Reagents and methods for esterification

Patent metadata

Field	Value
Publication number	US-12110523-B2
Application number	US-202117526816-A
Country	US
Kind code	B2
Filing date	Nov 15, 2021
Priority date	Apr 9, 2015
Publication date	Oct 8, 2024
Grant date	Oct 8, 2024

Abstract

Methods and reagents for esterification of biological molecules including proteins, polypeptides and peptides. Diazo compounds of formula I:

where R is hydrogen, an alkyl, an alkenyl or an alkynyl, R A represents 1-5 substituents on the indicated phenyl ring and R M is an organic group. R M includes a label, a cell penetrating group, a cell targeting group, or a reactive group or latent reactive group for reaction to bond to a label, a cell penetrating group, or a cell targeting group, among other organic groups useful for esterification of biological molecules. Also provided are diazo compounds which are bifunctional and trifunctional coupling reagents as well as reagents for the synthesis of compounds of formula I.

First claim

We claim:

1. A compound of formula I:

or salts thereof,
where:
R is hydrogen, an alkyl group, alkenyl group, or an alkynyl group;
R M is M or -L-M, where M is a polymer or a non-polymeric organic group, having from 1 to 100 carbon atoms and optionally nitrogen, oxygen or sulfur atoms, and -L is a divalent linker moiety having from 1-30 carbon atoms and optionally nitrogen, oxygen or sulfur atoms; and
R A represents 1 to 3 non-hydrogen substituents on the phenyl ring, wherein the non-hydrogen substituents are selected from the group consisting of alkyl, cycloalkyl, alkoxy, cycloalkoxy, aryl, arylalkyl, haloalkyl, haloalkoxy, heterocyclyl and R P —CO—NH—, where the alkyl, cycloalkyl, alkoxy, cycloalkoxy, aryl, arylalkyl and heterocyclyl groups are optionally substituted;
wherein optional substitution is substitution with one or more groups selected from oxo, thiox, -sulfhydryl, hydroxy, alkyl, alkoxy, alkenyl, alkenyloxy, alkynyl, alkynyloxy, aryl, aryloxy, heteroaryl, heteroaryloxy, carbocyclyl, carbocyclyloxy, heterocyclyl, heterocyclyloxy, alkylthio, alkenylthio, alkynylthio, arylthio, thioheteroaryl, thiocarbocyclyl, thioheterocyclyl, —CORs, —COH, —OCORs, —OCOH, —CO—ORs, —CO—OH, —CO—O—CO-Rs, —CON(Rs) 2 , —CONHRs, —CONH 2 , —NRs-CORs, —NHCORs, —NHRs, —N(Rs) 2 , —O—SO 2 -Rs, —SO 2 -Rs, —SO 2 —NHRs, —SO 2 —N(Rs) 2 , —NRs-SO 2 -Rs, —NH—SO 2 -Rs, —NRsCO—N(Rs) 2 , —NH—CO—NHRs, —O—PO(ORs) 2 , —O—PO(ORs)(N(Rs) 2 ), —O—PO(N(Rs) 2 ) 2 , —N—PO(ORs) 2 , —N—PO(ORs)(N(Rs) 2 ), —P(Rs) 2 , —B(OH) 2 , —B(OH)(ORs), and —B(ORs) 2 , where each Rs independently is an alkyl, alkenyl, alkynyl, aryl, heteroaryl, carbocyclyl, or heterocyclyl group or two Rs within the same substituent can together form a carbocyclic or heterocyclic ring having 3 to 10 ring atoms.

2. The compound of claim 1 , wherein R A represents ring substitution having one non-hydrogen group at the para ring or meta ring position on the phenyl ring.

3. The compound of claim 2 , wherein R A is an optionally substituted alkyl or alkoxy group at the para position on the phenyl ring.

4. The compound of claim 3 , wherein R A is substituted with a group selected from —CORs, —COH, —OCORs, —OCOH, —CO—ORs, —CO—OH, —CO—O—CO-Rs, wherein Rs is an alkyl, alkenyl, or alkynyl group.

5. The compound of claim 4 , wherein R A is substituted with a —CO—O—CO-Rs group, wherein Rs is an alkyl group.

6. The compound of claim 1 , wherein R A is an alkyl or alkoxy group substituted with a —CO—O—CO-Rs group, wherein Rs is an alkyl group.

