Compositions, culture media, cell populations, methods for producing cells, and therapeutic drugs

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

[Technical Field]

[0001] The technical field of this disclosure relates to compositions, culture media, cell populations, methods for producing cells, cells, and therapeutic agents used in cell culture. This application claims priority based on Japanese Patent Application No. 2024-009804, filed in Japan on January 25, 2024, and the contents of that application are incorporated herein by reference. The contents of WO2024 / 210126, filed on April 2, 2024, and published on October 10, 2024, are incorporated herein by reference. The information presented here is taken from the article "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 on May 5, 2016. [Background technology]

[0002] Hematopoietic stem cells are stem cells that can differentiate into blood cells. Transplanting hematopoietic stem cells into patients with cancer, immunodeficiency, and other diseases is a known treatment method for these conditions.

[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 a starting population of cells, EPCR + The procedure involves selecting and culturing a population of cells. EPCR is a hematopoietic stem cell marker, also known as CD201.

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

[0005] [Patent Document 1] WO2017 / 205977 [Patent Document 2] WO2021 / 149799 [Patent Document 3] WO2024 / 210126 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] Conventional methods were insufficient for efficiently obtaining human hematopoietic stem cells. Furthermore, there were few reports on albumin-free culture methods for human hematopoietic stem cells, limiting the available culture media. [Means for solving the problem]

[0007] The researchers investigated hematopoietic stem cell culture methods during their studies. As a result, they found that human CB CD34 cells could be cultured in an albumin-free medium with a specific composition different from conventional methods. + By culturing the cells, we were able to successfully proliferate hematopoietic stem cells. Furthermore, the resulting culture fraction was CD201 + Or CD90 +A culture fraction enriched with hematopoietic stem cells, all of which are hematopoietic stem cell markers. The present inventors completed the present disclosure based on this finding.

[0008] The present disclosure has the following aspects. [1] A composition (1) containing a KDM5 degrader as component (A) and an LSD1 inhibitor as component (B), or A composition (2) containing a KDM5 degrader as component (A), wherein In composition (2), the component (A) comprises at least one component selected from the group consisting of a compound represented by the following general formula (I) and its salt, and a compound represented by the following general formula (II) and its salt. Composition (2). [Chemical formula] 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, X is -(CH2CH2O) s -, -(CH2CH2CH2O) s -, -(CH(CH3)CH2O) s -, or -(CH2) t O-, and s is a number from 1 to 8, and t is a number from 2 to 10. [Chemical formula] 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 is composition (1), The composition according to [1], wherein component (A) comprises at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof. [ka] 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t It is a group represented by O-, where s is a number from 1 to 8 and t is a number from 2 to 10. [ka] 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. [3] In the above 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) 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. 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) above, 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -, or -(CH2) 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. The composition according to [1], wherein 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. [5] The composition according to [1], wherein component (A) is at least one 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. [ka] In general formula (I-1), L 1 X is a linear alkylene group having 3 to 10 carbon atoms. 1 ha-(CH2CH2O) s - is a group represented by -, where s is a number from 1 to 5, and in the general formula (I-2), L 2 X is a linear alkylene group having 3 to 10 carbon atoms. 2 ha-(CH2CH2O) s - or - (CH2) 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, and in the general formula (I-3), L 3 X is a linear alkylene group having 3 to 10 carbon atoms. 3 ha-(CH2CH2O) s - or - (CH2) 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 Z is a linear alkyl group having 1 to 3 carbon atoms. 1This 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 aforementioned component (A) is, In the general formula (I-1), L 1 X is a linear alkylene group with 5 carbon atoms, 1 ga-(CH2CH2O) s A compound with a group represented by - and s is 3. In the general formula (I-1), L 1 This is a linear alkylene group with 8 carbon atoms, X 1 ga-(CH2CH2O) s A compound with a group represented by - and s is 3. In the general formula (I-2), L 2 X is a linear alkylene group with 5 carbon atoms, 2 ga-(CH2CH2O) s A compound with a group represented by - and where the number of s is 3, In the general formula (I-2), L 2 X is a linear alkylene group with 5 carbon atoms, 2 ga-(CH2) t A compound with a group represented by O-, where t is the number 6. In the general formula (I-3), L 3 X is a linear alkylene group with 5 carbon atoms, 3 ga-(CH2CH2O) s A compound where the group is represented by - and s is the number 3, In the general formula (I-3), L 3 X is a linear alkylene group with 5 carbon atoms, 3 ga-(CH2) t A compound with a group represented by O-, where t is the number 6, or In the above general formula (II-1), R 1 is a methyl group, Z 1 The composition according to [5], wherein is an unsubstituted, straight-chain alkylene group having 9 carbon atoms. [7] The aforementioned composition is composition (1), The composition according to [1], comprising at least one component selected from the group consisting of tranylcypromine and salts thereof. [8] The aforementioned composition is composition (1), The composition according to [6], comprising at least one component selected from the group consisting of tranylcypromine and salts thereof. [9] A culture medium comprising the composition described in any one of items [1] to [8].

[10] The culture medium according to [9], wherein the culture medium is a culture medium for 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 culture medium.

[13] The culture medium is CD34 + cells, CD201 + cells, CD90 + and CD41 + The culture medium according to [9], comprising at least one type of cell selected from the group consisting of the cells.

[14] The culture medium is CD201 + cells, CD90 + and CD41 + The culture medium according to [9], comprising a cell population enriched with at least one cell species selected from the group consisting of the cells of [1].

[15] A composition used for culturing cells obtained by culturing them in the culture medium described in [9].

[16] The composition according to

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

[17] Cells obtained by culturing in the aforementioned medium are CD201 + Or CD41 + The composition described in

[15] , which is a cell of

[15] .

[18] A population of cells obtained by culturing in the culture medium described in [9].

[19] The cell population described in

[18] includes human hematopoietic stem cells.

[20] The aforementioned cell population is CD201 + Or CD41 + A population of cells, including the cells of

[18] . [twenty one] [9] A therapeutic agent selected from the group consisting of cancer therapeutic agents, immunodeficiency therapeutic agents, and multiple myeloma therapeutic agents, comprising cells obtained by culturing in the culture medium described above. [twenty two] A method for producing cells, comprising the step of culturing cells using the culture medium described in [9]. [twenty three] The cell production method according to

[22] , wherein the culturing step comprises obtaining a culture medium from the cells and the composition. [twenty four] A method for producing cells according to

[22] , comprising the step of culturing the cells cultured in the aforementioned culturing step using another culture medium. [twenty five] The method for producing cells according to

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

[26] The cultured cells mentioned above are CD201 + Or CD41 + A method for producing the cells described in

[22] , which is a cell population containing human hematopoietic stem cells.

[27] A population of cells produced by the method described in

[22] .

[28] The cell population described in

[27] includes human hematopoietic stem cells.

[29] The aforementioned cell population is CD201 + Or CD41 + A population of cells, including the cells of

[27] .

[30]

[22] A therapeutic agent selected from the group consisting of cancer therapies, immunodeficiency therapies, and multiple myeloma therapies, comprising cells obtained using the method described in

[22] .

[31] Use of a drug comprising at least one component selected from the group consisting of compounds represented by the following formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof, for the production of a culture medium for hematopoietic stem cell proliferation. [ka] 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t It is a group represented by O-, where s is a number from 1 to 8 and t is a number from 2 to 10. [ka] 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 drug containing at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof, as a drug for hematopoietic stem cell proliferation culture media. [ka] 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t It is a group represented by O-, where s is a number from 1 to 8 and t is a number from 2 to 10. [ka] 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 any one of the compositions described in [1] to [8] for the purpose of proliferating hematopoietic stem cells.

[34] The use described in

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

[35] Use of any one of the compositions described in [1] to [8] as a drug for the culture medium of hematopoietic stem cells for differentiation into megakaryocytes.

[36] Use of any one of the compositions described in [1] to [8] for the production of a drug for a culture medium for hematopoietic stem cells to differentiate into megakaryocytes.

[37] A composition for hematopoietic stem cell proliferation, as described in any one of [1] to [8].

[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 aforementioned tatami mat, CD34 + cells, CD201 + cells, CD90 + and CD41 + The composition according to any one of

[37] to

[40] , comprising at least one type of cell selected from the group consisting of the cells.

