Bodipy structure fluorescence probes for diverse biological applications

Patent metadata

Field	Value
Publication number	US-9423396-B2
Application number	US-201214367596-A
Country	US
Kind code	B2
Filing date	Dec 20, 2012
Priority date	Dec 22, 2011
Publication date	Aug 23, 2016
Grant date	Aug 23, 2016

Abstract

The present invention is directed to a fluorescence compound represented by structural Formula (I), with specificity to neural stem cells: (I), or a pharmaceutically acceptable salt thereof. The variables for structural Formula (I) are defined herein. Also described are methods for detection of beta cells, pancreatic islets and microglia cells, comprising using a compound of structural Formula (I) or pharmaceutically acceptable salts thereof. Compounds of structural Formula (I) can also differentiate healthy pancreatic islet cells from diabetic pancreatic islet cells.

First claim

What is claimed is:

1. A compound represented by structural Formula (I) or pharmaceutically acceptable salts thereof:

wherein:
R 1 is hydrogen or -COR 3 ;
R 2 is (C 6 -C 16 )aryl, (C 3 -C 10 )heteroaryl, (C 1 -C 6 )alkyl, (C 1 -C 6 )cycloalkyl, (C 2 -C 6 )alkenyl, or C≡CH;
R 2 is optionally substituted with 1-5 substituents independently selected from (C 1 -C 6 )alkyl, halogen, amino, cyano, —COOH, halo(C 1 -C 6 )alkyl, hydroxy(C 0 -C 6 )alkyl, (C 6 -C 10 )aryl, (C 3 -C 10 )heteroaryl, halo(C 6 -C 10 )aryl, hydroxy(C 6 -C 10 )aryl, (C 1 -C 6 )alkoxy, halo(C 1 -C 6 )alkoxy, (C 6 -C 16 )aryloxy, (C 3 -C 8 )cycloalkyl, halo(C 6 -C 10 )aryl(C 1 -C 6 )alkoxy, (C 1 -C 6 )alkoxy(C 6 -C 10 )aryl(C 1 -C 6 )alkoxy, nitro, (C 0 -C 6 )alkyl(C 6 -C 10 )aryl(C 0 -C 6 )alkoxy, (C 5 -C 10 )heterocycle, —OCHF 2 , —OCF 3 , —SCF 3 , —OBn, cyano(C 1 -C 6 )alkylene, (C 1 -C 6 )alkoxyamino, (C 6 -C 10 )aryl(C 2 -C 6 )alkenyl, (C 2 -C 6 )alkenyl(C 1 -C 6 )alkoxy, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkenyl(C 6 -C 10 )aryl, —N((C 0 -C 6 )alkyl)((C 1 -C 6 )alkyl), —N((C 1 -C 6 )alkyleneOH)((C 1 -C 6 )alkyleneOH), —N((C 0 -C 6 )alkyl)((C 1 -C 6 )alkyleneOH), —N((C 1 -C 6 )alkyleneOCO(C 1 -C 6 )alkyl)((C 1 -C 6 )alkyleneOCO(C 1 -C 6 )alkyl), —NCO(C 1 -C 6 )alkyl, —NPh 2 , —OPh(halogen) 0-3 , —OPhO(C 1 -C 6 )alkyl, —OPhO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkoxy, —O(C 1 -C 6 )alkyl(C 6 -C 10 )aryl, —O(C 2 -C 6 )alkenyl, —O(C 2 -C 6 )alkyleneN(CH 3 ) 2 , (C 0 -C 6 )alkylCOO(C 1 -C 6 )alkyl, —B(OH) 2 or —S(C 1 -C 6 )alkyl;
and wherein any of the substituents selected from (C 1 -C 6 )alkyl, (C 1 -C 6 )alkoxy, (C 6 -C 10 )aryl, (C 6 -C 16 )aryloxy or (C 5 -C 10 )heteroaryl is further optionally substituted with 1-4 substituents selected from halogen, (C 1 -C 6 )alkyl, halo(C 1 -C 6 )alkyl, amino, nitro, cyano, hydroxy(C 0 -C 6 )alkyl, (C 1 -C 6 )alkoxy, —COO(C 0 -C 6 )alkyl, or —CHO; and
R 3 is (C 1 -C 15 )alkyl, (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 6 -C 10 )aryl or (C 5 -C 10 )heteroaryl, wherein R 3 is optionally substituted with 1-4 substituents independently selected from halogen, amino, cyano or hydroxyl.

