Rapid and efficient bioorthogonal ligation reaction and boron-containing heterocycles useful in conjunction therewith

• US 10,435,418 B2
• Filed: 09/11/2017
• Issued: 10/08/2019
• Est. Priority Date: 04/23/2014
• Status: Active Grant

• Current Assignee: The Research Foundation for The State University of New York (State University of New York)
• Original Assignee: Research Foundation For The State University O
• Inventors: Tuttle, Susan Bane, Dilek, Ozlem, Mukherjee, Kamalika
• Primary Examiner(s): Balasubramanian, Venkataraman
• Application Number:
  US15/700,911
• Publication Number:
  US 20180065993A1
• Time in Patent Office: 757 Days
• Field of Search: 544229, 544 69
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/69 Boron compounds
• CPC Class Codes: C01B 35/10 Compounds containing boron and oxygen C01B35/06 takes precedence
• CPC Class Codes: C01B 35/146 Compounds containing boron and nitrogen, e.g. borazoles ammonium tetrafluoborates C01B35/063; ammonium borates C01B35/12
• CPC Class Codes: C07F 5/02 Boron compounds
• CPC Class Codes: C07F 5/025 Boronic and borinic acid compounds
• 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: C07K 14/765 Serum albumin, e.g. HSA
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/40 against enzymes

Abstract

A reaction method comprising combining a carbonyl-substituted arylboronic acid or ester and an α-effect amine in aqueous solution at a temperature between about −5 C to 55 C, and a pH between 2 and 8 to produce an adduct. A process is also provided comprising: contacting a boron compound having a boron atom bonded to a sp² hybridized carbon conjugated with a cis-carbonyl, the boron having at least one labile substituent, with an α-effect amine, in a solvent for a time sufficient to form an adduct, which may proceed to further products.

Claims

1. A process comprising:

(a) providing;

(1) a boron compound having a boron atom covalently bonded to an sp² hybridized carbon conjugated with a cis-carbonyl, selected from the group consisting of;

2. The process according to claim 1, wherein the solvent comprises an aqueous medium, the contacting being performed at temperatures between about −

5°

C. to 55°

C., and at a pH between 2 and 8, and wherein the boron compound, the α

-effect amine, and the solvent are bioorthogonal.

3. The process according to claim 1, wherein the α

-effect amine comprises an amine group bonded to an oxygen which is reactive with the boron compound.

4. The process according to claim 1, wherein the α

-effect amine comprises an amine group bonded to a nitrogen which is reactive with the boron compound.

5. The process according to claim 1, wherein the boron compound comprises a carbonyl-substituted arylboronic acid or ester.

6. The process according to claim 1, wherein the adduct undergoes a molecular rearrangement after forming an initial covalent bond between the cis-carbonyl carbon and the α

-effect amine.

7. The process according to claim 1, wherein:

R₂ is selected from the group consisting of;

H, CH₃; and

R₃ is selected from the group consisting of OH, O-alkyl, O-alkylbromide, O-alkylamine, O-alkylamide, O-alkylthiol, O-alkylthioester, alkylamine, alkylamide and alkylbromide.

8. The process according to claim 1, wherein the α

-effect amine is selected from the group consisting of alpha-hydrazides of tyrosine, phenylalanine, alanine, beta-alanine, glycine, dimethylglycine, and CBz-serine.

9. The process according to claim 1, wherein the α

-effect amine is selected from the group consisting of;

a hydrazine;

a semicarbazide, a thiosemicarbazide;

a hydrazide, a thiohydrazide, a hydroxylamine, an O-alkylhydroxylamine, and an O-arylhydroxylamine.

10. The process according to claim 1, wherein the boron compound comprises a carbonyl substituted arylboronic acid selected from the group consisting of:

an ortho formyl phenylboronic acid or ester derivative;

an ortho ketone phenylboronic acid or ester derivative;

an ortho aldehyde phenylboronic acid ester derivative of an amino acid;

a ketone phenylboronic acid or ester derivative of an amino acid;

an ortho aldehyde phenylboronic acid derivatized with a reactive functional group which is orthogonal with respect to the boron compound and the α

-effect amine; and

a ketone phenylboronic acid derivatized with a reactive functional group which is orthogonal with respect to the boron compound and the α

-effect amine.

11. The process according to claim 1, wherein the liquid solvent is an aqueous medium having a pH of about 6-8, and the spontaneous formation of the adduct is substantially complete within a period of less than about 10 minutes at a temperature of about 0-42°

C.

12. The process according to claim 1, wherein the adduct comprises a boron compound selected from the group consisting of:

13. The process according to claim 12, wherein the further product comprises a dehydration product selected from the group consisting of:

14. The process according to claim 12, wherein R₄ is selected from the group consisting of:

H, CH₃, CH₂CH₃, CH₂Ph, p-COOH Ph, o-NH₂Ph, o-OH Ph, COH, COCH₃, COCH₂Ph, COPh, CO-coumarin, and CONH₂.

15. A boron compound comprising a boron atom bonded to a sp² hybridized carbon conjugated with a cis-carbonyl and a biomolecule, selected from the group consisting of:

16. The boron compound according to claim 15, wherein the adduct is selected from the group consisting of:

(i)

17. The boron compound according to claim 15, wherein the adduct is selected from the group consisting of:

18. The boron compound according to claim 15, wherein the boron compound comprises a carbonyl substituted arylboronic acid selected from the group consisting of:

an ortho formyl phenylboronic acid or ester derivative;

an ortho ketone phenylboronic acid or ester derivative;

an ortho aldehyde phenylboronic acid ester derivative of an amino acid;

a ketone phenylboronic acid or ester derivative of an amino acid;

an ortho aldehyde phenylboronic acid derivatized with a reactive functional group which is orthogonal with respect to the boron compound and the α

-effect amine; and

a ketone phenylboronic acid derivatized with a reactive functional group which is orthogonal with respect to the boron compound and the α

-effect amine.

19. The boron compound according to claim 15, provided as a kit in combination with the α

-effect amine.

20. A process comprising:

combining;

(1) a biorthogonal compound having a boron atom covalently bonded to an sp² hybridized carbon conjugated with a cis-carbonyl, selected from the group consisting of;