Biocompatible nanoparticles with aggregation induced emission characteristics as fluorescent bioprobes and methods of using the same for in vitro and in vivo imaging

Patent metadata

Field	Value
Publication number	US-10753941-B2
Application number	US-201214342074-A
Country	US
Kind code	B2
Filing date	Sep 3, 2012
Priority date	Sep 1, 2011
Publication date	Aug 25, 2020
Grant date	Aug 25, 2020

Abstract

The development of fluorescent bioprobes comprising organic fluorescent compounds that exhibit aggregation induced emission (AIE) properties, methods of producing the same, and their practical applications for in vitro and in vivo bioimaging.

First claim

We claim:

1. A fluorescent bioprobe comprising fluorogen-loaded nanoparticles comprising a fluorogen that exhibits aggregation induced emission properties, wherein the fluorogen comprises one or more chromophores conjugated with one or more aggregation induced emission fluorophores; wherein the fluorogen-loaded nanoparticles have a fluorescence emission; and wherein the fluorogen comprises a backbone structure and the backbone structure is:

wherein each R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 and R 21 is independently H, alkyl, unsaturated alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or alkoxy.

2. The fluorescent bioprobe of claim 1 , wherein any one of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , and R 21 further comprises a terminal functional group independently selected from N 3 , NH 2 , COOH, NCS, SH, alkyne, N-Hydroxysuccinimide ester, a maleimide, a hydrazide, a nitrone group, —CHO, —OH, a halide, or a charged ionic group; wherein a peptide independently selected from a biorecognition peptide or a cell penetrating peptide is conjugated to the terminal functional group.

3. The fluorescent bioprobe of claim 1 , wherein the fluorogen-loaded nanoparticles further comprise a biocompatible polymer matrix.

4. The fluorescent bioprobe of claim 3 , wherein the biocompatible polymer matrix is animal serum albumin, 1,2-distearoyle-sn-glycero-3-phosphoethanolamine, polyethylene glycol, or polyfluorene vinylene, or mixtures thereof.

5. The fluorescent bioprobe of any claim 4 , wherein the biocompatible polymer matrix is BSA, DSPE-PEG, or DSPE-PEG-Folate.

6. The fluorescent bioprobe of claim 1 , wherein the fluorescence emission of the fluorogen loaded nanoparticles is further amplified by applying one or more of: (a) a conjugated polymer as a fluorescence resonance energy transfer donor or (b) an arginine-glycine-aspartic acid peptide as a bio-recognition reagent functionalized on a surface of the nanoparticle.

7. A fluorescent bioprobe of claim 1 , wherein the fluorogen-loaded nanoparticles are 1 nm to 100,000 nm in size.

8. The fluorescent bioprobe of claim 3 , wherein the fluorogen-loaded nanoparticles are BSA, DSPE-PEG, DSPE-PEG-Folate or DSPE-PEG-NH 2 encapsulated nanoparticles.

9. A method for preparing the fluorescent bioprobe of claim 3 comprising loading the fluorogen with the biocompatible polymer matrix by:
(a) preparing a solution comprising an organic solvent and the fluorogen;
(b) preparing an aqueous solution of a biocompatible polymer;
(c) mixing the solution comprising the organic solvent and the fluorogen with the aqueous solution together and sonicating; and
(d) removing the organic solvent to form the fluorogen-loaded nanoparticles.

10. The method of claim 9 , wherein the fluorogen-loaded nanoparticles are fabricated with any molecule that can specifically target cancer cells or can amplify the fluorescence imaging.

11. A method of cellular imaging comprising contacting target cells with the fluorescent bioprobe of claim 1 and detecting the fluorescent bioprobe by cellular imaging.

12. The method of claim 11 , further comprising determining whether a tumor or cancer cells are present.

13. The method of claim 12 , wherein in vitro cellular imaging is conducted using biological imaging samples selected from MCF-7 cells, HT-29 cancer cells, or HeLa cancer cells; or wherein in vivo cellular imaging is conducted using ICR mice bearing tumors as the biological imaging sample.

14. The fluorescent bioprobe of claim 1 , wherein the fluorogen comprises the following backbone structure:

15. The fluorescent bioprobe of claim 14 , wherein R 1 , R 2 and R 3 are each hydrogen.

Assignees

• Tang Benzhong
• Qin Wei
• Liu Jianzhao
• Chen Sijie
• Kwok Tsz Kin
• Liu Bin
• Li Kai
• Ding Dan
• Shi Haibin
• Geng jun long
• Sun Jingzhi
• Qin Anjun
• Zhao Qiuli
• Univ Hong Kong Sci & Tech
• Nat Univ Singapore

Inventors

• Tang Benzhong
• Qin Wei
• Liu Jianzhao
• Chen Sijie
• Kwok Tsz Kin
• Liu Bin
• Li Kai
• Ding Dan
• Shi Haibin
• Geng jun long
• Sun Jingzhi
• Qin Anjun
• Zhao Qiuli

Classifications

• G01N33/575

  for cancer · CPC title

• A61K49/0054

  Macromolecular compounds, i.e. oligomers, polymers, dendrimers · CPC title

• C09K11/06

  containing organic luminescent materials · CPC title

• A61K49/0021 Primary

  the fluorescent group being a small organic molecule · CPC title

• A61K49/0093

  Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle · CPC title