Composition, culture medium, cell population, cell production method, and therapeutic drug

Abstract

The present invention provides: a composition (1) comprising a KDM5 decomposing agent constituting component (A), and an LSD1 inhibitor constituting component (B); or a composition (2) comprising the KDM5 decomposing agent constituting component (A). In the composition (2), the component (A) comprises at least one substance selected from the group consisting of compounds represented by general formula (I), salts thereof, compounds represented by general formula (II), and salts thereof.

Description

Compositions, media, cell populations, cell production methods, and therapeutic agents

[0001] The technical field of the present disclosure relates to compositions for cell culture, media, cell populations, cell production methods, cells, and therapeutic agents. This application claims priority to Japanese Patent Application No. 2024-009804, filed January 25, 2024, the contents of which are incorporated herein by reference. The content of WO 2024 / 210126, filed April 2, 2024, and published October 10, 2024, is incorporated herein by reference. The content of "Aryl hydrocarbon receptor-dependent enrichment of a megakaryocytic precursor with a high potential to produce proplatelets," Catherine Strassel et al., Blood (2016) 127 (18): 2231-2240, published May 5, 2016, is incorporated herein by reference.

[0002] Hematopoietic stem cells are stem cells that can differentiate into blood cells. Transplantation of hematopoietic stem cells into patients with diseases such as cancer and immunodeficiency is known as a treatment for these diseases.

[0003] Umbilical cord blood (CB) is known as a source of hematopoietic stem cells. For example, Patent Document 1 describes a method for obtaining hematopoietic stem cells from human umbilical cord blood (hereinafter also referred to as "human CB"). Specifically, human CB CD34 ＋ From the starting population of cells, EPCR ＋ Selection and culturing of populations of cells is described. EPCR is a hematopoietic stem cell marker, also known as CD201.

[0004] Furthermore, methods for culturing hematopoietic stem cells have been studied. Generally, in culturing hematopoietic stem cells, albumin-containing media have been used to induce cell division. However, albumin has problems such as the inhibition of stable undifferentiation and biological contamination. Therefore, research on albumin-free culture methods has been conducted. For example, Patent Document 2 describes the culture of human hematopoietic stem cells by adding PVA or Soluplus (registered trademark) instead of albumin. Patent Document 3 describes a method for producing megakaryocytes (cells that become platelets) from hematopoietic progenitor cells, which is characterized by culturing hematopoietic progenitor cells in a medium containing a pyrimidoindole derivative.

[0005] WO2017 / 205977WO2021 / 149799WO2024 / 210126

[0006] Conventional methods have not been sufficient for efficiently obtaining human hematopoietic stem cells. Furthermore, there have been few reports on albumin-free culture methods for human hematopoietic stem cells, and available media are limited.

[0007] In the course of their research, the present inventors have investigated a method for culturing hematopoietic stem cells. As a result, they have discovered that human CB CD34 cells can be cultured in an albumin-free medium with a specific composition different from conventional methods. ＋ By culturing the cells, hematopoietic stem cells were successfully proliferated. Furthermore, the obtained culture fraction contained CD201 ＋ or CD90 ＋ (all hematopoietic stem cell markers) was a culture fraction enriched for hematopoietic stem cells. The present inventors have completed the present disclosure based on this finding.

[0008] The present disclosure has the following aspects: [1] A composition (1) comprising a KDM5 degrader as component (A) and an LSD1 inhibitor as component (B), or a composition (2) comprising a KDM5 degrader as component (A), wherein component (A) comprises at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof: In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10. In general formula (II), R is a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms. [2] The composition according to [1], wherein the composition is composition (1), and the component (A) contains at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof: In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10. In the general formula (II), R is a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms. [3] In the general formula (I), A is a single bond or a phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ The composition according to [1], wherein in the general formula (II), R is a hydrogen atom or a linear alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 18 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 18 carbon atoms. [4] In the general formula (I), A is a single bond, a p-phenylene group, or an m-phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ - or -(CH ２ ) ｔThe composition according to [1], wherein component (A) is a group represented by the general formula (II) below: O-, s is a number from 1 to 5, and t is a number from 2 to 10, and in general formula (II), R is a linear alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 3 to 16 carbon atoms, or a substituted or unsubstituted branched alkylene group having 3 to 16 carbon atoms. [5] The composition according to [1], wherein component (A) is at least one selected from the group consisting of a compound represented by the following general formula (I-1) and a salt thereof, a compound represented by the following general formula (I-2) and a salt thereof, a compound represented by the following general formula (I-3) and a salt thereof, and a compound represented by the following general formula (II-1) and a salt thereof. In general formula (I-1), L １ is a linear alkylene group having 3 to 10 carbon atoms, and X １ Ha-(CH ２ CH ２ O) ｓ -, s is a number of 1 to 5, and in general formula (I-2), L ２ is a linear alkylene group having 3 to 10 carbon atoms, and X ２ Ha-(CH ２ CH ２ O) ｓ - or - (CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10; and in general formula (I-3), L ３ is a linear alkylene group having 3 to 10 carbon atoms, and X ３ Ha-(CH ２ CH ２ O) ｓ - or - (CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10, and in general formula (II-1), R １ is a linear alkyl group having 1 to 3 carbon atoms, and Z １ is a substituted or unsubstituted linear alkylene group having 3 to 14 carbon atoms, or a substituted or unsubstituted branched alkylene group having 3 to 14 carbon atoms. [6] The component (A) is １ is a linear alkylene group having 5 carbon atoms, and X １ Ga-(CH２ CH ２ O) ｓ -, and s is 3; １ is a linear alkylene group having 8 carbon atoms, and X １ Ga-(CH ２ CH ２ O) ｓ -, and s is 3; ２ is a linear alkylene group having 5 carbon atoms, and X ２ Ga-(CH ２ CH ２ O) ｓ -, and s is the number 3; ２ is a linear alkylene group having 5 carbon atoms, and X ２ Ga-(CH ２ ) ｔ a compound in which L is a group represented by O— and t is the number 6; ３ is a linear alkylene group having 5 carbon atoms, and X ３ Ga-(CH ２ CH ２ O) ｓ -, and s is the number 3; ３ is a linear alkylene group having 5 carbon atoms, and X ３ Ga-(CH ２ ) ｔ O—, and t is the number 6, or １ is a methyl group, and Z １is an unsubstituted linear alkylene group having 9 carbon atoms. [7] The composition according to [1], wherein the composition is composition (1), and component (B) comprises at least one component selected from the group consisting of tranylcypromine and salts thereof. [8] The composition according to [6], wherein the composition is composition (1), and component (B) comprises at least one component selected from the group consisting of tranylcypromine and salts thereof. [9] A culture medium comprising the composition according to any one of [1] to [8].

[10] The culture medium according to [9], wherein the culture medium is a culture medium for culturing human hematopoietic stem cells.

[11] The culture medium according to [9], wherein the culture medium is a serum-free medium.

[12] The culture medium according to [9], wherein the culture medium is an albumin-free medium.

[13] The culture medium is a culture medium for culturing CD34 ＋ cells, CD201 ＋ cells, CD90 ＋ and CD41 ＋

[14] The medium according to [9], comprising at least one type of cell selected from the group consisting of cells of CD201. ＋ cells, CD90 ＋ and CD41 ＋

[15] A composition used for culturing cells obtained by culturing in the medium according to [9].

[16] The composition according to

[15] , wherein the cells obtained by culturing in the medium are human hematopoietic stem cells.

[17] The composition according to

[15] , wherein the cells obtained by culturing in the medium are human hematopoietic stem cells.

[18] The composition according to

[18] , wherein the cells obtained by culturing in the medium are human hematopoietic stem cells. ＋ or CD41 ＋

[18] A cell population obtained by culturing in the medium according to [9].

[19] The cell population according to

[18] , which comprises human hematopoietic stem cells.

[20] The cell population, which is a CD201 ＋ or CD41 ＋

[21] At least one therapeutic agent selected from the group consisting of a cancer therapeutic agent, an immunodeficiency therapeutic agent, and a multiple myeloma therapeutic agent, comprising cells obtained by culturing in the medium according to [9].

[22] A method for producing cells, comprising a step of culturing cells using the medium according to [9].

[23] The culturing step comprises obtaining a medium from the cells and the composition.

[24] The method for producing cells according to

[22] , comprising a step of culturing the cells cultured in the culturing step using another medium.

[25] The method for producing cells according to

[22] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[26] The method for producing cells according to

[22] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[27] The method for producing cells according to

[27] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[28] The method for producing cells according to

[28] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[29] The method for producing cells according to

[29] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[30] The method for producing cells according to

[30] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[31] The method for producing cells according to

[31] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[32] The method for producing cells according to

[31] , wherein the cultured cells are a cell population containing human hematopoietic stem cells.

[33] The method for producing cells according to

[31] , wherein the cultured cells are a cell population containing human hematopoietic stem cells. ＋ or CD41 ＋

[27] A cell population produced by the method of

[22] .

[28] The cell population of

[27] , which comprises human hematopoietic stem cells.

[29] The cell population, which is a cell population encoding CD201 ＋ or CD41 ＋

[30] At least one therapeutic agent selected from the group consisting of a therapeutic agent for cancer, a therapeutic agent for immunodeficiency, and a therapeutic agent for multiple myeloma, comprising cells obtained by the method according to

[22] .

[31] Use of a drug comprising at least one component selected from the group consisting of a compound represented by the following formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof, for producing a medium for expanding hematopoietic stem cells. In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O)ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10. In general formula (II), R is a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

[32] Use of a pharmaceutical agent comprising at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof, as a pharmaceutical agent for a culture medium for proliferation of hematopoietic stem cells. In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10. In general formula (II), R is a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

[33] Use of the composition according to any one of [1] to [8] for expanding hematopoietic stem cells.

[34] The use according to

[33] , wherein the hematopoietic stem cells are for differentiation into megakaryocytes.

[35] Use of the composition according to any one of [1] to [8] as a medium agent for expanding hematopoietic stem cells for differentiation into megakaryocytes.

[36] Use of the composition according to any one of [1] to [8] for producing a medium agent for expanding hematopoietic stem cells for differentiation into megakaryocytes.

[37] The composition according to any one of [1] to [8], wherein the composition is for expanding hematopoietic stem cells.

[38] The composition according to

[37] , wherein the hematopoietic stem cells are for differentiation into megakaryocytes.

[39] The composition according to

[37] or

[38] , wherein the composition does not contain serum.

[40] The composition according to any one of

[37] to

[39] , wherein the composition is albumin-free.

[41] The composition contains CD34 ＋ cells, CD201 ＋ cells, CD90 ＋ and CD41 ＋

[42] The composition according to any one of

[37] to

[40] , comprising at least one cell selected from the group consisting of CD201 ＋ cells, CD90 ＋ and CD41 ＋ The composition according to any one of

[37] to

[41] , comprising a cell population enriched for at least one type of cell selected from the group consisting of cells of

[0009] According to the present disclosure, compositions and media for use in cell culture can be provided. Furthermore, according to the present disclosure, cell populations that can be transplanted into patients with diseases such as cancer and immunodeficiency and methods for producing the same can be provided. Furthermore, according to the present disclosure, therapeutic agents containing the cell populations can be provided.

