Synthetic nanostructures including nucleic acids and/or other entities

Patent metadata

Field	Value
Publication number	US-9216155-B2
Application number	US-201414215569-A
Country	US
Kind code	B2
Filing date	Mar 17, 2014
Priority date	Jan 19, 2010
Publication date	Dec 22, 2015
Grant date	Dec 22, 2015

Abstract

Articles, compositions, kits, and methods relating to nanostructures, including synthetic nanostructures, are provided. Certain embodiments described herein include structures having a core-shell type arrangement; for instance, a nanostructure core may be surrounded by a shell including a material, such as a lipid bilayer, and may include other components such as oligonucleotides. In some embodiments, the structures, when introduced into a subject, can be used to deliver nucleic acids and/or can regulate gene expression. Accordingly, the structures described herein may be used to diagnose, prevent, treat or manage certain diseases or bodily conditions. In some cases, the structures are both a therapeutic agent and a diagnostic agent.

First claim

What is claimed is:

1. A method for inhibiting gene expression comprising:
delivering a structure to a subject or a biological sample in an effective amount for inhibiting gene expression in the subject or biological sample, the structure comprising a nanostructure core; a shell comprising a lipid surrounding and attached to the nanostructure core or a hydrophobic shell surrounding the nanostructure core; and an oligonucleotide adapted to regulate gene expression associated with at least a portion of the shell, wherein the structure is adapted to sequester cholesterol, wherein the oligonucleotide inhibits gene expression of a target by at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% under physiological conditions and at a concentration where a nonsense or sense control has little or no effect, wherein the oligonucleotide is a cholesterol-oligonucleotide adsorbed to a surface of the shell.

2. A method for promoting cellular uptake of an oligonucleotide comprising:
delivering a oligonucleotide structure to a subject or a biological sample in an effective amount for promoting cellular uptake of the oligonucleotide in the subject or biological sample, the structure comprising a nanostructure core; a shell comprising a lipid surrounding and attached to the nanostructure core or a hydrophobic shell surrounding the nanostructure core; and an oligonucleotide adapted to regulate gene expression associated with at least a portion of the shell, wherein the structure is adapted to sequester cholesterol wherein the structure promotes the cellular uptake of the oligonucleotide, wherein the oligonucleotide is electrostatically physiosorbed to a surface of the shell.

3. The method of claim 2 , wherein the oligonucleotides have greater than 80%, 85%, 90%, 95%, 97%, or 99% cellular uptake.

4. The method of claim 1 , wherein the structure has low endosomal sequestration.

5. The method of claim 4 , wherein the low endosomal sequestration refers to at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% of the structures found within in the cytoplasm of the cell.

6. The method of claim 1 , wherein the structure has a binding constant to cholesterol, Kd ,of less than or equal to about 10 mM.

7. The method of claim 1 , wherein the lipid bilayer comprises 50-200 phospholipids.

8. The method of claim 1 , wherein at least a portion of the lipid bilayer is covalently bound to the core.

9. The method of claim 1 , wherein at least a portion of the lipid bilayer is physisorbed to the core.

10. The method of claim 1 , further comprising a protein associated with at least a portion of the structure.

11. The method of claim 1 , wherein the structure has a largest cross-sectional dimension of less than or equal to about 50 nm, 35 nm, or 30 nm.

12. The method of claim 1 , wherein the structure comprises a core that is hollow or at least partially hollow.

13. The method of claim 1 , wherein the nanostructure core comprises a metal, or is substantially formed from a metal.

14. The method of claim 13 , wherein the nanostructure core comprises gold.

15. The method of claim 1 , wherein the oligonucleotide is selected from the group consisting of an oligonucleotide adapted to reduce intracellular miR-210 levels, an anti-survivin oligonucleotide and an Apo B-100 siRNA.

16. The method of claim 1 , wherein the structure is administered in a single or divided dose according to a dosing schedule.

17. The method of claim 1 , wherein the oligonucleotides have a length of about 8 to about 500 nucleotides or base pairs in length, between about 10 to about 200 nucleotides or base pairs in length, about 10 to about 150 nucleotides or base pairs in length, about 10 to about 100 nucleotides or base pairs in length, about 10 to about 75 nucleotides or base pairs in length, or about 10 to about 50 nucleotides or base pairs in length.

18. The method of claim 1 , wherein the oligonucleotide is single stranded.

19. The method of claim 1 , wherein the oligonucleotide is double stranded.

20. The method of claim 1 , wherein the oligonucleotide comprises antisense DNA, siRNA, or microRNA.

21. The method of claim 2 , wherein the oligonucleotide modulates expression of a target gene by at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% when contacted with a biological sample or patient under physiological conditions and at a concentration where a nonsense or sense control has little or no effect.

22. The method of claim 1 , wherein the oligonucleotide is adsorbed onto a portion of the shell.

23. The method of claim 22 , wherein the oligonucleotide is adsorbed to an inner portion, outer portion, interior portion of the shell and/or combinations thereof.

24. The method of claim 1 , wherein the oligonucleotide is attached to the surface of the core through an intervening layer such as a passivating layer.

25. The method of claim 1 , wherein the subject has cancer or the biological cell is associated with a cancer.

26. The method of claim 25 , wherein the cancer is prostate cancer.

Assignees

• Univ Northwestern

Inventors

• Thaxton C Shad
• Mirkin Chad A
• Mcmahon Kaylin M
• Tripathy Sushant
• Mutharasan Raja Kannan
• Leander David M
• Luthi Andrea

Classifications

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P7/04

  Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title