprising said polynucleotide encoding said luciferase, and a transformant comprising said polynucleotide encoding said luciferase.

10. The kit of claim 9 , wherein said luciferase is a 19 kDa Oplophorus luciferase.

11. The kit of claim 9 , 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.

12. The kit of claim 9 , 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 luciferin as a substrate.

13. A method for measuring luminescence contacting the compound of claim 1 with a luciferase.

14. The method of claim 13 , wherein said luciferase is a 19 kDa Oplophorus luciferase.

15. The method of claim 13 , 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.

16. 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;

contacting said host cell with the compound of claim 1 ; and

detecting luminescence, thereby assaying the activity of said target promoter.

17. The method of claim 16 , wherein said luciferase encoded by said polynucleotide is a 19 kDa Oplophorus luciferase.

18. The method of claim 16 , 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 ID NO: 1 and having luminescence catalytic activity with luciferin as a substrate.

19. The method of claim 16 , wherein the 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 luciferin as a substrate.