Method for labeling of aldehyde containing target molecules

Patent metadata

Field	Value
Publication number	US-11236386-B2
Application number	US-201916598685-A
Country	US
Kind code	B2
Filing date	Oct 10, 2019
Priority date	Apr 12, 2017
Publication date	Feb 1, 2022
Grant date	Feb 1, 2022

Abstract

The present description relates to a method for binding to a target molecule having an aldehyde compound derived from N-(2-aminoethyl)pyrrole, which compound also has a moiety of interest, to compounds (conjugates) obtained by this method, having both the target molecule and the moiety of interest and to novel substances derived from N-(2-aminoethyl)pyrrole. In one embodiment, the compound has the formula of Formula IIwherein R1, R2 and R3 independently are H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, wherein R4, R5, R6, R7 and R8 independently are H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or -LM, wherein L is absent or is a linker and M is a moiety of interest selected from a nucleotide, an oligonucleotide, a peptide, a label, a cytotoxic agent, a partner of a binding pair and a functional group, wherein two of R4, R5, R6, R7, and R8 optionally are linked to form a substituted or unsubstituted cycloalkyl or a substituted or unsubstituted heterocycloalkyl, and T is a target molecule selected from the group consisting of a solid phase, a polypeptide, a protein, a carbohydrate, a nucleotide and a nucleic acid, with the proviso that at least one of R4, R5, R6, R7 or R8 is -LM.

First claim

The invention claimed is:

1. A method of producing a compound according to Formula II, the method comprising; reacting a compound of Formula I with a target comprising an aldehyde group as given below

thereby obtaining a compound according to Formula II, Formula II:

wherein R1, R2 and R3 independently are H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, wherein R4, R5, R6, R7 and R8 independently are H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or -LM, wherein L is absent or is a linker, and when L is a linker, the linker L is bound to M via an ester, an ether, a thioether, an amine, an amide, a disulfide, a carbonate, a carbamate, an alkene, a triazole, a dihydropyridazine or a phosphodiester bond, wherein M is selected from the group consisting of a nucleotide, an oligonucleotide, a peptide, a label, a cytotoxic agent, a partner of a binding pair and a functional group, wherein two of R4, R5, R6, R7, and R8 optionally are linked to form a substituted or unsubstituted cycloalkyl or a substituted or unsubstituted heterocycloalkyl, and wherein T is a target molecule selected from the group consisting of a solid phase, a polypeptide, a protein, a carbohydrate, a nucleotide and a nucleic acid, with the proviso that at least one of R4, R5, R6, R7 or R8 is -LM.

2. The method according to claim 1 , wherein in a compound of Formula I or Formula II, R1, R2 and R3 independently are H or methyl.

3. The method according to claim 1 , wherein in a compound of Formula I or Formula II, the moiety M is selected from the group consisting of a nucleotide, an oligonucleotide, a fluorescent or a luminescent label, a cytotoxic agent, biotin or a biotin analogue, digoxin or a digoxin analogue, and maleimide.

4. The method according to claim 1 , wherein in a compound of Formula I or Formula II, the target molecule T is a polypeptide, a protein, or a nucleic acid.

5. The method according to claim 1 , wherein in a compound of Formula I or Formula II, the target molecule T is an antibody.

6. The method according to claim 1 , wherein in a compound of Formula I or Formula II, the linker L is absent or selected from substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or an oligopeptide and, if present, comprises an amide linkage, an ester linkage, an alkene or a triazole.

7. The method according to claim 1 , wherein in a compound of Formula I or Formula II, the linker L is selected from substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl or an oligopeptide.

8. A compound of Formula II

wherein
R1, R2 and R3 independently are H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, wherein R4, R5, R6, R7 and R8 independently are H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or -LM, wherein L is absent or is a linker, and when L is a linker, the linker L is bound to M via an ester, an ether, a thioether, an amine, an amide, a disulfide, a carbonate, a carbamate, an alkene, a triazole, a dihydropyridazine or a phosphodiester bond, wherein M is selected from the group consisting of a nucleotide, an oligonucleotide, a peptide, a label, a cytotoxic agent, a partner of a binding pair and a functional group, wherein two of R4, R5, R6, R7, and R8 optionally are linked to form a substituted or unsubstituted cycloalkyl or a substituted or unsubstituted heterocycloalkyl, and
T is a target molecule selected from the group consisting of a solid phase, a polypeptide, a protein, a carbohydrate, a nucleotide and a nucleic acid,
with the proviso that at least one of R4, R5, R6, R7 or R8 is -LM.

Assignees

• Roche Diagnostics Operations Inc

Inventors

• Bergmann Frank
• Oelschlaegel Tobias
• Pomplun Sebastian Johannes

Classifications

• G01N2333/59

  Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH] · CPC title

• C07D207/327

  Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals · CPC title

• C07H21/04

  with deoxyribosyl as saccharide radical · CPC title

• C07D207/452

  with hydrocarbon radicals, substituted by hetero atoms, directly attached to the ring nitrogen atom · CPC title

• C07D295/13

  to an acyclic saturated chain · CPC title