Nanotubes as carriers of nucleic acids into cells

Patent metadata

Field	Value
Publication number	US-10344300-B2
Application number	US-201514662334-A
Country	US
Kind code	B2
Filing date	Mar 19, 2015
Priority date	Jan 4, 2011
Publication date	Jul 9, 2019
Grant date	Jul 9, 2019

Abstract

The present invention is directed to transfection complexes of rosette nanotubes and one or more nucleic acids.

First claim

What is claimed is:

1. A method of delivering one or more nucleic acids into a cell comprising contacting the cell with a complex of the one or more nucleic acids and a structure formed from the assembly of one or more compounds in aqueous media having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof in a manner to deliver the complex into the cell.

2. A complex comprising one or more nucleic acids and a structure formed from the assembly of compounds having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof.

3. The method of claim 1 wherein the aliphatic moiety is a straight chain or a branched alkyl group being saturated or unsaturated.

4. The complex of claim 2 wherein the aliphatic moiety is a straight chain or a branched alkyl group being saturated or unsaturated.

5. The method of claim 1 wherein R 2 is the linker group (CH 2 ) n where n is 1, 2, 3 or 4 and Y is lysine.

6. A method of delivering one or more nucleic acids into a cell comprising contacting the cell with a complex of the one or more nucleic acids and a structure formed from the assembly of one or more compounds in aqueous media having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an a-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof in a manner to deliver the complex into the cell, wherein the one or more nucleic acids include DNA, RNA, a gene, a gene fragment, an exon, an intron, intergenic DNA, heterochromatic DNA, messenger RNA, transfer RNA, interference RNA, dsRNA, ssRNA, saRNA, siRNA, miRNA, ribosomal RNA, a ribozyme, cDNA, a recombinant polynucleotide, a branched polynucleotide, a plasmid, a vector, isolated DNA of a sequence, isolated RNA of a sequence, a nucleic acid probe, a molecular beacon or a primer.

7. A method of delivering one or more nucleic acids into a cell comprising contacting the cell with a complex of the one or more nucleic acids and a structure formed from the assembly of one or more compounds in aqueous media having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof in a manner to deliver the complex into the cell, wherein the one or more nucleic acids includes siRNA having between about 10 to about 30 nucleic acids in length.

8. A method of delivering one or more nucleic acids into a cell comprising contacting the cell with a complex of the one or more nucleic acids and a structure formed from the assembly of one or more compounds in aqueous media having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof in a manner to deliver the complex into the cell, wherein the cell includes an animal, plant or bacterial cell.

9. The method of claim 1 wherein the linker group is (CH 2 ) n where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

10. The complex of claim 2 wherein the linker group is (CH 2 ) n where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

11. A method of making a complex comprising mixing together one or more compounds having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof
and one or more nucleic acids in aqueous media to form a stable complex of the one or more compounds and the one or more nucleic acids in aqueous media.

12. A method of making a complex comprising mixing together one or more compounds having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof
and one or more nucleic acids in aqueous media to form a stable complex of the one or more compounds and the one or more nucleic acids in aqueous media, wherein the linker group is (CH 2 ) n , where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

13. A method of making a complex comprising mixing together one or more compounds having the formula

wherein X is CH or nitrogen; R 2 is hydrogen or a linker group; Y is absent when R 2 is hydrogen or is an amino acid or polypeptide having an amino group covalently bound to an α-carbon of the amino acid and the amino group is covalently bound to the linker group R 2 ; and R 1 is hydrogen or an aliphatic moiety; and salts thereof
and one or more nucleic acids in aqueous media to form a stable complex of the one or more compounds and the one or more nucleic acids in aqueous media, wherein the mixture is sonicated to form the stable complex in aqueous media.

14. The method of claim 11 wherein the mixture is sonicated and heated to boiling to form the stable complex in aqueous media.

Assignees

• Univ Brown
• Rhode Island Hospital
• Univ Alberta
• Nat Res Council Canada

Inventors

• Chen Yunpeng
• Chen Qian
• Webster Thomas J
• Fenniri Hicham
• Hemraz Usha Devi

Classifications

• C12Q1/6841

  In situ hybridisation · CPC title

• C12N15/115

  Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers} · CPC title

• C12N2320/30

  Special therapeutic applications · CPC title

• A61K9/0092

  Hollow drug-filled fibres, tubes of the core-shell type, coated fibres, coated rods, microtubules or nanotubes · CPC title

• A61K31/675

  having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate · CPC title