Conjugated oligomer-based macrocycles for live cell imaging
View Patent ↗A phenyleneethynylene macrocycle (PEMC) is constructed that is a cyclic or polycyclic oligo (oligophenyleneethynylene-co-polyamine) where a monodispersed oligophenyleneethynylene is coupled with a polyamine. The PEMC is formed by coupling a monodispersed di-aldehyde end-capped oligophenyleneethynylene with a polyamine to form a macrocyclic imine and reducing the imine to an amine. The PEMC is useful at entering living cells for the fluorescence imaging of the living cells without toxicity to the cells.
1. A phenyleneethynylene macrocycle (PEMC), comprising a cyclic or polycyclic (oligophenyleneethynylene-co-diamine), the oligophenyleneethynylene being a monodispersed dimer, trimer, tetramer, or pentamer, and the diamine comprising the structure:
where X is:
(CR 2 ) x where x is 1-3 and each R is independently H, C 1 -C 6 alkyl, or phenyl;
(CR 2 ) y C 6 H z R 4-z (CR 2 ) y , where y is 0-2, z is 0-4, and each R is independently H, C 1 -C 6 alkyl, aryl, or NHR′ 2 , where R′ is C 1 -C 6 alkyl; or
(CH 2 )[O(CH 2 ) 2 ] n (CH 2 ) where n is 1 to 4, and
where X is optionally interrupted one or more times with a disulphide group, an ester group, an acetal group, or a ketal group.
2. The PEMC according to claim 1 , wherein the oligophenyleneethynylene is of the structure:
where n is 0 to 4 and R is independently H, C 1 -C 3 alkyl, or C 1 -C 3 alkoxy where any of the carbons may be unsubstituted or substituted with: an amine or protected amine comprising a substituent; a OH or protected hydroxy comprising a substituent; a SH or protected thiol comprising a substituent; an amide comprising a substituent; or a guanidine or protected guanidine comprising a substituent; wherein the substituent comprises: an C 2 -C 5 alkylene oxide dimer, trimer, tetramer, or pentamer; an unprotected or protected C 2 -C 5 alkylene imine dimer, trimer, tetramer, or pentamer; or N-substituted C 1 -C 3 alkyl alkylene imine dimer, trimer, tetramer, or pentamer.
3. A PEMC comprising a cyclic or polycyclic (oligophenyleneethynylene-co-triamine), wherein the triamine comprises the structure:
where Y is:
((CR 2 ) x ) 3 CR where x is independently 0-3 and each R is independently H, C 1 -C 6 alkyl, or phenyl;
((CR 2 ) y ) 3 C 6 H z R 3-z where y is 0-2, z is 0-3, and each R is H, C 1 -C 6 alkyl, aryl, or NHR′ 2 ,
where R′ is C 1 -C 6 alkyl; or
(CH 2 )[O(CH 2 ) 2 ] n (CH 2 ) where n is 1 to 4 and
where Y is optionally interrupted one or more times with a disulphide group, an ester group, an acetal group, ora ketal group.
4. A PEMC comprising a cyclic or polycyclic (oligophenyleneethynylene-co-tetraamine), wherein is the tetraamine comprises the structure:
where Z is:
((CR 2 ) x ) 4 C where x is independently 0-3 and each R is independently H, C 1 -C 6 alkyl, or phenyl;
((CR 2 ) y ) 4 C 6 H z R 2-z where y is 0-2, z is 0-2, and each R is H, C 1 -C 6 alkyl, aryl, or NHR′ 2 , where R′ is C 1 -C 6 alkyl; or
(CH 2 )[O(CH 2 ) 2 ](CH 2 ) where n is 1 to 4 and
where Z is optionally interrupted one or more times with a disulphide group, an ester group, an acetal group, or a ketal group.
5. The PEMC according to claim 1 , wherein the cyclic (oligophenyleneethynylene-co-diamine) is a unimacrocycle (2/1).
6. The PEMC according to claim 3 , wherein the polycyclic (oligophenyleneethynylene-co-triamine) is a trimacrocycle (3/3) or a tetramacrocycle (3/6).
