# US 2025/0382402 A1 · Lipid nanodiscs solubilized through poly(acrylic acid-co-styrene) copolymers

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- Number: US 2025/0382402 A1
- Document ID: US20250382402A1
- Status: Published application
- Kind: A1
- Application number: 19/248,144
- Filed: 2025-06-24
- Published: 2025-12-18
- Earliest claimed: 2019-06-21
- Assignees: The Board of Trustees of the Leland Stanford Junior University; The Regents of the University of California; Aarhus University
- Applicants: The Board of Trustees of the Leland Stanford Junior University; The Regents of the University of California; Aarhus University
- Inventors: Eric A. Appel; Anton Smith; Henriette E. Autzen; Yifan Cheng
- Web page: https://patentgrep.com/patent/US20250382402A1

## CPC

- C08F 212/08: Styrene
- A61K 9/5169: {Proteins, e.g. albumin, gelatin}
- C08F 220/06: Acrylic acid; Methacrylic acid; Metal salts or ammonium salts thereof
- C08L 25/08: Copolymers of styrene (C08L29/08, C08L35/06, C08L55/02 take precedence)
- C08L 33/02: Homopolymers or copolymers of acids; Metal or ammonium salts thereof
- C08F 2438/03: 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]
- C08F: MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C08L: COMPOSITIONS OF MACROMOLECULAR COMPOUNDS (compositions based on polymerisable monomers C08F, C08G; artificial filaments or fibres D01F; textile treating compositions D06)

## 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.

## Claims

### Claim 1

A method of producing a nanodisc assembly, the method comprising:
incubation of 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 form a nanodisc assembly.

### Claim 2 (depends on 1)

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

### Claim 3 (depends on 1)

The method 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 %.

### Claim 4 (depends on 1)

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

### Claim 5 (depends on 1)

The method of claim 1, wherein the copolymer is a diblock copolymer described by formula (I):
wherein:
Z1 is selected from OR1, and NR22, wherein R1 is selected from hydrogen, alkyl, and substituted alkyl, and each R2 is independently selected from hydrogen, alkyl, and substituted alkyl;
X1 and Y1 are each independently a terminal group; and
n and m are each independently an integer from 15-100.

### Claim 6 (depends on 5)

The method of claim 5, wherein Z1 is OR1 and R1 is hydrogen.

### Claim 7 (depends on 5)

The method of claim 5, wherein Z1 is OR1 and R1 is alkyl or substituted alkyl.

### Claim 8 (depends on 5)

The method of claim 5, wherein Z1 is NR22 and each R2 is selected from hydrogen, alkyl and substituted alkyl.

### Claim 9 (depends on 5)

The method of claim 5, wherein one R2 is hydrogen and the other R2 is alkyl or substituted alkyl.

### Claim 10 (depends on 5)

The composition of claim 5, wherein X1 is selected from alkyl, substituted alkyl, nitrile, hydroxy, carboxyl, and halogen.

### Claim 11 (depends on 5)

The method of claim 5, wherein X1 is of the formula (X-I):
wherein:
R3 and R4 are each independently selected from hydrogen, alkyl, and substituted alkyl.

### Claim 12 (depends on 5)

The method of claim 5, wherein Y1 is selected from, alkyl, substituted alkyl, nitrile, hydroxy, carboxyl, halogen, thiol, substituted thiol, acyl, and substituted acyl.

### Claim 13 (depends on 5)

The method of claim 5, wherein Y1 is of the formula (Y-I) or (Y-II):
wherein:
R5 and R6 are each independently selected from hydrogen, alkyl, and substituted alkyl; and
X2 and X3 are each independently selected from S and O.

### Claim 14 (depends on 13)

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

### Claim 15 (depends on 1)

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

### Claim 16 (depends on 1)

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

### Claim 17 (depends on 1)

The method of claim 1, wherein the copolymer is described by formula (II):
wherein:
R1, R3, and R4 are each independently selected from hydrogen, alkyl, and substituted alkyl;
Y1 is selected from:
wherein:
R5 and R6 are each independently selected from hydrogen, alkyl, and substituted alkyl;
X2 and X3 are each independently selected from S and O; and
n and m are each independently an integer from 15-100.

### Claim 18 (depends on 1)

The method of claim 1, wherein the copolymer is described by the formula (III) or (IV):

### Claim 18 (depends on 1)

The method of claim 1, wherein the nanodisc assembly comprises membrane proteins.

### Claim 19

A method of solubilizing a membrane protein in an aqueous solution, the method comprising forming a nanodisc assembly of:
a lipid bilayer having one or more said membrane proteins embedded therein; and
a copolymer, wherein the copolymer comprises monomer units of styrene, and monomer units selected from acrylic acid and an acrylic acid derivative.

### Claim 20

A method of solubilizing a hydrophobic constituent in an aqueous solution, the method comprising forming a nanodisc assembly of:
a lipid;
a hydrophobic constituent; and
a copolymer, wherein the copolymer comprises monomer units of styrene, and monomer units selected from acrylic acid and an acrylic acid derivative.