Production method for t cells or nk cells, medium for culturing t cells or nk cells, method for culturing t cells or nk cells, method for maintaining undifferentiated state of undifferentiated t cells, and growth-accelerating agent for t cells or nk cells

Patent metadata

Field	Value
Publication number	US-2022017867-A1
Application number	US-201917311001-A
Country	US
Kind code	A1
Filing date	Dec 6, 2019
Priority date	Dec 6, 2018
Publication date	Jan 20, 2022
Grant date	—

Abstract

An object of the present invention is to provide a method for producing T cells or NK cells, a culture medium for culturing T cells or NK cells, a method for culturing T cells or NK cells, a method for maintaining the undifferentiated state of undifferentiated T cells, and a growth promoter for T cells or NK cells, which are capable of efficiently proliferating T cells or NK cells and maintaining the state of the cells (for example, undifferentiated property).The present invention relates to a method for producing T cells or NK cells including culturing T cells or NK cells in a culture medium containing a bisbenzylisoquinoline alkaloid represented by formula (X-1) or formula (X-2) or a compound resulting from cleavage of one ether bond thereof or a pharmaceutically acceptable salt thereof, and the like.

First claim

1 . A method for producing T cells or NK cells, comprising culturing T cells or NK cells in a culture medium containing a bisbenzylisoquinoline alkaloid represented by the following formula (X-1) or formula (X-2) or a compound resulting from cleavage of one ether bond thereof or a pharmaceutically acceptable salt thereof:

R 1 and R′ 1 are each independently H or straight-chain or branched-chain alkyl having 1 to 10 carbon atoms,
R 2 , R′ 2 , R 3 , R′ 3 , R 4 , R′ 4 , R 5 , and R′ 5 are each independently H, acyl, straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, and the alkyl may be interrupted by an O, N, or S heteroatom, or R 2 and R 3 , R 4 and R 5 , R′ 2 and R′ 3 , or R′ 4 and R′ 5 together represent O or S,
R 6 , R′ 6 , R 7 , and R′ 7 are each independently H, acyl, or straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, and the alkyl may be interrupted by an O, N, or S heteroatom, or R 6 and R 7 or R′ 6 and R′ 7 together represent O or S, and
X 1 , X 2 , X 3 , and X 4 may be the same or different, and are each independently H, hydroxy, straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, alkoxy, acyloxy, or sulfonyloxy, and each may further have a substituent, n 1 and n 3 are each independently an integer of 0 to 3, n 2 and n 4 are each independently an integer of 0 to 4, and these may be linked to each other to form a ring.

2 . The production method according to claim 1 , wherein the T cells or the NK cells are T cells or NK cells for immune cell therapy.

3 . The production method according to claim 1 , wherein the T cells or the NK cells are peripheral blood-derived, primary hematopoietic stem/progenitor cell-derived, or pluripotent stem cell-derived cells.

4 . The production method according to claim 1 , wherein the T cells are undifferentiated T cells.

5 . The production method according to claim 1 , wherein the T cells express at least one of CD4 and CD8.

6 . The production method according to claim 1 , wherein the T cells are cells selected from the group consisting of helper T cells, regulatory T cells, cytotoxic T cells, naive T cells, memory T cells, and terminal effector T cells.

7 . The production method according to claim 6 , wherein the regulatory T cells express FoxP3.

8 . The production method according to claim 6 , wherein the memory T cells are stem cell memory T cells, central memory T cells, or effector memory T cells.

9 . The production method according to claim 1 , wherein the NK cells are immature NK cells or mature NK cells.

10 . The production method according to claim 1 , wherein the bisbenzylisoquinoline alkaloid or the compound resulting from cleavage of one ether bond thereof or the pharmaceutically acceptable salt thereof is at least one selected from the group consisting of berbamine, (+)-berbamine, E6-berbamine, cepharanthine, and a pharmaceutically acceptable salt thereof.

11 . The production method according to claim 1 , wherein the concentration of the bisbenzylisoquinoline alkaloid or the compound resulting from cleavage of one ether bond thereof or the pharmaceutically acceptable salt thereof in the culture medium is from 0.1 nM to 10 μM.

12 . The production method according to claim 1 , wherein the culture medium further contains a MAPK cascade inhibitor.

13 . The production method according to claim 1 , wherein feeder cells are not contained in the culture medium.

