Electrochemical deblocking solution for electrochemical oligomer synthesis on an electrode array

• US 9,267,213 B1
• Filed: 04/30/2014
• Issued: 02/23/2016
• Est. Priority Date: 03/25/2005
• Status: Active Grant

• Current Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Original Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Inventors: Maurer, Karl, Cooper, John J
• Primary Examiner(s): Olson, Eric
• Application Number:
  US14/266,595
• Time in Patent Office: 664 Days
• Field of Search: None
• US Class Current: 1/1
• 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 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: C07B 31/00 Reduction in general
• CPC Class Codes: C07B 33/00 Oxidation in general
• CPC Class Codes: C07B 41/02 of hydroxy or O-metal groups
• CPC Class Codes: C07B 41/06 of carbonyl groups
• CPC Class Codes: C07C 309/30 of six-membered aromatic rings substituted by alkyl groups
• CPC Class Codes: C07C 39/08 Dihydroxy benzenes; Alkylated derivatives thereof
• CPC Class Codes: C07C 50/00 Quinones for quinone methides, see unsaturated ketones with a keto group being part of a ring
• CPC Class Codes: C07C 50/04 Benzoquinones, i.e. C6H4O2
• CPC Class Codes: C07D 213/16 containing only one pyridine ring
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C25B 3/00 Electrolytic production of organic compounds
• CPC Class Codes: C25B 3/23 Oxidation halogenation C25B3/27
• CPC Class Codes: C25B 3/25 Reduction
• CPC Class Codes: C25B 3/29 Coupling reactions
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

There is disclosed an electrochemical deblocking solution for use on an electrode microarray. There is further disclosed a method for electrochemical synthesis on an electrode array using the electrochemical deblocking solution. The solution and method are for removing acid-labile protecting groups for synthesis of oligonucleotides, peptides, small molecules, or polymers on a microarray of electrodes while substantially improving isolation of deblocking to active electrodes. The method comprises applying a voltage or a current to at least one electrode of an array of electrodes. The array of electrodes is covered by the electrochemical deblocking solution.

Claims

1. An electrochemical deblocking solution comprising:

an organic base including N,N-diisopropylethylamine, a benzoquinone and a hydroquinone;

tetrabutylammonium hexafluorophosphate as an organic salt; and

a solvent including acetonitrile and an alcohol.

2. The electrochemical deblocking solution of claim 1, where the solvent has approximately 1% to 99% acetonitrile and approximately 1% to 99% isopropanol.

3. The electrochemical deblocking solution of claim 1, where the N,N-diisopropylethylamine is approximately 0.01 to 200 mM of N,N-diisopropylethylamine.

4. The electrochemical deblocking solution of claim 1, where the benzoquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl benzoquinone.

5. The electrochemical deblocking solution of claim 1, where the hydroquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl hydroquinone.

6. The electrochemical deblocking solution of claim 1, where the tetrabutylammonium hexafluorophosphate is approximately 0.05 to 5 M tetrabutylammonium hexafluorophosphate.

7. The electrochemical deblocking solution of claim 6, where the N,N-diisopropylethylamine is approximately 0.01 to 200 mM of N,N-diisopropylethylamine, the hydroquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl hydroquinone and the benzoquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl benzoquinone.

8. An electrochemical deblocking solution comprising:

an organic base including a lutidine, a benzoquinone and a hydroquinone;

tetraethylammonium p-toluenesulfonate as an organic salt; and

a solvent selected from the group consisting of two or more of an alcohol, acetonitrile and dichloromethane.

9. The electrochemical deblocking solution of claim 8 for use on a prepared surface, where the lutidine is not bound to the prepared surface.

10. The electrochemical deblocking solution of claim 8, where the solvent has approximately 10% to 99% acetonitrile and approximately 1% to 90% methanol.

11. The electrochemical deblocking solution of claim 8, where the solvent has approximately 1% to 99% dichloromethane and approximately 1% to 99% isopropanol.

12. The electrochemical deblocking solution of claim 8, where the lutidine is approximately 0.01 to 200 mM of dimethyl pyridine.

13. The electrochemical deblocking solution of claim 8, where the benzoquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl benzoquinone.

14. The electrochemical deblocking solution of claim 8, where the hydroquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl hydroquinone.

15. The electrochemical deblocking solution of claim 8, where the tetraethylammonium p-toluenesulfonate is approximately 0.1 mM to 5 M tetraethylammonium p-toluenesulfonate.

16. The electrochemical deblocking solution of claim 15, where the lutidine is approximately 0.01 to 200 mM of dimethyl pyridine, the hydroquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl hydroquinone and the benzoquinone is approximately 0.1 to 100 mM 2,5-di-tert-butyl benzoquinone.

17. The electrochemical deblocking solution of claim 10, where the tetraethylammonium p-toluenesulfonate is dissolved in the acetonitrile and methanol solvent to form a first solution and the lutidine provides basicity to quench the acidity of the tetraethylammonium p-toluenesulfonate in acetonitrile and methanol solution.

18. The electrochemical deblocking solution of claim 11, where the tetraethylammonium p-toluenesulfonate is dissolved in the dichloromethane and isopropanol solvent to form a first solution and the lutidine provides basicity to quench the acidity of the tetraethylammonium p-toluenesulfonate in dichloromethane and isopropanol solution.

19. An electrochemical deblocking solution for use on an electrode microarray comprising:

approximately 0.01 to 200 mM of a pyridine derivative or isomer thereof, where the pyridine derivative or isomer thereof is not bound to the electrode microarray;

approximately 0.1 to 100 mM 2,5-di-tert-butyl benzoquinone;

approximately 0.1 to 100 mM 2,5-di-tert-butyl hydroquinone;

approximately 0.1 mM to 5 M tetraethylammonium p-toluenesulfonate; and

a solvent having approximately 10% to 99% acetonitrile and approximately 1% to 90% methanol.

20. The electrochemical deblocking solution of claim 19, where the pyridine derivative or isomer thereof is selected from the group consisting of