[0010] 1 shows the expression analysis results of the cell population in Example 1. FIG. 1 shows the expression analysis results of the cell population in Example 2. FIG. 1 shows the expression analysis results of the cell population in Example 3. FIG. 1 shows the expression analysis results of the cell population in Example 4. FIG. 2 shows the expression analysis results of the cell population in Example 5. FIG. 3 shows the expression analysis results of the cell population in Example 6. FIG. 4 shows the expression analysis results of the cell population in Example 7. FIG. 5 shows the expression analysis results of the cell population in Example 8. FIG. 6 shows the expression analysis results of the cell population in Example 9. FIG. 7 shows the expression analysis results of the cell population in Example 10. FIG. 8 shows the expression analysis results of the cell population in Example 11. FIG. 9 shows the expression analysis results of the cell population in Example 12. FIG. 11 shows the expression analysis results of the cell population in Example 13. FIG. 12 shows the expression analysis results of the cell population in Example 14. FIG. 13 shows the expression analysis results of the cell population in Example 15. FIG. 14 shows the expression analysis results of the cell population in Example 16. FIG. 17 shows the expression analysis results of the cell population in Example 18. FIG. 19 shows the expression analysis results of the cell population in Example 20. FIG. 21 shows the expression analysis results of the cell population in Example 21. FIG. 2 shows the results of expression analysis of the cell population in Example 22. FIG. 2 shows the results of expression analysis of the cell population in Example 23. FIG. 2 shows the results of expression analysis of the cell population in Example 24. FIG. 2 shows the results of expression analysis of the cell population in Example 25. FIG. 2 shows the results of expression analysis of the cell population in Example 26. FIG. 2 shows the results of expression analysis of the cell population in Example 27. FIG. 2 shows the results of expression analysis of the cell population in Example 28. The results of expression analysis of the mature megakaryocytes (CD41 ＋ CD42b ＋ 1 shows the ratio of mature megakaryocytes (CD41 cells) to the cell population after 14 days of culture in Example 30. ＋ CD42b ＋ Figure 1 shows the ratio of erythrocytes (cells) to total erythrocytes (cells).

[0011] Hereinafter, embodiments of the present disclosure will be described in detail. Note that similar content will be omitted where appropriate to avoid repetition. Unless otherwise specified, an "alkyl group" is intended to include linear, branched, and cyclic monovalent saturated hydrocarbon groups. The same applies to alkyl groups in alkoxy groups. Unless otherwise specified, an "alkylene group" is intended to include linear, branched, and cyclic divalent saturated hydrocarbon groups. Unless otherwise specified, an "alkenylene group" is intended to include linear, branched, and cyclic divalent unsaturated hydrocarbon groups. Examples of "halogen atom" include a fluorine atom, chlorine atom, bromine atom, and iodine atom. The term "substituted or unsubstituted" refers to a case where a hydrogen atom (-H) is substituted with a monovalent substituent, and a case where a methylene group (-CH ２-) or methine group (>CH- or ═CH-) is substituted (replaced) with a divalent substituent. Examples of "monovalent substituent" include an alkyl group, an alkoxy group, a halogen atom, a halogenated alkyl group, a hydroxy group, a carboxy group, an amino group, and a nitro group. As the alkyl group as a substituent, an alkyl group having 1 to 5 carbon atoms is preferred, and a methyl group, an ethyl group, a propyl group, an n-butyl group, or a tert-butyl group is most preferred. As the alkoxy group as a substituent, an alkoxy group having 1 to 5 carbon atoms is preferred, and a methoxy group, an ethoxy group, an n-propoxy group, an iso-propoxy group, an n-butoxy group, or a tert-butoxy group is more preferred, and a methoxy group or an ethoxy group is most preferred. As the halogen atom as a substituent, a fluorine atom or an iodine atom is preferred. Examples of halogenated alkyl groups as substituents include alkyl groups having 1 to 5 carbon atoms, such as methyl, ethyl, propyl, n-butyl, and tert-butyl, in which some or all of the hydrogen atoms have been substituted with the halogen atoms. The "divalent substituent" includes -O-, -C(=O)-O-, -O-C(=O)-, -C(=O)-, -O-C(=O)-O-, -C(=O)-NH-, -C(=O)-NH-C(=O)-, -N=, -NH-, -NH-C(=NH)- (H may be substituted with a substituent such as a hydrocarbon group, an acyl group, or an alkoxyalkyl group), -S-, and -S(=O). ２ -, -S(=O) ２ -O-, general formula -Y ２１ -O-Y ２２ -, -Y ２１ -O-, -Y ２１ -C(=O)-O-, -C(=O)-O-Y ２１ -, -[Y ２１ -C(=O)-O] ｍ” -Y ２２ -, -Y ２１ -OC(=O)-Y ２２ -or-Y ２１ -S(=O) ２ -O-Y ２２ -, wherein Y ２１ and Y ２２are each independently a substituted or unsubstituted divalent hydrocarbon group, O is an oxygen atom, and m″ is an integer of 0 to 3.

[0012] (1) Composition According to one embodiment of the present disclosure, there is provided a composition (1) comprising a KDM5 degrader as component (A) and an LSD1 inhibitor as component (B), or a composition (2) comprising a KDM5 degrader as component (A), wherein component (A) comprises at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof (hereinafter, compositions (1) and (2) may be collectively referred to simply as the “composition”).

[0013]

[0014] In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10.

[0015]

[0016] In general formula (II), R represents a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z represents a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

[0017] In the general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear or branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10.

[0018] As the A, a single bond, a phenylene group, a naphthylene group, and the like are preferred; a single bond, a 1,2-phenylene group, a 1,3-phenylene group (m-phenylene group), a 1,4-phenylene group (p-phenylene group), a 1,2-naphthylene group, a 1,4-naphthylene group, a 1,5-naphthylene group, a 2,3-naphthylene group, a 2,6-naphthylene group, and the like are more preferred; a single bond, a 1,3-phenylene group (m-phenylene group), and a 1,4-phenylene group (p-phenylene group) are even more preferred.

[0019] L is preferably a linear alkylene group having 3 to 10 carbon atoms, more preferably a linear alkylene group having 4 to 9 carbon atoms, and even more preferably a linear alkylene group having 5 to 8 carbon atoms.

[0020] The X may be -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ A group represented by -O- is preferred; ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH２ O) ｓ - or -(CH ２ ) ｔ A group represented by O- is more preferred. The s is preferably a number of 1 to 5, more preferably 2 to 4, and even more preferably 2 or 3. The t is preferably a number of 2 to 10, more preferably 3 to 8, and even more preferably 4 to 7.

[0021] In the general formula (I), A is a single bond or a phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is —(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is preferable that A is a single bond, a p-phenylene group, or an m-phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ - or -(CH ２ ) ｔ It is more preferable that the group is represented by O—, where s is a number from 1 to 5 and t is a number from 2 to 10.

[0022] In general formula (II), R is preferably a linear or branched alkyl group having 1 to 6 carbon atoms, more preferably a linear or branched alkyl group having 1 to 3 carbon atoms, still more preferably a methyl group or an ethyl group, and particularly preferably a methyl group. Z is preferably a linear or branched alkylene group having 2 to 20 carbon atoms, more preferably a linear or branched alkylene group having 3 to 18 carbon atoms, still more preferably a linear or branched alkylene group having 5 to 15 carbon atoms, particularly preferably an n-hexylene group, an n-heptylene group, an n-octylene group, an n-nonylene group, an n-decylene group, or an n-undecylene group, and most preferably an n-octylene group, an n-nonylene group, or an n-decylene group.

[0023] In the composition (1), it is preferable that the component (A) contains at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof:

[0024]

[0025] In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ It is a group represented by O—, in which s is a number from 1 to 8, and t is a number from 2 to 10.

[0026]

[0027] In general formula (II), R represents a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z represents a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

[0028] In the general formula (I), A is a single bond or a phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is —(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ O-, s is a number from 1 to 5, and t is a number from 2 to 10, and in the general formula (II), R is a hydrogen atom or a linear alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 18 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 18 carbon atoms.

[0029] In the general formula (I), A is a single bond, a p-phenylene group, or an m-phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is —(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ - or -(CH ２ ) ｔ O-, s is a number from 1 to 5, and t is a number from 2 to 10, and in the general formula (II), R is a linear alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 3 to 16 carbon atoms, or a substituted or unsubstituted branched alkylene group having 3 to 16 carbon atoms.

[0030] The component (A) is preferably at least one compound selected from the group consisting of compounds represented by the following general formula (I-1) and salts thereof, compounds represented by the following general formula (I-2) and salts thereof, compounds represented by the following general formula (I-3) and salts thereof, and compounds represented by the following general formula (II-1) and salts thereof:

[0031]

[0032] In general formula (I-1), L １ is a linear alkylene group having 3 to 10 carbon atoms, and X １ Ha-(CH ２ CH ２ O) ｓ -, s is a number of 1 to 5, and in general formula (I-2), L ２ is a linear alkylene group having 3 to 10 carbon atoms, and X ２ Ha-(CH ２ CH ２ O) ｓ - or - (CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10; and in general formula (I-3), L ３ is a linear alkylene group having 3 to 10 carbon atoms, and X ３ Ha-(CH ２ CH ２ O) ｓ - or - (CH ２ ) ｔ It is a group represented by O—, where s is a number from 1 to 5, and t is a number from 2 to 10. In general formula (II-1), R １ is a linear alkyl group having 1 to 3 carbon atoms, and Z １ is preferably a substituted or unsubstituted linear alkylene group having 3 to 14 carbon atoms, or a substituted or unsubstituted branched alkylene group having 3 to 14 carbon atoms.

[0033] The component (A) is, in the general formula (I-1), L １ is a linear alkylene group having 5 carbon atoms, and X １ Ga-(CH ２ CH ２ O) ｓ-, and s is 3; １ is a linear alkylene group having 8 carbon atoms, and X １ Ga-(CH ２ CH ２ O) ｓ -, and s is 3; ２ is a linear alkylene group having 5 carbon atoms, and X ２ Ga-(CH ２ CH ２ O) ｓ -, and s is the number 3; ２ is a linear alkylene group having 5 carbon atoms, and X ２ Ga-(CH ２ ) ｔ a compound in which L is a group represented by O— and t is the number 6; ３ is a linear alkylene group having 5 carbon atoms, and X ３ Ga-(CH ２ CH ２ O) ｓ -, and s is the number 3; ３ is a linear alkylene group having 5 carbon atoms, and X ３ Ga-(CH ２ ) ｔ O—, and t is the number 6, or １ is a methyl group, and Z １ is an unsubstituted linear alkylene group having 9 carbon atoms.

