Detection of biological molecules using boronate-based chemical amplification and optical sensors

• US 6,002,954 A
• Filed: 11/21/1996
• Issued: 12/14/1999
• Est. Priority Date: 11/22/1995
• Status: Expired due to Term

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America), Medtronic Minimed Incorporated (Medtronic Plc), Regents of the University of California (University of California)
• Original Assignee: Minimed, Inc. (Medtronic Plc), Regents of the University of California (University of California)
• Inventors: Peyser, Thomas A., Satcher, Joe H. Jr., Lane, Stephen M., Mastrototaro, John Joseph, Van Antwerp, William Peter, Darrow, Christopher B., Harder, Jennifer
• Primary Examiner(s): WINAKUR, ERIC FRANK
• Application Number:
  US08/749,366
• Time in Patent Office: 1,118 Days
• Field of Search: 600/310, 600/317, 600/342, 600/322
• US Class Current: 600/317
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/1455 using optical sensors, e.g. spectral photometrical oximeters
• CPC Class Codes: A61B 5/1459 invasive, e.g. introduced into the body by a catheter A61B5/1464 takes precedence
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose

Abstract

Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal.

Claims

1. An implantable amplification system comprising a polymer matrix and an amplification component contained in said matrix, said amplication component producing a polyhydroxylated analyte signal upon interrogation by an optical system, wherein said amplification component requires intramolecular electron transfer for production of said signal and comprise a compound of the formula ##STR2## wherein:

D¹ is a dye selected from the group consisting of fluorescent dyes, luminescent dyes and colorimetric dyes;

R¹, R³ and R⁴ are each independently substituents which alter the electronic properties of the groups to which they are attached or are functional groups which can form covalent linkages to the surrounding polymer matrix;

each R² is a member independently selected from the group consisting of hydrogen and C₁ -C₄ alkyl;

each L¹ and L² is a linking group having from zero to four contiguous atoms selected from the group consisting of carbon, oxygen, nitrogen, sulfur and phosphorus;

Z is a heteroatom selected from the group consisting of nitrogen, sulfur, oxygen and phosphorus; and

x is an integer from zero to four.

2. An implantable amplification system in accordance with claim 1, wherein said polymer matrix is a biocompatible polymer matrix.

3. An implantable amplification system in accordance with claim 1, wherein R¹, R³ and R⁴ are each members independently selected from the group consisting of hydrogen, hydroxy, acyl, C₁ -C₄ alkoxy, halogen, thiol, sulfonic acid, sulfonamide, sulfinic acid, nitro, cyano, carboxylic acid, a C₁ -C₁₂ alkyl group, a substituted C₁ -C₁₂ alkyl group, a C₁ -C₁₂ alkenyl group, a substituted C₁ -C₁₂ alkenyl group, a C₁ -C₁₂ alkynyl group, a substituted C₁ -C₁₂ alkynyl group, aryl, substituted aryl, arylalkyl, substituted arylalkyl, amine, and substituted amine, wherein said substituents are selected from the group consisting of hydroxy, acyl, aryl, C₁ -C₄ alkoxy, halogen, thiol, sulfonic acid, amines, sulfonamide, sulfinic acid, nitro, cyano, carboxamide and carboxylic acid.

4. An implantable amplification system in accordance with claim 1, wherein D¹ is a long wavelength fluorescent dye having an emission wavelength of at least about 450 nm.

5. An implantable amplification system in accordance with claim 1, wherein D¹ is a short wavelength fluorescent dye having an emission wavelength of from about 320 to about 450 nm.

6. An implantable amplification system in accordance with claim 4, further comprising a detectable calibration fluorophore contained in said matrix, said calibration fluorophore providing a second signal which does not interfere with the signal produced by said compound.

7. An implantable amplification system in accordance with claim 1, wherein D¹ is selected from the group consisting of fluoresceins, coumarins, oxazines, xanthenes, cyanines, metal complexes and polyaromatic hydrocarbons which produce a fluorescent signal.

8. An implantable amplification system in accordance with claim 1, wherein said compound is covalently attached to said polymer matrix.

9. An implantable amplification system in accordance with claim 8, wherein said covalent attachment is made through a functional group present in R¹.

10. An implantable amplification system in accordance with claim 8, wherein said covalent attachment is made through a functional group present in R³.

11. An implantable amplification system in accordance with claim 8, wherein said covalent attachment is made through a functional group present in R⁴.

12. An implantable amplification system in accordance with claim 1, wherein said polymer matrix comprises a polymer prepared from a reaction mixture of:

(a) a diisocyanate, said diisocyanate comprising about 50 mol % of the reactants in said mixture; and

(b) a hydrophilic polymer which is a member selected from the group consisting of a hydrophilic polymer diol, a hydrophilic polymer diamine and combinations thereof.

