Delivery of agents using interfering nanoparticles
View Patent ↗Provided are compositions and methods for delivery of therapeutic agents, such as chemically stabilized antisense oligonucleotides useful in RNA silencing. The compositions include interfering nanoparticles (iNOPs) associated with one or more agents. Several functional iNOP derivatives are provided which allow for targeted delivery of agents to specific cell types as well as exhibiting reduced cellular toxicity.
1. A composition comprising a nanotransporter interfering nanoparticle-7 (iNOP-7) having a polylysine dendrimer generation 4 (LDG4) core conjugated to a lipid functional group having the following structure:
wherein the iNOP-7 is functionalized with any of functional surface groups A-I by being conjugated to iNOP-7 through an amide bond formed via an NH 2 group of the iNOP7:
wherein A forms iNOP-7E; B forms iNOP-7LE; C forms iNOP-7DS; D forms iNOP-7His; E-forms iNOP-7Bio; F forms iNOP-7AD; G forms iNOP-7PEG; H forms iNOP-7A; and I forms iNOP-7Lac, wherein the iNOP-7 is associated with a sense strand oligonucleotide molecule.
2. The composition of claim 1 , wherein the oligonucleotide comprises DNA.
3. The composition of claim 1 , wherein the oligonucleotide comprises RNA.
4. The composition of claim 3 , wherein the RNA is selected from the group consisting of microRNA mimic, anti-microRNA, dsRNA, siRNA, or shRNA.
5. The composition of claim 3 , wherein the RNA is chemically modified.
6. The composition of claim 5 , wherein the chemical modification comprises a 2′-O—F, 2′-Ome, 2′MOE, 2′-H, 2′-amino, 4-thioU or 6-thioG modification of one or more nucleotides, introduction of one or more phosphorothioate linkages, introduction of one or more locked nucleotides, or a combination thereof.
7. The composition of claim 1 , wherein the oligonucleotide is about 5-60 nucleotides in length.
8. The composition of claim 7 , wherein the oligonucleotide is about 7-36 nucleotides in length.
9. A method of delivering an oligonucleotide to a cell comprising contacting the cell with a nanotransporter interfering nanoparticle-7 (iNOP-7) associated with a sense oligonucleotide, the iNOP-7 having a polylysine dendrimer generation 4 (LDG4) core conjugated to a lipid functional group having the following structure:
wherein the iNOP-7 is functionalized with any of functional surface groups A-I by being conjugated to iNOP-7 through an amide bond formed via an NH 2 group of the iNOP7:
wherein A forms iNOP-7E; B forms iNOP-7LE; C forms iNOP-7DS; D forms iNOP-7His; E-forms iNOP-7Bio; F forms iNOP-7AD; G forms iNOP-7PEG; H forms iNOP-7A; and I forms iNOP-7Lac.
10. The method of claim 9 , wherein the oligonucleotide comprises DNA.
11. The method of claim 9 , wherein the oligonucleotide comprises RNA.
12. The method of claim 11 , wherein the RNA is selected from the group consisting of microRNA mimic, anti-microRNA, dsRNA, siRNA, or shRNA.
13. The method of claim 11 , wherein the RNA is chemically modified.
14. The method of claim 13 , wherein the chemical modification comprises a 2′-O—F, 2′-Ome, 2′MOE, 2′-H, 2′-amino, 4-thioU or 6-thioG modification of one or more nucleotides, introduction of one or more phosphorothioate linkages, introduction of one or more locked nucleotides, or a combination thereof.
15. The method of claim 9 , wherein the oligonucleotide is about 5-60 nucleotides in length.
16. The method of claim 15 , wherein the oligonucleotide is about 7-36 nucleotides in length.
17. The method of claim 9 , wherein the method is performed in vivo.
18. The method of claim 9 , wherein the method is performed in vitro.
19. The method of claim 9 , wherein the cell is mammalian.
20. The method of claim 19 , wherein the mammal is a human, mouse, rat, dog, monkey or ape.
21. The method of claim 9 , wherein the cell is an embryonic stem cell or induced pluripotent stem cell.