Polymer nanoparticle and DNA nanostructure compositions and methods for non-viral delivery

Patent metadata

Field	Value
Publication number	US-12433910-B2
Application number	US-202418816696-A
Country	US
Kind code	B2
Filing date	Aug 27, 2024
Priority date	Dec 3, 2020
Publication date	Oct 7, 2025
Grant date	Oct 7, 2025

Abstract

The invention relates to polymer nanoparticle and DNA nanostructure delivery compositions for non-viral delivery, and methods therefor. More particularly, the invention relates to polymer nanoparticle delivery compositions, such as reversible addition-fragmentation chain transfer (RAFT) polymer compositions, and DNA nanostructure delivery compositions, such as DNA origami compositions, for the delivery of more than one payload, or for the delivery of a nucleic acid construct payload of 3 kB or more, and methods therefor.

First claim

What is claimed is:

1. A composition comprising a polymer nanoparticle and a payload, wherein the polymer nanoparticle comprises a RAFT block copolymer that comprises:
a first block prepared from one or more monomer units, and having a molecular weight (M n ) in the range of about 20 kDa to about 80 kDa and a degree of polymerization in the range of about 20 to about 400;
a second block prepared from one or more monomer units covalently attached to the first block, and having a molecular weight (M n ) in the range of about 5 kDa to about 80 kDa and a degree of polymerization in the range of about 10 to about 500; and
wherein the RAFT block copolymer has one or more of an overall molecular weight (M n ) in the range of about 25 kDa to about 160 kDa, and overall degree of polymerization in the range of about 30 to about 900, and
wherein the payload comprises a nucleic acid.

2. The composition of claim 1 , wherein the payload is chemically bonded to the polymer nanoparticle.

3. The composition of claim 1 , wherein the payload is complexed with the polymer nanoparticle.

4. The composition of claim 3 , wherein the payload is complexed with the polymer nanoparticle through electrostatic interactions.

5. The composition of claim 1 , wherein the polymer nanoparticle has a size in the range of about of about 10 to about 60 nm.

6. The composition of claim 1 , wherein the first block is prepared from one or more monomer units selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

7. The composition of claim 6 , wherein the second block is prepared from one or more monomer units selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

8. The composition of claim 6 , wherein the second block is a random copolymer prepared from two different monomer units independently selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

9. The composition of claim 6 , wherein the second block is a random copolymer prepared from three different monomer units independently selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

10. The composition of claim 1 , wherein the first block is prepared from one of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, or methyl methacrylate.

11. The composition of claim 10 , wherein the second block is prepared from one or more monomer units selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

12. The composition of claim 10 , wherein the second block is a random copolymer prepared from two different monomer units independently selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

13. The composition of claim 10 , wherein the second block is a random copolymer prepared from three different monomer units independently selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

14. The composition of claim 1 , wherein the second block is prepared from one or more monomer units selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

15. The composition of claim 1 , wherein the second block is a random copolymer prepared from two different monomer units independently selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

16. The composition of claim 1 , wherein the second block is a random copolymer prepared from three different monomer units independently selected from the group consisting of 2-dimethylaminoethyl acrylate, 2-(diethylamino) ethyl methacrylate, 2-(diisopropylamino) ethyl methacrylate, butyl methacrylate, ethyl acrylic acid, propyl acrylic acid, (hydroxyethyl) methacrylate, and methyl methacrylate.

17. The composition of claim 1 , wherein the first block comprises a first capping unit of the formula

wherein * represents a point of covalent attachment to the first block.

18. The composition of claim 1 , wherein the second block comprises a second capping unit of the formula

wherein * represents a point of covalent attachment to the second block, and R is-SC 2 -C 12 alkyl or C 6 H 5 .

19. The composition of claim 1 , wherein the nucleic acid is a DNA or an RNA.

20. The composition of claim 19 , wherein the nucleic acid is complexed with the polymer nanoparticle through electrostatic interactions.

21. The composition of claim 19 , wherein the nucleic acid is a DNA.

22. The composition of claim 21 , wherein the DNA is complexed with the polymer nanoparticle through electrostatic interactions.

23. The composition of claim 19 , wherein the nucleic acid is an RNA.

24. The composition of claim 23 , wherein the RNA is complexed with the polymer nanoparticle through electrostatic interactions.

25. The composition of claim 19 , wherein the nucleic acid has a size of 3 kB-kB or more.

26. The composition of claim 19 , wherein the nucleic acid has size of 6.5 kB or more.

27. The composition of claim 19 , wherein the nucleic acid has a size of 8.5 kB or more.

28. The composition of claim 1 , wherein the payload comprises one or more of the components of the CRISPR RNP system.

29. The composition of claim 28 , wherein the one or more of the components of the CRISPR RNP system includes a CRISPR-associated enzyme, a short guide RNA (sgRNA), and a donor DNA strand.

30. The composition of claim 29 , wherein the CRISPR-associated enzyme i

Assignees

• Battelle Memorial Institute

Inventors

• Duong Anthony D
• Gupta Cherry
• Sims Jr Kenneth R
• Huk Danielle
• Lilly Jacob
• Kute Stephanie M
• Koeris Mike

Classifications

• C08F2438/03

  Use of a di- or tri-thiocarbonylthio compound, e.g. di- or tri-thioester, di- or tri-thiocarbamate, or a xanthate as chain transfer agent, e.g . Reversible Addition Fragmentation chain Transfer [RAFT] or Macromolecular Design via Interchange of Xanthates [MADIX] · CPC title

• C08F293/005

  using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• C12N2310/11

  Antisense · CPC title

• B82Y35/00

  Methods or apparatus for measurement or analysis of nanostructures · CPC title