CLICK CHEMISTRY ON HETEROGENEOUS CATALYSTS

• US 20120100633A1
• Filed: 04/08/2010
• Published: 04/26/2012
• Est. Priority Date: 04/09/2009
• Status: Active Grant

• Current Assignee: Baseclick GmbH (BASF SE)
• Original Assignee: Baseclick GmbH (BASF SE)
• Inventors: Manetto, Antonio, Gramlich, Philipp Mathias Edwin, Warncke, Simon
• Granted Patent: US 8,883,516 B2
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: B01J 21/18 Carbon
• CPC Class Codes: B01J 23/72 Copper
• CPC Class Codes: C07D 249/04 1,2,3-Triazoles; Hydrogenated 1,2,3-triazoles

Abstract

The present invention relates to new methods and reagents for coupling molecules by a Click reaction using a heterogeneous catalyst system. Further, the present invention refers to novel devices for carrying out Click reactions

Claims

1-16. -16. (canceled)

17. A method of coupling a first molecule to a second molecule by a Click reaction, comprising contacting the first molecule comprising a first Click functional group, which is a Click-reactive unsaturated group, and the second molecule comprising a second complementary Click functional group, which is a Click-reactive 1,3-dipolar group capable of reacting with the first Click-functional group by a Click reaction, in the presence of a heterogeneous catalyst under conditions wherein the Click reaction between the first and second molecule occurs wherein one of the first and second molecules is immobilized on a solid carrier, wherein one of the first and second molecules is immobilized on the heterogeneous catalyst.

18. The method of claim 17, wherein one of the first and second molecules is a biomolecule.

19. The method of claim 18, wherein a dry solid carrier pre-impregnated with one of the first and second molecules is provided, which is contacted with a liquid medium comprising the other molecule.

20. The method of claim 17, wherein the first molecule is a biomolecule selected from the group consisting of nucleosides, nucleotides, nucleic acids, amino acids, peptides, saccharides and lipids.

21. The method of claim 20, wherein the biomolecule is a nucleic acid.

22. The method of claim 17, wherein the second molecule is selected from

(i) a reporter molecule, optionally a dye;

(ii) an affinity molecule;

(iii) a solid phase;

(iv) a biomolecule, optionally a protein or a lipid;

(v) a linker or a spacer, optionally comprising an aliphatic or cycloaliphatic group, an aromatic or heteroaromatic group, an alkene group, an alkyne group, and/or a polymeric group, optionally a polyethylene glycol group; and

(vi) a pharmaceutical compound or group, a photoactive group, a redox active group, and/or a recognition site.

23. The method of claim 17, wherein the Click-reactive unsaturated group is an alkyne group, and wherein the Click-reactive 1,3-dipolar group is an azide group.

24. The method of claim 17, wherein the heterogeneous catalyst is a heterogeneous Cu catalyst.

25. The method of claim 24, wherein the heterogeneous Cu catalyst is a Cu-C catalyst.

26. The method of claim 17, wherein the Click reaction is carried out in the presence of a molecule able to interact with the heterogeneous system increasing the rate of the Click reaction, particularly in the presence of an amine, optionally in amounts of up to 10% (v/v), 1% (v/v) or 0.1% (v/v), wherein the amine is preferably a primary, secondary or tertiary amine optionally having an aromatic, heteroaromatic, aliphatic, cycloaliphatic, unsaturated or fully saturated backbone.

27. The method of claim 26, wherein the amine is triethylamine.

28. The method of claim 17, wherein (i) the reaction temperature is between about 4 and about 80°

C., optionally between about 10 and about 40°

C. and/or (ii) the reaction time is between about 1 min and about 8 h, optionally between about 1 min and about 10 min, optionally between about 2 min and about 5 min, or between about 10 min and 8 h, and/or wherein the reaction volume is from about 0.1 to about 1000 μ

l.

29. The method of claim 17, wherein the reaction is carried out in a pipette tip, a spin column or a syringe.

30. The method of claim 17, further comprising a separation/purification step.

31. A device having at least one reaction chamber, comprising a heterogeneous catalyst for a Click-reaction and optionally a further solid carrier material, wherein a partner of the Click-reaction is immobilized on said heterogeneous catalyst and/or on said further solid carrier material, wherein the device is preferably a pipette tip or a spin column.

33. A reagent kit for a Click reaction, comprising the device of claim 31, a first molecule, and second molecule, wherein the first molecule comprises a first Click-functional group, which is a Click-reactive unsaturated group, and the second molecule comprises a second complementary Click-functional group, which is a Click-reactive 1,3-dipolar group capable of reacting with the first Click-functional group by a Click reaction.

32. A device having at least a reaction chamber, comprising the heterogeneous catalyst for a Click-reaction and one of the partners for the Click-reaction immobilized thereon, preferably a partner comprising a Click-reactive 1,3 dipolar group.

34. A method for detecting an analyte in a sample comprising:

(i) providing a device having at least one reaction chamber, wherein the reaction chamber comprises a heterogeneous catalyst for carrying out a Click-reaction, and optionally a further solid carrier material, wherein a partner of the Click-reaction is immobilized on said heterogeneous catalyst and/or on said further solid carrier material;

(ii) contacting in said reaction chamber a first molecule, which is a first Click partner, with a second molecule, which is a second Click partner, in the presence of the heterogeneous catalyst under conditions wherein a Click-reaction between the first and second molecule occurs to form a coupling product, wherein the second molecule preferably comprises a reporter group or a reporter precursor group,

(iii) optionally converting reporter precursor groups to reporter groups,

(iv) contacting the coupling product with the sample within an integrated apparatus under conditions which allow detection of the analyte, and

(v) detecting the analyte qualitatively and/or quantitatively, optionally by detecting said reporter group.