[0034] Among them, the compound represented by the general formula (I) and its salts are preferably a compound represented by the general formula (I) in which A is an m-phenylene group, L is an n-pentylene group (a linear alkylene group having 5 carbon atoms), and X is -(CH ２ ) ｔA compound in which R is a group represented by O- and t is 6 (hereinafter also referred to as "compound 126") or a salt thereof is preferred. As the compound represented by general formula (II) and a salt thereof, a compound in which R is a methyl group and Z is an n-nonylene group (a linear alkylene group having 9 carbon atoms) (hereinafter also referred to as "compound 135") or a salt thereof is preferred.

[0035] The composition is preferably composition (1), and the component (B) contains at least one component selected from the group consisting of tranylcypromine and its salts represented by the following general formula (B-1) or (B-2): Of these, tranylcypromine and its salts are more preferably tranylcypromine and its salts represented by the following general formula (B-2), and even more preferably tranylcypromine hydrochloride represented by the following general formula (B-2-1):

[0036]

[0037]

[0038]

[0039] Salts of the compounds include all pharmacologically acceptable salts. All pharmacologically acceptable salts are preferably low-toxicity, water-soluble salts. Examples of suitable salts include acid addition salts (e.g., inorganic acid salts [e.g., hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, nitrate, etc.], organic acid salts [e.g., acetate, trifluoroacetate, lactate, tartrate, oxalate, fumarate, maleate, benzoate, citrate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, isethionate, glucuronate, gluconate, etc.], salts with acidic natural amino acids [e.g., aspartic acid, glutamic acid, etc.], etc.).

[0040] The compound represented by the general formula (I) can be produced by a method similar to the production method described in Japanese Patent Application No. 2023-143583. The compound represented by the general formula (II) can be produced by a method similar to the production method described in ChemMedChem. 2021 May 18; 16 (10): 1609-1618.

[0041] The composition (1) can be produced by mixing the KDM5 decomposer (component (A)), the LSD1 inhibitor (component (B)), and, if necessary, other optional base components. The composition (2) can be produced by mixing, as the KDM5 decomposer (component (A)), at least one component selected from the group consisting of the compound represented by the general formula (I) above and a salt thereof, and the compound represented by the general formula (II) above and a salt thereof, and, if necessary, other components. Examples of the other components include the optional base components described below.

[0042] In the composition (1), the content of the compound (I) of the component (A) is preferably 0.1 to 100 μmol / L, more preferably 0.3 to 30 μmol / L, and even more preferably 1 to 10 μmol / L, relative to the total amount (1 L) of the composition. In the composition (1), the content of the compound (II) of the component (A) is preferably 0.1 to 100 μmol / L, more preferably 0.3 to 30 μmol / L, and even more preferably 1 to 10 μmol / L, relative to the total amount (1 L) of the composition. In the composition (1), the content of the component (B) is preferably 0.1 to 100 μmol / L, more preferably 0.3 to 30 μmol / L, and even more preferably 1 to 10 μmol / L, relative to the total amount (1 L) of the composition. In the composition (2), the content of compound (I) of component (A) is preferably 0.1 to 100 μmol / L, more preferably 0.3 to 30 μmol / L, and even more preferably 1 to 10 μmol / L, relative to the total amount (1 L) of the composition. In the composition (2), the content of compound (II) of component (A) is preferably 0.1 to 100 μmol / L, more preferably 0.3 to 30 μmol / L, and even more preferably 1 to 10 μmol / L, relative to the total amount (1 L) of the composition. When the composition contains albumin, the content of albumin is preferably 0.1 to 20% by mass, more preferably 0.5 to 10% by mass, and even more preferably 1 to 4% by mass, relative to the total mass of the composition.

[0043] Here, the composition may be a culture medium. The culture medium may be a culture medium for culturing human hematopoietic stem cells. The composition may further contain hematopoietic stem cells. By using this composition, CD201 ＋ or CD90 ＋ For example, in the Examples below, this composition is used to efficiently produce a culture fraction rich in human CB CD34 cells. ＋ As a result of culturing the cells, CD201 ＋ Cells or CD90 ＋It is described that a cell-enriched culture fraction was obtained. Furthermore, according to one embodiment of the present disclosure, a method for producing cells, a method for culturing cells, a method for producing a medium, and a method for producing CD201 using this composition are also disclosed. ＋ or CD90 ＋ A method for concentrating cells of the present invention and a container containing the composition are provided.

[0044] (2) Culture Medium According to one embodiment of the present disclosure, a culture medium containing the composition (1) above is provided. Using this culture medium, CD201 ＋ or CD90 ＋ This allows efficient production of a culture fraction containing a large number of cells.

[0045] The medium is preferably a medium for culturing human hematopoietic stem cells. ＋ When the cells were cultured, CD201 ＋ or CD90 ＋ Since the culture fraction containing a large number of human CB CD41 cells can be efficiently produced, the culture fraction is preferable as a medium for culturing human hematopoietic stem cells. ＋ When the cells were cultured, CD41 ＋ Since the culture fraction containing a large number of human CB CD41 cells can be efficiently produced, the culture fraction is preferred as a medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets. ＋ 34 ＋ When the cells were cultured, CD41 ＋ and CD34 ＋ Since the culture fraction containing a large number of CB CD41 cells can be efficiently produced, it is more preferable as a medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets. ＋ The cells are not limited to those mentioned above, and hematopoietic stem cells derived from peripheral blood, bone marrow, or pluripotent stem cells (including iPS cells and embryonic stem cells) may be differentiated into megakaryocytes or platelets.

[0046] The medium is preferably a serum-free medium.

[0047] The medium is preferably an albumin-free medium.

[0048] The medium contains CD34 ＋ cells, CD201 ＋ cells, CD90＋ and CD41 ＋ It is preferable that the cell contains at least one type of cell selected from the group consisting of CD34 ＋ cells, CD201 ＋ cells, CD90 ＋ The medium for expanding human hematopoietic stem cells is preferably one containing at least one type of cell selected from the group consisting of CD41. ＋ The medium contains at least one type of cell selected from the group consisting of the above cells, making it preferable as a medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets.

[0049] The medium contains CD201 ＋ cells and CD90 ＋ The medium preferably contains a cell population enriched for at least one type of cell selected from the group consisting of CD41. ＋ It is preferable that the culture medium contains a cell population enriched for the cells of the above group. Therefore, it is preferable as a culture medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets.

[0050] The cells obtained by culturing in the medium may be used for culturing in a separately prepared, unused medium.

[0051] The cell population obtained by culturing in the medium is preferably human hematopoietic stem cells, and ＋ Preferably, the cell population for human hematopoietic stem cell proliferation is a cell population containing cells of the formula: ＋ A cell population containing the above cells is preferred for use in differentiating into human megakaryocytes or human platelets or for expanding human hematopoietic stem cells.

[0052] The medium preferably contains the cells and the composition, and the medium is preferably obtained from the cells and the composition.

[0053] The medium contains CD201 ＋ cells and CD90 ＋and the number of hematopoietic stem cells after 10 days of culture using the composition is preferably greater than the number of hematopoietic stem cells at the start of culture using the composition. ＋ Preferably, the composition contains human hematopoietic stem cells containing cells of the formula (I), and the number of human hematopoietic stem cells after 10 days of culture using the composition is greater than the number of human hematopoietic stem cells at the start of culture using the composition. Note that the number of days of culture is not limited to 10 days, and may be from a few days (about 5 days) to about 3 weeks.

[0054] The medium contains CD201 ＋ cells and CD90 ＋ and [CD201 after 10 days of culture using the composition]. ＋ the number of CD201 cells at the start of culture using the composition] - [the number of CD201 cells at the start of culture using the composition ＋ The number of cells calculated by [the number of CD90 cells after 10 days of culture using the composition] is ＋ the number of cells]−[the number of CD90 cells at the time of starting culture using the composition ＋ The number of cells in the medium is preferably greater than the number of cells calculated by the formula [number of cells in the medium]. ＋ and [a CD41 after 10 days of culture using the composition]. ＋ [Number of cells] / [Number of CD41 cells at the start of culture using the composition ＋ The ratio of [number of CD41 cells] to total number of CD41 cells is preferably 10 or more. ＋ can be obtained.

[0055] In this specification, the number of cells can be measured using flow cytometry.

[0056] (3) Method According to one embodiment of the present disclosure, there is provided a method comprising the step of contacting cells with the composition of (1) above. This method is applicable to, for example, a method for producing cells, a method for culturing cells, a method for detecting CD201 ＋, CD90 ＋ , or CD41 ＋ The present invention also includes a method for concentrating cells.

[0057] (4) Production Method According to one embodiment of the present disclosure, there is provided a method for producing cells, comprising a culture step of culturing cells using a medium containing the composition of (1) above. The cells may be human cells. Using this method, it is possible to produce CD201 ＋ , CD90 ＋ , or CD41 ＋ The method for producing cells may include, prior to the culturing step, a step of obtaining a medium from the composition of (1) above and the cells.

[0058] (5) Production Method According to one embodiment of the present disclosure, CB CD34 cells are produced using a medium containing the composition of (1) above. ＋ A method for producing cells is provided, which includes a culturing step of culturing cells. ＋ or CD90 ＋ According to another embodiment of the present disclosure, a culture fraction enriched in CB CD41 cells can be efficiently produced using a medium containing the composition (1) above. ＋ A method for producing cells is provided, which includes a culturing step of culturing cells. ＋ or CD34 ＋ This allows efficient production of a culture fraction containing a large number of cells.

[0059] (6) Production Method According to one embodiment of the present disclosure, CD201 is produced using a medium containing the composition of (1) above. ＋ or CD90 ＋ The present invention provides a method for producing cells, which comprises culturing the cells of the present invention. ＋ or CD90 ＋ According to another embodiment of the present disclosure, a medium containing the composition (1) can be used to effectively grow CD41 cells. ＋ The present invention provides a method for producing cells, which comprises culturing the cells of the present invention. ＋ or CD34 ＋The cells can be grown well.

[0060] (7) Production Method According to one embodiment of the present disclosure, there is provided a cell production method comprising a culture step of culturing human hematopoietic stem cells using a medium containing the composition of (1) above. Using this method, human hematopoietic stem cells can be successfully proliferated. According to one embodiment of the present disclosure, there is provided a cell production method comprising a culture step of culturing human megakaryocytes or human platelets using a medium containing the composition of (1) above. Using this method, human megakaryocytes or human platelets can be successfully proliferated.

[0061] (8) Production Method According to one embodiment of the present disclosure, there is provided a method for producing the composition and a medium according to (1) above. The medium obtained by this method can be used to produce CD201. ＋ or CD90 ＋ According to another embodiment of the present disclosure, there are provided a method for producing the composition and a method for producing a medium as described above in (1). The medium obtained by this method can be used to efficiently produce a culture fraction containing a large amount of CD41 cells. ＋ or CD34 ＋ This allows efficient production of a culture fraction containing a large number of cells.

