Palladium mediated reaction of organic disulfides with mercurated nucleic acid components

• US 4,983,727 A
• Filed: 04/11/1988
• Issued: 01/08/1991
• Est. Priority Date: 04/11/1988
• Status: Expired due to Fees

• Current Assignee: Center For Innovation & Business Development Foundation
• Original Assignee: University of North Dakota School of Engineering & Mines Foundation
• Inventors: Bergstrom, Donald E., Jenson, Jeffrey A.
• Primary Examiner(s): Rollins, John W.
• Assistant Examiner(s): Wilson, James O.
• Application Number:
  US07/179,751
• Time in Patent Office: 1,002 Days
• Field of Search: 536/23, 536/28, 536/29, 536/4.1, 536/120
• US Class Current: 536/28.53
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/16 Purine radicals

Abstract

A method of preparing C-5 substituted nucleotides by reacting a C-5 posit mercurated nucleoside with an alkyl or aryl disulfide in the presence of palladium(II) ion. The resulting new method for attaching linker arms to nucleic acids (DNA,RNA) provides a wide ranging reaction far more useful than those heretofore developed, and allows preparation of modified components with linking reactive functionality that allows usefulness as probes, therapeutics, and for DNA sequencing.

Claims

1. A process of preparing a C-5 substituted nucleoside useful as linking units for probes, therapeutics and nucleotide sequencing, said process comprising:

reacting a C-5 mecuri-substituted nucleic acid moiety of the formula;

##STR3## wherein X is a soluble organic or inorganic ion, with a disulfide of the formula (RS)₂,

wherein R is a C₁ to C₂₀ alkyl, aryl, alkylaryl, alkoxyalkyl, alkyl carboxyl, aklylcarboxamido, alkyl carbonyl or a C₁ to C₂₀ functionally substituted alkyl group,

said reaction occurring in the presence of a mediating effective amount of palladium(II) ion source to provide a C-5 substituted nucleoside having a C-5 linker arm attached which will not interfere with the normal Watson-Crick hydrogen bonding process.

2. The process of claim 1 wherein X is an organic ion.

3. The process of claim 1 wherein X is an inorganic ion.

4. The process of claim 3 wherein X is a halide ion.

5. The process of claim 2 wherein R is a functionally substituted alkyl group.

6. The process of claim 2 wherein R is a C₁ to C₁₀ alkyl.

7. The process of claim 1 wherein the palladium(II) ion source is a palladium(II) salt.

8. The process of claim 7 wherein the source of palladium(II) is dilithium tetrachloro palladate.

9. The process of claim 1 wherein said reaction is conducted using an excess of alkyl disulfide over the equimolar stoichiometric amounts.

10. The process of claim 1 wherein said reaction is conducted in the presence of an organic solvent.

11. The process of claim 1 wherein said reaction is conducted in an aqueous medium.

12. The process of claim 1 wherein the C-5 substituent is a fluorescent marker.

13. The process of claim 1 wherein said C-5 substituent is biotin.

14. The process of claim 1 wherein C-5 substituent is a hapten.

15. The process of claim 1 wherein said reaction is conducted at a temperature within the range of from 0°

C. up to about 65°

C.

16. The process of claim 10 wherein said solvent is methanol.

17. The process off claim 1 wherein R is a C₁ to C₁₀ functional substituted alkyl group.