Ruthenium-lumazine energy transfer systems

• US 5,278,043 A
• Filed: 01/22/1991
• Issued: 01/11/1994
• Est. Priority Date: 01/25/1990
• Status: Expired due to Fees

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Bannwarth, Wilhelm, Muller, Francis
• Primary Examiner(s): Hill, Jr., Robert J.
• Assistant Examiner(s): WANG, GIAN
• Application Number:
  US07/643,313
• Time in Patent Office: 1,085 Days
• Field of Search: 546/10, 424/9, 424/8, 424/12, 422/55, 422/56, 435/6, 435/52, 435/53, 435/91, 435/191-194, 436/501-504, 436/537, 436/546, 436/172, 436/800, 536/27
• US Class Current: 536/23.1
• CPC Class Codes: C07D 475/02 with an oxygen atom directly attached in position 4
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

Energy-transfer systems which can be used, inter alia, for measuring distances within or between different molecules are described, comprising derivatives of lumazine and ruthenium, in particular derivatives of DNA or RNA sequences.

Claims

1. An energy-transfer system consisting of two organic compounds, one of which is a chromophore of a lumazine type and the other, with which it interacts, is a ruthenium (Ru) complex.

2. The energy-transfer system of claim 1, wherein the chromophore of the lumazine type is a lumazine derivative of the formula ##STR7## in which R₁ and R₂ each represent an H atom, an optionally substituted C_1-10 -alkyl group, 1'"'"'-ribosyl, 1'"'"'-(2'"'"'-deoxyribosyl) or the radical of an analogous hydroxyl compound;

R₃ represents an H atom or represents an optionally substituted C_1-10 -alkyl group; and

R₄ represents an optionally substituted C_1-10 -alkyl group;

1'"'"'-ribosyl, 1'"'"'-(2'"'"'-deoxyribosyl) or the radical of an analogous hydroxyl compound, or of the formula ##STR8## in which R₅ and R₆ each represent an optionally substituted C_1-10 -alkyl group; and

R₇ and R₈ represent 1'"'"'-ribosyl, 1'"'"'-(2'"'"'-deoxyribosyl) or the radical of an analogous hydroxyl compound.

3. The energy-transfer system of claim 1 wherein the ruthenium complex is a compound of the formula

4. The energy-transfer system of claim 3 wherein A represents an alkylene group substituted with sulphonamide, thioether, ether, carboxyl or carboxamide functionalities, and X represents an aldehyde, carboxyl, hydroxyl, amino or thiocyanate group, halogen or a phosphite or phosphate group or a modified phosphate group.

5. The energy-transfer system of claim 2, wherein the chromophore of the lumazine type is a lumazine derivative of formula I in which R₁ and R₂ each represent an H atom, an optionally substituted C_1-10 -alkyl group, 1'"'"'-ribosyl, 1'"'"'-(2'"'"'-deoxyribosyl) or the radical of an analogous hydroxyl compound;

R₃ represents an H atom or represents an optionally substituted C_1-10 -alkyl group; and

R₄ represents an optionally substituted C_1-10 -alkyl group, 1'"'"'-ribosyl, 1'"'"'-(2'"'"'-deoxyribosyl) or the radical of an analogous hydroxyl compound.

6. The energy-transfer system of claim 2, wherein the chromophore of the lumazine type is a lumazine derivative of the formula II in which R₅ and R₆ represent an optionally substituted C_1-10 -alkyl group;

and R₇ and R₈ represent 1'"'"'-ribosyl, 1'"'"'-(2'"'"'-deoxyribosyl) or the radical of an analogous hydroxyl compound.

7. The energy-transfer system according to any of claims 1-6, characterised in that one or more chromophores of the lumazine type are incorporated at the end of a DNA or RNA sequence or within a DNA or RNA sequence and the ruthenium complex is of the formula III in which the charge-transfer units L₁, L₂ and L₃ are identical or different and represent optionally substituted bipyridyl, bathophenanthroline or benzobathophenanthroline groups, said ruthenium complex being bound to the same or different DNA or RNA sequence as the lumazine.

8. Energy-transfer system of claim 7 of the following formulae

Ru²⁺ L₁ L₂ L₃ - Lu-SEQ ID No:

1,

Ru²⁺ L₁ L₂ L₃ - GTLU-SEQ ID No;

2,

Ru²⁺ L₁ L₂ L₃ - GTTGALU-SEQ ID No. 3,

Ru²⁺ L₁ L₂ L₃ - GTTGACAALU-SEQ ID No;

4 or

Ru²⁺ L₁ L₂ L₃ - LuLuLuLuLu-SEQ ID No;

1

in which the ruthenium complex of the general formula III is linked via a stable phosphodiester linkage to the DNA or RNA, and the chromophores of the lumazine type are 1-(2'"'"'-deoxy-α

-D-ribofuranosyl)-6,7-dimethyl-lumazine [Lu].