Methods of intracellular peptide delivery
Methods and kits for transporting heterologous compounds into cells, particularly mammalian cells, are disclosed. The method involves the pretreatment of the cell with a polycation, cationic polymer and/or cationic peptide before transportation of the heterologous compound into the cell. The methods are particularly applicable to the transport of peptides and proteins.
1. A method of transporting a peptide into an isolated mammalian cell comprising: pretreating the isolated mammalian cell with a composition comprising at least one compound comprising a recurring unit represented by a formula selected from the group consisting of formula (I) and (II); administering the peptide to the pretreated cell, and transporting the peptide into the pretreated mammalian cell, wherein the peptide has 2 to 50 amino acids and wherein formula (I) and (II) are represented by:
wherein A comprises at least one acetal group;
wherein B is selected from the group consisting of —CH—, —CH(CH 3 )—, CH 2 CH—, CH 2 C(CH 3 )—, —CH(CH 3 )CH—, and —CHCH(CH 3 )CH(CH 3 )—;
wherein Z is selected from the group consisting of C(O)OR 1 , C(O)SR 1 , C(O)NR 1 R 2 , and VU;
wherein V is a linker group; wherein U is selected from the group consisting of poly(ethyleneimine) (PU), poly(propyleneimine) (PPI), poly(lysine), PAMAM dendrimer, octaamine dendrimer, hexadecaamine dendrimer, enhancer, and targeting receptor;
wherein R 1 and R 2 are each individually selected from the group consisting of hydrogen, C 1 to C 10 alkyl, and C 6 to C 10 aryl;
wherein D is a linkage selected from the group consisting of carboxylic amide, carboxylic ester, urea, and urethane; and
wherein G is selected from the group consisting of C 4 to C 20 alkyl, C 6 -C 10 aryl, and —(OCH 2 CH 2 ) n —, where n is in the range of 1 to about 250.
2. The method of claim 1 , wherein U is poly(ethyleneimine).
3. The method of claim 2 in which the poly(ethyleneimine) has a molecular weight in the range of about 200 to about 100,000 Daltons.
4. The method of claim 2 in which V is ——C(O)NH——.
5. The method of claim 1 , wherein the cell is an oocyte.
6. The method of claim 1 in which U is poly(lysine).
7. The method of claim 6 in which the poly(lysine) has a molecular weight in the range of about 200 to about 50,000 Daltons.
8. The method of claim 1 in which Z is VU.
9. The method of claim 1 , wherein U is selected from the group consisting of polyethyleneimine, enhancer and targeting receptor.
10. The method of claim 9 in which U is a poly(ethylenimine) having a molecular weight in the range of about 200 to about 100,000 Daltons.
11. The method of claim 9 in which U is an enhancer selected from the group consisting of lipid, cholesterol, lipoprotein, fatty acid, lysosomotrophic agent, and fusogenic agent.
12. The method of claim 9 in which U is a targeting receptor selected from the group consisting of transferrin, antibody, antibody fragment, galactose, and mannose.