Zinc indicators for cellular imaging
View Patent ↗The present disclosure provides compounds of the formula: wherein the variables are defined herein. The present disclosure also provides methods of imaging Zn 2+ within granules in cells, such as pancreatic α-, β-, and δ-cells. The present disclosure also provides methods of sorting cells comprising the use of the compounds of the present disclosure.
1. A compound of the formula:
wherein:
X is —O—;
R 1 and R 1 ′ are each independently hydrogen; or
alkyl (C≤12) , alkenyl (C≤12) , alkynyl (C≤12) , acyl (C≤12) , or a substituted version of any of these groups; or
R 1 and R 1 ′ are taken together and is alkanediyl (C2-7) , substituted alkanediyl (C2-7) , -alkanediyl (C1-3) -O-alkanediyl (C1-3) -, or substituted -alkanediyl (C1-3) -O-alkanediyl (C1-3) -;
R 4 and R 4 ′ are each independently hydrogen; or
alkyl (C≤12) , alkenyl (C≤12) , alkynyl (C≤12) , acyl (C≤12) , or a substituted version of any of these groups; or
R 4 and R 4 ′ are taken together and is alkanediyl (C2-7) , substituted alkanediyl (C2-7) , -alkanediyl (C1-3) -O-alkanediyl (C1-3) -, or substituted -alkanediyl (C1-3) -O-alkanediyl (C1-3) -;
R 2 , R 3 , R 5 , and R 6 are each independently hydrogen or halo;
R 7 is hydrogen, alkyl (C≤12) , substituted alkyl (C≤12) , acyl (C≤12) , or substituted acyl (C≤12) ;
A 1 and A 2 are each independently heteroaryl (C≤12) or substituted heteroaryl (C≤12) ;
m is 1, 2, or 3;
n is 1, 2, 3, or 4; and
o is 1, 2, 3, or 4; or
a compound of the formula:
wherein:
X, R 7 , A 1 , A 2 , m, n, and o are as defined above;
wherein a chemical group is used with the “substituted” modifier, one or more hydrogen atom has been replaced, independently at each instance, by —OH, —F, —Cl, —Br, —I, —NH 2 , —NO 2 , —CO 2 H, —CO 2 CH 3 , —CN, —SH, —OCH 3 , —OCH 2 CH 3 , —C(O)CH 3 , —NHCH 3 , —NHCH 2 CH 3 , —N(CH 3 ) 2 , —C(O)NH 2 , —C(O)NHCH 3 , —C(O)N(CH 3 ) 2 , —OC(O)CH 3 , —NHC(O)CH 3 , —S(O) 2 OH, or —S(O) 2 NH 2 ;
or a zinc complex or a salt of either of these formulae.
2. The compound of claim 1 , wherein the compound is a compound of formula (I) or a zinc complex or a salt thereof.
3. The compound of claim 1 , wherein the compound is further defined as:
wherein:
R 1 and R 1 ′ are each independently hydrogen; or
alkyl (C≤12) , alkenyl (C≤12) , alkynyl (C≤12) , acyl (C≤12) , or a substituted version of any of these groups; or
R 1 and R 1 ′ are taken together and is alkanediyl (C2-7) , substituted alkanediyl (C2-7) , -alkanediyl (C1-3) -O-alkanediyl (C1-3) -, or substituted -alkanediyl (C1-3) -O-alkanediyl (C1-3) -;
R 4 and R 4 ′ are each independently hydrogen; or
alkyl (C≤12) , alkenyl (C≤12) , alkynyl (C≤12) , acyl (C≤12) , or a substituted version of any of these groups; or
R 4 and R 4 ′ are taken together and is alkanediyl (C2-7) , substituted alkanediyl (C2-7) , -alkanediyl (C1-3) -O-alkanediyl (C1-3) -, or substituted -alkanediyl (C1-3) -O-alkanediyl (C1-3) -;
R 7 is hydrogen, alkyl (C≤12) , substituted alkyl (C≤12) , acyl (C≤12) , or substituted acyl (C≤12) ;
A 1 and A 2 are each independently heteroaryl (C≤12) or substituted heteroaryl (C≤12) ;
m is 1, 2, or 3;
n is 1, 2, 3, or 4; and
o is 1, 2, 3, or 4;
or a zinc complex or a salt thereof.
4. The compound according to claim 1 , wherein the compound is further defined as:
wherein:
R 1 and R 1 ′ are each independently hydrogen; or
alkyl (C≤12) , alkenyl (C≤12) , alkynyl (C≤12) , acyl (C≤12) , or a substituted version of any of these groups; or
R 1 and R 1 ′ are taken together and is alkanediyl (C2-7) , substituted alkanediyl (C2-7) , -alkanediyl (C1-3) -O-alkanediyl (C1-3) -, or substituted -alkanediyl (C1-3) -O-alkanediyl (C1-3) -;
R 4 and R 4 ′ are each independently hydrogen; or
alkyl (C≤12) , alkenyl (C≤12) , alkynyl (C≤12) , acyl (C≤12) , or a substituted version of any of these groups; or
R 4 and R 4 ′ are taken together and is alkanediyl (C2-7) , substituted alkanediyl (C2-7) , -alkanediyl (C1-3) -O-alkanediyl (C1-3) -, or substituted -alkanediyl (C1-3) -O-alkanediyl (C1-3) -;
R 7 is hydrogen, alkyl (C≤12) , substituted alkyl (C≤12) , acyl (C≤12) , or substituted acyl (C≤12) ;
A 1 and A 2 are each independently heteroaryl (C≤12) or substituted heteroaryl (C≤12) ;
m is 1, 2, or 3;
n is 1, 2, 3, or 4; and
o is 1, 2, 3, or 4;
or a zinc complex or a salt thereof.
5. The compound according to claim 1 , wherein R 7 is hydrogen.
6. The compound according to claim 1 , wherein m is 1.
7. The compound according to claim 1 , wherein n is 1.
8. The compound according to claim 1 , wherein o is 2.
9. The compound according to claim 1 , wherein A 1 is heteroaryl (C≤12) .
10. The compound according to claim 1 , wherein A 2 is heteroaryl (C≤12) .
11. The compound according to claim 1 , wherein R 1 is hydrogen.
12. The compound according to claim 1 , wherein R 1 is alkyl (C≤12) or substituted alkyl (C≤12) .
13. The compound according to claim 1 , wherein R 1 ′ is hydrogen.
14. The compound according to claim 1 , wherein R 1 ′ is alkyl (C≤12) or substituted alkyl (C≤12) .
15. The compound according to claim 1 , wherein R 4 is hydrogen.
16. The compound according to claim 1 , wherein R 4 is alkyl (C≤12) or substituted alkyl (C≤12) .
17. The compound according to claim 1 , wherein R 4 ′ is hydrogen.
18. The compound according to claim 1 , wherein R 4 ′ is alkyl (C≤12) or substituted alkyl (C≤12) .
19. The compound according to claim 1 , wherein the compound is further defined as:
or a zinc complex or a salt thereof.
20. The zinc complex according to claim 19 , where the zinc complex is further defined as:
21. A method of detecting zinc ion (Zn 2+ ) in a cell comprising:
a) contacting the cell with a compound according to claim 1 ; and
b) detecting fluorescence of the said compound following binding of zinc ion by said compound.
22. A method of characterizing a cell comprising:
a) contacting the cell with a compound according to claim 1 ; and
b) detecting fluorescence using flow cytometry.
23. A method of sorting cells comprising:
a) contacting the cells with a compound according to claim 1 ; and
b) sorting the cells by fluorescence-activated cell sorting (FACS).