Electrochemical deblocking solution for electrochemical oligomer synthesis on an electrode array

• US 10,006,131 B1
• Filed: 02/22/2016
• Issued: 06/26/2018
• 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, Jr., John J.
• Primary Examiner(s): Olson, Eric
• Application Number:
  US15/050,377
• Time in Patent Office: 855 Days
• Field of Search: None
• 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 a lutidine, a benzoquinone and a hydroquinone;

tetraethylammonium p-toluenesulfonate as an organic salt; and

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

2. The electrochemical deblocking solution of claim 1, where the solvent is between:

a lower limit of approximately 10% acetonitrile; and

an upper limit of approximately 99% acetonitrile.

3. The electrochemical deblocking solution of claim 1, where the solvent has approximately 10% to 99% acetonitrile.

4. The electrochemical deblocking solution of claim 1, where the solvent is between:

a lower limit of approximately 1% dichloromethane; and

an upper limit of approximately 99% dichloromethane.

5. The electrochemical deblocking solution of claim 1, where the solvent is between:

a lower limit of approximately 1% isopropanol; and

an upper limit of approximately 99% isopropanol.

6. The electrochemical deblocking solution of claim 1, where the lutidine is between:

a lower limit of approximately 0.01 mM dimethyl pyridine; and

an upper limit of approximately 200 mM dimethyl pyridine.

7. The electrochemical deblocking solution of claim 1, where the benzoquinone is between:

a lower limit of approximately 0.1 mM; and

an upper limit of approximately 100 mM.

8. The electrochemical deblocking solution of claim 7, where the hydroquinone is between:

a lower limit of approximately 0.1 mM; and

an upper limit of approximately 100 mM.

9. The electrochemical deblocking solution of claim 1, where the tetraethylammonium p-toluenesulfonate is approximately is between:

a lower limit of approximately 0.1 mM; and

an upper limit of approximately 5 M.

10. The electrochemical deblocking solution of claim 9, where the lutidine is between:

a lower limit of approximately 0.01 mM; and

an upper limit of approximately 200 mM.

11. The electrochemical deblocking solution of claim 9, where the hydroquinone is between:

a lower limit of approximately 0.1 mM; and

an upper limit of approximately 100 mM.

12. The electrochemical deblocking solution of claim 9, where the benzoquinone is between:

a lower limit of approximately 0.1 mM; and

an upper limit of approximately 100 mM.

13. The electrochemical deblocking solution of claim 1, where the tetraethylammonium p-toluenesulfonate is dissolved in the solvent to form a first solution and the lutidine provides basicity to quench acidity of the tetraethylammonium p-toluenesulfonate in solution.

14. The electrochemical deblocking solution of claim 1, where the tetraethylammonium p-toluenesulfonate is dissolved in the solvent to form a tetraethylammonium p-toluenesulfonate solution and the lutidine provides basicity to quench acidity of the tetraethylammonium p-toluenesulfonate solution.

15. A method of electrochemical deblocking an oligomer comprising the steps of:

covering an array of electrodes with a deblocking solution including;

lutidine, benzoquinone, hydroquinone and tetraethylammonium p-toluenesulfonate; and

one or more solvents selected from the group consisting of an alcohol, acetonitrile and dichloromethane; and

applying one or both a voltage and a current to at least one electrode of the array of electrodes.

16. The method of claim 15, where lutidine is not bound to the array of electrodes.

17. The method of claim 15, where the one or more solvents is between:

a lower limit of approximately 10% acetonitrile; and

an upper limit of approximately 99% acetonitrile.

18. The method of claim 15, where the one or more solvents is between:

a lower limit of approximately 1% methanol; and

an upper limit of approximately 90% methanol.

19. The method of claim 15, where the one or more solvents is between:

a lower limit of approximately 1% dichloromethane; and

an upper limit of approximately 99% dichloromethane.

20. A kit for electrochemical deblocking comprising:

an array of electrodes adapted to connect with one or both a voltage and a current source to apply one or both a voltage and a current to at least one electrode of the array of electrodes;

a deblocking solution including;

lutidine, benzoquinone, hydroquinone and tetraethylammonium p-toluenesulfonate; and

one or more solvents selected from the group consisting of an alcohol, acetonitrile and dichloromethane; and

instructions for one or both preparing the deblocking solution and applying one or both a voltage and a current to at least one electrode of the array of electrodes.