Non-nucleoside 1,3-diol reagents for labeling synthetic oligonucleotides

• US 5,451,463 A
• Filed: 06/03/1993
• Issued: 09/19/1995
• Est. Priority Date: 08/28/1989
• Status: Expired due to Term

• Current Assignee: Clontech Laboratories Incorporated
• Original Assignee: Clontech Laboratories Incorporated
• Inventors: Nelson, Paul S., Kent, Mark A.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): Kunz, Gary L.
• Application Number:
  US08/071,937
• Time in Patent Office: 838 Days
• Field of Search: 536/25.3, 536/25.32, 501/33, 560/155, 428/402, 546/25, 546/102, 548/304.1, 549/223, 549/227, 552/544, 558/93, 558/168, 558/191, 558/192, 568/933
• US Class Current: 428/402
• 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: Y10T 428/2982 Particulate matter [e.g., sphere, flake, etc.]

Abstract

A trifunctional reagent possessing a primary hydroxyl, a secondary hydroxyl, and a primary amino group. This reagent is useful in solid phase oligonucleotide synthesis for the convenient labeling of the 3'"'"'-terminus. The secondary hydroxyl may be an phosphoramidite derivative permitting the attachment to the solid phase support. The reporter molecule may be attached to the trifunctional molecule prior to the completion of the oligonucleotide synthesis or after the oligonucleotide is cleaved from the support. A preferred species of this trifunctional molecule is shown below.

Claims

1. A reagent for use in oligonucleotide synthesis having the following structure:

##STR6## wherein n is from 0 to about 15;

R₁ is selected from the group consisting of dimethoxytrityl (DMT) and monomethoxytrityl;

R₂ is selected from the group consisting of controlled pore glass (CPE);

alkylamine controlled pore glass, wherein alkyl is 1 to 50 carbon atoms;

polystyrene;

divinylbenzene;

##STR7## and salts thereof, wherein R₅ and R₆ are independently selected from the group consisting of C_3-10 branched alkyl, C_1-12 unbranched alkyl, and cyclic hydrocarbons; and

Y is a beta-cyanoethyl group;

X₁ is a functional group selected from the group consisting of oxygen, sulfur, or --NH;

R₃ is a reporter molecule or protecting group selected from the group consisting of biotin, rhodamine, Fmoc, fluorescein, dinitrophenyl, cholesterol, and acridine.

2. The reagent, according to claim 1, wherein R₂ is selected from the group consisting of ##STR8## and salts thereof, wherein R₅ and R₆ are independently selected from the group consisting of C_3-10 branched alkyl and C_1-12 unbranched alkyl and cyclic hydrocarbons, and Y is a beta-cyanoethyl group.

3. The reagent, according to claim 1, wherein R₂ is CPG or alkylamine CPG, wherein alkyl is 1 to 50 carbon atoms.

4. The reagent, according to claim 1, wherein R₂ is polystyrene or divinylbenzene.

5. The reagent for use in oligonucleotide synthesis, according to claim 1, having the following structure:

##STR9##

7. The reagent for use in oligonucleotide synthesis, according to claim 1, having the following structure:

##STR11##

8. The reagent for use in oligonucleotide synthesis, according to claim 1, having the following structure:

##STR12##

9. The reagent for use in oligonucleotide synthesis, according to claim 1, having the following structure:

##STR13##

10. The reagent, according to claim 1, having the structure:

##STR14##

11. The reagent, according to claim 1, having the structure:

##STR15##

12. The reagent, according to claim 1, having the structure:

##STR16##

13. The reagent, according to claim 1, having the structure:

##STR17##

14. The reagent, according to claim 1, having the structure:

##STR18##

15. The reagent for use in oligonucleotide synthesis, according to claim 1, having the following structure:

##STR19##

6. A reagent for use in oligonucleotide synthesis having the following structure:

##STR10##