[42] The aforementioned composition is CD201 + cells, CD90 + and CD41 + The composition according to any one of

[37] to

[41] , comprising a cell population in which at least one type of cell selected from the group consisting of cells is concentrated. [Effects of the Invention]

[0009] According to this disclosure, compositions and culture media for use in cell culture can be provided. Furthermore, this disclosure provides a cell population that can be transplanted into patients with diseases such as cancer and immunodeficiency, as well as a method for producing the same. Furthermore, according to this disclosure, therapeutic agents comprising the aforementioned cell population can be provided. [Brief explanation of the drawing]

[0010] [Figure 1] This figure shows the expression analysis results of the cell population in Example 1. [Figure 2] This figure shows the expression analysis results of the cell population in Example 2. [Figure 3] This figure shows the expression analysis results of the cell population in Example 3. [Figure 4] This figure shows the expression analysis results of the cell population in Example 4. [Figure 5] This figure shows the expression analysis results of the cell population in Example 5. [Figure 6] This figure shows the expression analysis results of the cell population in Example 6. [Figure 7]This figure shows the expression analysis results of the cell population in Example 7. [Figure 8] This figure shows the expression analysis results of the cell population in Example 8. [Figure 9] This figure shows the results of the expression analysis of the cell population in Example 9. [Figure 10] This figure shows the expression analysis results of the cell population in Example 10. [Figure 11] This figure shows the expression analysis results of the cell population in Example 11. [Figure 12] This figure shows the expression analysis results of the cell population in Example 12. [Figure 13] This figure shows the expression analysis results of the cell population in Example 13. [Figure 14] This figure shows the expression analysis results of the cell population in Example 14. [Figure 15] This figure shows the expression analysis results of the cell population in Example 15. [Figure 16] This figure shows the expression analysis results of the cell population in Example 16. [Figure 17] This figure shows the expression analysis results of the cell population in Example 17. [Figure 18] This figure shows the expression analysis results of the cell population in Example 18. [Figure 19] This figure shows the expression analysis results of the cell population in Example 19. [Figure 20] This figure shows the expression analysis results of the cell population in Example 20. [Figure 21] This figure shows the expression analysis results of the cell population in Example 21. [Figure 22] This figure shows the expression analysis results of the cell population in Example 22. [Figure 23] This figure shows the expression analysis results of the cell population in Example 23. [Figure 24] This figure shows the expression analysis results of the cell population in Example 24. [Figure 25] This figure shows the expression analysis results of the cell population in Example 25. [Figure 26] This figure shows the expression analysis results of the cell population in Example 26. [Figure 27] This figure shows the expression analysis results of the cell population in Example 27. [Figure 28] This figure shows the expression analysis results of the cell population in Example 28. [Figure 29] This figure shows the ratio of mature megakaryocytes (CD41+CD42b+ cells) to the cell population after 7 days of culture in the cell population in Example 29. [Figure 30] This figure shows the ratio of mature megakaryocytes (CD41+CD42b+ cells) to the cell population after 14 days of culture in Example 30. [Modes for carrying out the invention]

[0011] The embodiments of this disclosure will be described in detail below. To avoid repetition and unnecessary complexity, similar content will be omitted from explanation as appropriate. Unless otherwise specified, "alkyl group" includes linear, branched, and cyclic monovalent saturated hydrocarbon groups. The same applies to alkyl groups within alkoxy groups. Unless otherwise specified, the term "alkylene group" includes linear, branched, and cyclic divalent saturated hydrocarbon groups. Unless otherwise specified, the term "alkenylene group" includes linear, branched, and cyclic divalent unsaturated hydrocarbon groups. Examples of "halogen atoms" include fluorine, chlorine, bromine, and iodine atoms. The term "substituted or unsubstituted" includes both cases where a hydrogen atom (-H) is substituted with a monovalent substituent and cases where a methylene group (-CH2-) or methine group (>CH-, or =CH-) is substituted with a divalent substituent. "Monovalent substituents" include alkyl groups, alkoxy groups, halogen atoms, alkyl halides, hydroxyl groups, carboxyl groups, amino groups, nitro groups, and the like. As the alkyl group as a substituent, an alkyl group having 1 to 5 carbon atoms is preferable, and a methyl group, an ethyl group, a propyl group, an n-butyl group, and a tert-butyl group are most preferable. As the alkoxy group as a substituent, an alkoxy group having 1 to 5 carbon atoms is preferable, a methoxy group, an ethoxy group, an n-propoxy group, an iso-propoxy group, an n-butoxy group, and a tert-butoxy group are more preferable, and a methoxy group and an ethoxy group are most preferable. As the halogen atom as a substituent, a fluorine atom and an iodine atom are preferable. As the halogenated alkyl group as a substituent, a group in which part or all of the hydrogen atoms of an alkyl group having 1 to 5 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an n-butyl group, a tert-butyl group, etc., are substituted with the above halogen atoms can be mentioned. The "divalent substituent" refers to -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, an alkoxyalkyl group, etc.), -S-, -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 -O-C(=O)-Y 22 - or -Y 21 -S(=O)2-O-Y 22 -represented group [where Y 21 and Y 22 are each independently a substituted or unsubstituted divalent hydrocarbon group, O is an oxygen atom, and m” is an integer from 0 to 3.], etc. can be mentioned.

[0012] (1) Composition According to one embodiment of the present disclosure, a composition (1) comprising a KDM5 degrading agent as component (A) and an LSD1 inhibitor as component (B), or a composition (2) comprising a KDM5 degrading agent as component (A), wherein in composition (2), component (A) comprises at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof (hereinafter, composition (1) and composition (2) are collectively referred to simply as "composition").

[0013] [ka]

[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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t It is a group represented by O-, where s is a number from 1 to 8 and t is a number from 2 to 10.

[0015] [ka]

[0016] 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.

[0017] In general formula (I), A is a single-bonded, 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t It is a group represented by O-, where s is a number from 1 to 8 and t is a number from 2 to 10.

[0018] As for A, single bonds, phenylene groups, and naphthylene groups are preferred; single bonds, 1,2-phenylene groups, 1,3-phenylene groups (m-phenylene groups), 1,4-phenylene groups (p-phenylene groups), 1,2-naphthylene groups, 1,4-naphthylene groups, 1,5-naphthylene groups, 2,3-naphthylene groups, and 2,6-naphthylene groups are more preferred; single bonds, 1,3-phenylene groups (m-phenylene groups), and 1,4-phenylene groups (p-phenylene groups) are even more preferred.

[0019] The 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 aforementioned X is -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t A group represented by O- is preferred; -(CH2CH2O) s -,-(CH2CH2CH2O) s -, or -(CH2) t A group represented by O- is more preferred. The aforementioned s is preferably a number from 1 to 5, more preferably 2 to 4, and even more preferably 2 or 3. The aforementioned t is preferably a number between 2 and 10, more preferably 3 to 8, and even more preferably 4 to 7.

[0021] In the above 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t The group is represented as O-, and preferably s is a number from 1 to 5, and t is a number from 2 to 10. In the general formula (I) above, 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -, or -(CH2) t The group is represented as O-, and it is more preferable that 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, even more preferably a methyl group or an ethyl group, and particularly preferably a methyl group. The aforementioned 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, even more preferably a linear or branched alkylene group having 5 to 15 carbon atoms, particularly preferably an n-hexylene group, n-heptylene group, n-octylene group, n-nonylene group, n-decylene group, or n-undecylene group, and most preferably an n-octylene group, n-nonylene group, or n-decylene group.

[0023] In the above composition (1), It is preferable that component (A) contains at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof.

[0024] [ka]

[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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) t It is a group represented by O-, where s is a number from 1 to 8 and t is a number from 2 to 10.

[0026] [ka]

[0027] 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.

[0028] In the above 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -,-(CH(CH3)CH2O) s -, or -(CH2) 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 the above general formula (II), it is preferable that the composition is such that 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) above, 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 -(CH2CH2O) s -,-(CH2CH2CH2O) s -, or -(CH2) 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 the general formula (II) described above, it is preferable that the composition is such that 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] Preferably, the composition is one selected from the group consisting of compounds represented by the following general formula (I-1) and their salts, compounds represented by the following general formula (I-2) and their salts, compounds represented by the following general formula (I-3) and their salts, and compounds represented by the following general formula (II-1) and their salts.

