Polymers functionalized with fluorescent boronate motifs and methods for making them

• US 20020197724A1
• Filed: 02/14/2002
• Published: 12/26/2002
• Est. Priority Date: 02/15/2001
• Status: Active Grant

• Current Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Minimed Incorporated (Medtronic Plc)
• Inventors: Walsh, Joseph C., Noronha, Glenn, Ponder, Bill C., Heiss, Aaron M., Vachon, David J., Reilly, Jonathan, Cochran, Brooks
• Granted Patent: US 6,927,246 B2
• Time in Patent Office: Days
• US Class Current: 436/95
• CPC Class Codes: C07F 5/025 Boronic and borinic acid compounds
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

Improved polymer matrices which incorporate fluorescent biosensor molecules as well as methods of making and using these polymer matrices are described. Such matrices can be used in fluorescent biosensors and biosensor systems, including those which are used in the detection of polyhydroxylated analytes such as glucose. The properties of the polymer matrices of the invention renders biosensors utilizing such matrices particularly well-suited for detecting and measuring in-vivo glucose concentrations.

Claims

1. A polymer composition comprising a fluorescent boronic acid of the general formula:

2. The polymer composition of claim 1, wherein the polymer composition further includes an additional polymer that is coupled to the polymer matrix after polymerization;

and wherein the additional polymer enhances the biocompatibility, swellability or hydrophllicity of the polymer composition.

3. The polymer composition of claim 2, wherein the polymer matrix is a block copolymer.

4. The polymer composition of claim 2, wherein the additional polymer is grafted on to the polymer matrix.

5. The polymer composition of claim 2, wherein the additional polymer is a polyethyleneoxide or polyethyleneoxide-polypropyleneoxide compound.

6. The polymer composition of claim 2, wherein the polymer matrix is crosslinked.

7. The polymer composition of claim 6, wherein the polymer matrix is crosslinked with polyethyleneoxide or polyethyleneoxide-polypropyleneoxide compounds.

8. The polymer composition of claim 1, wherein the nitrogen atom in the fluorescent boronic acid is covalently coupled to the polymer matrix after polymerization via the group designated R³.

9. The polymer composition of claim 8, wherein the atoms that link the fluorescent boronic acid to the polymer matrix of the polymer composition enhance the solubility of the polymer composition.

10. The polymer composition of claim 1, wherein the polymerized matrix is a polyvinylalcohol.

11. The polymer composition of claim 1, wherein the polymerized matrix is a polystyrene.

12. The polymer composition of claim 11, wherein the tether that links the polymer matrix (PM) to the fluorescent boronic acid (FBA) includes the following atoms:

PM-CO-NH-FBA;

PM-SO₂—

NH-FBA;

PM-CO—

NH-FBA;

PM-COO-FBA;

PM-NH—

COO-FBA;

PM-NH—

CO—

N-FBA or PM-NH—

SO₂-FBA, wherein C denotes carbon, N denotes nitrogen, 0 denotes oxygen, S denotes sulfur and H denotes hydrogen.

13. The polymer composition of claim 1, wherein the fluorophore is highly soluble in water.

14. The polymer composition of claim 13, wherein the fluorophore is Nile Blue.

16. The polymer composition of claim 15, wherein the polymer composition further includes an additional polymer that is covalently coupled to the polymerized matrix;

and wherein the additional polymer enhances the solubility of the polymer composition.

17. The polymer composition of claim 16, wherein the polymerized matrix is a block copolymer.

18. The polymer composition of claim 16, wherein the additional polymer is grafted on to the polymerized matrix.

19. The polymer composition of claim 16, wherein the additional polymer is a polyethyleneoxide or polyethyleneoxide-polypropyleneoxide compound.

20. The polymer composition of claim 16, wherein the polymerized matrix is crosslinked.

21. The polymer composition of claim 20, wherein the polymerized matrix is crosslinked with polyethyleneoxide or polyethyleneoxide-polypropyleneoxide compounds.

22. The polymer composition of claim 15, wherein the nitrogen atom in the fluorescent boronic acid is covalently coupled to the polymerized matrix via the group designated R³.

23. The polymer composition of claim 22, wherein the atoms that link the fluorescent boronic acid to the polymerized matrix of polymer composition enhance the solubility of the polymer composition.