14 . The production method according to claim 1 , wherein the culturing period is 50 days or more.

15 . T cells or TK cells produced by the production method according to claim 1 .

16 . An immune cell therapeutic agent, comprising the T cells or the TK cells according to claim 15 .

17 . A culture medium for culturing T cells or NK cells, comprising a bisbenzylisoquinoline alkaloid represented by the following formula (X-1) or formula (X-2) or a compound resulting from cleavage of one ether bond thereof or a pharmaceutically acceptable salt thereof:

R 1 and R′ 1 are each independently H or straight-chain or branched-chain alkyl having 1 to 10 carbon atoms,
R 2 , R′ 2 , R 3 , R′ 3 , R 4 , R′ 4 , R 5 , and R′ 5 are each independently H, acyl, straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, and the alkyl may be interrupted by an O, N, or S heteroatom, or R 2 and R 3 , R 4 and R 5 , R′ 2 and R′ 3 , or R′ 4 and R′ 5 together represent O or S,
R 6 , R′ 6 , R 7 , and R′ 7 are each independently H, acyl, or straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, and the alkyl may be interrupted by an O, N, or S heteroatom, or R 6 and R 7 or R′ 6 and R′ 7 together represent O or S, and
X 1 , X 2 , X 3 , and X 4 may be the same or different, and are each independently H, hydroxy, straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, alkoxy, acyloxy, or sulfonyloxy, and each may further have a substituent, n 1 and n 3 are each independently an integer of 0 to 3, n 2 and n 4 are each independently an integer of 0 to 4, and these may be linked to each other to form a ring.

18 . The culture medium according to claim 17 , wherein the T cells or the NK cells are peripheral blood-derived, primary hematopoietic stem/progenitor cell-derived, or pluripotent stem cell-derived cells.

19 . The culture medium according to claim 17 , wherein the bisbenzylisoquinoline alkaloid or the compound resulting from cleavage of one ether bond thereof or the pharmaceutically acceptable salt thereof is at least one selected from the group consisting of berbamine, (+)-berbamine, E6-berbamine, cepharanthine, and a pharmaceutically acceptable salt thereof.

20 . The culture medium according to claim 17 , wherein the concentration of the bisbenzylisoquinoline alkaloid or the compound resulting from cleavage of one ether bond thereof or the pharmaceutically acceptable salt thereof is from 0.1 nM to 10 μM.

21 . The culture medium according to claim 17 , further comprising a MAPK cascade inhibitor.

22 . A method for culturing T cells or NK cells, comprising culturing T cells or NK cells in a culture medium containing a bisbenzylisoquinoline alkaloid represented by the following formula (X-1) or formula (X-2) or a compound resulting from cleavage of one ether bond thereof or a pharmaceutically acceptable salt thereof:

R 1 and R′ 1 are each independently H or straight-chain or branched-chain alkyl having 1 to 10 carbon atoms,
R 2 , R′ 2 , R 3 , R′ 3 , R 4 , R′ 4 , R 5 , and R′ 5 are each independently H, acyl, straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, and the alkyl may be interrupted by an O, N, or S heteroatom, or R 2 and R 3 , R 4 and R 5 , R′ 2 and R′ 3 , or R′ 4 and R′ 5 together represent O or S,
R 6 , R′ 6 , R 7 , and R′ 7 are each independently H, acyl, or straight-chain or branched-chain alkyl having 1 to 10 carbon atoms, and the alkyl may be interrupted by an O, N, or S heteroatom, or R 6 and R 7 or R′ 6 and R′ 7 together represent O or S, and
X 1 , X 2 , X 3 , and X 4 may be the same or different, and are each independently H, h

Assignees

• Kirin Holdings Kk
• Univ Kyoto

Inventors

• Takayanagi Shinichiro
• Kunisato Atsushi
• CHUGANJI Sayaka
• Fukumoto Ken
• NAKAZONO Kazuki
• Kaneko Shin
• Kawai Yohei

Classifications

• C07D471/18 Primary

  Bridged systems · CPC title

• C12N5/0636

  T lymphocytes · CPC title

• C12N5/0646 Primary

  Natural killers cells [NK], NKT cells · CPC title

• C12N5/0637

  Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg · CPC title

• C12N2501/999

  Small molecules not provided for elsewhere · CPC title