[0031] [ka]

[0032] In general formula (I-1), L 1 X is a linear alkylene group having 3 to 10 carbon atoms. 1 ha-(CH2CH2O) s - is a group represented by -, where s is a number from 1 to 5, and in the general formula (I-2), L 2 X is a linear alkylene group having 3 to 10 carbon atoms. 2 ha-(CH2CH2O) s - or - (CH2) 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, and in the general formula (I-3), L 3 X is a linear alkylene group having 3 to 10 carbon atoms. 3 ha-(CH2CH2O) s - or - (CH2) tThe group is represented as O-, and preferably s is a number from 1 to 5, and t is a number from 2 to 10. In general formula (II-1), R 1 Z is a linear alkyl group having 1 to 3 carbon atoms. 1 It is preferable that the group 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.

[0033] The aforementioned component (A) is, In the general formula (I-1), L 1 X is a linear alkylene group with 5 carbon atoms, 1 ga-(CH2CH2O) s A compound with a group represented by - and s is 3. In the general formula (I-1), L 1 This is a linear alkylene group with 8 carbon atoms, X 1 ga-(CH2CH2O) s A compound with a group represented by - and s is 3. In the general formula (I-2), L 2 X is a linear alkylene group with 5 carbon atoms, 2 ga-(CH2CH2O) s A compound with a group represented by - and where the number of s is 3, In the general formula (I-2), L 2 X is a linear alkylene group with 5 carbon atoms, 2 ga-(CH2) t A compound with a group represented by O-, where t is the number 6. In the general formula (I-3), L 3 X is a linear alkylene group with 5 carbon atoms, 3 ga-(CH2CH2O) s A compound where the group is represented by - and s is the number 3, In the general formula (I-3), L 3 X is a linear alkylene group with 5 carbon atoms, 3 ga-(CH2) t A compound with a group represented by O-, where t is the number 6, or In the above general formula (II-1), R 1 is a methyl group, Z 1The composition is preferably an unsubstituted, straight-chain alkylene group having 9 carbon atoms.

[0034] In particular, the compound represented by general formula (I) and its salts are as follows: in formula (I), A is an m-phenylene group, L is an n-pentylene group (a linear alkylene group with 5 carbon atoms), and X is -(CH2) t A compound represented by the group O- and where t is 6 (hereinafter also referred to as "compound 126") or a salt thereof is preferred. As for the compound represented by general formula (II) and its salt, a compound in which R is a methyl group and Z is an n-nonylene group (a linear alkylene group with 9 carbon atoms) (hereinafter also referred to as "compound 135") or a salt thereof is preferred.

[0035] Preferably, the composition is composition (1), and component (B) is a composition that includes at least one component selected from the group consisting of tranylcypromine represented by the following general formula (B-1) or (B-2) and its salts. Among these, tranylcypromine represented by the following general formula (B-2) and its salts are more preferred, and tranylcypromine hydrochloride represented by the following general formula (B-2-1) is even more preferred.

[0036] [ka]

[0037] [ka]

[0038] [ka]

[0039] The salts of the aforementioned 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 salts [e.g., hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, nitrate, etc.]), organic salts [e.g., acetate, trifluoroacetate, lactate, tartrate, oxalate, fumarate, maleate, benzoate, citrate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, isethionate, glucuronate, gluconate, etc.]), and salts with acidic natural amino acids [e.g., aspartic acid, glutamic acid, etc.]).

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

[0041] The composition (1) can be prepared by mixing a KDM5 degrading agent, which is component (A), an LSD1 inhibitor, which is component (B), and any other base component as needed. The composition (2) can be produced by mixing, as an option, at least one component selected from the group consisting of the compound represented by the general formula (I) and its salts, and the compound represented by the general formula (II) and its salts, as component (A) of the KDM5 decomposing agent, with other components. Other components include the following optional base components.

[0042] In the composition (1) described above, 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, based on the total amount of the composition (1 L). In the composition (1) described above, 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, based on the total amount of the composition (1 L). In the composition (1) described above, the content of 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, based on the total amount of the composition (1 L). In the composition (2) described above, 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, based on the total amount (1 L) of the composition. In the composition (2) described above, 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, based on the total amount (1 L) of the composition. If the composition contains albumin, the albumin content 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, based on the total mass of the composition.

[0043] Here, the composition may be a culture medium. The culture medium may be a culture medium for human hematopoietic stem cells. The composition may further contain hematopoietic stem cells. Using this composition, CD201 + Or CD90 + This allows for the efficient production of a culture fraction containing a large amount of human CB CD34 cells. For example, in the examples described later, this composition is used to produce human CB CD34 + As a result of culturing the cells, CD201 + Cells or CD90 + It is stated that a culture fraction in which cells were concentrated was obtained. Furthermore, according to one embodiment of this disclosure, a method for producing cells, a method for culturing cells, a method for producing a culture medium using this composition, and CD201 +Or CD90 + A method for concentrating cells and a container containing this composition are provided.

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

[0045] The culture medium is preferably a culture medium for human hematopoietic stem cells. Human CB CD34 + When cells are cultured, CD201 + Or CD90 + Because it can efficiently produce a culture fraction containing a large amount of these cells, it is preferable as a culture medium for human hematopoietic stem cells. Human CB CD41 + When cells are cultured, CD41 + Because it can efficiently produce a culture fraction containing a large amount of human cells, it is preferable as a culture medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets. Furthermore, human CB CD41 + 34 + When cells are cultured, CD41 + CD34 + Because it can efficiently produce a culture fraction containing a large amount of these cells, it is more preferable as a culture medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets. CB CD41 + The process is not limited to cells; megakaryocytes or platelets may be differentiated from hematopoietic stem cells derived from peripheral blood, bone marrow, or pluripotent stem cells (including iPS cells and embryonic stem cells).

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

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

[0048] The culture medium is CD34 + cells, CD201 + cells, CD90 +and CD41 + It is preferable to include at least one type of cell selected from the group consisting of the following cells. CD34 + cells, CD201 + cells, CD90 + It is preferable as a culture medium for human hematopoietic stem cell proliferation by containing at least one type of cell selected from the group consisting of the following cells. CD41 + It is preferable as a culture medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets by including at least one cell type selected from the group consisting of the following cells.

[0049] The culture medium is CD201 + cells and CD90 + It is preferable that the medium contains a cell population in which at least one type of cell selected from the group of cells is concentrated. This makes it preferable as a culture medium for human hematopoietic stem cell proliferation. The culture medium is CD41 + It is preferable that the culture medium contains a concentrated population of cells. This makes it suitable as a culture medium for human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets.

[0050] Cells cultured in the aforementioned medium may be used to culture them in a separately prepared, unused portion of the aforementioned medium.

[0051] The cell population obtained by culturing in the aforementioned medium is preferably human hematopoietic stem cells, and CD201 + A cell population containing the cells of CD41 is preferred for human hematopoietic stem cell proliferation, and CD41 + It is preferable that the cell population includes cells of the specified type for use in the proliferation of human hematopoietic stem cells for differentiation into human megakaryocytes or human platelets.

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

[0053] The culture medium is CD201 + cells and CD90+ The composition contains at least one hematopoietic stem cell selected from the group of cells, and it is preferable that the number of hematopoietic stem cells after culturing with the composition for 10 days is greater than the number of hematopoietic stem cells at the start of culturing with the composition. The culture medium is CD41 + The composition contains human hematopoietic stem cells, and it is preferable that the number of human hematopoietic stem cells after culturing with the composition for 10 days is greater than the number of human hematopoietic stem cells at the start of culturing with the composition. Furthermore, the incubation period is not limited to 10 days; it can range from a few days (approximately 5 days) to about 3 weeks.

[0054] The culture medium is CD201 + cells and CD90 + The composition comprises at least one hematopoietic stem cell selected from the group consisting of the following cells, and CD201 after being cultured for 10 days using the composition. + [Number of cells] - [CD201 at the start of culture using the above composition] + The number of cells calculated by [the number of cells of CD90 after culturing for 10 days using the above composition] is [the number of CD90 + [Number of cells] - [CD90 at the time of starting culture using the composition] + It is preferable that the number of cells is greater than the number of cells calculated by [the number of cells]. The culture medium is CD41 + The composition comprises at least one hematopoietic stem cell selected from the group consisting of the following cells, and CD41 after being cultured for 10 days using the composition. + [Number of cells] / [CD41 at the start of culture using the above composition] + The ratio expressed as [number of cells] is preferably 10 or more. This is because the total number of cells increases more than 10 times during 10 days of culture, thus increasing the number of CD41 + We can obtain cells from this.