2. The compound of claim 1 , wherein R 2 is (C 6 -C 10 )aryl.

3. The compound of claim 2 , wherein R 2 is phenyl, optionally substituted with 1-4 substituents selected from —C(CH 3 ) 3 and —O—(C 1 -C 3 )alkyl.

4. The compound of claim 3 , wherein the compound is represented by structural Formula (II):

5. The compound of claim 3 , wherein the compound is represented by structural Formula (III):

6. The compound of claim 3 , wherein the compound is represented by structural Formula (IV):

7. The compound of claim 3 , wherein the compound is represented by structural Formula (V):

8. The compound of claim 3 , wherein the compound is represented by structural Formula (VI):

9. A pharmaceutical composition comprising one or more pharmaceutically acceptable carriers and/or one or more pharmaceutically acceptable diluents and the compound of claim 1 .

10. A method for detecting beta-cells using image based screening, comprising:
a) contacting a sample comprising cells with a compound of structural Formula (I) or pharmaceutically acceptable salts thereof:

wherein:
R 1 is hydrogen or —COR 3 ;
R 2 is (C 6 -C 16 )aryl, (C 3 -C 10 )heteroaryl, (C 1 -C 6 )alkyl, (C 1 -C 6 )cycloalkyl, (C 2 -C 6 )alkenyl, or C≡CH;
R 2 is optionally substituted with 1-5 substituents independently selected from (C 1 -C 6 )alkyl, halogen, amino, cyano, —COOH, halo(C 1 -C 6 )alkyl, hydroxy(C 0 -C 6 )alkyl, (C 6 -C 10 )aryl, (C 3 -C 10 )heteroaryl, halo(C 6 -C 10 )aryl, hydroxy(C 6 -C 10 )aryl, (C 1 -C 6 )alkoxy, halo(C 1 -C 6 )alkoxy, (C 6 -C 16 )aryloxy, (C 3 -C 8 )cycloalkyl, halo(C 6 -C 10 )aryl(C 1 -C 6 )alkoxy, (C 1 -C 6 )alkoxy(C 6 -C 10 )aryl(C 1 -C 6 )alkoxy, nitro, (C 0 -C 6 )alkyl(C 6 -C 10 )aryl(C 0 -C 6 )alkoxy, (C 5 -C 10 )heterocycle, —OCHF 2 , —OCF 3 , —SCF 3 , —OBn, cyano(C 1 -C 6 )alkylene, (C 1 -C 6 )alkoxyamino, (C 6 -C 10 )aryl(C 2 -C 6 )alkenyl, (C 2 -C 6 )alkenyl(C 1 -C 6 )alkoxy, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkenyl(C 6 -C 10 )aryl, —N((C 0 -C 6 )alkyl)((C 1 -C 6 )alkyl), —N((C 1 -C 6 )alkyleneOH)((C 1 -C 6 )alkyleneOH), —N((C 0 -C 6 )alkyl)((C 1 -C 6 )alkyleneOH), —N((C 1 -C 6 )alkyleneOCO(C 1 -C 6 )alkyl)((C 1 -C 6 )alkyleneOCO(C 1 -C 6 )alkyl), —NCO(C 1 -C 6 )alkyl, —NPh 2 , —OPh(halogen) 0-3 , —OPhO(C 1 -C 6 )alkyl, —OPhO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkoxy, —O(C 1 -C 6 )alkyl(C 6 -C 10 )aryl, —O(C 2 -C 6 )alkenyl, —O(C 2 -C 6 )alkyleneN(CH 3 ) 2 , (C 0 -C 6 )alkylCOO(C 1 -C 6 )alkyl, —B(OH) 2 or —S(C 1 -C 6 )alkyl;
and wherein any of the substituents selected from (C 1 -C 6 )alkyl, (C 1 -C 6 )alkoxy, (C 6 -C 10 )aryl, (C 6 -C 16 )aryloxy or (C 5 -C 10 )heteroaryl is further optionally substituted with 1-4 substituents selected from halogen, (C 1 -C 6 )alkyl, halo(C 1 -C 6 )alkyl, amino, nitro, cyano, hydroxy(C 0 -C 6 )alkyl, (C 1 -C 6 )alkoxy, —COO(C 0 -C 6 )alkyl, or —CHO; and
R 3 is (C 1 -C 15 )alkyl, (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 6 -C 10 )aryl or (C 5 -C 10 )heteroaryl, wherein R 3 is optionally substituted with 1-4 substituents independently selected from halogen, amino, cyano or hydroxyl;
b) incubating the sample and the compound of step a) together for a period of time sufficient to stain the cells; and
c) analyzing the incubated stained cells by spectroscopy to detect a fluorescence signal, wherein the presence of a fluorescence signal is indicative of the presence of the beta cells.