7. A compound of formula I:

or salts thereof,
where:
R is hydrogen, an alkyl group, alkenyl group, or an alkynyl group;
R A represents 1-3 non-hydrogen substituents on the ring; and
R M is M or -L-M, where M is a polymer or a non-polymeric organic group, having from 1 to 100 carbon atoms and optionally nitrogen, oxygen or sulfur atoms, and -L is a divalent linker moiety having from 1-30 carbon atoms and optionally nitrogen, oxygen or sulfur atoms;
wherein R A is an alkyl or alkoxy group substituted with a —CO—O—CO-Rs group, wherein Rs is an alkyl group;
wherein R M is -L-M, where M is an alkyl, alkynyl, cycloalkyl, aryl, arylalkyl, heterocyclyl or heteroaryl group which is optionally substituted; and
wherein optional substitution is substitution with one or more groups selected from oxo, thiox, halogen, nitro, cyano, cyanate, azido, thiocyano, isocyano, isothiocyano, sulfhydryl, hydroxy, alkyl, alkoxy, alkenyl, alkenyloxy, alkynyl, alkynyloxy, aryl, aryloxy, heteroaryl, heteroaryloxy, carbocyclyl, carbocyclyloxy, heterocyclyl, heterocyclyloxy, alkylthio, alkenylthio, alkynylthio, arylthio, thioheteroaryl, thiocarbocyclyl, thioheterocyclyl, —CORs, —COH, —OCORs, —OCOH, —CO—ORs, —CO—OH, —CO—O—CO-Rs, —CON(Rs) 2 , —CONHRs, —CONH 2 , —NRs-CORs, —NHCORs, —NHRs, —N(Rs) 2 , —O—SO 2 -Rs, —SO 2 -Rs, —SO 2 —NHRs, —SO 2 —N(Rs) 2 , —NRs-SO 2 -Rs, —NH—SO 2 -Rs, —NRsCO—N(Rs) 2 , —NH—CO—NHRs, —O—PO(ORs) 2 , —O—PO(ORs)(N(Rs) 2 ), —O—PO(N(Rs) 2 ) 2 , —N—PO(ORs) 2 , —N—PO(ORs)(N(Rs) 2 ), —P(Rs) 2 , —B(OH) 2 , —B(OH)(ORs), and —B(ORs) 2 , where each Rs independently is an alkyl, alkenyl, alkynyl, aryl, heteroaryl, carbocyclyl, or heterocyclyl group or two Rs within the same substituent can together form a carbocyclic or heterocyclic ring having 3 to 10 ring atoms.

8. The compound of claim 7 , wherein L is a divalent linker having 1-6 carbon atoms and an —S—S— moiety.

9. The compound of claim 7 , wherein M is an optionally substituted furyl, pyridinyl, pyrazinyl, pyrimidinyl, quinolinyl, or purinyl group.

10. The compound of claim 7 , wherein M is a heteroarylthio group.

11. The compound of claim 7 , wherein M is a 2-pyridyldithio group.

12. The compound of claim 11 , wherein L is an alkenylene linker having 1-6 carbon atoms.

13. The compound of claim 1 , wherein R is hydrogen or a methyl group.

14. The compound of claim 13 , wherein R A is substituted with a —CO—O—CO-Rs group, and wherein Rs is an alkyl group.

15. The compound of claim 14 , wherein M is a 2-pyridyldithio group.

16. The compound of claim 15 , wherein L is an alkylene linker having 1-6 carbon atoms.

17. A method for esterifying one or more carboxylic acid groups in an organic or biological molecule which comprises contacting the organic or biological molecule with a compound of claim 1 .

18. A method for enhancing cellular uptake of an organic or biological molecule having one or more carboxylate groups which comprises esterifying the organic or biological molecule employing the method of claim 17 .

19. A compound of formula I:

or salts thereof,
where:
R is hydrogen, or an alkyl group;
R M is -L-M, where -L- is an alkylene linker having 1-6 carbon atoms and M is a 2-pyridyldithio group;
wherein R A is an alkyl or alkoxy group substituted with a —CO—O—CO-Rs group, wherein Rs is an alkyl group.

20. The compound or salt of claim 19 , wherein R is hydrogen, -L- is an alkylene linker having 2 carbon atoms, and R A is an alkoxy group substituted with a —CO—O—CO-Rs group, wherein Rs is an alkyl group.

21. A method for esterifying one or more carboxylic acid groups in an organic or biological molecule which comprises contacting the organic or biological molecule with a compound of claim 7 .

22. A method for enhancing cellular uptake of an organic or biological molecule having one or more carboxylate groups which comprises esterifying the organic or biological molecule employing the method of claim 21 .

23. A method for esterifying one or more carboxylic acid groups in an organic or biological molecule which comprises contacting the organic or biological molecule with a compound of claim 19 .

24. A method for enhancing cellular uptake of an organic or biological molecule having one or more carboxylate groups which comprises esterifying the organic or biological molecule employing the method of claim 23 .

Assignees

• Wisconsin Alumni Res Found

Inventors

• Raines Ronald T
• Mix Kalie

Classifications

• C07C245/18

  the carbon skeleton being further substituted by carboxyl groups · CPC title

• C07C269/06

  by reactions not involving the formation of carbamate groups · CPC title

• C07K1/13

  Labelling of peptides · CPC title

• C07D207/46

  with hetero atoms directly attached to the ring nitrogen atom · CPC title

• C07C319/12

  by reactions not involving the formation of mercapto groups · CPC title