[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, a method is provided which includes the step of bringing the composition of (1) above into contact with cells. This method is, for example, a method for producing cells, a method for culturing cells, CD201 + CD90 + , or CD41 + This includes a method for concentrating cells.

[0057] (4) Production method According to one embodiment of the present disclosure, a method for producing cells is provided, comprising a culture step of culturing cells using a culture medium containing the composition of (1) above. The cells may be human cells. Using this method, CD201 + CD90 + , or CD41 + This allows for the efficient production of culture fractions containing a large number of these cells. The method for producing the cells may include a step of obtaining a culture medium from the composition of (1) and the cells before the culture step.

[0058] (5) Production method According to one embodiment of the present disclosure, a culture medium containing the composition of (1) above is used to obtain CB CD34 + A method for producing cells is provided, which includes a cell culture step. Using this method, CD201 + Or CD90 + This allows for the efficient production of culture fractions containing a large number of these cells. According to other embodiments of the present disclosure, a culture medium containing the composition of (1) above is used to prepare CB CD41 + A method for producing cells is provided, which includes a cell culture step. Using this method, CD41 + or CD34 + This allows for the efficient production of culture fractions containing a large number of these cells.

[0059] (6) Production method According to one embodiment of the present disclosure, a culture medium containing the composition of (1) above is used to obtain CD201 + Or CD90 + A method for producing cells is provided, which includes a culture step for culturing the cells of CD201. +Or CD90 + It can promote the healthy proliferation of these cells. According to other embodiments of the present disclosure, a culture medium containing the composition of (1) above is used to produce CD41 + A method for producing cells is provided, which includes a culture step for culturing CD41 cells. Using this method, CD41 + or CD34 + It can promote the healthy proliferation of these cells.

[0060] (7) Production method According to one embodiment of the present disclosure, a method for producing cells is provided, comprising a culture step of culturing human hematopoietic stem cells using a culture 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, a method for producing cells is provided, comprising a culture step of culturing human megakaryocytes or human platelets using a culture 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, a method for producing the composition (1) and a method for producing a culture medium are provided. Using the culture medium obtained by this method, CD201 + Or CD90 + This allows for the efficient production of culture fractions containing a large number of these cells. According to other embodiments of the present disclosure, a method for producing the composition (1) and a method for producing a culture medium are provided. Using the culture medium obtained by this method, CD41 + or CD34 + This allows for the efficient production of culture fractions containing a large number of these cells.

[0062] (9) Production method According to one embodiment of the present disclosure, a hematopoietic stem cell marker is obtained from a composition containing a population of cells obtained by any of the methods described in (3) to (8) above. + The process of separating cells, and hematopoietic stem cell markers after separation. + The process of culturing cells, or hematopoietic stem cell markers after culture +A method for producing a cell population is provided, which includes a step of collecting cells. By using this method, more hematopoietic stem cells can be concentrated or more hematopoietic stem cells can be collected. Furthermore, by using this method, hematopoietic stem cells for differentiation into megakaryocytes or platelets can be more concentrated or more hematopoietic stem cells for differentiation into megakaryocytes or platelets can be collected.

[0063] (10) Culture method According to one embodiment of the present disclosure, a method for culturing cells is provided, which includes a culturing step of culturing cells in a medium containing the composition of (1) above. By using this method, a culture fraction rich in cells of CD201 + or CD90 + can be efficiently produced. According to another embodiment of the present disclosure, a method for culturing cells is provided, which includes a culturing step of culturing cells in a medium containing the composition of (1) above. By using this method, a culture fraction rich in cells of CD41 + or CD34 + can be efficiently produced.

[0064] (11) Concentration method According to one embodiment of the present disclosure, a method for concentrating cells of CD201 + or CD90 + is provided, which includes a culturing step of culturing cells in a medium containing the composition of (1) above. By using this method, a culture fraction rich in cells of CD201 + or CD90 + can be efficiently produced. According to one embodiment of the present disclosure, a method for concentrating cells of CD41 + or CD34 + is provided, which includes a culturing step of culturing cells in a medium containing the composition of (1) above. By using this method, a culture fraction rich in cells of CD41 + or CD34 + can be efficiently produced.

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

[0066] (13) Container According to one embodiment of the present disclosure, there is provided a container containing the composition of (1) above. By using this container, a culture fraction rich in cells of CD201 + or CD90 + can be efficiently produced. According to one embodiment of the present disclosure, there is provided a container containing the composition of (1) above. By using this container, a culture fraction rich in cells of CD41 + or CD34 + can be efficiently produced.

[0067] (14) Container According to one embodiment of the present disclosure, there is provided a container containing cells or a cell population obtained by each method described in any one of (3) to (12) above. This container can be used for hematopoietic stem cell transplantation. Before hematopoietic stem cell transplantation, a step of separating desired cells (for example, cells of a hematopoietic stem cell marker + ) from the cell population, a step of culturing the separated cells, a step of recovering the cultured cells, or a step of holding the recovered cells in the container may be performed.

[0068] (15) Cell According to one embodiment of the present disclosure, there is provided a cell or a cell population obtained by each method described in any one of (3) to (12) above. This cell or cell population can be used for hematopoietic stem cell transplantation. Before hematopoietic stem cell transplantation, a step of separating desired cells (for example, cells of a hematopoietic stem cell marker + ) from the cell population, a step of culturing the separated cells, a step of recovering the cultured cells, or a step of holding the recovered cells in the container may be performed.

[0069] (16) Cell According to one embodiment of the present disclosure, CD201 + or CD90+ A cell population containing 5% or more of the specified cells is provided. This cell population can be used to produce pharmaceutical compositions for hematopoietic stem cell transplantation. According to one embodiment of this disclosure, CD41 + or CD34 + A cell population containing 5% or more of the specified cells is provided. This cell population can be used to produce pharmaceutical compositions for hematopoietic stem cell transplantation.

[0070] The aforementioned cell population preferably includes human hematopoietic stem cells, and CD201 + Or CD41 + It is preferable that the cell population includes the cells of [specific cell type].

[0071] The aforementioned cell population is CD34 + cells, CD201 + cells, CD90 + and CD41 + It is preferable to include at least one type of cell selected from the group consisting of the following cells.

[0072] The aforementioned cell population is CD201 + cells, CD90 + and CD41 + It is preferable that the cell population contains an enriched population of at least one type of cell selected from the group consisting of the following cells.

[0073] The cell population obtained by culturing in the aforementioned medium may be used to cultivate in a separately prepared, unused portion of the aforementioned medium.

[0074] The aforementioned cell population is CD201 + cells, CD90 + and CD41 + The composition contains at least one hematopoietic stem cell selected from the group of cells, and it is preferable that the number of hematopoietic stem cells after culturing with the composition for 10 days is greater than the number of hematopoietic stem cells at the start of culturing with the composition.

[0075] The aforementioned cell population is CD201 + cells and CD90+ The composition comprises at least one hematopoietic stem cell selected from the group consisting of the following cells, and CD201 after being cultured for 10 days using the composition. + [Number of cells] - [CD201 at the start of culture using the above composition] + The number of cells calculated by [the number of cells of CD90 after culturing for 10 days using the above composition] is [the number of CD90 + [Number of cells] - [CD90 at the time of starting culture using the composition] + It is preferable that the number of cells is greater than the number of cells calculated by [the number of cells].

[0076] (17) Culture medium According to one embodiment of the present disclosure, CD201 in total cells in culture medium + CD90 + Or CD41 + A cell-containing culture medium is provided, containing a cell percentage of 5% or more. This medium can be used for the production of pharmaceutical compositions for hematopoietic stem cell transplantation.

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

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

[0079] (20) Composition According to one embodiment of the present disclosure, a pharmaceutical composition for use in hematopoietic stem cell transplantation or a method for producing the same is provided, comprising cells or a population of cells obtained by any of the methods described in (3) to (12) above. 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, a hematopoietic stem cell marker from the composition described in (19) above. + The process of separating cells, and hematopoietic stem cell markers after separation. + The process of culturing cells, or hematopoietic stem cell markers after culture + The process may further include a step of recovering the cells. In another aspect, according to one embodiment of the present disclosure, the cells or cell population described in (15) or (16) above, or a pharmaceutical composition containing such cells or cell population, is provided for use in hematopoietic stem cell transplantation. In another aspect, according to one embodiment of the present disclosure, the use of the cells or cell population described in (15) or (16) above in the production of a pharmaceutical composition for hematopoietic stem cell transplantation is provided.