24. The polymer composition of claim 15, wherein the polymerized matrix is a polyvinylalcohol.

25. The polymer composition of claim 15, wherein the polymerized matrix is a polystyrene.

26. The polymer composition of claim 25, wherein the tether that links the polymer matrix (PM) to the fluorescent boronic acid (FBA) includes the following atoms:

PM-CO—

NH-FBA;

PM—

SO₂-NH-FBA;

PM-CO—

NH-FBA;

PM-COO-FBA;

PM-NH—

COO-FBA;

PM-NH—

CO—

N-FBA or PM-NH—

SO₂-FBA, wherein C denotes carbon, N denotes nitrogen, O denotes oxygen, S denotes sulfur and H denotes hydrogen.

27. The polymer composition of claim 15, wherein the fluorophore is highly soluble in water.

28. The polymer composition of claim 27, wherein the fluorophore is Nile Blue.

30. The method of claim 29, further including the step of covalently coupling an additional polymer to the polymerized matrix;

and wherein the additional polymer enhances the solubility of the polymer composition.

31. The method of claim 30, wherein the additional polymer is grafted on to the polymerized matrix.

32. The method of claim 31, wherein the additional polymer is a polyethyleneoxide or polyethyleneoxide-polypropyleneoxide compound.

33. The method of claim 29, further including the step of covalently crosslinking the polymer composition.

34. The method of claim 33, wherein the polymer matrix is crosslinked with polyethyleneoxide or polyethyleneoxide-polypropyleneoxide compounds.

35. The method of claim 29, wherein the nitrogen atom in the fluorescent boronic acid is covalently coupled to the polymerized matrix via the group designated R³.

36. The method of claim 35, wherein the atoms that link the fluorescent boronic acid to the polymerized matrix of polymer composition enhance the solubility of the polymer composition.

37. The method of claim 29, wherein the polymerized matrix is a polyvinylalcohol.

38. The method of claim 29, wherein the polymerized matrix is a polystyrene.

39. The method of claim 38, wherein the atomic group that is used to covalently couple the fluorescent boronic acid (FBA) to the polymer matrix (PM) is:

HO—

(CH₂)-FBA;

NH_2—

(CH₂)-FBA or NH₂—

(CH₂)—

(O—

CH₂)-FBA wherein H is hydrogen, C is carbon, N is nitrogen, O is oxygen and n is an integer greater than or equal to 1.

40. The method of claim 38, wherein the tether that links the polymer matrix (PM) to the fluorescent boronic acid (FBA) includes the following atoms:

PM-CO—

NH-FBA;

PM-SO₂—

NH-FBA;

PM-CO—

NH-FBA;

PM-COO-FBA;

PM-NH—

COO-FBA;

PM-NH—

CO—

N-FBA or PM-NH—

SO₂-FBA, wherein C denotes carbon, N denotes nitrogen, O denotes oxygen, S denotes sulfut and H denotes hydrogen.

41. The method of claim 29, wherein the fluorophore is highly soluble in water.

15. A polymer composition comprising a fluorescent boronic acid and a reference fluorophore;

wherein the composition is produced by a process of covalently coupling the fluorescent boronic acid and the reference fluorophore to a polymerized matrix, wherein the fluorescent boronic acid has the general formula;

29. A method of making a polymer composition comprising a fluorescent boronic acid and a reference fluorophore, the method comprising the steps of:

covalently coupling the fluorescent boronic acid and the reference fluorophore to a polymerized matrix, wherein the fluorescent boronic acid has the general formula;

wherein;

F is a fluorophore;

N is a nitrogen atom;

B is a boron atom;

R¹ is selected ftom the functional group consisting of hydrogen, aliphatic and aromatic groups, wherein the functional group (R¹O)₂B is capable of binding glucose;

R², R³ and R⁴ are optional and independent hydrogen, aliphatic or aromatic groups, further functionalized aliphatic or aromatic groups or groups that are capable of forming a covalent linkage to the polymer matrix;

L¹ and L² are optional linking groups having from zero to four atoms selected from the group consisting of nitrogen, carbon, oxygen, sulfur and phosphorous; and

wherein the polymer composition including the covalently coupled reference fluorophore and the covalently coupled fluorescent boronic acid is soluble in an aqueous environment; and

the fluorescence of the polymer composition including the covalently coupled reference fluorophore and the covalendy coupled fluorescent boronic acid increases in the presence of bound glucose.

42. The method of claim 42, wherein the fluorophore is Nile Blue.

43. A method of coupling a fluorescent boronic acid (FBA) compound to a polymerized matrix (PM) to generate a polymerized matrix composition (PMC), wherein the fluorescent boronic acid compound has the general formula: