Ultra bright polymeric dyes with peptide backbones

Patent metadata

Field	Value
Publication number	US-11370922-B2
Application number	US-201716099643-A
Country	US
Kind code	B2
Filing date	May 10, 2017
Priority date	May 10, 2016
Publication date	Jun 28, 2022
Grant date	Jun 28, 2022

Abstract

Compounds useful as fluorescent or colored dyes are disclosed. The compounds have the following structure (I), including stereoisomers, salts and tautomers thereof, wherein R1, R2, R3, L1, L2, L3, L4, L5, M, m and n are as defined herein. Methods associated with preparation and use of such compounds are also provided.

First claim

What is claimed is:

1. A compound having the following structure (I):

or a stereoisomer, salt or tautomer thereof, wherein:
M is, at each occurrence, independently a fluorescent or colored moiety comprising two or more carbon-carbon double bonds and at least one degree of conjugation;
L 1 is at each occurrence, independently either: i) an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker; or ii) a linker comprising a functional group capable of formation by reaction of two complementary reactive groups;
L 2 , L 3 , L 4 and L 5 are, at each occurrence, independently optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linkers;
R 1 is, at each occurrence, independently a natural or unnatural amino acid side chain;
R 2 and R 3 are each independently H, —OH, —SH, —NH 2 , —CO 2 H, alkyl, alkylether, alkoxy, heteroalkyl, alkylaminyl, alkylcarbonyl, alkyloxycarbonyl, Q, a linker comprising a covalent bond to Q, a linker comprising a covalent bond to a targeting moiety, a linker comprising a covalent bond to an analyte molecule, a linker comprising a covalent bond to a solid support, a linker comprising a covalent bond to a solid support residue or a linker comprising a covalent bond to a further compound of structure (I), wherein the alkyl, alkylether, alkylaminyl, alkylcarbonyl and alkyloxycarbonyl are optionally substituted with hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether or thiophosphoalkylether, or combinations thereof; Q is, at each occurrence, independently a moiety comprising a reactive group capable of forming a covalent bond with an analyte molecule, a solid support or a complementary reactive group Q′;
m is, at each occurrence, independently an integer of zero or greater; and
n is an integer of one or greater.

2. The compound of claim 1 , wherein R 1 is, at each occurrence, independently H, alkyl, —CH 2 CO 2 − , —CH 2 CH 2 CO 2 − , —CH 2 CH 2 CH 2 CH 2 NH 3 + , —CH 2 CH 2 CH 2 NHC(═NH 2 + )NH 2 or imidazolyl.

3. The compound of claim 1 , wherein R 1 , L 4 and m are selected such that

has an amino acid sequence of (G) 10 , (GDGDGDGDGD) or (GKGKGKGKGK).

4. The compound of claim 1 , wherein R 1 , L 4 and m are selected such that

has an amino acid sequence capable of forming an α-helix or β-sheet secondary structure.

5. The compound of claim 4 , wherein the amino acid sequence is (GGEEFMLVYKFARKHGG) or (GGMSMVVSGG).

6. The compound of claim 1 , wherein L 4 and L 5 are absent at each occurrence.

7. The compound of claim 1 , wherein at least one occurrence of L 4 or L 5 , or both, has the following structure:

8. The compound of claim 1 , wherein at least one occurrence of L 4 or L 5 , or both, has the following structure:

9. The compound of claim 1 , wherein L′ is at each occurrence an alkylene or heteroalkylene linker.

10. The compound of claim 1 , wherein L 1 is at each occurrence a linker comprising a functional group capable of formation by reaction of two complementary reactive groups and for at least one occurrence of L 1 , the functional group can be formed by reaction of an aldehyde, oxime, hydrazone, alkyne, amine, azide, acylazide, acylhalide, nitrile, nitrone, sulfhydryl, disulfide, sulfonyl halide, isothiocyanate, imidoester, activated ester, ketone, α,β-unsaturated carbonyl, alkene, maleimide, α-haloimide, epoxide, aziridine, tetrazine, tetrazole, phosphine, biotin or thiirane functional group with a complementary reactive group.

11. The compound of claim 10 , wherein for at least one occurrence of L 1 , L 1 -M has the following structure:

wherein L 1a and L 1b are each independently optional linkers.

12. The compound of claim 10 , wherein for at least one occurrence of L 1 , L 1 -M has the following structure:

wherein L 1a and L 1b are each independently optional linkers.

13. The compound of claim 1 , wherein R 2 is —NH 2 .

14. The compound of claim 1 , wherein Q has one of the following structures:

wherein:
X is halo; and
EWG is an electron withdrawing group.

15. The compound of claim 1 , wherein one of R 2 or R 3 is a linker comprising a covalent bond to an analyte molecule, wherein the analyte molecule is a nucleic acid or a polymer thereof, amino acid or a polymer thereof, an enzyme, receptor, receptor ligand, antibody, glycoprotein, aptamer, prion or a linker comprising a covalent bond to a polymeric bead or non-polymeric bead.

16. The compound of claim 1 , wherein M is, at each occurrence, independently a dimethylaminostilbene, quinacridone, fluorophenyl-dimethyl-BODIPY, his-fluorophenyl-BODIPY, acridine, terrylene, sexiphenyl, porphyrin, benzopyrene, (fluorophenyl-dimethyl-difluorobora-diaza-indacene)phenyl, (bis-fluorophenyl-difluorobora-diaza-indacene)phenyl, quaterphenyl, bi-benzothiazole, ter-benzothiazole, bi-naphthyl, bi-anthracyl, squaraine, squarylium, 9, 10-ethynylanthracene or ter-naphthyl moiety.

17. The compound of claim 1 , wherein M, at each occurrence, independently has one of the following structures:

18. The compound of claim 1 , wherein the compound has one of the following structures:

19. A method of staining a sample, comprising adding to said sample the compound of claim 1 in an amount sufficient to produce an optical response when said sample is illuminated at an appropriate wavelength.

20. A compound having the following structure (II):

or a stereoisomer, salt or tautomer thereof, wherein:
G is, at each occurrence, independently a moiety comprising a reactive group capable of forming a covalent bond with a complementary reactive group;
L 1a , L 2 , L 3 , L 4 and L 5 are, at each occurrence, independently an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene or heteroalkynylene linker;
R 1 is, at each occurrence, independently a natural or unnatural amino acid side chain;
R 2 and R 3 are each independently H, —OH, —SH, —NH 2 , —CO 2 H, alkyl, alkylether, alkoxy, heteroalkyl, alkylaminyl, alkylcarbonyl, alkyloxycarbonyl, Q, a linker comprising a covalent bond to Q, a linker comprising a covalent bond to a targeting m

Assignees

• Sony Corp

Inventors

• Matray Tracy
• Singh Sharat
• Vanbrunt Michael

Classifications

• C09B69/109

  containing other specific dyes · CPC title

• C07K17/00

  Carrier-bound or immobilised peptides (carrier-bound or immobilised enzymes C12N11/00); Preparation thereof · CPC title

• C07K7/06

  having 5 to 11 amino acids · CPC title

• G01N33/582

  with fluorescent label · CPC title

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title