Lipid nanodiscs solubilized through poly(acrylic acid-co-styrene) copolymers

Patent metadata

Field	Value
Publication number	US-12378337-B2
Application number	US-202017618788-A
Country	US
Kind code	B2
Filing date	Jun 19, 2020
Priority date	Jun 21, 2019
Publication date	Aug 5, 2025
Grant date	Aug 5, 2025

Abstract

Provided herein are compositions including lipids and copolymers in the form of a nanodisc assembly. The subject copolymers include monomer units of styrene and monomer units selected from acrylic acid and an acrylic acid derivative. In certain cases, the copolymer is a copolymer of styrene and acrylic acid. Also provided herein, is an aqueous solution comprising the subject composition. Also provided herein, are methods for producing a nanodisc assembly, including incubation of a lipid and a subject copolymer. Further provided herein, are methods for solubilizing a membrane protein in an aqueous solution, wherein the method includes forming a nanodisc assembly of a lipid bilayer having one or more membrane proteins embedded therein, and a subject copolymer. Also provided are methods of solubilizing a hydrophobic constituent in an aqueous solution, including forming a nanodisc assembly of a lipid, a hydrophobic constituent, and a subject copolymer.

First claim

What is claimed is:

1. A composition comprising:
a lipid; and
a copolymer, wherein the copolymer comprises monomer units of styrene, and monomer units selected from acrylic acid, and an acrylic acid derivative;
wherein the lipid and copolymer are in the form of a nanodisc assembly.

2. The composition of claim 1 , wherein the acrylic acid derivative is selected from acrylate, acrylate ester, acrylamide, and N-substituted acrylamide.

3. The composition of claim 1 , wherein the copolymer comprises an acrylic acid, or an acrylic acid derivative content of from 30 molar % to 70 molar % or from 35 molar % to 50 molar %.

4. The composition of claim 1 , wherein the copolymer is a copolymer of styrene and acrylic acid, optionally with high regularity of alternating monomer units.

5. The composition of claim 1 , wherein the copolymer is a diblock copolymer described by formula (I):

wherein:
Z 1 is selected from OR 1 , and NR 2 2 , wherein R 1 is selected from hydrogen, sodium, potassium, alkyl, and substituted alkyl, and each R 2 is independently selected from hydrogen, alkyl, and substituted alkyl;
X 1 and Y 1 are each independently a terminal group; and
n and m are each independently an integer from 15-100.

6. The composition of claim 5 , wherein Z 1 is OR 1 and R 1 is hydrogen.

7. The composition of claim 5 , wherein Z 1 is OR 1 and R 1 is alkyl or substituted alkyl.

8. The composition of claim 5 , wherein Z 1 is NR 2 2 and each R 2 is selected from hydrogen, alkyl and substituted alkyl.

9. The composition of claim 8 , wherein one R 2 is hydrogen and the other R 2 is alkyl or substituted alkyl.

10. The composition of claim 5 , wherein X 1 is selected from alkyl, substituted alkyl, nitrile, hydroxy, carboxyl, and halogen.

11. The composition of claim 5 , wherein X 1 is of the formula (X-I):

wherein:
R 3 and R 4 are each independently selected from hydrogen, alkyl, and substituted alkyl.

12. The composition of claim 5 , wherein Y 1 is selected from, alkyl, substituted alkyl, nitrile, hydroxy, carboxyl, halogen, thiol, substituted thiol, acyl, and substituted acyl.

13. The composition of claim 5 , wherein Y 1 is of the formula (Y-I)

wherein:
R 5 and R 6 are each independently selected from hydrogen, alkyl, and substituted alkyl; and
X 2 and X 3 are each independently selected from S and O.

14. The composition of claim 13 , wherein the formula (Y-II) is of formula (Y-III):

15. The composition of claim 5 , wherein n and m are each independently an integer from 15-50.

16. The composition of claim 1 , wherein the molecular weight of the copolymer is from 2 kDa to 15 kDa or from 3 kDa to 8 kDa.

17. The composition of claim 1 , wherein the nanodisc assembly has a diameter from 5 to 40 nm.

18. The composition of claim 5 , wherein the copolymer is described by formula (II):

wherein:
R 1 , R 3 , and R 4 are each independently selected from hydrogen, alkyl, and substituted alkyl;
Y 1 is selected from:

wherein:
R 5 and R 6 are each independently selected from hydrogen, alkyl, and substituted alkyl;
X 2 and X 3 are each independently selected from S and O; and
n and m are each independently an integer from 15-100.

19. The composition of claim 18 , wherein the copolymer is described by the formula (III) or (IV):

20. The composition of claim 1 , further comprising a membrane protein.

21. An aqueous solution comprising a composition according to claim 1 .

Assignees

• Univ Leland Stanford Junior
• Univ California
• Univ Aarhus

Inventors

• Appel Eric A
• Smith Anton
• Autzen Henriette E
• Cheng Yifan

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

• A61K9/5169

  Proteins, e.g. albumin, gelatin · CPC title

• C08L33/02

  Homopolymers or copolymers of acids; Metal or ammonium salts thereof · CPC title

• C08L25/08

  Copolymers of styrene (C08L29/08, C08L35/06, C08L55/02 take precedence) · CPC title

• C08F220/06

  Acrylic acid; Methacrylic acid; Metal salts or ammonium salts thereof · CPC title