[0080] (21) Transplant method According to one embodiment of the present disclosure, a transplantation method is provided. This transplantation method may include, for example, the step of transplanting the cells or cell population described in (15) or (16) above, or a pharmaceutical composition containing such cells or cell population. This transplantation method allows for the effective engraftment of hematopoietic stem cells, thereby enabling the treatment of a disease.

[0081] (22) Method of using the cell population According to one embodiment of the present disclosure, at least one therapeutic agent can be provided, selected from the group consisting of cancer therapeutic agents, immunodeficiency therapeutic agents, and multiple myeloma therapeutic agents, including the aforementioned cell population. This therapeutic agent can be used by transplanting it into a subject, for example, according to the transplantation method described in (21) above. This therapeutic agent can effectively engraft hematopoietic stem cells, thereby enabling treatment of the disease.

[0082] (23) Method of using the compound According to one embodiment of the present disclosure, a medicament containing at least one compound selected from the group consisting of the compound represented by the above general formula (I) and the compound represented by the above general formula (II) can be used for producing a culture medium for hematopoietic stem cell proliferation. According to one embodiment of the present disclosure, a medicament containing at least one compound selected from the group consisting of the compound represented by the above general formula (I) and the compound represented by the above general formula (II) can be used as a medicament for a culture medium for hematopoietic stem cell proliferation. Preferably, the medicament contains at least one compound selected from the group consisting of Compound 126 and Compound 135. Other components that may be contained in the medicament include those described later as arbitrary base components.

[0083] In an embodiment of the present disclosure (including, for example, the above (3) to (12) or (21)), the method may include any one or more of the following steps (i) to (v). (i) A step of placing or holding the composition of (1) above in a container before culturing, (ii) A step of bringing the cells into contact with the composition in the container, (iii) A step of culturing the cells in the container, (iv) CD201 + , CD90 + Or a step of generating a cell population in which cells of CD41 + are concentrated, or (v) A step of generating a cell-containing culture medium in which the ratio of cells of CD201 + , CD90 + Or CD41 + is 5% or more with respect to the total cells in the culture medium. The method of an embodiment 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) A step of preparing a cell population containing hematopoietic stem cells, (vii) A step of preparing a cell population containing CB CD34 + cells, (viii) A step of separating CD34 + cells from the cell population, (ix) CD34 after separation +(x) A step of culturing the cells, (x) a step of producing the composition of (1) above before culturing, (xi) a step of incubating the cells in a culture medium, (xii) a step of seeding the cells into a culture medium in a container and incubating them, (xiii) a step of CD201 from the cell population + CD90 + Or CD41 + (xiv) A step of separating or recovering cells, (xv) A step of separating or recovering hematopoietic stem cells from the cell population, (xv) CD201 after separation or recovery + CD90 + Or CD41 + (xvi) A step of culturing the cells, (xvii) A step of culturing the hematopoietic stem cells after sorting or recovery, (xvii) CD201 after sorting or recovery and culture + , is CD90 + Or CD41 + (xviii) Steps to collect the cells, (xix) Steps to collect the hematopoietic stem cells after sorting or collection and culture, (xix) Steps to collect the CD201 + CD90 + Or CD41 + The process of preserving (e.g., cryopreserving) the cells, or (xx) CD201 after recovery + CD90 + Or CD41 + The process of transplanting cells. Employing one or more of these processes is considered a CD201 procedure. + CD90 + Or CD41 + This is useful for efficiently producing a culture fraction containing a large amount of cells. When two or more steps from embodiments of this disclosure (for example, including the methods of (3) to (12), (21) above, or the steps of (i) to (xx)) are adopted, the order is arbitrary and can be determined according to the desired operation. Each method or step of embodiments of this disclosure (for example, including the methods of (3) to (12), (21) above, or the steps of (i) to (xx)) may be carried out in vitro or ex vivo.

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

[0085] In embodiments of this 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 lists CD90 as a hematopoietic stem cell marker). CD90 includes a protein having the amino acid sequence of the protein indicated by NCBI RefSeq Accession number NP_001298089.1 or UniProt ID P04216. CD90 is sometimes referred to as Cluster of Differentiation 90.

[0086] In embodiments of this disclosure (including, for example, (1) to (23) above), CD41 is a membrane protein that is mainly expressed in megakaryocytes and platelets. CD34 + And CD41 + These cells are cells that are inclined to differentiate into megakaryocytes.

[0087] In embodiments of the present disclosure (including, for example, (1) to (23) above), 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 embodiments of the present disclosure (including, for example, (1) to (23) above), the LSD1 inhibitor preferably comprises at least one component (B) selected from the group consisting of tranylcypromine and salts thereof.

[0089] In embodiments of this disclosure (including, for example, (1) to (23) above), tranylcypromine includes the compound with CAS registry number 155-09-9 or PubChem CID 96025309. Trannylcypromine is sometimes referred to as 2-PCPA. Trannylcypromine contains (1R,2S)-2-phenylcyclopropane-1-amine. In this specification, tranylcypromine may be written as 2-PCPA for abbreviation, but these are interchangeable.

[0090] In embodiments of the present disclosure (including, for example, (1) to (23) above), the PI3K activator comprises an activator of phosphatidylinositol 3-kinase. The PI3K activator comprises a PI3K agonist, for example, 740Y-P or a salt thereof. 740Y-P comprises a compound with CAS registry number 1236188-16-1 or PubChem CID 90488730.

[0091] In embodiments of the present disclosure (including, for example, (1) to (23) above), culture includes incubating cells under conditions suitable for growth or maintenance. Culture may include the steps of contacting a culture medium and cells to produce a cell-containing composition, and incubating the cell-containing composition. The incubation step may be carried out with the cell-containing composition standing or agitated. Incubation may be carried out at approximately 37°C and in an atmosphere of approximately 5% CO2. Contact includes, for example, seeding cells into a culture medium or mixing cells with a culture medium. Culture may be carried out in an albumin-free or serum-free medium. Free includes the state in which the target component is not added to the medium, the state in which the medium does not contain the target component at all, or the state in which the medium does not contain the component at a concentration above the detection limit. Albumin-free includes being substantially albumin-free.

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

[0093] In embodiments of the present disclosure (including, for example, (1) to (23) above), culture may include growing hematopoietic stem cells under conditions sufficient for the maintenance or proliferation of hematopoietic stem cells. Growth may include, for example, growing hematopoietic stem cells tenfold or more from the start of culture. This multiplier may be 10, 50, 100, 200, 300, 400, or 500 times or more, and may be within the range of any two of these values.

[0094] The cells used for culture in embodiments of this disclosure (including, for example, (1) to (23) above) may be umbilical cord blood (CB) cells, peripheral blood cells, bone marrow cells, or a population of such cells. The cells include cells collected from CB, cells collected from peripheral blood, and cells collected from bone marrow. These cells or cell populations are CD34 + It may be a cell. For example, a cell is CB CD34 + Cells (CD34 derived from CB) + Cells), peripheral blood CD34 + Cells (CD34 derived from peripheral blood) + (Cells) or bone marrow CD34 + Cells (CD34 derived from bone marrow) + These may be cells. These cells may be purchased from a manufacturer or distributor (e.g., StemExpress or Lonza) and obtained from CB, peripheral blood, or bone marrow. + They may also be obtained by separating cells. In another embodiment of this disclosure, the cells used in culture or obtained in culture are stem cells, hematopoietic stem cells, or CD34 + CD201 + CD90 + Or CD41 +This includes cells of the following species. The cells used for culture or the cell population thereof may also be referred to as starting cells or starting cell population. In embodiments of this disclosure (including, for example, (1) to (23) above), the cells used for culture or the cells obtained by culture may be mammalian cells. Mammals include, for example, humans, monkeys, rodents (mice, hamsters, etc.), rabbits, dogs, cats, horses, cattle, sheep, pigs, goats, marmosets, etc. In embodiments of this disclosure (including, for example, (1) to (23) above), the cells may exist in the form of a cell population.

