Polymeric BODIPY dyes and methods for using the same

Patent metadata

Field	Value
Publication number	US-9758625-B2
Application number	US-201615059181-A
Country	US
Kind code	B2
Filing date	Mar 2, 2016
Priority date	Mar 12, 2015
Publication date	Sep 12, 2017
Grant date	Sep 12, 2017

Abstract

Polymeric BODIPY dyes including light harvesting BODIPY unit-comprising multichromophores are provided. In some embodiments, the dyes are polymeric tandem dyes that include a light harvesting BODIPY unit-comprising multichromophore and an acceptor chromophore covalently linked to the multichromophore in energy-receiving proximity therewith. The polymeric tandem dyes may be covalently linked to a specific binding member. Also provided are methods of evaluating a sample for the presence of a target analyte and methods of labelling a target molecule using compositions including the polymeric tandem dyes. Kits and systems for practicing the subject methods are also provided.

First claim

What is claimed is:

1. A water soluble polymeric tandem dye comprising:
a light harvesting multichromophore that is a conjugated polymer comprising π conjugated BODIPY units; and
an acceptor chromophore covalently linked to the multichromophore in energy-receiving proximity therewith.

2. The dye according to claim 1 , wherein the multichromophore comprises a BODIPY-comprising conjugated segment described by formula (I):

wherein:
B is a BODIPY unit;
M is a π conjugated co-monomer;
each L is independently selected from the group consisting of a terminal group, a π conjugated segment, a linker and a linked specific binding member; and
n is an integer of 2 to 100,000.

3. The dye according to claim 2 , wherein the BODIPY unit is described by the structure:

wherein:
R 1 , R 2 , R 3 and R 4 are each independently selected from H, an alkyl and a substituted alkyl;
R 5 is selected from the group consisting of an alkyl, a substituted alkyl, an aryl, a substituted aryl, a heteroaryl, a substituted heteroaryl, wherein R 5 is optionally substituted with a water solubilizing group; and
each R is selected from the group consisting of F, OH, H, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alkoxy, substituted alkoxy, alkynyl and substituted alkynyl.

4. The dye according to claim 1 , wherein the multichromophore is described by the formula (II):

wherein:
B 1 and B 2 are each independently a BODIPY unit;
each M 1 and each M 2 are independently a π conjugated co-monomer;
a, b, c, d, e and f are each independently 0, 1 or 2, wherein b+e≧1;
n and m are independently 0 or an integer from 1 to 100,000, wherein n+m≧1;
p is an integer from 1 to 100,000; and
each L 2 is independently selected from the group consisting of a terminal group, a π conjugated segment, a linker and a linked specific binding member.

5. The dye according to claim 4 , wherein at least one of B 1 , B 2 , M 1 and M 2 comprises -L 1 -C 1 , wherein L 1 is an optional linker and C 1 is the acceptor chromophore.

6. The dye according to claim 4 , wherein the dye is described by the formula (III):

wherein each L 1 is an optional linker and each C 1 is an acceptor chromophore.

7. The dye according to claim 4 , wherein the dye is described by the formula (IV):

wherein each L 1 is an optional linker and each C 1 is an acceptor chromophore.

8. The dye according to claim 4 , wherein the dye is described by the formula (V):

wherein each L 2 is an optional linker and each C 1 is an acceptor chromophore.

9. The dye according to claim 4 , wherein the dye is described by the formula (VI):

wherein each L 1 is an optional linker and each C 1 is an acceptor chromophore.

10. The dye according to claim 4 , wherein B 1 and B 2 are each independently described by the following structure:

wherein R 6 is an aryl or a heteroaryl, optionally substituted with one or more water solubilizing groups.

11. The dye according to claim 4 , wherein at least one L 2 is -L 3 -Z where L 3 is a linker and Z is a specific binding member.

12. The dye according to claim 4 , wherein each M 1 and M 2 are each independently described by the following structure:

wherein:
each R 7 is independently selected from the group consisting of an alkyl, a substituted alkyl, an aralkyl, a substituted aralkyl, a PEG moiety and -L 1 -C 1 .

13. The dye according to claim 4 , wherein:
M 1 is described by the following structure:

wherein each R 8 is a substituted aralkyl that comprises a water solubilizing group; and
M 2 is described by the following structure:

wherein R 9 is a substituted alkyl that comprises a water solubilizing group, and R 19 is -L 1 -C 1 .

14. A labelled specific binding member, comprising:
a polymeric tandem dye comprising:
a water soluble light harvesting BODIPY unit-comprising multichromophore;
an acceptor chromophore covalently linked to the multichromophore in energy-receiving proximity therewith; and

a specific binding member covalently linked to the multichromophore.

15. The labelled specific binding member according to claim 14 , wherein the multichromophore comprises a BODIPY-comprising conjugated segment described by formula (I):

wherein:
B is a BODIPY unit;
M is a π conjugated co-monomer;
each L is independently selected from the group consisting of a terminal group, a π conjugated segment, a linker and a linked specific binding member; and
n is an integer of 1 to 100,000.

16. The labelled specific binding member according to claim 14 , wherein the BODIPY unit is described by the structure:

wherein:
R 1 , R 2 , R 3 and R 4 are each independently selected from H, an alkyl and a substituted alkyl;
R 5 is selected from the group consisting of an alkyl, a substituted alkyl, an aryl, a substituted aryl, a heteroaryl and a substituted heteroaryl, wherein R 5 is optionally substituted with a water solubilizing group; and
each R is selected from the group consisting of F, OH, H, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alkoxy, substituted alkoxy, alkynyl and substituted alkynyl.

17. The labelled specific binding member according to claim 14 , wherein the multichromophore is described by the formula (II):

wherein:
B 1 and B 2 are each independently a BODIPY unit;
each M 1 and each M 2 are independently a π conjugated co-monomer;
a, b, c, d, e and f are each independently 0, 1 or 2, wherein b+e≧1;
n and m are independently 0 to 100,000, wherein n+m≧1; and
one L 2 group is a terminal group (G 1 ) and the other L 2 group is a linked specific binding member.

18. The labelled specific binding member according to claim 17 , wherein at least one of B 1 , B 2 , M 1 and M 2 comprises -L 1 -C 1 , wherein L′ is an

Assignees

• Becton Dickinson Co

Inventors

• Bartholomew Glenn P
• Gaylord Brent S
• Zhao Lucy

Classifications

• C09B69/109 Primary

  containing other specific dyes · CPC title

• C08G79/08 Primary

  a linkage containing boron · CPC title

• G01N33/542

  with steric inhibition or signal modification, e.g. fluorescent quenching · CPC title

• G01N33/582 Primary

  with fluorescent label · CPC title

• C09B69/105

  containing a methine or polymethine dye · CPC title