[0062] (9) Production Method According to one embodiment of the present disclosure, a hematopoietic stem cell marker is extracted from a composition containing a population of cells obtained by any of the methods described in (3) to (8) above. ＋ a step of separating the cells, and hematopoietic stem cell markers after separation ＋ A step of culturing the cells of the present invention or a hematopoietic stem cell marker after culturing the cells of the present invention ＋ The present invention provides a method for producing a cell population, comprising the step of recovering cells of the present invention. By using this method, it is possible to further concentrate hematopoietic stem cells or recover a larger amount of hematopoietic stem cells. Furthermore, by using this method, it is possible to further concentrate hematopoietic stem cells for differentiation into megakaryocytes or platelets or recover a larger amount of hematopoietic stem cells for differentiation into megakaryocytes or platelets.

[0063] (10) Culturing Method According to one embodiment of the present disclosure, there is provided a cell culturing method, which includes a culturing step of culturing cells in a medium containing the composition of (1) above. ＋ or CD90 ＋ According to another embodiment of the present disclosure, there is provided a cell culture method comprising a culture step of culturing cells in a medium containing the composition of (1) above. By using this method, it is possible to efficiently produce a culture fraction containing a large amount of CD41 cells. ＋ or CD34 ＋ This allows efficient production of a culture fraction containing a large number of cells.

[0064] (11) Concentration method According to one embodiment of the present disclosure, a method for enriching CD201, comprising culturing cells in a medium containing the composition of (1) above. ＋ or CD90 ＋ This method provides a method for enriching cells containing CD201. ＋ or CD90 ＋ According to one embodiment of the present disclosure, a culture fraction enriched in CD41 cells can be efficiently produced. ＋ or CD34 ＋ This method provides a method for enriching cells containing CD41 ＋ or CD34 ＋ This allows efficient production of a culture fraction containing a large number of cells.

[0065] (12) Concentration Method According to one embodiment of the present disclosure, there is provided a method for concentrating hematopoietic stem cells, comprising a culture step of culturing cells in a medium containing the composition described in (1) above. By using this method, a culture fraction enriched in hematopoietic stem cells can be efficiently produced.

[0066] (13) Container According to one embodiment of the present disclosure, a container containing the composition (1) above is provided. Using this container, CD201 ＋ or CD90 ＋ According to one embodiment of the present disclosure, a container containing the composition (1) above is provided. Using this container, a culture fraction containing a large amount of CD41 cells can be efficiently produced. ＋ or CD34 ＋This allows efficient production of a culture fraction containing a large number of cells.

[0067] (14) Container According to one embodiment of the present disclosure, there is provided a container containing cells or a cell population thereof obtained by the method according to any one of (3) to (12) above. This container can be used for hematopoietic stem cell transplantation. Prior to hematopoietic stem cell transplantation, desired cells (e.g., hematopoietic stem cell markers) can be isolated from the cell population. ＋ The method may also include a step of sorting the cells (the cells), a step of culturing the sorted cells, a step of recovering the cultured cells, or a step of retaining the recovered cells in the container.

[0068] (15) Cells According to one embodiment of the present disclosure, there is provided a cell or a cell population thereof obtained by the method according to any one of (3) to (12) above. The cell or the cell population thereof can be used for hematopoietic stem cell transplantation. Prior to hematopoietic stem cell transplantation, a desired cell (e.g., a hematopoietic stem cell marker) can be isolated from the cell population. ＋ The method may also include a step of sorting the cells (the cells), a step of culturing the sorted cells, a step of recovering the cultured cells, or a step of retaining the recovered cells in the container.

[0069] (16) Cells According to one embodiment of the present disclosure, CD201 ＋ or CD90 ＋ According to one embodiment of the present disclosure, a cell population is provided, which comprises 5% or more of CD41 cells. This cell population can be used to produce a pharmaceutical composition for hematopoietic stem cell transplantation. ＋ or CD34 ＋ The present invention provides a cell population comprising 5% or more of the above cells. This cell population can be used to produce a pharmaceutical composition for hematopoietic stem cell transplantation.

[0070] The cell population preferably comprises human hematopoietic stem cells, and ＋ or CD41 ＋ Preferably, the cell population comprises cells of the above formula (I).

[0071] The cell population is CD34 ＋ cells, CD201 ＋ cells, CD90 ＋ and CD41 ＋It is preferable that the cell contains at least one type of cell selected from the group consisting of the cells of

[0072] The cell population is CD201 ＋ cells, CD90 ＋ and CD41 ＋ Preferably, the cell population is enriched for at least one type of cell selected from the group consisting of:

[0073] The cell population obtained by culturing in the medium may be used for culturing in a separately prepared, unused medium.

[0074] The cell population is CD201 ＋ cells, CD90 ＋ and CD41 ＋ and the number of hematopoietic stem cells after 10 days of culture using the composition is preferably greater than the number of hematopoietic stem cells at the time of initiation of culture using the composition.

[0075] The cell population is CD201 ＋ cells and CD90 ＋ and [CD201 after 10 days of culture using the composition]. ＋ the number of CD201 cells at the start of culture using the composition] - [the number of CD201 cells at the start of culture using the composition ＋ The number of cells calculated by [the number of CD90 cells after 10 days of culture using the composition] is ＋ the number of cells]−[the number of CD90 cells at the time of starting culture using the composition ＋ It is preferable that the number of cells is larger than the number of cells calculated by [number of cells per 1000 cells].

[0076] (17) Culture Medium According to one embodiment of the present disclosure, CD201 relative to total cells in a culture medium ＋ , CD90 ＋ or CD41 ＋ The present invention provides a cell-containing medium in which the percentage of cells of the present invention is 5% or more. This medium can be used to produce a pharmaceutical composition for hematopoietic stem cell transplantation.

[0077] (18) Composition According to one embodiment of the present disclosure, there is provided the composition of (1) above for use in any of the methods of (3) to (12) above.

[0078] (19) Composition According to one embodiment of the present disclosure, there is provided the cell or cell population thereof according to (15) or (16) above and the composition according to (1) above. This composition contains CD201 ＋ , CD90 ＋ or CD41 ＋ Since it contains a large amount of these cells, it is useful for producing a pharmaceutical composition for use in hematopoietic stem cell transplantation.

[0079] (20) Composition According to one embodiment of the present disclosure, there is provided a pharmaceutical composition for use in hematopoietic stem cell transplantation, comprising cells or a cell population thereof obtained by the method described in any one of (3) to (12) above, or a method for producing the same. This production method may include, for example, a step of purifying hematopoietic stem cells from the cell population described in (15) or (16) above. This production method may include, for example, purifying hematopoietic stem cells from the composition described in (19) above by isolating hematopoietic stem cell markers. ＋ a step of separating the cells, and hematopoietic stem cell markers after separation ＋ A step of culturing the cells of the present invention or a hematopoietic stem cell marker after culturing the cells of the present invention ＋

[0023] In another aspect, according to an embodiment of the present disclosure, there is provided the cell or cell population according to (15) or (16) above, or a pharmaceutical composition comprising said cell or cell population, for use in hematopoietic stem cell transplantation. In another aspect, according to an embodiment of the present disclosure, there is provided use of the cell or cell population according to (15) or (16) above in the production of a pharmaceutical composition for hematopoietic stem cell transplantation.

[0080] (21) Transplantation Method According to one embodiment of the present disclosure, there is provided a transplantation method. This transplantation method may include, for example, a step of transplanting the cells or cell population described in (15) or (16) above, or a pharmaceutical composition containing the cells or cell population, into a subject. This transplantation method enables effective engraftment of hematopoietic stem cells, thereby treating a disease.

[0081] (22) Method of Using the Cell Population According to one embodiment of the present disclosure, at least one therapeutic agent selected from the group consisting of a cancer therapeutic agent, an immunodeficiency therapeutic agent, and a multiple myeloma therapeutic agent, comprising the cell population can be provided. This therapeutic agent can be used, for example, by transplanting it into a subject according to the transplantation method described in (21) above. This therapeutic agent allows hematopoietic stem cells to be effectively engrafted, thereby treating the disease.

[0082] (23) Method of Use of Compound According to one embodiment of the present disclosure, a pharmaceutical agent containing at least one compound selected from the group consisting of the compound represented by the general formula (I) above and the compound represented by the general formula (II) above can be used to produce a medium for proliferation of hematopoietic stem cells. According to one embodiment of the present disclosure, a pharmaceutical agent containing at least one compound selected from the group consisting of the compound represented by the general formula (I) above and the compound represented by the general formula (II) above can be used as a pharmaceutical agent for a medium for proliferation of hematopoietic stem cells. The pharmaceutical agent preferably contains at least one compound selected from the group consisting of compound 126 and compound 135. Other components that may be contained in the pharmaceutical agent include those described below as optional base components.

[0083] In embodiments of the present disclosure (including, for example, (3) to (12) or (21) above), the method may include any one or more of the following steps (i) to (v): (i) placing the composition of (1) above in a container or holding it in a container before culturing, (ii) contacting the cells with the composition in the container, (iii) culturing the cells in the container, and (iv) culturing the cells. ＋ , CD90 ＋ or CD41 ＋ or (v) generating a cell population enriched for cells of CD201 relative to the total cells in the culture medium. ＋ , CD90 ＋ or CD41 ＋The method of the present disclosure (including, for example, the above (3) to (12) or (21)) may include any one or more of the following steps (vi) to (xx): (vi) preparing a cell population containing hematopoietic stem cells; (vii) detecting CB CD34 cells; ＋ (viii) providing a cell population comprising CD34 cells; ＋ (ix) separating the separated CD34 cells; ＋ (x) producing the composition of (1) above before culturing; (xi) incubating the cells in a culture medium; (xii) seeding the cells in a medium in a container and incubating; (xiii) isolating CD201 from the cell population. ＋ , CD90 ＋ or CD41 ＋ (xiv) separating or recovering hematopoietic stem cells from the cell population; (xv) detecting CD201 after separation or recovery; ＋ , CD90 ＋ or CD41 ＋ (xvi) culturing the hematopoietic stem cells after sorting or recovery; (xvii) culturing the CD201 cells after sorting or recovery and recovery; ＋ , is CD90 ＋ or CD41 ＋ (xviii) recovering the hematopoietic stem cells after sorting or recovery and culture; (xix) recovering CD201 cells after recovery. ＋ , CD90 ＋ or CD41 ＋ (xx) storing (e.g., cryopreserving) the cells; or (xx) storing (e.g., cryopreserving) the cells after collection of CD201. ＋ , CD90 ＋ or CD41 ＋ Transplanting cells of the present invention into a subject. ＋ , CD90 ＋ or CD41 ＋This method is useful for efficiently producing a culture fraction enriched in cells of the present disclosure. When two or more steps from the embodiments of the present disclosure (e.g., the above methods (3) to (12), (21), or steps (i) to (xx)) are employed, the order can be arbitrary and can be determined depending on the desired operation. Each method or step from the embodiments of the present disclosure (e.g., the above methods (3) to (12), (21), or steps (i) to (xx)) may be performed in vitro or ex vivo.