[0095] In embodiments of the present disclosure (including, for example, (1) to (23) above), the cell population includes, for example, a plurality of cells produced by cell division. In embodiments of the present disclosure, CD201 is added to the total cells in the culture medium after cultivation. + CD90 + Or CD41 + The percentage of 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, and may be within the range of any two of these values. This percentage may be, for example, 5-50%, 15-40%, or 25-40%. CD201 + The cell percentage 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. CD90 + The cell percentage is preferably 13% or more, and more preferably 15% or more, in order to efficiently obtain hematopoietic stem cells. In the embodiments of this disclosure, CD201 in the cell population after culture + Or CD90 + The proportion of cells may be five times or more compared to before culture. This multiple may be, for example, 10, 20, 50, 100, or 200 times or more, and may be within the range of any two of these values. CD41 + The cell proportion is preferably 13% or more, and more preferably 15% or more, in order to differentiate into megakaryocytes. The cell proportion may also be determined by the culture time and the proportion of cells when cultured, as described later.

[0096] In embodiments of this disclosure (including, for example, (1) to (23) above), hematopoietic stem cells include stem cells capable of differentiating into hematopoietic cells. Hematopoietic stem cells can be collected from the umbilical cord, bone marrow, placenta, and peripheral blood. Human hematopoietic stem cells are CD34 + It contains cells. Human hematopoietic stem cells are CD34 + It may also be CD38-. Human hematopoietic stem cells are CD34- + CD201 + CD90 + CD34 + CD201 + or CD34 + CD41 + It may also be the case that the human hematopoietic stem cell marker is CD34. + CD201 + CD90 + Or CD41 + This includes hematopoietic stem cells, including long-term hematopoietic stem cells.

[0097] In embodiments of the present disclosure (including, for example, (1) to (23) above), the cell culture time 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 longer, and may be within the 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. The culture includes the time for incubating the culture medium containing the cells.

[0098] In embodiments of the present disclosure (including, for example, (1) to (23) above), the cell seeding density during culture may be 1000 cells / mL. This value may be 1000, 3000, 5000, 10000, 30000, 50000, 70000, 100000, or 150000 or more, and may be within the range of any two of these values.

[0099] In embodiments of the present disclosure (including, for example, (1) to (23) above), the composition or culture medium includes a culture medium used for cell culture. The base components of the culture medium may be any common base components, and its composition is not limited. The base components of the culture medium may include, for example, amino acids, peptides, proteins, inorganic salts, vitamins, minerals, or carbon sources (e.g., glucose). The base components of the culture 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 culture medium may include, for example, a basic culture medium for cell culture. Basic culture media include, for example, 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), KnockOut DMEM / F-12 medium (Invitrogen), Stemline hematopoietic stem cell proliferation medium (Sigma), SYN serum-free medium (SYN H, SYN B) (AbCys SA), SPE IV medium (AbCys SA), and MyeloCult medium (StemCell This includes HPG serum-free medium (Lonza Technologies), UltraCULTURE medium (Lonza), Opti-MEM medium (Gibco Invitrogen, etc.), MEM medium (Gibco Invitrogen, etc.), MEMα (Gibco Invitrogen, etc.), DMEM medium (Gibco Invitrogen, etc.), IMDM medium (Wako Pure Chemical, etc.), PRMI1640 medium (Gibco Invitrogen, etc.), Ham F-12 medium (Gibco, etc.), RD medium, etc.The culture medium is preferably albumin-free (e.g., GenBank registry number AAN17825.1) or serum-free. In this case, stable inhibition of undifferentiation and problems of biological contamination can be suppressed. The culture medium may contain Soluplus® (CAS registry number 402932-23-4). Soluplus mainly consists of a graft polymer of N-vinylcaprolactam-vinyl acetate copolymer and oxirane. The culture medium may contain antibiotics (e.g., penicillin or streptomycin). The culture medium contains hematopoietic stem cells and CD201. + cells, CD90 + cells, CD41 + The medium may include cells, including CB cells. The culture medium may be a medium for culturing these cells. The culture medium may be a liquid medium or a solid medium. Culturing cells using a culture medium includes culturing cells in a culture medium.

[0100] In embodiments of this disclosure (including, for example, (1) to (23) above), the composition or culture medium is CB CD34 + The composition or culture medium may further contain cells. By incubation, the composition or medium will produce CD201 + CD90 + Or CD41 + The composition or culture medium contains cells that are concentrated.

[0101] In embodiments of the present disclosure (including, for example, (1) to (23) above), the concentration of 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 higher, and may be within the 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. This concentration is preferably 2 μmol / L or higher, more preferably 3 μmol / L or higher, and even more preferably 4 μmol / L or higher, in order to efficiently obtain hematopoietic stem cells.

[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 higher, and may be within the 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. This concentration is preferably 0.03 μmol / L or higher, more preferably 0.05 μmol or higher, and even more preferably 0.08 μmol / L or higher, in order to efficiently obtain hematopoietic stem cells.

[0103] In embodiments of the present disclosure (including, for example, (1) to (23) above), the concentration of UM729 or its salt in the 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 higher, and may be within the 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. This concentration is preferably 0.3 μmol / L or higher, more preferably 0.5 μmol / L or higher, and even more preferably 0.8 μmol / L or higher, in order to efficiently obtain hematopoietic stem cells.

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

[0105] In embodiments of the present disclosure (including, for example, (1) to (23) above), the composition or culture medium may contain, as other protein components, a protein component in 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, and may be within the 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, an Flt3 ligand.

[0106] In embodiments of the present disclosure (including, for example, (1) to (23) above), the composition or culture medium may contain modified polyalkylene glycol as another additive 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, and may be within the range of any two of these values. This concentration may be, for example, 0.001 to 0.1% (w / v), 0.005 to 0.05% (w / v), 0.005 to 0.02% (w / v), 0.008 to 0.02% (w / v), or 0.008 to 0.015% (w / v). Modified polyalkylene glycol may be, for example, a polyalkylene glycol modified with a copolymer of polyvinylcaprolactam block and polyvinyl acetate block. Polyalkylene glycol modified with a copolymer of polyvinylcaprolactam block and polyvinyl acetate block may be, for example, polyethylene glycol modified with a copolymer of polyvinylcaprolactam block and polyvinyl acetate block. Modified polyalkylene glycol may be, for example, a polyvinylcaprolactam-polyvinylacetate-polyethylene glycol graft copolymer. Modified polyalkylene glycol may be, for example, a graft copolymer obtained from (i) N-vinylcaprolactam, (ii) vinyl acetate, and (iii) polyether. The polyether may be polyethylene glycol. Modified polyalkylene glycol may be, for example, a compound obtained by free radical polymerization of a mixture of (i) to (iii) above. 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%, and may be within the range of any two of these values. (ii) may be, for example, 15, 20, 25, 30, or 35 wt%, and may be within the range of any two of these values.(iii) may be, for example, 10, 13, 15, 20, 25, or 30 wt%, and may be within the range of any two of these values. The above (i) to (iii) may total 100 wt%. The modified polyalkylene glycol may be, for example, the compound represented by the following structural formula (S) or a salt thereof.

[0107] [ka]

[0108] Here, the wt% of l, m, and n may be, for example, 40-60 wt%, 15-35 wt%, and 10-30 wt%, respectively. The wt% of l may be, for example, 40, 45, 50, 55, 57, or 60, and may be within the range of any two of these values. The wt% of m may be, for example, 15, 20, 25, 30, or 35, and may be within the range of any two of these values. The wt% of n may be, for example, 10, 13, 15, 20, 25, or 30, and may be within the range of any two of these values. The sum of the above wt%s of l, m, and n may be 100 wt%. The wt%s of l, m, and n may be, for example, about 57, about 30, and about 13 wt%, respectively. The degrees of polymerization of l, m, and n may be, for example, 60-160, 470-1110, and 480-1130, respectively. The degree of polymerization of l may be, for example, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or 160, and may be within the 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, and may be within the 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, and may be within the range of any two of these values. The degree of polymerization may also be the average degree of polymerization. The mass-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, and may be within the range of 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 also 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. 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. In addition, the components (i) to (iii) above may be components described in these documents.

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

[0110] In embodiments of the present disclosure (including, for example, (1) to (23) above), the recovery step may include, for example, transferring the desired cells or cell population from a container containing cells or cell populations to another container. Recovery may include a separation or purification step. Recovery may be carried out in a sterile environment.

