# US 2019/0194376 A1 · Block copolymer and surface treatment agent using same

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- Number: US 2019/0194376 A1
- Document ID: US20190194376A1
- Status: Published application
- Kind: A1
- Application number: 16/322,361
- Filed: 2017-07-28
- Published: 2019-06-27
- Earliest claimed: 2016-08-03
- Assignees: Tosoh Corporation
- Applicants: Tosoh Corporation
- Inventors: Yukie MAEJIMA; Satoru KONDOU; Shinya IMATOMI; Satoru YAMADA; Hiroyuki ITO
- Web page: https://patentgrep.com/patent/US20190194376A1

## CPC

- C08F 293/005: {using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent}
- C08F 299/024: {the unsaturation being in acrylic or methacrylic groups}
- C08F 20/18: with acrylic or methacrylic acids
- C08F 20/56: Acrylamide; Methacrylamide
- C08F 293/00: Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule (on to polymers modified by introduction of unsaturated end groups C08F290/02)
- C08G 61/025: {Polyxylylenes}
- C08G 81/02: at least one of the polymers being obtained by reactions involving only carbon-to-carbon unsaturated bonds
- C08G 81/024: {Block or graft polymers containing sequences of polymers of C08C or C08F and of polymers of C08G}
- C12M 3/00: Tissue, human, animal or plant cell, or virus culture apparatus
- C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)}
- 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]
- C08G 2261/126: block
- C08G 2261/1424: containing ether groups, including alkoxy
- C08G 2261/143: Side-chains containing nitrogen
- C08G 2261/76: crosslinking
- C12N 2533/30: Synthetic polymers
- C08F: MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08G: MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention addresses the problem of providing a block copolymer which is useful as a surface treatment agent for cell culture substrates, said surface treatment agent enabling cell separation in a short period of time. The above-mentioned problem is solved by a block copolymer that includes the following blocks (A), (B) and (C):(A) a temperature-responsive polymer block that has a lower critical solution temperature (LCST) within the range of from 0° C. to 50° C. with respect to water (B) a hydrophilic polymer block that does not have an LCST within the range of from 0° C. to 50° C., while having an HLB value within the range of from 9 (inclusive) to 20 (exclusive) (C) a hydrophobic polymer block that does not have an LCST within the range of from 0° C. to 50° C., while having an HLB value within the range of from 0 (inclusive) to 9 (exclusive).

## Claims

### Claim 1

A block copolymer comprising the following blocks (A), (B), and (C):
(A) a temperature-responsive polymer block having a lower critical solution temperature (LCST) in water in the range of 0° C. to 50° C.;
(B) a hydrophilic polymer block which does not have an LCST in the range of 0° C. to 50° C. and which has an HLB value (as determined by the Griffin method) in the range of 9 to 20; and
(C) a hydrophobic polymer block which does not have an LCST in the range of 0° C. to 50° C. and which has an HLB value (as determined by the Griffin method) in the range from 0 to less than 9.

### Claim 2 (depends on 1)

The block copolymer according to claim 1, wherein the block (A) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (1):
where R2 is a hydrogen atom or methyl group, R3 and R are each independently a hydrogen atom, C1-6 hydrocarbon group, C2-4 hydrocarbon group which may be substituted with a C1-2 alkyloxy group, C2-4 hydrocarbon group which may be substituted with fluorine, furfuryl group, or tetrahydrofurfuryl group, and R3 and R4 may be connected to form a pyrrolidine ring, piperidine ring or morpholine ring.

### Claim 3 (depends on 1)

The block copolymer according to claim 1, wherein the block (A) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (2):
where R5 is a hydrogen atom or methyl group, R6 is a hydrogen atom or C1-4 hydrocarbon group, and r is an integer from 1 to 10.

### Claim 4 (depends on 1)

The block copolymer according to claim 1, wherein the block (A) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (3):
where R7 is a hydrogen atom or methyl group and R8 is a C1s hydrocarbon group.

### Claim 5 (depends on 1)

The block copolymer according to claim 1, wherein the block (B) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (4):
where R9 is a hydrogen atom or methyl group, R10 is a C1-10 alkylene group, R11 is a C1-4 divalent hydrocarbon group, R12, R13, and R14 are each independently a hydrogen atom, methyl group, or ethyl group, and A1 is a divalent bond selected from the group consisting of an ester bond, amide bond, urethane bond, and ether bond.

### Claim 6 (depends on 1)

The block copolymer according to claim 1, wherein the block (B) is a polymer comprising at least one repeating unit from among the repeating units represented by the following Formula (5):
where R15 is a hydrogen atom or methyl group and R16 is —(CH2CH2O)i—(CH2O)j—(CH2CH(CH3)O)k—R17 (where R17 is a hydrogen atom or C1-10 alkyl group, i is an integer from 1 to 30, and j and k are each independently an integer from 1 to 30).