[0084] In the embodiments of the present disclosure (including, for example, (1) to (23) above), CD201 is a protein known as a hematopoietic stem cell marker (for example, WO2017 / 205977 describes the identification of human CB CD34 using CD201 as a marker). ＋ (It is described that hematopoietic stem cells were obtained from these cells.) CD201 includes a protein having the amino acid sequence of a protein represented by NCBI RefSeq Accession number NP_006395.2 or UniProt ID Q9UNN8. CD201 is also referred to as cluster of differentiation 201 or EPCR.

[0085] In embodiments of the present disclosure (including, for example, (1) to (23) above), CD90 is a protein known as a hematopoietic stem cell marker (for example, Rix et al., Front Physiol. 2022 Sep 30;13:1009160 describes CD90 as a hematopoietic stem cell marker). CD90 includes proteins having the amino acid sequence of the protein represented by NCBI RefSeq Accession number NP_001298089.1 or UniProt ID P04216. CD90 is also sometimes referred to as Cluster of Differentiation 90.

[0086] In the embodiments of the present disclosure (including, for example, (1) to (23) above), CD41 is a membrane protein that is mainly expressed in megakaryocytes and platelets. ＋ and CD41 ＋ These cells are cells that are inclined to differentiate into the megakaryocytic lineage.

[0087] In embodiments of the present disclosure (including, for example, the above (1) to (23)), the KDM5 decomposing agent preferably comprises at least one component (A) selected from the group consisting of compounds represented by general formula (I) and salts thereof, and compounds represented by general formula (II) and salts thereof.

[0088] In the embodiments of the present disclosure (including, for example, the above (1) to (23)), the LSD1 inhibitor preferably contains at least one component (B) selected from the group consisting of tranylcypromine and salts thereof.

[0089] In embodiments of the present disclosure (including, for example, the above (1) to (23)), tranylcypromine includes the compound having CAS Registry Number 155-09-9 or PubChem CID 96025309. Tranylcypromine is also referred to as 2-PCPA. Tranylcypromine includes (1R,2S)-2-phenylcyclopropan-1-amine. In this specification, tranylcypromine may be referred to as 2-PCPA for shorthand, but the terms may be used interchangeably.

[0090] In embodiments of the present disclosure (including, for example, (1) to (23) above), the PI3K activator includes an activator of phosphatidylinositol 3-kinase. PI3K activators include agonists of PI3K, such as 740Y-P or a salt thereof. 740Y-P includes the compound with CAS registration number 1236188-16-1 or PubChem CID 90488730.

[0091] In embodiments of the present disclosure (including, for example, (1) to (23) above), culturing includes incubating cells under conditions suitable for growth or maintenance. Culturing may include a step of contacting a medium and cells to produce a cell-containing composition, and a step of incubating the cell-containing composition. The incubation step may be performed while the cell-containing composition is left standing or stirred. Incubation may be performed at about 37°C and about 5% CO ２The contacting may be carried out under an ambient atmosphere. The contacting may include, for example, seeding the cells in a medium or mixing the cells with a medium. The culture may be carried out in an albumin-free or serum-free medium. "Free" includes a state in which the target component is not added to the medium, a state in which the medium is completely free of the target component, or a state in which the medium does not contain the component at a concentration above the detection limit. "Albumin-free" includes a state in which the medium is substantially free of albumin.

[0092] In embodiments of the present disclosure (including, for example, (1) to (23) above), the culture may be performed in the presence of a cell adhesion factor (e.g., fibronectin). The culture may be performed under conditions that allow contact between the hematopoietic stem cells and the cell adhesion factor. For example, the culture may be performed with the inside (e.g., bottom) of the culture vessel coated with the cell adhesion factor.

[0093] In embodiments of the present disclosure (including, for example, (1) to (23) above), culturing may include expanding hematopoietic stem cells under conditions sufficient for the maintenance or proliferation of hematopoietic stem cells. Expansion may include, for example, expanding hematopoietic stem cells by 10-fold or more from the start of culture. This expansion may be 10-, 50-, 100-, 200-, 300-, 400-, or 500-fold or more, or may be within a range of any two of these values.

[0094] In the embodiments of the present disclosure (including, for example, (1) to (23) above), the cells used for culture may be cord blood (CB) cells, peripheral blood cells, bone marrow cells, or cell populations thereof. The cells include cells collected from CB, cells collected from peripheral blood, and cells collected from bone marrow. These cells or cell populations may be CD34 ＋ The cells may be, for example, CB CD34 ＋ Cells (CB-derived CD34 ＋ cells), peripheral blood CD34 ＋ Cells (CD34 derived from peripheral blood) ＋ cells) or bone marrow CD34 ＋ cells (bone marrow-derived CD34 ＋ These cells may be purchased from manufacturers or distributors (e.g., StemExpress or Lonza) or extracted from CB, peripheral blood, or bone marrow.＋ In another embodiment of the present disclosure, the cells used for culture or the cells obtained by culture are stem cells, hematopoietic stem cells, or CD34 ＋ , CD201 ＋ , CD90 ＋ or CD41 ＋ The cells used for culturing or the cell population thereof may also be referred to as starting cells or a starting cell population. In embodiments of the present disclosure (including, for example, (1) to (23) above), the cells used for culturing or the cells obtained by culturing may be mammalian cells. Mammals include, for example, humans, monkeys, rodents (mice, hamsters, etc.), rabbits, dogs, cats, horses, cows, sheep, pigs, goats, marmosets, etc. In embodiments of the present disclosure (including, for example, (1) to (23) above), the cells may exist in the form of a cell population.

[0095] In the embodiments of the present disclosure (including, for example, the above (1) to (23)), the cell population includes, for example, a plurality of cells generated by cell division. ＋ , CD90 ＋ or CD41 ＋ The percentage of cells containing CD201 may be, for example, 5% or more. This percentage may be, for example, 2, 3, 4, 5, 8, 10, 15, 20, 25, 30, 35, 40, 45, or 50% or more, or may be within a range of any two of these values. This percentage may be, for example, 5-50%, 15-40%, or 25-40%. ＋ In order to efficiently obtain hematopoietic stem cells, the percentage of CD90 cells is preferably 10% or more, more preferably 15% or more, and even more preferably 25% or more. ＋ In order to efficiently obtain hematopoietic stem cells, the percentage of CD201 cells is preferably 13% or more, more preferably 15% or more. ＋ or CD90 ＋ The proportion of CD41 cells may be 5 times or more compared to before culture. This fold increase may be, for example, 10, 20, 50, 100, or 200 times or more, or may be within a range of any two of these values.＋ The percentage of cells is preferably 13% or more, more preferably 15% or more, in order to differentiate into megakaryocytes. The percentage of cells may be the percentage when the cells are cultured for the culture time described below.

[0096] In the embodiments of the present disclosure (including, for example, (1) to (23) above), hematopoietic stem cells include stem cells that can differentiate into blood cells. Hematopoietic stem cells can be collected from the umbilical cord, bone marrow, placenta, and peripheral blood. Human hematopoietic stem cells express CD34 ＋ Human hematopoietic stem cells contain CD34 ＋ Human hematopoietic stem cells may be CD34 ＋ CD201 ＋ CD90 ＋ , CD34 ＋ CD201 ＋ or CD34 ＋ CD41 ＋ The human hematopoietic stem cell marker may be CD34. ＋ , CD201 ＋ , CD90 ＋ or CD41 ＋ Hematopoietic stem cells include long-term hematopoietic stem cells.

[0097] In embodiments of the present disclosure (including, for example, (1) to (23) above), the culture time of the cells may be, for example, 0.5, 1, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 17, 18, 20, 30, or 35 days or more, or may be within a range of any two of these values. This culture time may be, for example, 1 to 20 days, 3 to 17 days, 4 to 16 days, or 5 to 14 days. Culturing includes the time for incubating a medium containing the cells.

[0098] In embodiments of the present disclosure (including, for example, (1) to (23) above), the seeding density of cells during culture may be 1,000 cells / mL. This value may be 1,000, 3,000, 5,000, 10,000, 30,000, 50,000, 70,000, 100,000, or 150,000 or more, or may be within a range of any two of these values.

[0099] In the embodiments of the present disclosure (including, for example, (1) to (23) above), the composition or medium comprises a medium used for culturing cells. The base components of the medium may be any common base components, and the composition is not limited. The base components of the medium may include, for example, amino acids, peptides, proteins, inorganic salts, vitamins, minerals, or a carbon source (e.g., glucose). The base components of the medium may include, for example, GlutaMAX, L-alanine-L-glutamine dipeptide, L-glutamine, ITS-X, insulin, transferrin (apo), sodium selenite, ethanolamine, or Flt3 ligand. The medium may include, for example, a basal medium for cell culture. Examples of basal media include S-clone SF-3 medium, F12 medium, StemSpan (Stem Cell technologies), STEMα (STEM ALPHA), StemPro-34 serum-free medium (Gibco Invitrogen), StemPro MSC serum-free medium (Invitrogen), HSC-CFU medium (Miltenyl Biotech), S-Clone serum-free medium (SF-02, SF-03, CM-B, SF-B) (Sanko Junyaku), HPGM medium (Sanko Junyaku), AIM V medium (Invitrogen), Marrow MAX bone marrow medium (Invitrogen), and KnockOut DMEM / F-12 medium (Invitrogen), Stemline hematopoietic stem cell growth medium (Sigma), SYN serum-free medium (SYN H, SYN B) (AbCys SA), SPE IV medium (AbCys SA), MyeloCult medium (StemCell Technologies), HPG serum-free medium (Lonza), UltraCULTURE medium (Lonza), Opti-MEM medium (Gibco Invitrogen and others), MEM medium (Gibco Invitrogen and others), MEMα (Gibco Invitrogen and others), DMEM medium (Gibco Invitrogen and others), IMDM medium (Wako Pure Examples of suitable medium include RPMI1640 medium (Gibco Invitrogen and others), Ham F-12 medium (Gibco and others), and RD medium.The medium is preferably albumin-free (e.g., GenBank accession number AAN17825.1) or serum-free. In this case, stable inhibition of undifferentiated state and suppression of problems with biological contamination can be achieved. The medium may contain Soluplus (registered trademark) (CAS accession number 402932-23-4). Soluplus is primarily composed of an N-vinylcaprolactam-vinyl acetate copolymer and a graft polymer of oxirane. The medium may also contain antibiotics (e.g., penicillin or streptomycin). The medium is suitable for hematopoietic stem cells, CD201. ＋ cells, CD90 ＋ cells, CD41 ＋ The medium may include cells, such as CB cells. The medium may be a medium for culturing these cells. The medium may be a liquid medium or a solid medium. Culturing cells using a medium includes culturing the cells in the medium.