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

[0112] In embodiments of this disclosure (including, for example, (1) to (23) above), the pharmaceutical composition may be prepared by mixing cells (e.g., hematopoietic stem cells) with one or more pharmaceutically acceptable carriers by any method known in the art of pharmaceuticals. The pharmaceutical composition may contain stabilizers, buffers, or pH adjusters. The dosage, administration interval, method of administration, and route of administration are not particularly limited and can be appropriately selected depending on the patient's age, weight, symptoms, target organ, etc. The pharmaceutical composition preferably contains a therapeutically effective amount or an effective amount of hematopoietic stem cells that exert the desired effect. In one embodiment of this disclosure, a therapeutically effective amount includes the amount necessary for clinically observed improvement or suppression of symptoms in the patient. In one embodiment of this disclosure, pharmaceutically acceptable includes a state suitable for use in proportion to a reasonable benefit-risk ratio, within the bounds of reasonable medical judgment.

[0113] In embodiments of the present disclosure (including, for example, (1) to (23) above), the subject (including the patient) includes one or more non-human mammals (e.g., mice, guinea pigs, hamsters, rats, mice, rabbits, pigs, sheep, goats, cattle, horses, cats, dogs, marmosets, monkeys, or chimpanzees). The patient may also be a patient diagnosed with a disease (e.g., cancer or immunodeficiency) or a patient in need of treatment for a disease.

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

[0115] In embodiments of the present disclosure (including, for example, (1) to (23) above), the salt is not particularly limited and includes, for example, inorganic salts or organic salts (see, for example, “Bharate et al., Drug Discov Today. 2021 Feb;26(2):384-398.” or “Berge et al., J Pharm Sci. 1977 Jan;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, etc. 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, etc. 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 this disclosure, "pharmaceutically acceptable" includes a form having reasonable benefit for pharmaceutical use.

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

[0117] All publications cited herein are referred to in their entirety. In this specification, “or” is used when “at least one” of the items listed in the text can be adopted. The same applies to “or.” For example, CD201 + Or CD90 + CD201 + CD90 + CD201 + and CD90 + This includes any of the following. Where it is specified herein that “within the range of two values” includes the two values ​​themselves. Where it is specified herein that “A to B” includes A and B. Where it is specified herein that “(1) to (23) above” includes references to 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 component (A) in (1) above, a compound represented by the following formula (III) or (IV) may be used. [ka]

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

[0120] [ka]

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

[0122] As the compound represented by formula (III) above, the compound represented by the following formula (NPC-4034) is preferred.

[0123] [ka]

[0124] As the compound represented by formula (IV) above, the compound represented by the following formula (T-31) is preferred.

[0125] [ka]

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

[0127] While embodiments of the present disclosure have been described above, these are merely examples of forms that may be included in the present disclosure, and the present disclosure is not limited to these, and various other configurations may be adopted. Furthermore, the present disclosure may be adopted in combination or independently of the configurations or features described in the embodiments above.

[0128] <Effects and Actions> According to this disclosure, compositions and culture media for use in cell culture can be provided. Specifically, CD34 + cells, CD201 + cells, CD90 + , and CD41 + It is possible to proliferate a cell population containing these cells. [Examples]

[0129] The present disclosure is not limited to the following examples, which will be explained further below. In the examples, compound 126 (TIA01-126) is a compound represented by formula (I), where A is an m-phenylene group, L is an n-pentylene group (a linear alkylene group with 5 carbon atoms), and X is -(CH2) tThe group is represented by O-, and t is 6. Compound 135 (TIA02-135) is a compound represented by formula (II), where R is a methyl group and Z is an n-nonylene group (a linear alkylene group with 9 carbon atoms).

[0130] <Example 1> Figure 1 shows the results of analyzing human umbilical cord blood CD34+ cells cultured with the compound 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)] using flow cytometry. The vertical axis shows the CD34+ lineage marker-cell percentage. The horizontal axis shows the compounds, with each representing a combination of 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, 2PCPA: 5.0 μmol / L 2PCPA, and DMSO: 0.1% DMSO.

[0131] <Example 2> Figure 2 shows the results of analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with compounds for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker-cell proliferation rate. The horizontal axis represents the compounds, specifically TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with compounds for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker - CD201+CD45RA- rate in cells. The horizontal axis represents the compounds, specifically TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with a compound for 10 days using flow cytometry. The vertical axis represents the number of CD34+ lineage markers -CD201+CD45RA-. The horizontal axis represents the compounds, specifically TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with compounds for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker - CD90+CD45RA- rate in cells. The horizontal axis represents the compounds, specifically TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with compounds for 10 days using flow cytometry. The vertical axis represents the number of CD34+ lineage markers - CD90 + CD45RA - cells. The horizontal axis represents the compounds, specifically TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA, TIA02 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker-cell percentage. The horizontal axis represents the compounds, and they are as follows: 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 analyzing human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker-cell proliferation rate. The horizontal axis represents the compounds, specifically 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with a compound for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker - CD201+CD45RA- rate in the cells. The horizontal axis shows the compounds, and each represents the following: TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA; TIA02 / 2PCPA: combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA; NCP / 2PCPA: combination of 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA; T-31 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with a compound for 10 days using flow cytometry. The vertical axis shows the CD34+ lineage marker, cell CD201+, and CD45RA, respectively, and cell number. The horizontal axis shows the compounds, and each represents the following: TIA01 / 2PCPA: combination of 5.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA; TIA02 / 2PCPA: combination of 1.0 μmol / L TIA02-135 and 5.0 μmol / L 2-PCPA; NCP / 2PCPA: combination of 1.0 μmol / L NCP-4034 and 5.0 μmol / L 2-PCPA; T-31 / 2PCPA: 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 analyzing human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker - the percentage of CD90+CD45RA- cells in the cells. The horizontal axis represents the compounds, and they are as follows: TIA01 / 2PCPA: 5.0 μmol / L TIA01-126 in combination with 5.0 μmol / L 2-PCPA; TIA02 / 2PCPA: 1.0 μmol / L TIA02-135 with 5.0 μmol / L 2-PCPA; NCP / 2PCPA: 1.0 μmol / L NCP-4034 with 5.0 μmol / L 2-PCPA; T-31 / 2PCPA: 1.0 μmol / L T-31 with 5.0 μmol / L 2-PCPA; and 2PCPA: 5.0 μmol / L 2PCPA.

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

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

[0144] <Example 15> Figure 15 shows the results of analyzing human umbilical cord blood CD34+ cells cultured with compounds in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the CD34+ lineage marker - CD201+CD45RA- rate in cells. The horizontal axis represents the compounds, specifically 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 analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with compounds for 10 days using flow cytometry. The vertical axis represents the number of CD34+ lineage markers -CD201+CD45RA-. The horizontal axis represents the compounds, specifically TIA01: 5.0 μmol / L TIA01-126, TIA02: 1.0 μmol / L TIA02-135, NCP: 1.0 μmol / L NCP-4034, T-31 / 2PCPA: combination of 1.0 μmol / L T-31 and 5.0 μmol / L 2-PCPA, and 2PCPA: 5.0 μmol / L 2PCPA.

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

[0148] <Example 19> Figure 19 shows the results of analyzing human umbilical cord blood CD34+ cells cultured for 12 days with compounds 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)] using flow cytometry. The vertical axis shows the CD34+ lineage marker-cell percentage. The horizontal axis shows the compounds, with DMSO: 0.1% DMSO and TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA being used in combination.

[0149] <Example 20> Figure 20 shows the results of analyzing human umbilical cord blood CD34+ cells cultured in albumin-based medium with compounds for 12 days using flow cytometry. The vertical axis represents the CD34+ lineage marker and cell number. The horizontal axis represents the compounds, with DMSO: 0.1% DMSO and TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA being used in combination.

[0150] <Example 21> Figure 21 shows the results of analyzing human umbilical cord blood CD34+ cells cultured in albumin-based medium with compounds for 12 days using flow cytometry. The vertical axis represents the CD34+ lineage marker - the CD201+CD45RA- rate in the cells. The horizontal axis represents the compounds, with DMSO: 0.1% DMSO and TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA being used in combination.

[0151] <Example 22> Figure 22 shows the results of analyzing human umbilical cord blood CD34+ cells cultured in albumin-based medium with compounds for 12 days using flow cytometry. The vertical axis represents the number of CD34+ lineage markers -CD201+CD45RA-. The horizontal axis represents the compounds, with DMSO: 0.1% DMSO and TIA01 / 2PCPA: 4.0 μmol / L TIA01-126 and 5.0 μmol / L 2-PCPA being used in combination.

