Process for preparing polynucleotides

• US 5,153,319 A
• Filed: 03/31/1986
• Issued: 10/06/1992
• Est. Priority Date: 03/31/1986
• Status: Expired due to Fees

• Current Assignee: University Patents, Inc.
• Original Assignee: University Patents, Inc.
• Inventors: Caruthers, Marvin H., Matteucci, Mark D.
• Primary Examiner(s): Michl, Paul R.
• Application Number:
  US06/846,453
• Time in Patent Office: 2,381 Days
• Field of Search: 536/22, 536/23, 536/24, 536/25, 536/26, 536/27, 536/28, 536/29
• US Class Current: 536/25.3
• CPC Class Codes: A61K 47/52 the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00686 Automatic
• CPC Class Codes: B01J 2219/00698 Measurement and control of process parameters
• CPC Class Codes: B01J 2219/00722 Nucleotides
• 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: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

New and useful intermediate nucleotides bound to an inorganic polymer support, including the preparation thereof, and processes for the conversion to oligonucleotides which are especially useful for the synthesis of polynucleotides, particularly ribonucleic (RNA) and deoxyribonucleic (DNA) acids, are described.

Claims

1. A process for the production of oligonnucleotides which comprises:

(a) converting an inorganic silica into a coupling agent-silica,

(b) combining the coupling agent-silica with a blocked nucleoside to obtain a blocked nucleoside-modified support; and

(c) removing the blocking group from the blocked nucleoside-modified support to provide for the coupling of nucleotides to the nucleoside-modified support.

2. A process for the production of oligonucleotides which comprise:

(a) treating an inorganic silica with a coupling agent which agent has a reactive group capable of reacting with a reactive group carried on a nucleoside; and

(b) combining the treated silica with a nucleoside or a blocked nucleoside to obtain a nucleoside-modified support or a blocked nucleoside-modified support.

3. A process for the production of oligonucleotides which comprises:

(a) providing an inorganic silica;

(b) converting the silica into an amino-silica;

(c) providing a nucleoside

(d) combining the amino-silica and the nucleoside to obtain a nucleoside-modified support; and

(e) attaching nucleotides to the nucleoside-modified support.

4. A process for the support of oligonucleotides which comprises:

(a) converting a silica gel into a coupling agent-silica gel;

(b) combining the coupling agent-silica gel with a blocked nucleoside to obtain a blocked nucleoside-modified support; and

(c) removing the blocking group from the blocked nucleoside-modified support to provide for the coupling of nucleotides to the nucleoside-modified support.

5. A process for the production of oligonucleotides which comprise:

(a) treating a silica gel with a coupling agent which agent has a reactive group capable of reacting with a reactive group carried on a nucleoside; and

(b) combining the treated silica gel with a nucleoside or a blocked nucleoside to obtain a nucleoside-modified support or a blocked nucleoside-modified support.

6. A process for the production of oligonucleotides which comprises:

(a) providing a silica gel;

(b) converting the silica gel into an amino-silica gel;

(c) providing a nucleoside

(d) combining the amino-silica gel and the nucleoside to obtain a nucleoside-modified support; and

(e) attaching nucleotides to the nucleoside-modified support.

7. A process for the production of oligonucleotides which comprises:

(a) providing a silica gel support material;

(b) converting said silica gel to a derivatize silica gel containing aminoalkyl or hydroxyalkyl groups;

(c) providing a nucleoside modified to contain the half ester of a dicarboxylic acid; and

(d) combining the nucleoside ester with the derivatized silica gel to obtain a nucleoside modified silica gel support.

8. A process according to claim 7 in which the derivatized silica gel contains aminoalkyl groups.

9. A process according to claim 8 in which the dicarboxylic acid is succinic acid.

10. A process according to claim 8 in which the succinic anhydride is use in step (c).

11. A process for the production of oligonucleotides which

(a) providing a silica gel support material;

(b) converting the silica gel to a derivatized silica

(c) reacting the derivatized silica gel with a

(d) condensing the free carboxy groups on silica with a freehydroxy group of a nucleoside to obtain a nucleoside modified silica gel support.

12. A process according to claim 11 in which the derivatized silica contains aminoalkyl groups.

13. A process according to claim 11 in which the dicarboxylic acid is succinic acid.

14. A process for production of oligonucleotides which comprises the step of condensing the 3'"'"'-OH or 5'"'"'-OH group of a nucleoside or oligonucleotide covalently linked to an inorganic polymer by a coupling agent through the 5'"'"'-O- or 3'"'"'-O-, respectively, of said nucleoside or oligonucleotide with a phosphite compound of the formula:

##STR5## wherein R is a blocking group;

B is a nucleosides or deoxynucleoside base;

A is H or OR;

R₁ is lower alkyl; and

X is formed by the removal of the hydrogen atom from a ring nitrogen of a nitrogen herterocyclic compound which contains unsaturated bonds in the ring structure.

15. The process according to claim 14 wherein the phosphite compound is of the formula:

##STR6## wherein substituents R, R₁, X, A and B are previously defined and said condensation occurs through the 5'"'"'-OH of said nucleoside or oligonucleotide.

16. The process according to claim 14 in which the heterocyclic compound is triazole.

17. The process according to claim 14 in which the heterocyclic compound is tetrazole.