[0100] In the embodiments of the present disclosure (including, for example, (1) to (23) above), the composition or medium contains CB CD34 ＋ The composition or medium may further contain CD201 cells. ＋ , CD90 ＋ or CD41 ＋ The present invention includes a composition or medium in which the cells of the present invention are enriched.

[0101] In embodiments of the present disclosure (including, for example, (1) to (23) above), the concentration of the PI3K activator in the composition or culture medium may be, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, or 50 μmol / L or more, or may be within a range of any two of these values. This concentration may be, for example, 1 to 25 μmol / L, 2 to 20 μmol / L, or 3 to 10 μmol / L. In order to efficiently obtain hematopoietic stem cells, this concentration is preferably 2 μmol / L or more, more preferably 3 μmol / L or more, and even more preferably 4 μmol / L or more.

[0102] In embodiments of the present disclosure (including, for example, (1) to (23) above), the concentration of the TPO receptor agonist in the composition or culture medium may be, for example, 0.01, 0.03, 0.05, 0.08, 0.1, 0.12, 0.15, 0.2, 0.3, 0.5, 1, or 2 μmol / L or more, or may be within a range of any two of these values. This concentration may be, for example, 0.01 to 0.3 μmol / L, 0.05 to 0.2 μmol / L, or 0.08 to 0.15 μmol / L. In order to efficiently obtain hematopoietic stem cells, this concentration is preferably 0.03 μmol / L or more, more preferably 0.05 μmol / L or more, and even more preferably 0.08 μmol / L or more.

[0103] In embodiments of the present disclosure (including, for example, (1) to (23) above), the concentration of UM729 or a salt thereof in a composition or culture medium may be, for example, 0.1, 0.3, 0.5, 0.8, 1, 1.3, 1.5, 1.8, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, or 30 μmol / L or more, or may be within a range of any two of these values. This concentration may be, for example, 0.1 to 3 μmol / L, 0.5 to 2 μmol / L, or 0.8 to 1.5 μmol / L. In order to efficiently obtain hematopoietic stem cells, this concentration is preferably 0.3 μmol / L or more, more preferably 0.5 μmol / L or more, and even more preferably 0.8 μmol / L or more.

[0104] In embodiments of the present disclosure (including, for example, (1) to (23) above), the concentration of 2-PCPA or a salt thereof in a composition or culture medium may be, for example, 1, 3, 5, 8, 10, 13, 15, 18, 20, 30, 40, or 50 μmol / L or more, or may be within a range of any two of these values. This concentration may be, for example, 1 to 30 μmol / L, 5 to 20 μmol / L, or 8 to 15 μmol / L. To efficiently obtain hematopoietic stem cells, this concentration is preferably 3 μmol / L or more, more preferably 5 μmol / L or more, and even more preferably 8 μmol / L or more. 2-PCPA or a salt thereof may be, for example, tranylcypromine hydrochloride.

[0105] In embodiments of the present disclosure (including, for example, the above (1) to (23)), the composition or medium may contain, as another protein component, a protein component at a concentration of 1 ng / mL or more. This concentration may be, for example, 1, 5, 8, 10, 15, 20, 30, or 50 ng / mL or more, or may be within a range of any two of these values. This concentration may be, for example, 1 to 30 μmol / L, 5 to 20 μmol / L, or 8 to 12 μmol / L. The other protein component may be, for example, Flt3 ligand.

[0106] In embodiments of the present disclosure (including, for example, (1) to (23) above), the composition or culture medium may contain a modified polyalkylene glycol as another added component. The concentration of the modified polyalkylene glycol may be, for example, 0.001, 0.005, 0.008, 0.01, 0.012, 0.015, 0.02, 0.03, 0.05, 0.1, 0.2, 0.5, 1, 2, or 5% (w / v) or more, or may be within a range of any two of these values. This concentration may be, for example, 0.001-0.1% (w / v), 0.005-0.05% (w / v), 0.005-0.02% (w / v), 0.008-0.02% (w / v), or 0.008-0.015% (w / v). The modified polyalkylene glycol may be, for example, a polyalkylene glycol modified with a copolymer of a polyvinyl caprolactam block and a polyvinyl acetate block. The polyalkylene glycol modified with a copolymer of a polyvinyl caprolactam block and a polyvinyl acetate block may be, for example, a polyethylene glycol modified with a copolymer of a polyvinyl caprolactam block and a polyvinyl acetate block. The modified polyalkylene glycol may be, for example, a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer. The modified polyalkylene glycol may be, for example, a graft copolymer obtained from (i) N-vinyl caprolactam, (ii) vinyl acetate, and (iii) a polyether. The polyether may be polyethylene glycol. The modified polyalkylene glycol may be, for example, a compound obtained by free radical polymerization of a mixture of the above (i) to (iii). For example, (i) may be 40-60 wt%, (ii) may be 15-35 wt%, and (iii) may be 10-30 wt%. (i) may be, for example, 40, 45, 50, 55, 57, or 60 wt%, or may be within a range between any two of these values. (ii) may be, for example, 15, 20, 25, 30, or 35 wt%, or may be within a range between any two of these values.(iii) may be, for example, 10, 13, 15, 20, 25, or 30 wt %, or may be within a range of any two of these values. The total of (i) to (iii) may be 100 wt %. The modified polyalkylene glycol may be, for example, a compound represented by the following structural formula (S) or a salt thereof.

[0107]

[0108] Here, the wt% of l, m, and n may be, for example, 40 to 60 wt%, 15 to 35 wt%, and 10 to 30 wt%, respectively. The wt% of l may be, for example, 40, 45, 50, 55, 57, or 60 wt%, or may be within a range of any two values ​​therein. The wt% of m may be, for example, 15, 20, 25, 30, or 35 wt%, or may be within a range of any two values ​​therein. The wt% of n may be, for example, 10, 13, 15, 20, 25, or 30 wt%, or may be within a range of any two values ​​therein. The wt% of l, m, and n may total 100 wt%. The wt% of l, m, and n may be, for example, about 57 wt%, about 30 wt%, and about 13 wt%, respectively. The degrees of polymerization of l, m, and n may be, for example, 60 to 160, 470 to 1110, and 480 to 1130, respectively. The degree of polymerization of l may be, for example, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or 160, or may be within a range of any two of these values. The degree of polymerization of m may be, for example, 470, 500, 550, 600, 650, 700, 750, 780, 800, 850, 900, 950, 1000, 1050, or 1110, or may be within a range of any two of these values. The degree of polymerization of n may be, for example, 480, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, or 1130, or may be within a range between any two of these values. The degree of polymerization may be an average degree of polymerization. The weight average molecular weight (Mw) of the modified polyalkylene glycol may be, for example, 70,000 g / mol or more. This value may be, for example, 70,000, 80,000, 90,000, 100,000, 120,000, 140,000, 150,000, 160,000, or 170,000 g / mol, or may be within a range between any two of these values. This value may be, for example, 70,000 to 170,000 g / mol, 80,000 to 160,000 g / mol, 90,000 to 140,000 g / mol, 100,000 to 130,000 g / mol, or 110,000 to 125,000 g / mol. This value may be approximately 118,000 g / mol.The modified polyalkylene glycol may be, for example, Soluplus. Soluplus is a compound having the structure of formula (S) above. The l, m, and n of Soluplus may have the wt% or degree of polymerization described above. The method for producing the modified polyalkylene glycol may be, for example, the method described in US 2008 / 0293828 A1, US 2010 / 0204425 A1, or US 2018 / 0305636 A1. Furthermore, the components (i) to (iii) above may be the components described in these documents.

[0109] In the embodiments of the present disclosure (including, for example, (1) to (23) above), the container includes a culture container or a medical container. A culture container may be used for culture, and a medical container may be used for hematopoietic stem cell transplantation. The culture container may be, for example, a plate-type, petri dish-type, flask-type, or bottle-type. The medical container may be, for example, a bottle-type, bag-type, or syringe-type.

[0110] In embodiments of the present disclosure (including, for example, (1) to (23) above), the recovering step may include, for example, transferring the desired cells or cell population from a container containing the cells or cell population to another container. The recovering step may include a sorting or purification step. The recovering step may be performed in a sterile environment.

[0111] In the embodiments of the present disclosure (including, for example, the above (1) to (23)), the composition includes, for example, a medium or an additive. The medium is, for example, a medium for hematopoietic stem cell culture or CB CD34 ＋ The additive may be a medium for cell culture. The additive may be an additive for providing additional nutrients to the basal medium.

[0112] In embodiments of the present disclosure (including, for example, (1) to (23) above), pharmaceutical compositions may be prepared by mixing cells (e.g., hematopoietic stem cells) with one or more pharmaceutically acceptable carriers and using any method known in the technical field of pharmaceuticals. The pharmaceutical composition may also contain a stabilizer, a buffer, or a pH adjuster. The dosage, administration interval, administration method, and administration route are not particularly limited and can be appropriately selected depending on the age and weight of the patient, symptoms, target organ, and the like. The pharmaceutical composition preferably contains a therapeutically effective amount, or an amount of hematopoietic stem cells effective to exert a desired effect. In one embodiment of the present disclosure, a therapeutically effective amount includes the amount necessary to clinically observe improvement or suppression of symptoms in a patient. In one embodiment of the present disclosure, being pharmaceutically acceptable includes being suitable for use at a reasonable benefit / risk ratio within the scope of sound medical judgment.

[0113] In embodiments of the present disclosure (including, for example, (1) to (23) above), the subject (including a patient) includes a human or a non-human mammal (e.g., one or more of a mouse, guinea pig, hamster, rat, mouse, rabbit, pig, sheep, goat, cow, horse, cat, dog, marmoset, monkey, or chimpanzee). The patient may also be a patient diagnosed with a disease (e.g., cancer or an immunodeficiency) or a patient in need of treatment for a disease.

[0114] In the embodiments of the present disclosure (including, for example, (1) to (23) above), enrichment includes increasing the proportion of a particular type of cell in a culture medium or a cell population. ＋ , CD90 ＋ or CD41 ＋ The cell-enriched culture fraction was found to be approximately 100% CD201 relative to the total number of cells in the medium. ＋ , CD90 ＋ or CD41 ＋ The percentage of the number of CD201 cells may be, for example, 5% or more. This percentage may be, for example, 2, 3, 4, 5, 8, 10, 15, 20, 25, 30, 35, 40, 45, or 50% or more, or may be within a range of any two of these values. This percentage may be, for example, 8-50%, 15-40%, or 25-40%. ＋In order to efficiently obtain hematopoietic stem cells, the percentage of CD90 cells is preferably 10% or more, more preferably 15% or more, and even more preferably 25% or more. ＋ The percentage of cells is preferably 13% or more, more preferably 15% or more, in order to efficiently obtain hematopoietic stem cells. The culture fraction contains a cell-containing composition obtained by culturing cells. ＋ The percentage of cells is preferably 10% or more, more preferably 15% or more. The culture fraction includes a fraction obtained after cell culture but before selection or purification treatment.