7. The PEMC according to claim 1 , wherein the cyclic (oligophenyleneethynylene-co-diamine) is;
where: X is CH 2 OCH 2 CH 2 OCH 2 , C 6 H 4 , or CH 2 OCH 2 ;
R is OCH 3 , or OCH 2 CH 2 O CH 2 CH 2 NH 2 .
8. The PEMC according to claim 1 , wherein the amines of the cyclic (oligophenyleneethynylene-co-diamine) are in the form of an ammonium ion paired with an anion.
9. The PEMC according to claim 8 , wherein the anion is a trifluoroacetate, acetate, or a halide.
10. A method of preparing a PEMC according to claim 1 , comprising:
providing a monodispersed di-aldehyde end-capped oligophenyleneethynylene;
providing a diamine;
combining the monodispersed di-aldehyde end-capped oligophenyleneethynylene with the diamine to form a macrocyclic imine; and
reducing the imine to an amine to form a cyclic or polycyclic (oligophenyleneethynylene-co-diamine).
11. The method according to claim 10 , wherein the di-aldehyde end-capped oligophenyleneethynylene is of the structure:
where n is 0 to 4 and R is independently H, C 1 -C 3 alkyl, or C 1 -C 3 alkoxy wherein any of the carbons may be unsubstituted or substituted with: an amine or protected amine comprising a substituent; a OH or protected hydroxy comprising a substituent; a SH or protected thiol comprising a substituent; an amide comprising a substituent; or a guanidine or protected guanidine comprising a substituent; wherein the substituent comprises: an C 2 -C 5 alkylene oxide dimer, trimer, tetramer, or pentamer; an unprotected or protected C 2 -C 5 alkylene imine dimer, trimer, tetramer, or pentamer; or N-substituted C 1 -C 3 alkyl alkylene imine dimer, trimer, tetramer, or pentamer.
12. The method according to claim 10 , wherein the polyamine is a diamine of the structure:
where X is:
(CR 2 ) x where x is 1-3 and each R is independently H, C 1 -C 6 alkyl, or phenyl;
(CR 2 ) y C 6 H z R 4-z (CR 2 ) y , where y is 0-2, z is 0-4, and each R is independently H, C 1 -C 6 alkyl, aryl, or NHR′ 2 , where R′ is C 1 -C 6 alkyl; or
(CH 2 )[O(CH 2 ) 2 ] n (CH 2 ) where n is 1 to 4 and
where X is optionally interrupted one or more times with a disulphide group, an ester group, an acetal group, or a ketal group.
13. The method according to claim 10 , wherein the reducing comprises combining with NaBH 4 .
14. The method according to claim 10 , further comprising protonating with an acid.
15. The method according to claim 14 , wherein the acid is trifluoroacetic acid, acetic acid, or a hydrogen halide.
16. A method of intracellular labeling, comprising providing an aqueous solution of a protonated PEMC according to claim 1 to plurality of live cells, wherein the oligophenyleneethynylene comprises at least one side chain with a protonated amine, where upon irradiation the PEMC fluoresces from within the live cells.
17. The PEMC according to claim 4 , wherein the polycyclic oligo (oligophenyleneethynylene-co-tetraamine) is a tetramacrocycle (4/4); or a hexamacrocycle (4/6).
18. A method of preparing a PEMC according to claim 3 , comprising:
providing a monodispersed di-aldehyde end-capped oligophenyleneethynylene;
providing a triamine;
combining the monodispersed di-aldehyde end-capped oligophenyleneethynylene with the triamine to form a macrocyclic imine; and
reducing the imine to an amine to form a cyclic or polycyclic (oligophenyleneethynylene-co-triamine).
19. A method of preparing a PEMC according to claim 4 , comprising:
providing a monodispersed di-aldehyde end-capped oligophenyleneethynylene;
providing a tetraamine;
combining the monodispersed di-aldehyde end-capped oligophenyleneethynylene with the tetraamine to form a macrocyclic imine; and
reducing the imine to an amine to form a cyclic or polycyclic (oligophenyleneethynylene-co-tetraamine).
20. A method of intracellular labeling, comprising providing an aqueous solution of a protonated PEMC according to claim 3 to plurality of live cells, wherein the oligophenyleneethynylene comprises at least one side chain with a protonated amine, wherein upon irradiation the PEMC fluoresces from within the live cells.