[0152] <Example 23> Figure 23 shows the results of analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with a compound for 10 days using flow cytometry. The vertical axis represents the percentage of CD41+CD34+ cells among all viable cells. The horizontal axis shows the compounds and their content, 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, and 2PCPA_5: 5.0 μmol / L 2-PCPA.

[0153] <Example 24> Figure 24 shows the results of analyzing human umbilical cord blood CD34+ cells cultured with a compound in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the number of CD41+CD34+ cells. The horizontal axis represents the compound and its content, which are the same as in Figure 23.

[0154] <Example 25> Figure 25 shows the results of analyzing human umbilical cord blood CD34+ cells cultured with a compound in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the percentage of CD41+CD34+ cells among all viable cells. The horizontal axis represents the compound and its content, with the same type and content of compound as in Figure 23, but with the addition of 5.0 μmol / L 2-PCPA. 2PCPA_5: represents 5.0 μmol / L 2-PCPA.

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

[0156] <Example 27> Figure 27 shows the results of analyzing human umbilical cord blood CD34+ cells cultured in Soluplus-based medium with a compound for 10 days using flow cytometry. The vertical axis represents the percentage of CD41+CD34+ cells among all viable cells. The horizontal axis shows the compound and its content, with the following combinations: P3_T1: 3 μmol / L 2-PCPA and 1.0 μmol / L T1A02-135, P3_T3: 3 μmol / L 2-PCPA and 3 μmol / L T1A02-135, P4_T1: 4.0 μmol / L 2-PCPA and 1.0 μmol / L T1A02-135, P4_T2: 4.0 μmol / L 2-PCPA and 2 μmol / L T1A02-135, P4_T3: 4.0 μmol / L 2-PCPA and 3 μmol / L T1A02-135, P5_T1: 5.0 μmol / L 2-PCPA and 1.0 μmol / L T1A02-135, P5_T2: The following shows the combined use of 5.0 μmol / L 2-PCPA and 2 μmol / L T1A02-135, and P5_T3 shows the combined use of 5.0 μmol / L 2-PCPA and 3 μmol / L T1A02-135.

[0157] <Example 28> Figure 28 shows the results of analyzing human umbilical cord blood CD34+ cells cultured with a compound in Soluplus-based medium for 10 days using flow cytometry. The vertical axis represents the number of CD41+CD34+ cells. The horizontal axis represents the compound and its content, which are the same as in Figure 27.

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

[0159] <Example 30> Figure 30 shows the ratio of mature megakaryocytes (CD41+CD42b+ cells) to the cell population when cells obtained by culturing human umbilical cord blood CD34+ cells with the compound in Soluplus-based medium for 14 days (proliferation rate 27.0 times, CD34 positivity rate 73.4%, viability rate 88.1%, CD41 positivity rate 15.2%) were cultured for 14 days in megakaryocyte differentiation medium (StemSpan SFEM, StemCell Technologies, catalog number ST-09600). From the above results, it was found that this disclosure provides compositions and culture media for use in cell culture. [Industrial applicability]

[0160] According to this disclosure, compositions and culture media for use in cell culture can be provided. Furthermore, this disclosure provides a cell population that can be transplanted into patients with diseases such as cancer and immunodeficiency, as well as a method for producing the same. Furthermore, according to this disclosure, therapeutic agents comprising the aforementioned cell population can be provided.

Claims

1. CD34 comprising the following composition (1) or composition (2) ＋ cells, CD201 ＋ cells, CD90 ＋ The cells of, and CD41 ＋ A culture medium for the growth of cells selected from a group consisting of the following cells: A composition (1) comprising a component selected from (A) a KDM5 degrading agent and (B) tranylcypromine and its salts, or (A) A composition (2) containing a KDM5 decomposing agent, Here, the component (A) in compositions (1) and (2) includes at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts 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, 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 - represented group, s is a number from 1 to 8, and t is a number from 2 to 10; 【Chemistry 2】 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. Component (A) in compositions (1) and (2) comprises at least one component selected from the group consisting of compounds represented by the general formula (I) and salts thereof, and compounds represented by the general formula (II) and salts thereof. In the general formula (I) above, 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 a group represented by O-, where s is a number from 1 to 5, and t is a number from 2 to 10. The culture medium according to claim 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.

3. Component (A) in compositions (1) and (2) comprises at least one component selected from the group consisting of compounds represented by the general formula (I) and salts thereof, and compounds represented by the general formula (II) and salts thereof. In the general formula (I) above, 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 a group represented by O-, where s is a number from 1 to 5, and t is a number from 2 to 10. The culture medium according to claim 1, wherein 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.

4. Component (A) in compositions (1) and (2) comprises at least one component selected from the group consisting of compounds represented by the general formula (I) and salts thereof, and compounds represented by the general formula (II) and salts thereof. The culture medium according to claim 1, wherein the component (A) is at least one 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: 【Transformation 3】 In general formula (I-1), L １ X is a linear alkylene group having 3 to 10 carbon atoms. １ ha- (CH ２ CH ２ O) ｓ It is a base represented by -, where s is a number from 1 to 5, and in formula (I-2), L ２ X is a linear alkylene group having 3 to 10 carbon atoms. ２ 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, and in formula (I-3), L ３ X is a linear alkylene group having 3 to 10 carbon atoms. 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 Z is a linear alkyl group having 1 to 3 carbon atoms. １ This 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 above-mentioned component (A) is, In the general formula (I-1), L １ X is a linear alkylene group with 5 carbon atoms, １ ga- (CH ２ CH ２ O) ｓ A compound with a group represented by - and s = 3, In the general formula (I-1), L １ This is a linear alkylene group with 8 carbon atoms, X １ ga- (CH ２ CH ２ O) ｓ A compound with a group represented by - and s = 3, In the general formula (I-2), L ２ X is a linear alkylene group with 5 carbon atoms, ２ ga- (CH ２ CH ２ O) ｓ A compound in which the group is represented by - and the number of s is 3, In the general formula (I-2), L ２ X is a linear alkylene group with 5 carbon atoms, ２ ga- (CH ２ ) ｔ A compound with a group represented by O-, where t is the number 6. In the general formula (I-3), L ３ X is a linear alkylene group with 5 carbon atoms, ３ ga- (CH ２ CH ２ O) ｓ A compound in which the group is represented by - and s is the number 3, In the general formula (I-3), L ３ X is a linear alkylene group with 5 carbon atoms, ３ ga- (CH ２ ) ｔ A compound having a group represented by O- and where t is a number 6, or In the above general formula (II-1), R １ is a methyl group, Z １ The culture medium according to claim 4, wherein is an unsubstituted, linear, nine-carbon alkylene group.

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

7. The culture medium according to claim 1, wherein the culture medium is a culture medium for hematopoietic stem cells.

8. The culture medium according to claim 7, wherein the hematopoietic stem cells are for differentiation into megakaryocytes, and the culture medium is used for culturing hematopoietic stem cells during the differentiation process into megakaryocytes.

9. The culture medium according to claim 1 or 7, which is a culture medium for concentrating CD90+ cells.

10. The culture medium is CD34 ＋ The culture medium according to claim 1, which is a culture medium for cells.

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

12. A method for producing cells, comprising a step of culturing cells selected from the group consisting of CD34 ＋ cells, CD201 ＋ cells, CD90 ＋ cells, and CD41 ＋ cells using the medium according to claim 1.

13. A method for producing a culture fraction concentrated with CD90+ cells, comprising the 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.

14. Using the culture medium described in claim 1, CD34 ＋ cells, CD201 ＋ cells, CD90 ＋ The cells of, and CD41 ＋ 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 the step of culturing cells selected from a group consisting of the following cells.

15. A compound comprising at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof, Components selected from tranylcypromine and its salts, Use of drugs containing the following for the production of hematopoietic stem cell growth media: 【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 It is a group represented by O-, where s is a number from 1 to 8, and t is a number from 2 to 10; 【Transformation 5】 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.

16. Use of a drug containing at least one component selected from the group consisting of compounds represented by the following general formula (I) and salts thereof, and compounds represented by the following general formula (II) and salts thereof, as a drug for hematopoietic stem cell proliferation culture media: 【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 It is a group represented by O-, where s is a number from 1 to 8, and t is a number from 2 to 10; 【Transformation 7】 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.

Images