[0115] In the embodiments of the present disclosure (including, for example, (1) to (23) above), the salt is not particularly limited and includes, for example, an inorganic salt or an organic salt (see, for example, "Bharate et al., Drug Discov Today. 2021 February; 26(2): 384-398." or "Berge et al., J Pharm Sci. 1977 January; 66(1): 1-19."). Salts include, for example, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Metal salts include, for example, alkali metal salts (sodium salts, potassium salts, etc.), alkaline earth metal salts (calcium salts, magnesium salts, barium salts, etc.), aluminum salts, and the like. Salts with organic bases include, for example, salts with trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, etc. Salts with inorganic acids include, for example, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. Salts with organic acids include, for example, salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, mesylic acid, tosylic acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. Salts with basic amino acids include, for example, salts with arginine, lysine, ornithine, etc. Salts with acidic amino acids include, for example, salts with aspartic acid, glutamic acid, etc. Salts include pharmaceutically acceptable salts. In one embodiment of the present disclosure, pharmaceutically acceptable includes forms that have reasonable benefits for pharmaceutical use.

[0116] In the embodiments of the present disclosure (including, for example, (1) to (23) above) ＋ includes the cells being found to be positive for the molecule identified by the term immediately preceding it.

[0117] All publications cited herein are incorporated by reference in their entirety. In this specification, "or" is used when "at least one or more" of the items listed in the sentence can be employed. The same applies to "or." For example, CD201＋ or CD90 ＋ is CD201 ＋ , CD90 ＋ , CD201 ＋ and CD90 ＋ In this specification, when it is stated that a range is "within a range of two values," the range also includes the two values ​​themselves. In this specification, "A to B" includes A and B. In this specification, "the above (1) to (23)" includes reference to any one or more of (1), (2), (3), (4), (5), (6), (7), (8), (9), (10), (11), (12), (13), (14), (15), (16), (17), (18), (19), (20), (21), (22), or (23).

[0118] Furthermore, as the KDM5 decomposing agent, which is the component (A) in the above (1), a compound represented by the following formula (III) or (IV) may be used.

[0119] In formula (III), R １ is a straight-chain alkyl group having 1 to 6 carbon atoms.

[0120]

[0121] In formula (IV), R ２ is a straight-chain alkyl group having 1 to 6 carbon atoms.

[0122] The compound represented by formula (III) is preferably a compound represented by the following formula (NPC-4034).

[0123]

[0124] The compound represented by formula (IV) is preferably a compound (T-31) represented by the following formula:

[0125]

[0126] These compounds can be used under the same conditions as the compounds represented by formulas (I) and (II).

[0127] Although the embodiments of the present disclosure have been described above, these are merely examples of embodiments that may be included in the present disclosure, and the present disclosure is not limited to these, and various configurations other than those described above may also be employed. Furthermore, the present disclosure may employ the respective configurations or features described in the above embodiments in combination or independently.

[0128] <Actions and Effects> According to the present disclosure, a composition and a medium for use in cell culture can be provided. ＋ cells, CD201 ＋ cells, CD90 ＋ , and CD41 ＋ A cell population comprising the cells can be expanded.

[0129] The present disclosure will be further explained below with reference to examples, but is not limited thereto. In the examples, compound 126 (TIA01-126) is a compound represented by formula (I), in which A is an m-phenylene group, L is an n-pentylene group (a linear alkylene group having 5 carbon atoms), and X is -(CH ２ ) ｔ O-, and t is 6. Compound 135 (TIA02-135) is a compound represented by formula (II), in which R is a methyl group and Z is an n-nonylene group (a linear alkylene group having 9 carbon atoms).

[0130] Example 1 Figure 1 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds for 10 days in Soluplus-based medium [IMDM (Life Technologies) supplemented with 0.01% Soluplus (BASF), 1% Insulin-Transferrin-Selenium-Ethanolamine (ITSX; Life Technologies), 1% Penicillin-Streptomycin (P / S; Life Technologies), 0.5 μmol / L 740Y-P, 0.1 μmol / L Butyzamide, and 10 ng / mL recombinant FMS-like-Tyrosine kinase 3 Ligand (FLT3L; PeproTech)]. The vertical axis indicates the percentage of CD34+Linge marker-positive cells. The horizontal axis shows the compounds, with TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, 2PCPA: 5.0 μmol / L 2PCPA, and DMSO: 0.1% DMSO.

[0131] Example 2 Figure 2 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the proliferation rate of CD34+ Lineage marker- cells. The horizontal axis represents the compound. TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA; TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA; 2PCPA: 5.0 μmol / L 2PCPA; and DMSO: 0.1% DMSO.

[0132] Example 3 Figure 3 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the CD201+CD45RA- ratio among CD34+Linge marker- cells. The horizontal axis shows compounds, with TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, 2PCPA: 5.0 μmol / L 2PCPA, and DMSO: 0.1% DMSO.

[0133] Example 4 Figure 4 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the number of CD34+Linge marker-CD201+CD45RA- cells. The horizontal axis represents the compounds, with TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, 2PCPA: 5.0 μmol / L 2PCPA, and DMSO: 0.1% DMSO.

[0134] Example 5 Figure 5 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the CD90+CD45RA- ratio among CD34+Linge marker- cells. The horizontal axis shows the compounds, with TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, 2PCPA: 5.0 μmol / L 2PCPA, and DMSO: 0.1% DMSO.

[0135] Example 6 Figure 6 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the number of CD34+Lingering marker-CD90+CD45RA- cells. The horizontal axis represents the compounds, with TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, 2PCPA: 5.0 μmol / L 2PCPA, and DMSO: 0.1% DMSO.

[0136] <Example 7> Figure 7 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD34+Linage marker- cells. The horizontal axis shows the compounds, with TIA01 / 2PCPA: 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, NCP / 2PCPA: 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA, T-31 / 2PCPA: 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0137] <Example 8> Figure 8 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the proliferation rate of CD34+Linge marker- cells. The horizontal axis shows the compounds, with TIA01 / 2PCPA: 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, NCP / 2PCPA: 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA, T-31 / 2PCPA: 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0138] <Example 9> Figure 9 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD201+CD45RA- cells among CD34+Linage marker- cells. The horizontal axis shows the compounds, and the following are shown: TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA; TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA; NCP / 2PCPA: a combination of 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA; T-31 / 2PCPA: a combination of 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA; and 2PCPA: 5.0 μmol / L 2PCPA.

[0139] Example 10 Figure 10 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the number of CD34+Linage marker- cells and CD201+CD45RA- cells. The horizontal axis shows the compounds, and the following are shown: TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA; TIA02 / 2PCPA: a combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA; NCP / 2PCPA: a combination of 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA; T-31 / 2PCPA: a combination of 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA; and 2PCPA: 5.0 μmol / L 2PCPA.

[0140] Example 11 Figure 11 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD90+CD45RA- cells among CD34+Linage marker- cells. The horizontal axis shows the compounds, with TIA01 / 2PCPA: a combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, NCP / 2PCPA: 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA, T-31 / 2PCPA: 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0141] <Example 12> Figure 12 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the number of CD34+Linage marker-CD90+CD45RA- cells. The horizontal axis shows the compounds, with TIA01 / 2PCPA: 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA, NCP / 2PCPA: 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA, T-31 / 2PCPA: 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0142] Example 13 Figure 13 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the percentage of CD34+ Lineage marker-cells. The horizontal axis represents the compounds: TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: 1.0 μmol / L T-31 in combination with 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0143] Example 14 Figure 14 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the proliferation rate of CD34+ Lineage marker- cells. The horizontal axis shows the compounds: TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: 1.0 μmol / L T-31 in combination with 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0144] Example 15 Figure 15 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the CD201+CD45RA- ratio among CD34+Linge marker- cells. The horizontal axis shows the compounds: TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: 1.0 μmol / L T-31, and 2PCPA: 5.0 μmol / L 2PCPA.

[0145] Example 16 Figure 16 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the number of CD34+Linge marker-CD201+CD45RA- cells. The horizontal axis represents the compounds: TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: 1.0 μmol / L T-31 in combination with 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0146] Example 17 Figure 17 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD201+CD45RA- cells among CD34+Linge marker- cells. The horizontal axis shows the compounds: TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: 1.0 μmol / L T-31, and 2PCPA: 5.0 μmol / L 2PCPA.

[0147] Example 18 Figure 18 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the number of CD34+Linge marker-CD201+CD45RA- cells. The horizontal axis represents the compounds: TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: 1.0 μmol / L T-31 in combination with 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

[0148] Example 19 Figure 19 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds for 12 days in albumin-based medium [StemSpan SFEM (STEMCELL Technologies) supplemented with 100 ng / mL recombinant stem cell factor (SCF, PeproTech), 50 ng / mL recombinant TPO (PeproTech), 100 ng / mL FLT3L, and 10 μg / mL low-density lipoprotein (LDL; STEMCELL Technologies)]. The vertical axis represents the percentage of CD34+ lineage marker- negative cells. The horizontal axis represents the compound. DMSO: 0.1% DMSO, TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 in combination with 5.0 μmol / L 2-PCPA, respectively.

[0149] Example 20 Figure 20 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in an albumin-based medium for 12 days. The vertical axis shows the number of CD34+ Lineage marker-cells. The horizontal axis shows the compound. DMSO: 0.1% DMSO, TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 in combination with 5.0 μmol / L 2-PCPA, respectively.

[0150] Example 21 Figure 21 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in an albumin-based medium for 12 days. The vertical axis shows the CD201+CD45RA- ratio among CD34+Lingage marker- cells. The horizontal axis shows the compound. DMSO: 0.1% DMSO, TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 in combination with 5.0 μmol / L 2-PCPA, respectively.

[0151] Example 22 Figure 22 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in an albumin-based medium for 12 days. The vertical axis shows the number of CD34+Lingering marker-CD201+CD45RA- cells. The horizontal axis shows the compound. DMSO: 0.1% DMSO, TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 in combination with 5.0 μmol / L 2-PCPA, respectively.

[0152] <Example 23> Figure 23 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD41+CD34+ cells among all live cells. The horizontal axis shows the compounds and their contents, respectively: C7_0.1: 0.1 μmol / L TIA02-135, C7_1.0: 1.0 μmol / L TIA02-135, C7_1.0: 1.0 μmol / L TIA02-135, C8_0.1: 0.1 μmol / L NPC-4034, C8_1.0: 1.0 μmol / L NPC-4034, C9_0.1: 0.1 μmol / L T-31, C9_1.0: 1.0 μmol / L T31, C6_5: 5.0 μmol / L TIA01-126, 2PCPA_5: 5.0 μmol / L 2-PCPA.

