# US 11,111,375 B2 · Compositions and methods for generating oligodendrocyte precursors

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- Number: US 11,111,375 B2
- Document ID: US11111375B2
- Status: Granted
- Kind: B2
- Application number: 15/758,231
- Filed: 2016-10-04
- Published: 2018-09-06
- Granted: 2021-09-07
- Earliest claimed: 2015-10-05
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Gonçalo Rodrigues; David V. Schaffer
- Web page: https://patentgrep.com/patent/US11111375B2

## CPC

- C08L 51/006: {grafted on to block copolymers containing at least one sequence of polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds}
- C07D 471/04: Ortho-condensed systems
- C12N 5/0018: {Culture media for cell or tissue culture (media for specific animal cell type C12N5/06)}
- C12N 5/0622: {Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells}
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- C08L 2203/02: for biomedical use
- C12N 2501/385: of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
- C12N 2501/41: Hedgehog proteins; Cyclopamine (inhibitor)
- C12N 2501/415: Wnt; Frizzeled
- C12N 2506/02: from embryonic cells
- C12N 2506/45: from artificially induced pluripotent stem cells
- C12N 2539/10: Coating allowing for selective detachment of cells, e.g. thermoreactive coating
- C08L: COMPOSITIONS OF MACROMOLECULAR COMPOUNDS (compositions based on polymerisable monomers C08F, C08G; artificial filaments or fibres D01F; textile treating compositions D06)
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)

## Abstract

The present disclosure provides a three-dimensional culture system comprising a biocompatible polymer and a combination of factors in an amount effective to induce differentiation of oligodendrocyte precursors and/or oligodendrocytes from pluripotent stem cells. The present disclosure provides methods of generating oligodendrocyte precursors and/or oligodendrocytes using a three-dimensional culture system of the present disclosure. The present disclosure provides methods to treat neurological diseases and demyelinating diseases.

## Claims

### Claim 1

A three-dimensional culture system comprising:
a) biocompatible thermoresponsive polymer that forms a hydrogel at 37° C.; and
b) a combination of factors that promote differentiation of oligodendrocyte precursors from pluripotent stem cells in 20 days or less, wherein the combination of factors consists of a Sonic hedgehog (Shh) signaling pathway agonist and retinoic acid (RA); and
wherein the thermoresponsive polymer comprises:
an N-isopropylacrylamide co-monomer;
an alkyl [meth]acryl[ate/amide] co-monomer, wherein the alkyl is a lower alkyl group;
a PEG acrylamide co-monomer; and
a modifying acryl[ate/amide] co-monomer comprising a linked functional group or a linked modifying agent,
wherein the thermoresponsive polymer is described by formula (II):
wherein:
a, b, c and d are molar fractions of the co-monomers, wherein a >0.8, 0.1>b>0, and 0.2>c>0;
PEGn is a polyethylglycol polymer;
Z2 is a functional group or a linked modifying agent;
L is a linker;
R1 is a lower alkyl;
G1 and G2 are each independently selected from a polymer segment, a terminal group, a linker and a linked modifying agent.

### Claim 2 (depends on 1)

The culture system of claim 1, wherein:
the Shh signaling pathway agonist is 3,4-dichloro-N-(cis-4-(methylamino)cyclohexyl)-N-(3-pyridin-4-ylbenzyl)benzo[b]thiophene-2-carboxamide, an Shh polypeptide, an Shh protein-polymer conjugate, or a compound of the formula:

### Claim 3 (depends on 1)

The culture system of claim 1, comprising a pluripotent stem cell.

### Claim 4 (depends on 1)

The three-dimensional culture system of claim 1, wherein the Shh signaling pathway agonist is a small molecule.

### Claim 5 (depends on 1)

The three-dimensional culture system of claim 1, wherein the alkyl [meth]acryl[ate/amide] co-monomer is an isobutyl methacrylate co-monomer.

### Claim 6 (depends on 1)

The three-dimensional culture system of claim 1, wherein R1 is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, isopropyl, isobutyl, isopentyl, tert-butyl, cyclopropyl, and cyclobutyl.

### Claim 7 (depends on 1)

The three-dimensional culture system of claim 1, wherein Z2 is a chemoselective functional group.

### Claim 8 (depends on 1)

The three-dimensional culture system of claim 1, wherein the thermoresponsive polymer is described by the formula (IV):

### Claim 9 (depends on 1)

The three-dimensional culture system of claim 1, wherein G1 and G2 are each independently selected from a terminal group, a linker and a linked modifying agent.

### Claim 10 (depends on 1)

The three-dimensional culture system of claim 1, wherein G1, G2 or Z2 comprise a linked hyaluronic acid that is linked via conjugation to the carboxylic acid group of a hyaluronic acid monomer.

### Claim 11 (depends on 1)

The three-dimensional culture system of claim 1, wherein G1, G2 or Z2 comprise the following structure:

### Claim 12 (depends on 11)

The three-dimensional culture system of claim 11, wherein Z2 comprises the following structure:

### Claim 13 (depends on 1)

The three-dimensional culture system of claim 1, wherein G1 and/or G2 comprises the following structure: