IP Library Granted Patent US 10,364,440
Granted Patent B2
US 10,364,440 · App. 13/977,138 · Granted Jul 30, 2019

Nanotubes as carriers of nucleic acids into cells

Inventors: Thomas J. Webster (Barrington, RI); Qian Chen (Barrington, RI); Yupeng Chen (Mansfield, MA); Hicham Fenniri (Edmonton, CA); Usha Devi Hemraz (Edmonton, CA)
Assignees: Brown University; Rhode Island Hospital; The Governors of the University of Alberta; National Research Council of Canada
C12N15/87A61K9/0092A61K31/675A61K31/713A61K48/005A61K48/0075C12N15/113C12N15/115C12Q1/6841B82Y5/00C12N2310/12C12N2310/14C12N2310/141C12N2320/30
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Quick Facts
Patent No.
US 10,364,440
App. No.
13/977,138
Granted
Jul 30, 2019
Kind
B2
Abstract

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

Claims (49)

1. A method of delivering one or more nucleic acids into a cell comprising contacting the cell with a complex of a rosette nanotube and the one or more nucleic acids in a manner to deliver the complex into the cell, wherein the one or more nucleic acids are released from the complex after entry into the cell.

2. The method of claim 1 wherein the nanotube is formed from 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 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.

4. The method of claim 1 wherein the one or more nucleic acids includes siRNA having between about 10 to about 30 nucleic acids in length.

5. The method of claim 1 wherein the cell is an animal, plant or bacterial cell.

6. A method of treating an individual requiring treatment comprising administering a complex of a rosette nanotube and one or more nucleic acids to the individual in a manner to deliver the one or more nucleic acids into cells within the individual and wherein the cells either express the one or more nucleic acids in a therapeutic manner or the one or more nucleic acids inhibit expression of one or more proteins within the cells in a therapeutic manner.

7. A method of making a complex comprising

mixing together rosette nanotubes and one or more nucleic acids in aqueous media under conditions which cause the rosette nanotubes to combine with the one or more nucleic acids to form a stable complex in aqueous media.

8. A product made by the process comprising mixing together rosette nanotubes and one or more nucleic acids in aqueous media under conditions which cause the rosette nanotubes to combine with the one or more nucleic acids to form a stable complex in aqueous media.

9. A complex comprising a rosette nanotube including one or more nucleic acids.

10. A complex comprising one or more nucleic acids and a rosette nanotube formed from the self 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.

11. A complex comprising one or more nucleic acids and a rosette nanotube formed from the self assembly in aqueous media of 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 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.

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

13. The complex of claim 10 wherein the aliphatic moiety is a straight chain or a branched alkyl group being saturated or unsaturated.

14. The complex of claim 11 wherein the aliphatic moiety is a straight chain or a branched alkyl group being saturated or unsaturated.

15. 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,

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

17. The method of claim 11 wherein the linker group is (CH 2 ) n where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

18. A method of making a complex comprising mixing together 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, such alkyl, straight or branched chain, saturated or unsaturated; and salts thereof,

and one or more nucleic acids in aqueous media and sonicating the mixture to cause the compounds to combine with the one or more nucleic acids to form a stable complex in aqueous media.

19. The method of claim 18 wherein the linker group is (CH 2 ) n where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

20. The method of claim 18 wherein the mixture is sonicated and heated to boiling.

21. The method of claim 20 wherein the linker group is (CH 2 ) n where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

22. The method of claim 2 wherein R 2 is hydrogen.

23. The method of claim 1 wherein the complex is formed by sonicating and heating a nucleic acid and a compound 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.

24. The method of claim 23 wherein R 2 is hydrogen.

25. The method of claim 2 wherein R 2 is —CH 2 CH 2 NH 3 + , —CH 2 CH 2 CH 2 CH 2 NH 3 + , or —CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 NH 3 + and Y is absent.

26. The method of claim 23 wherein R 2 is —CH 2 CH 2 NH 3 + , —CH 2 CH 2 CH 2 CH 2 NH 3 + , or —CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 NH 3 + and Y is absent.

27. A method of delivering one or more nucleic acids into a cell comprising contacting the cell with a complex of a rosette nanotube and the one or more nucleic acids in a manner to deliver the complex into the cell.

28. The method of claim 27 wherein the nanotube is formed from 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.

29. The method of claim 27 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.

30. The method of claim 27 wherein the one or more nucleic acids includes siRNA having between about 10 to about 30 nucleic acids in length.

31. The method of claim 27 wherein the cell is an animal, plant or bacterial cell.

32. The method of claim 28 wherein the aliphatic moiety is a straight chain or a branched alkyl group being saturated or unsaturated.

33. The method of claim 27 wherein the linker group is (CH 2 ) n where n is an integer of 1, 2, 3, or 4, (CH 2 ) 3 CO,

34. The method of claim 28 wherein R 2 is hydrogen.

35. The method of claim 27 wherein the complex is formed by sonicating and heating a nucleic acid and a compound 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.

36. The method of claim 35 wherein R 2 is hydrogen.

37. The method of claim 28 wherein R 2 is —CH 2 CH 2 NH 3 + , —CH 2 CH 2 CH 2 CH 2 NH 3 + , or —CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 NH 3 + and Y is absent.

38. The method of claim 35 wherein R 2 is —CH 2 CH 2 NH 3 + , —CH 2 CH 2 CH 2 CH 2 NH 3 + , or —CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 NH 3 + and Y is absent.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 6, 2017
From: HEMRAZ, USHA DEVI
To: THE GOVERNORS OF THE UNIVERSITY OF ALBERTA
Reel/Frame 044318/0364 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 6, 2017
From: FENNIRI, HICHAM
To: NATIONAL RESEARCH COUNCIL OF CANADA
Reel/Frame 044318/0470 →
CONFIRMATORY LICENSE Recorded Sep 16, 2014
From: BROWN UNIVERSITY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 033753/0416 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 25, 2014
From: WEBSTER, THOMAS J.; CHEN, QIAN; CHEN, YUPENG
To: BROWN UNIVERSITY; RHODE ISLAND HOSPITAL
Reel/Frame 032289/0706 →
Continuity (2)
Provisional Application 61429555 · Jan 4, 2011
Related Publication 20140171482A1 · Jun 19, 2014
Cited By (2)
US 12,214,043 US 12,534,468