11. A method for detecting microglia cells, comprising:
a) contacting a sample comprising cells with a compound of structural Formula (I) or a pharmaceutically acceptable salt thereof:

wherein:
R 1 is hydrogen or —COR 3 ;
R 2 is (C 6 -C 16 )aryl, (C 3 -C 10 )heteroaryl, (C 1 -C 6 )alkyl, (C 1 -C 6 )cycloalkyl, (C 2 -C 6 )alkenyl, or C≡CH;
R 2 is optionally substituted with 1-5 substituents independently selected from (C 1 -C 6 )alkyl, halogen, amino, cyano, —COOH, halo(C 1 -C 6 )alkyl, hydroxy(C 0 -C 6 )alkyl, (C 6 -C 10 )aryl, (C 3 -C 10 )heteroaryl, halo(C 6 -C 10 )aryl, hydroxy(C 6 -C 10 )aryl, (C 1 -C 6 )alkoxy, halo(C 1 -C 6 )alkoxy, (C 6 -C 16 )aryloxy, (C 3 -C 8 )cycloalkyl, halo(C 6 -C 10 )aryl(C 1 -C 6 )alkoxy, (C 1 -C 6 )alkoxy(C 6 -C 10 )aryl(C 1 -C 6 )alkoxy, nitro, (C 0 -C 6 )alkyl(C 6 -C 10 )aryl(C 0 -C 6 )alkoxy, (C 5 -C 10 )heterocycle, —OCHF 2 , —OCF 3 , —SCF 3 , —OBn, cyano(C 1 -C 6 )alkylene, (C 1 -C 6 )alkoxyamino, (C 6 -C 10 )aryl(C 2 -C 6 )alkenyl, (C 2 -C 6 )alkenyl(C 1 -C 6 )alkoxy, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkenyl(C 6 -C 10 )aryl, —N((C 0 -C 6 )alkyl)((C 1 -C 6 )alkyl), —N((C 1 -C 6 )alkyleneOH)((C 1 -C 6 )alkyleneOH), —N((C 0 -C 6 )alkyl)((C 1 -C 6 )alkyleneOH), —N((C 1 -C 6 )alkyleneOCO(C 1 -C 6 )alkyl)((C 1 -C 6 )alkyleneOCO(C 1 -C 6 )alkyl), —NCO(C 1 -C 6 )alkyl, —NPh 2 , —OPh(halogen) 0-3

Assignees

• Nat Univ Singapore
• Agency Science Tech & Res

Inventors

• Chang Young-Tae
• Lee Sung Chan
• Kang Nam-Young
• Yun Seong Wook
• Leong Cheryl Kit Mun
• Ha Hyung Ho

Classifications

• C07F5/022 Primary

  without C-boron linkages · CPC title

• G01N33/5306 Primary

  Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding · CPC title

• G01N33/582

  with fluorescent label · CPC title

• G01N2458/00

  Labels used in chemical analysis of biological material · CPC title

• A61K49/0021

  the fluorescent group being a small organic molecule · CPC title