[0153] Example 24 Figure 24 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the number of CD41+CD34+ cells, and the horizontal axis shows the compound and its content, as in Figure 23.

[0154] Example 25 Figure 25 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD41+CD34+ cells among all live cells. The horizontal axis shows the compound and its concentration. Each compound was used in combination with the same type and concentration as in Figure 23, but with the addition of 5.0 μmol / L of 2-PCPA. 2PCPA_5: 5.0 μmol / L 2-PCPA.

[0155] Example 26 Figure 26 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis represents the number of CD41+CD34+ cells. The horizontal axis represents the compound and its concentration. As with Figure 25, the same compounds and concentrations as in Figure 23 were used in combination with 5.0 μmol / L of 2-PCPA. 2PCPA_5: 5.0 μmol / L 2-PCPA.

[0156] <Example 27> Figure 27 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the percentage of CD41+CD34+ cells among all live cells. The horizontal axis indicates the compounds and their contents. P3_T1: 3 μmol / L 2-PCPA in combination with 1.0 μmol / L T1A02-135, P3_T3: 3 μmol / L 2-PCPA in combination with 3 μmol / L T1A02-135, P4_T1: 4.0 μmol / L 2-PCPA in combination with 1.0 μmol / L T1A02-135, P4_T2: 4.0 μmol / L 2-PCPA in combination with 2 μmol / L T1A02-135, P4_T3: 4.0 μmol / L 2-PCPA in combination with 3 μmol / L T1A02-135, P5_T1: 5.0 μmol / L 2-PCPA in combination with 1.0 μmol / L T1A02-135. P5_T1: combination of 5.0 μmol / L 2-PCPA and T1A02-135, P5_T2: combination of 5.0 μmol / L 2-PCPA and 2 μmol / L T1A02-135, P5_T3: combination of 5.0 μmol / L 2-PCPA and 3 μmol / L T1A02-135.

[0157] Example 28 Figure 28 shows the results of flow cytometry analysis of human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days. The vertical axis shows the number of CD41+CD34+ cells, and the horizontal axis shows the compound and its content, as in Figure 27.

[0158] Example 29 Figure 29 shows the ratio of mature megakaryocytes (CD41+CD42b+ cells) to the cell population when human umbilical cord blood CD34+ cells were cultured in Soluplus-based medium with a compound for 14 days, and the resulting cells (growth rate: 27.0-fold, CD34 positivity rate: 73.4%, viability rate: 88.1%, CD41 positivity rate: 15.2%) were cultured for 7 days using a megakaryocyte differentiation medium (StemCell Technologies, StemSpan SFEM, catalog number ST-09600).

[0159] Example 30 Figure 30 shows the ratio of mature megakaryocytes (CD41+CD42b+ cells) to the cell population when human umbilical cord blood CD34+ cells were cultured in Soluplus-based medium with a compound for 14 days (growth rate: 27.0-fold, CD34 positivity rate: 73.4%, viability rate: 88.1%, CD41 positivity rate: 15.2%) and then cultured for 14 days using megakaryocyte differentiation medium (StemCell Technologies, StemSpan SFEM, catalog number ST-09600). These results demonstrate that the present disclosure provides a composition and medium for cell culture.

[0160] According to the present disclosure, compositions and media for use in cell culture can be provided. Furthermore, according to the present disclosure, cell populations that can be transplanted into patients with diseases such as cancer and immunodeficiency and methods for producing the same can be provided. Furthermore, according to the present disclosure, therapeutic agents containing the cell populations can be provided.

Claims

1. A medium for culturing cells selected from the group consisting of CD34 + cells, CD201 + cells, CD90 + cells, and CD41 + cells, comprising the following composition (1) or composition (2): A composition (1) comprising a KDM5 decomposer as component (A) and an LSD1 inhibitor as component (B), or (2) a composition containing a KDM5 decomposing agent as component (A); The component (A) in the compositions (1) and (2) contains at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof: 【Chemistry 1】 In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ A group represented by O—, in which s is a number from 1 to 8 and t is a number from 2 to 10; 【Chemistry 2】 In general formula (II), R represents a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z represents a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

2. The component (A) in the compositions (1) and (2) comprises at least one component selected from the group consisting of the compound represented by the general formula (I) and a salt thereof, and the compound represented by the general formula (II) and a salt thereof, In the general formula (I), A is a single bond or a phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is —(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10; In the general formula (II), R is a hydrogen atom or a linear alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 2 to 18 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 18 carbon atoms. The medium according to claim 1.

3. The component (A) in the compositions (1) and (2) comprises at least one component selected from the group consisting of the compound represented by the general formula (I) and a salt thereof, and the compound represented by the general formula (II) and a salt thereof, In the general formula (I), A is a single bond, a p-phenylene group, or an m-phenylene group, L is a linear alkylene group having 3 to 10 carbon atoms, and X is —(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ - or -(CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10; In the general formula (II), R is a linear alkyl group having 1 to 6 carbon atoms, and Z is a substituted or unsubstituted linear alkylene group having 3 to 16 carbon atoms, or a substituted or unsubstituted branched alkylene group having 3 to 16 carbon atoms. The medium according to claim 1.

4. The component (A) in the compositions (1) and (2) comprises at least one component selected from the group consisting of the compound represented by the general formula (I) and a salt thereof, and the compound represented by the general formula (II) and a salt thereof, The medium according to claim 1, wherein the component (A) is at least one selected from the group consisting of a compound represented by the following general formula (I-1) and a salt thereof, a compound represented by the following general formula (I-2) and a salt thereof, a compound represented by the following general formula (I-3) and a salt thereof, and a compound represented by the following general formula (II-1) and a salt thereof: 【Transformation 3】 In general formula (I-1), L １ is a linear alkylene group having 3 to 10 carbon atoms, and X １ Ha-(CH ２ CH ２ O) ｓ -, s is a number of 1 to 5, and in formula (I-2), L ２ is a linear alkylene group having 3 to 10 carbon atoms, and X ２ Ha-(CH ２ CH ２ O) ｓ - or - (CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10; and in formula (I-3), L ３ is a linear alkylene group having 3 to 10 carbon atoms, and X ３ Ha-(CH ２ CH ２ O) ｓ - or - (CH ２ ) ｔ O—, s is a number from 1 to 5, and t is a number from 2 to 10; In general formula (II-1), R １ is a linear alkyl group having 1 to 3 carbon atoms, and Z １ is a substituted or unsubstituted linear alkylene group having 3 to 14 carbon atoms, or a substituted or unsubstituted branched alkylene group having 3 to 14 carbon atoms.

5. The component (A) is In the general formula (I-1), L １ is a linear alkylene group having 5 carbon atoms, and X １ Ga-(CH ２ CH ２ O) ｓ -, and s is 3; In the general formula (I-1), L １ is a linear alkylene group having 8 carbon atoms, and X １ Ga-(CH ２ CH ２ O) ｓ -, and s is 3; In the general formula (I-2), L ２ is a linear alkylene group having 5 carbon atoms, and X ２ Ga-(CH ２ CH ２ O) ｓ -, and s is the number 3; In the general formula (I-2), L ２ is a linear alkylene group having 5 carbon atoms, and X ２ Ga-(CH ２ ) ｔ a compound in which t is a group represented by O— and t is the number 6; In the general formula (I-3), L ３ is a linear alkylene group having 5 carbon atoms, and X ３ Ga-(CH ２ CH ２ O) ｓ -, and s is the number 3; In the general formula (I-3), L ３ is a linear alkylene group having 5 carbon atoms, and X ３ Ga-(CH ２ ) ｔ O—, and t is the number 6, or In the general formula (II-1), R １ is a methyl group, and Z １ The medium according to claim 4, wherein is an unsubstituted linear alkylene group having 9 carbon atoms.

6. The culture medium described in claim 1, wherein the culture medium contains composition (2), and the (A) component contains at least one component selected from compounds represented by general formula (I) and salts thereof.

7. The composition is composition (1), 2. The medium according to claim 1, wherein the component (B) comprises at least one component selected from the group consisting of tranylcypromine and salts thereof.

8. A medium for culturing cells selected from the group consisting of CD34 + cells, CD201 + cells, CD90 + cells, and CD41 + cells, comprising a composition (1) containing (A) a KDM5 degrading agent as a component, and (B) an LSD1 inhibitor as a component.

9. The culture medium described in claim 8, wherein the component (B) includes at least one component selected from the group consisting of tranylcypromine and its salts.

10. The culture medium described in claim 1 or 9, wherein the culture medium is a culture medium for culturing hematopoietic stem cells.

11. The culture medium described in claim 10, wherein the hematopoietic stem cells are for differentiation into megakaryocytes.

12. The medium according to claim 1 or 9, wherein the medium is a medium for culturing CD34 + cells.

13. The medium according to claim 1 or 9, comprising a cell population obtained by culturing cells selected from the group consisting of CD34+ cells, CD201+ cells, CD90+ cells, and CD41+ cells in the medium according to claim 1 or 9.

14. A method for producing cells, comprising the step of culturing cells selected from the group consisting of CD34 + cells, CD201 + cells, CD90 + cells, and CD41 + cells using the medium of claim 1 or 9.

15. A method for producing at least one therapeutic agent selected from the group consisting of cancer therapeutic agents, immunodeficiency therapeutic agents, and multiple myeloma therapeutic agents, comprising a step of culturing cells selected from the group consisting of CD34 + cells, CD201 + cells, CD90 + cells, and CD41 + cells using the culture medium described in claim 1 or 9.

16. At least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof, an LSD1 inhibitor; Use of a drug comprising the following for producing a medium for expanding hematopoietic stem cells: 【Chemistry 4】 In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ A group represented by O—, in which s is a number from 1 to 8 and t is a number from 2 to 10; 【Transformation 5】 In general formula (II), R represents a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z represents a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

17. Use of a drug containing at least one component selected from the group consisting of a compound represented by the following general formula (I) and a salt thereof, and a compound represented by the following general formula (II) and a salt thereof, as a drug for a medium for proliferation of hematopoietic stem cells: 【Transformation 6】 In general formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear alkylene group having 1 to 13 carbon atoms or a substituted or unsubstituted branched alkylene group having 1 to 13 carbon atoms, and X is -(CH ２ CH ２ O) ｓ -, -(CH ２ CH ２ CH ２ O) ｓ -, -(CH(CH ３ ) CH ２ O) ｓ - or -(CH ２ ) ｔ A group represented by O—, in which s is a number from 1 to 8 and t is a number from 2 to 10; 【Transformation 7】 In general formula (II), R represents a hydrogen atom, a substituted or unsubstituted linear alkyl group having 1 to 6 carbon atoms, or a substituted or unsubstituted branched alkyl group having 1 to 6 carbon atoms, and Z represents a substituted or unsubstituted linear alkylene group having 2 to 20 carbon atoms, or a substituted or unsubstituted branched alkylene group having 2 to 20 carbon atoms.

Images