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

Patent metadata

Field	Value
Publication number	US-10435418-B2
Application number	US-201715700911-A
Country	US
Kind code	B2
Filing date	Sep 11, 2017
Priority date	Apr 23, 2014
Publication date	Oct 8, 2019
Grant date	Oct 8, 2019

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 sp 2 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.

First claim

What is claimed is:

1. A process comprising:
(a) providing:
(1) a boron compound having a boron atom covalently bonded to an sp 2 hybridized carbon conjugated with a cis-carbonyl, selected from the group consisting of:

wherein:
X 1 , X 2 are independently selected from groups that can hydrolyze from the boron to yield boronic acid; and
R 1 , R 2 , and R 3 are independently selected from the group consisting of hydrogen, organic ligands, and heterorganic ligands and optionally joined in one or more rings,
wherein X 1 , X 2 , R 1 , R 2 , and R 3 exclude α-effect amines, and
wherein the boron compound is adapted to selectively and spontaneously react with an α-effect amine at in a liquid solvent, a temperature of less than 55° C., and at a pH above 2, to at least form an adduct by formation of a covalent bond between the boron compound and the α-effect amine; and
(2) an α-effect amine comprising an amino acid, and having an amine group bonded to an oxygen, nitrogen, or sulfur; and

(b) contacting the boron compound with the α-effect amine in the liquid solvent at the temperature of less than 55° C. and at the pH above 2, to spontaneously form the adduct.

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:
R 2 is selected from the group consisting of: H, CH 3 ; and
R 3 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:

and a further product thereof formed through at least one of dehydration, interaction with the solvent, and interaction with a reactive heteroatom in the solvent, wherein:
X 3 is O or N; and
R 4 is an alkyl, aryl, or heteroatom containing group.

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

wherein X 3 is N, and
X 4 is selected from the group consisting of alkyl, aryl, heteroalkyl, heteroaryl, hydroxyl, and water.

14. The process according to claim 12 , wherein R 4 is selected from the group consisting of: H, CH 3 , CH 2 CH 3 , CH 2 Ph, p-COOH Ph, o-NH 2 Ph, o-OH Ph, COH, COCH 3 , COCH 2 Ph, COPh, CO-coumarin, and CONH 2 .

15. A boron compound comprising a boron atom bonded to a sp 2 hybridized carbon conjugated with a cis-carbonyl and a biomolecule, selected from the group consisting of:

wherein:
X 1 , X 2 are independently selected from groups that can hydrolyze from the boron to yield boronic acid; and
R 1 , R 2 , and R 3 are independently selected from the group consisting of hydrogen, organic ligands, and heterorganic ligands and optionally joined in one or more rings, at least one of R 1 , R 2 , and R 3 being the biomolecule;
wherein X 1 , X 2 , R 1 , R 2 , and R 3 exclude α-effect amines, the boron compound being selectively and spontaneously reactive to form at least a covalently bonded adduct by formation of a covalent bond between the boron compound and an α-effect amine comprising an amine group bonded to an oxygen, nitrogen, or sulfur, and non-reactive to form a covalently bonded adduct with a non α-effect amine, in an aqueous solution at pH 7 at 25° C.,
wherein the biomolecule comprises at least one of an amino acid, a peptide, a nucleotide, a carbohydrate, and a drug.

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

(ii) a further product formed from (I) formed through dehydration:

wherein X 3 is N, and X 4 is selected from the group consisting of alkyl, aryl, heteroalkyl, heteroaryl, hydroxyl and water,
(iii) a further product formed from (I) through interaction with the solvent; and
(iv) a further product formed from (I) through interaction with a reactive heteroatom in the solvent.

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

wherein:
R 2 is H or CH 3 ;
R 3 and R 6 are independently selected from the group consisting of alkyl or OR, wherein R is selected from the group consisting of alkyl, heteroalkyl, heteroaryl, alkylamine, alkylthiol, alkylbromide, arylbromide, C 2 -C 6 alkanoyl, CH 2 Ar or

Assignees

• Univ New York State Res Found
• The Research Foundation For The State Univ O

Inventors

• Tuttle Susan Bane
• Dilek Ozlem
• Mukherjee Kamalika

Classifications

• C07K16/40

  against enzymes · CPC title

• C01B35/146

  Compounds containing boron and nitrogen, e.g. borazoles (ammonium tetrafluoborates C01B35/063; ammonium borates C01B35/12) · CPC title

• A61K31/69

  Boron compounds · CPC title

• C01B35/10

  Compounds containing boron and oxygen (C01B35/06 takes precedence) · CPC title

• C07F5/02

  Boron compounds · CPC title