FLUOROGENIC HYDRAZINE-SUBSTITUTED COMPOUNDS

• US 20100291547A1
• Filed: 01/23/2009
• Published: 11/18/2010
• Est. Priority Date: 01/24/2008
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Chen, Aimei, Kang, Hee Chol, Gee, Kyle
• Granted Patent: US 8,772,487 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: C07D 311/82 Xanthenes
• CPC Class Codes: C07D 471/06 Peri-condensed systems
• CPC Class Codes: C07F 5/022 without C-boron linkages
• CPC Class Codes: C07F 9/5442 Aromatic phosphonium compounds (P-C aromatic linkage)
• CPC Class Codes: C07F 9/65522 condensed with carbocyclic rings or carbocyclic ring systems
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C09K 11/06 containing organic luminescent materials
• CPC Class Codes: C09K 2211/10 Non-macromolecular compounds
• CPC Class Codes: C09K 2211/1007 Non-condensed systems
• CPC Class Codes: C09K 2211/1014 bridged by heteroatoms, e.g. N, P, Si or B
• CPC Class Codes: C09K 2211/1022 bridged by heteroatoms, e.g. N, P, Si or B
• CPC Class Codes: C09K 2211/1033 with oxygen
• CPC Class Codes: C09K 2211/1055 with other heteroatoms
• CPC Class Codes: C09K 2211/1088 containing oxygen as the only heteroatom
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/64 involving ketones
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]

Abstract

The present disclosure is directed to fluorogenic schiff base-forming dyes capable of detecting analytes containing aldehyde and ketone groups. The dyes contain nucleophilic hydrazinyl appendages and are capable of binding and detecting analytes in situ.

Claims

1. A compound of Formula I:

2. The compound of claim 1, wherein R¹ and R^1a are both H.

3. The compound of claim 1, wherein R¹ is alkyl or substituted alkyl;

and R^1a is H.

4. The compound of claim 1, wherein R³ hydrazinyl or substituted hydrazinyl.

5. The compound of claim 1, wherein R³ is methoxy or hydroxy.

6. The compound of claim 1, wherein R³ is amino.

7. The compound of claim 1, wherein R², R³ and R⁴ are taken together to form a fused heterocyclyl group or substituted fused heterocyclyl group of the formula:

8. The compound of claim 1, wherein R⁴, R⁵, R⁶, R⁷, R⁸ and R¹⁰ are all H.

9. The compound of claim 1, wherein R², R⁴, R⁵, R⁶, R⁷, R⁸ and R¹⁰ are all H.

10. The compound of claim 1, wherein R⁹ is —

SO₃⁻

.

11. The compound of claim 1, wherein R⁹ is —

CO₂⁻

.

12. The compound of claim 1, wherein X is H.

13. The compound of claim 1, wherein X is an analyte.

14. The compound of claim 13, wherein the analyte is bound through an imine.

15. The compound of claim 13, wherein the analyte is selected from the group consisting of an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a drug, a lipid, and a synthetic polymer.

16. The compound of claim 13, wherein the analyte is malondialdehyde or 4-hydroxynonenal.

17. The compound of claim 13, wherein the analyte comprises an aldehyde group.

18. The compound of claim 1, wherein R³ is analyte substituted hydrazinyl.

19. The compound of claim 18, wherein the analyte is bound through an imine.

20. The compound of claim 18, wherein the analyte is selected from the group consisting of an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a drug, a lipid, and a synthetic polymer.

21. The compound of claim 18, wherein the analyte comprises a carbonyl group.

22. The compound of claim 18, wherein the analyte comprises an aldehyde group.

23. The compound of claim 1, wherein R³ is methoxy and R⁹ is —

CO₂⁻

.

24. A salt of the compound of claim 1.

43. The compound of claim 1, wherein one of Z¹ or Z² is selected from the group consisting of a xanthene, an indole and a borapolyazaindacine.

44. The compound of claim 1, wherein Z¹ or Z² is:

45. The compound of claim 44, wherein R¹⁵ is ═

O.

46. The compound of claim 44, wherein R¹⁵ is ═

⁽⁺⁾NR¹⁶R¹⁷.

47. The compound of claim 46, wherein R¹⁶ and R¹⁷ are methyl.

48. The compound of claim 44, wherein R², R⁴ and R¹⁰ are H.

49. The compound of claim 44, wherein R³ is amino or substituted amino.

50. The compound of claim 44, wherein R³ is hydroxy.

51. The compound of claim 44, wherein R³ is —

N(CH₃)₂.

52. The compound of claim 44, wherein R², R³ and R⁴ are taken together to form a fused heterocyclyl group or substituted fused heterocyclyl group of the following structure:

25-42. -42. (canceled)

53-71. -71. (canceled)

72. A composition comprising:

(a) an analyte; and

(b) a compound of Formula I;

73. The composition of claim 72, further comprising a buffer solution.

74. The composition of claim 72, wherein the analyte is selected from the group consisting of:

an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a hapten, a psoralen, a drug, a hormone, a lipid, a lipid assembly, a synthetic polymer, a biotin-binding protein, a polymeric microparticle, a biological cell and a virus.

75. A method for determining the presence of an analyte of interest in a sample, wherein the method comprises:

contacting the sample with a compound having Formula I;

76. The method of claim 75, wherein the analyte and the compound are connected by a covalent bond.

77. The method of claim 75, wherein the compound becomes fluorescent after formation of the complex.

78. The method of claim 76, wherein the analyte is bound to the compound through an imine.

79. The method of claim 75, wherein the analyte is selected from the group consisting of:

an amino acid, a peptide, a protein, a carbohydrate, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a hapten, a psoralen, a drug, a hormone, a lipid, a lipid assembly, a synthetic polymer, a biotin-binding protein, a polymeric microparticle, a biological cell and a virus.

80. The method of claim 75, wherein the analyte is malondialdehyde or 4-hydroxynonenal.

81. The method of claim 75, wherein the analyte comprises an aldehyde group.

82. The method of claim 75, wherein the sample comprises live cells, intracellular fluids, extracellular fluids, biological fluids, biological fermentation media, environmental sample, industrial samples, proteins, peptides, buffer solutions biological fluids or chemical reactors.

83. The method of claim 75, wherein the sample comprises blood cells, immune cells, cultured cells, muscle tissue, neurons, extracellular vesicles;

vascular tissue, blood fluids, saliva, urine;

water, soil, waste water, sea water;

pharmaceuticals, foodstuffs or beverages.

84. The method of claim 75, wherein the sample is immobilized on a polymeric membrane, within a polymeric gel, on a microparticle, on a microarray, on a silicon chip, on a glass slide, on a microwell plate, and on a microfluidic chip.

85. A kit for detecting an analyte in a sample, wherein the kit comprises:

a compound of Formula I that binds the analyte;

86. The kit of claim 85, further comprising instructions for covalently bonding the compound to the analyte.

87. The kit of claim 85, further comprising one or more of the following:

a buffering agent, a purification medium, a vial comprising the analyte, or an organic solvent.

88-118. -118. (canceled)