Rapid design, build, test, and learn technologies for identifying and using non-viral carriers

Patent metadata

Field	Value
Publication number	US-12031128-B2
Application number	US-202217715784-A
Country	US
Kind code	B2
Filing date	Apr 7, 2022
Priority date	Apr 7, 2021
Publication date	Jul 9, 2024
Grant date	Jul 9, 2024

Abstract

The disclosure relates to barcoded polymer nanoparticles for in vivo screening and for in vivo therapeutic delivery, and methods therefor. More particularly, the invention relates to polymer nanoparticles, such as reversible addition-fragmentation chain transfer (RAFT) polymer compositions, associated with polynucleotide barcodes, for therapeutic delivery, and for high throughput in vivo screening of drug delivery nanoparticles.

First claim

What is claimed is:

1. A composition comprising:
a) a non-viral delivery vehicle comprising one or more nanoparticle forming polymers, and
b) a nucleic acid construct, wherein the one or more nanoparticle forming polymers are RAFT block copolymers comprising
i) a first terminus comprising a first capping unit derived from a first chain transfer agent in a RAFT copolymerization process, said first chain transfer agent comprising a first reactive functional unit;
ii). a first block prepared from one or more monomer units covalently attached to the first reactive functional unit, and having a molecular weight (M n ) in the range of about 1 kDa to about 200 kDa and a degree of polymerization in the range of about 10 to about 2500;
iii). optionally 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 1 kDa to about 200 kDa and a degree of polymerization in the range of about 20 to about 2000; and
iv) a second terminus comprising a second capping unit derived from the first or a second chain transfer agent.

2. The composition of claim 1 , wherein the non-viral delivery vehicle has one or more of an overall molecular weight (M n ) in the range of about 25 kDa to about 60 kDa, an overall degree of polymerization in the range of about 700 to about 900, a target size in the range of about of about 10 to about 60 nm, and a maximum corona-to-core ratio (CCR) of about 1.5 to about 3.5.

3. 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.

4. 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.

5. 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.

6. 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.

7. 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.

8. The composition of claim 1 , wherein the second block is a random copolymer prepared from 2-dimethylaminoethyl acrylate, butyl methacrylate, and propyl acrylic acid; or 2-dimethylaminoethyl acrylate and butyl methacrylate; or 2-dimethylaminoethyl acrylate, butyl methacrylate, and ethyl acrylic acid.

9. The composition of claim 1 , wherein each chain transfer agent is independently selected from the group consisting of bis(carboxymethyl)trithiocarbonate, bis(2-amino-2-oxoethyl) trithiocarbonate, bis[4-(2-hydroxyethoxycarbonyl)benzyl]trithiocarbonate, 4-cyano-4-(ethylsulfanylthiocarbonyl)sulfanylvpentanoic acid, 4-cyano-4-((phenylcarbonothioyl)thio)pentanoic acid, and 4-cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanoic acid.

10. The composition of claim 1 , wherein the first capping unit is of the formula

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

11. The composition of claim 1 , wherein the second capping unit is 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 .

12. A composition of claim 1 wherein
said 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;
said second block comprises a random copolymer prepared from 2-dimethylaminoethyl acrylate, butyl methacrylate, and propyl acrylic acid; or 2-dimethylaminoethyl acrylate and butyl methacrylate; or 2-dimethylaminoethyl acrylate, butyl methacrylate, and ethyl acrylic acid; and
each chain transfer agent is independently selected from the group consisting of bis(carboxymethyl)trithiocarbonate, bis(2-amino-2-oxoethyl) trithiocarbonate, bis[4(2-hydroxyethoxycarbonyl)benzyl] trithiocarbonate, 4-cyano-4-(ethylsulfanylthiocarbonyl) sulfanylvpentanoic acid, 4-cyano-4-((phenylcarbonothioyl)thio)pentanoic acid, and 4-cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl] pentanoic acid.

13. The composition of claim 12 , wherein the first capping unit is of the formula

wherein * represents a point of covalent attachment to the first block; and the second capping unit is 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 .

Assignees

• Battelle Memorial Institute

Inventors

• Duong Anthony D
• Huk Danielle J
• Gupta Cherry
• Sims Jr Kenneth R
• Koeris Michael S
• Shank Zachary R
• Colbert Ashlee J
• Mccue Andrea D
• Schmitz Emma K
• Hillrich Caleb T
• Miller Shannon D
• Hoy Joanna L

Classifications

• C12N15/88

  using microencapsulation, e.g. using {amphiphile} liposome vesicle · CPC title

• C12Q2525/301

  Hairpin oligonucleotides · CPC title

• C12Q2525/313

  Branched oligonucleotides · CPC title

• C12Q1/682

  Signal amplification · CPC title

• C12Q1/6816

  characterised by the detection means (C12Q1/6804 takes precedence) · CPC title