Cell transfecting formulations of small interfering RNA related compositions and methods of making and use
Compositions incorporating small interfering ribonucleic acid (siRNA) and certain lipid-conjugated polyamide compound-based delivery vehicles that are particularly useful in the delivery siRNA and other polynucleotides to cells. Also, methods of making and using the compositions.
1. A method of inhibiting expression of a target gene in a subject, the method comprising administering to the subject a composition comprising:
a small interfering ribonucleic acid (siRNA) in a pharmaceutically acceptable delivery vehicle, wherein the delivery vehicle comprises a lipid-cationic peptoid conjugate of the formula:
L-X—[N(CH 2 CH 2 NH 2 )CH 2 (C═O)—N(CH 2 CH 2 R)CH 2 (C═O)—N(CH 2 CH 2 R)CH 2 (C═O)] 3 —NH 2
and positional isomers
where
L is selected from a non-sterol lipid moiety comprising at least one fatty alkyl or alkenyl chain between about 8 and 24 carbon atoms in length and a sterol moiety, wherein when L is a non-sterol lipid moiety, the peptoid conjugate is a lipitoid and when L is a sterol moiety, the peptoid conjugate is a cholesteroid;
each group R is independently selected from alkyl, aminoalkyl, and aralkyl, and
X is selected from the group consisting of a direct bond, an oligopeptide, a substantially linear alkyl chain from 2 to about 30 bonds in length, and a substantially linear chain from 2 to about 30 bonds in length consisting of alkyl bonds and one or more linkages selected from the group consisting of ester, amide, carbonate, carbamate, disulfide, peptide, and ether; wherein the delivery vehicle comprises a combination of lipitoid and cholesteroid peptoid conjugates; and
wherein one strand of the siRNA duplex has a nucleotide sequence comprised in an mRNA derived from the target gene.
2. The method of claim 1 , wherein the lipitoid:cholesteroid ratio is about 5:1 to 1:5.
3. The method of claim 2 , wherein the lipitoid:cholesteroid ratio is about 1:1 to 1:3.
4. The method of claim 3 , wherein the lipitoid is DMPE(NaeNmpeNmpe) 3 and the cholesteroid is Chol-β-ala-(NaeNmpeNmpe) 3 .
5. The method of claim 1 , wherein the siRNA comprises the sequence represented by SEQ ID NO: 1 with two nucleotide 3′-overhangs and phosphodiester link throughout.
6. The method of claim 1 , wherein the subject is a mammalian cell or cell line.
7. The method of claim 1 , wherein the subject is a mammal.
8. The method of claim 7 , wherein the subject is a human.