### Claim 7 (depends on 1)

The block copolymer according to claim 1, wherein the block (B) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (6):
where R19 is a hydrogen atom or methyl group, R2 is a C1-10 alkylene group, R21 is a C1-4 alkylene group, R22 and R23 are each independently a hydrogen atom or C1-4 hydrocarbon group, A2 is a divalent bond selected from the group consisting of an ester bond, amide bond, urethane bond, and ether bond, and X is a sulfonic acid group, carboxyl group, or phosphoric acid group.

### Claim 8 (depends on 1)

The block copolymer according to claim 1, wherein the block (B) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (7):
where R24, R25, and R26 are each independently a hydrogen atom or methyl group.

### Claim 9 (depends on 1)

The block copolymer according to claim 1, wherein the block (B) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (8):
where R2 is a hydrogen atom or methyl group, R29 is a C2-7 alkylene group, R30 and R31 are each independently a hydrogen atom, methyl group, or ethyl group, and A3 is a divalent bond selected from the group consisting of an ester bond, amide bond, urethane bond, and ether bond.

### Claim 10 (depends on 1)

The block copolymer according to claim 1, wherein the block (B) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (9):
where R28 is a hydrogen atom or methyl group, R29 is a C2-7 alkylene group, R30 and R31 are each independently a hydrogen atom, methyl group, or ethyl group, R32 is a C1-4 hydrocarbon group or C2-4 hydrocarbon group which may be substituted with a hydroxyl group or C1-2 alkyloxy group, A3 is a divalent bond selected from the group consisting of an ester bond, amide bond, urethane bond, and ether bond, and X− is a halide ion, hydroxide ion, or acetate ion.

### Claim 11 (depends on 1)

The block copolymer according to claim 1, wherein the block (C) is a polymer comprising at least one repeating group from among repeating groups represented by the following Formula (10):
where R33 is a hydrogen atom or methyl group, and Y is a hydrogen atom, chlorine atom, acetoxy group, nitrile group, or C6-30 aromatic hydrocarbon group.

### Claim 12 (depends on 1)

The block copolymer according to claim 1, wherein the block (C) is a polymer comprising at least one repeating unit from among repeating units represented by the following Formula (11):
where R is a hydrogen atom or methyl group, R35 is a C1-30 hydrocarbon group, and Z is a divalent bond selected from the group consisting of an ester bond, amide bond, urethane bond, and ether bond.

### Claim 13 (depends on 1)

The block copolymer according to claim 1, wherein block (A), block (B), and block (C) constituting the block copolymer have the following mol % (a) to (c) with respect to the total thereof, respectively:
(a) the ratio of block (A) is 25 mol % to 85 mol %;
(b) the ratio of block (B) is 2 mol % to 50 mol %; and
(c) the ratio of block (C) is 10 mol % to 70 mol %.

### Claim 14 (depends on 1)

The block copolymer according to claim 1, wherein the number average molecular weight (Mn) of the block copolymer is in the range of 3,000 to 1,000,000.

### Claim 15 (depends on 1)

The block copolymer according to claim 1, comprising at least one bond via a spacer between blocks (A), (B), and (C), wherein at least one of the bonds via the spacer is a divalent bond comprising at least one bond from among divalent bonds represented by the following Formulae (12) and (13):
where R1 is a hydrogen atom or C1-20 hydrocarbon group.

### Claim 16 (depends on 1)

A method for the production of the block copolymer according to claim 1, comprising the following steps (1) to (3):
(1) producing any one of the blocks from among blocks (A), (B), and (C);
(2) producing a partial block copolymer comprising the block produced in step (1) and, connected thereto, one of the blocks from among blocks (A), (B), and (C) except the block produced in step (1); and
(3) producing a block copolymer comprising the partial block copolymer produced in step (2) and, connected thereto, the block among blocks (A), (B), and (C) which does not constitute a block copolymer comprising the partial block copolymer produced in step (2).

### Claim 17 (depends on 1)

A surface treatment agent for substrates, comprising the block copolymer according to claim 1.

### Claim 18 (depends on 17)

A membrane comprising the surface treatment agent according to claim 17 applied to a substrate.

### Claim 19 (depends on 18)

A substrate for cell culture having a surface coated with the membrane according to claim 18.

### Claim 20 (depends on 19)

A cell culture method, comprising culturing, using the cell culture substrate according to claim 19, a cell at a temperature higher than the LCST of the temperature-responsive polymer block, and after cell proliferation, lowering the temperature below the LCST to detach proliferated cells from the substrate.