13. An implantable amplification system in accordance with claim 12, wherein said polymer matrix further comprises an outer hydrogel coating, wherein said hydrogel is formed from a reaction mixture of:

(a) a diisocyanate, said diisocyanate comprising about 50 mol % of the reactants in said mixture;

(b) a hydrophilic polymer which is a member selected from the group consisting of a hydrophilic polymer diol, a hydrophilic polymer diamine and combinations thereof; and

optionally;

(c) a chain extender,

said hydrogel having a water pickup of from about 120% to about 400% by weight.

14. An implantable amplification system in accordance with claim 12, wherein said reaction mixture further comprises:

(c) a siloxane polymer having amino, hydroxyl or carboxylic acid functional groups at the chain termini.

15. An implantable amplification system in accordance with claim 12, wherein said reaction mixture further comprises:

(d) a chain extender.

16. A method for quantifying the amount of a polyhydroxylated analyte in an individual, said method comprising:

(a) interrogating a subcutaneously implanted amplification system of claim 1 with an energy source to provide an excited amplification system which produces an energy emission corresponding to said amount of said polyhydroxylated analyte; and

(b) detecting said emission to thereby quantify the amount of said polyhydroxylated analyte in said individual.

17. A method in accordance with claim 16, wherein said energy source is a laser diode, LED or other optical source.

18. A method in accordance with claim 16, wherein said polyhydroxylated analyte is glucose.

19. A method in accordance with claim 16, wherein D¹ is a long wavelength fluorescent dye having an emission wavelength of at least about 450 nm.

20. A method in accordance with claim 19 wherein said amplification system further comprises a detectable calibration fluorophore contained in said matrix, said calibration fluorophore providing a second signal which does not interfere with the signal produced by said compound.

21. A method in accordance with claim 16, wherein D¹ is selected from the group consisting of fluorescein, coumarins, oxazines, xanthenes, cyanines, metal complexes and polyaromatic hydrocarbons which produce a fluorescent signal.

22. A method in accordance with claim 16, further comprising:

providing a polymeric matrix wherein said compound of Formula I is covalently attached thereto.

23. A biosensor for measuring the amount of a polyhydroxylated analyte in vivo, said sensor comprising:

(a) an implantable amplification system of claim 1 comprising a biocompatible polymer matrix and an amplification component contained therein which produce a polyhydroxylated analyte signal upon interrogation by an optical source, wherein said amplification component does not require resonance energy transfer for production of said signal and wherein said signal corresponds to said amount of said polyhydroxylated analyte; and

(b) an optical system comprising said optical source and a detector which detects said signal thereby measuring the in vivo amounts of said analyte.

24. A biosensor in accordance with claim 23, wherein said optical source is a member selected from the group consisting of a LED, an incandescent source, a fluorescent source, an electroluminescent source and a laser diode.

25. A biosensor in accordance with claim 23, wherein said optical system further comprises at least one filter and wherein said optical source is a LED.

26. A biosensor in accordance with claim 23, wherein said detector comprises a diode array spectrometer.

27. A biosensor in accordance with claim 23, wherein said optical system comprises a fiber optic.

28. A biosensor in accordance with claim 27, wherein said amplification system is at the terminus of said fiber optic.

29. A biosensor in accordance with claim 23, wherein said optical system is coated with a biocompatible polymer.

30. A biosensor in accordance with claim 23, further comprising an insulin pump which is activated by a signal from said detector, wherein said optical system and said insulin pump are coated with a biocompatible polymer.

31. An implantable amplification system comprising a biocompatible polymer matrix and an amplification component which produces a polyhydroxylated analyte signal upon interrogation by an optical system, wherein said amplification component requires photoelectron transfer for production of said signal and comprise an arylboronic acid moiety.

32. An implantable amplification system in accordance with claim 31 wherein said arylboronic acid moiety is attached to an amine-functionalized dye molecule.

33. An implantable amplification system in accordance with claim 32, wherein said dye molecule is selected from the group consisting of anthracene, rhodamine and fluorescein.

34. An implantable amplification system in accordance with claim 32, wherein said arylboronic acid moiety attached to an amine-functionalized dye molecule is entrapped within said biocompatible matrix.

35. An implantable amplification system in accordance with claim 31, wherein said arylboronic acid moiety has the following general formula:

##STR3## wherein;

D¹ is a dye selected from the group consisting of fluorescent dyes, luminescent dyes and calorimetric dyes;

R¹, R³ and R⁴ are each independently substituents which alter the electronic properties of the groups to which they are attached or are functional groups which can form covalent linkages to the surrounding polymer matrix;

each R² is a member independently selected from the group consisting of hydrogen and C₁ -C₄ alkyl;

each L¹ and L² is a linking group having from zero to four contiguous atoms selected from the group consisting of carbon, oxygen, nitrogen, sulfur and phosphorus;

Z is a heteroatom selected from the group consisting of nitrogen, sulfur, oxygen and phosphorus; and

x is an integer from zero to four.