Maturation promoter for stem cell-derived non-mature nerve cells, maturation promotion method, proliferation inhibitor, and proliferation suppression method

Abstract

The present invention pertains to: a maturation promoter for stem cell-derived non-mature nerve cells, the maturation promoter being characterized by containing at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof; and a method for promoting maturation of stem cell-derived non-mature nerve cells, the method being characterized by making stem cell-derived non-mature nerve cells coexist with the maturation promoter.

Description

Maturation promoter, maturation promoter method, proliferation inhibitor, and proliferation inhibitor method for stem cell-derived immature nerve cells

[0001] Related Applications This specification includes the contents of the specification of Japanese Patent Application No. 2024-131286 (filed August 7, 2024), which is the priority basis of this application. Technical Field The present invention relates to a maturation promoter, a maturation promoting method, a proliferation inhibitor, and a proliferation inhibitor method for stem cell-derived immature nerve cells.

[0002] The usefulness of cell transplantation in regenerative medicine for neurological disorders such as Parkinson's disease and cerebrovascular disorders is expected. In these cell transplantation methods, pluripotent stem cells such as iPS cells (induced pluripotent stem cells) or ES cells (embryonic stem cells) are used, and methods have been attempted in which the pluripotent stem cells are differentiated into target cells before transplantation. However, differentiated and mature target cells are vulnerable to environmental stress, and there is a problem of low intracerebral engraftment rate after transplantation. Therefore, in recent years, it has become clear that the use of progenitor cells that have a certain degree of differentiation directionality determined before terminal differentiation for transplantation is effective in terms of safety and efficacy. As an example, the present inventors have induced differentiation of human cerebral organoids mimicking the cerebral cortical layer structure in cerebrovascular disorders (Non-Patent Document 1), and by transplanting the progenitor cells into the brain of a cynomolgus monkey, they have confirmed the engraftment and differentiation of the transplanted cells (Non-Patent Document 2).

[0003] Sakaguchi et al, Stem Cell Reports 2019 Jun 27;13(3):458-473. doi: 10.1016 / j.stemcr.2019.05.029Kitahara et al, Stem Cell Reports 2020 Jul 16;15(2):467-481. doi: 10.1016 / j.stemcr.2020.06.016

[0004] Furthermore, research into cerebral organoids transplanted into the brain has revealed that some progenitor cells contained in the organoids continue to proliferate within the brain as progenitor cells. Therefore, promoting the maturation of progenitor cells and inhibiting their proliferation within the brain after transplantation is considered to be effective in improving therapeutic effects.

[0005] Therefore, an object of the present invention is to provide a novel agent that promotes the maturation of immature nerve cells derived from stem cells.

[0006] In a first aspect, the present invention provides the following [1] to

[22] . [1] A maturation promoter for stem cell-derived non-mature neurons, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. [2] The maturation promoter according to [1], wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons. [3] The maturation promoter according to [1] or [2], wherein the compound comprises asenapine or a salt thereof. [4] The maturation promoter according to [1] or [2], wherein the compound comprises entacapone or a salt thereof. [5] The maturation promoter according to [1] or [2], wherein the compound comprises asenapine or a salt thereof and entacapone or a salt thereof. [6] The maturation promoter according to any of [1] to [5], which is administered to a subject receiving or who has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue fragment containing stem cell-derived non-mature neurons (e.g., an organoid, preferably a cerebral organoid). [7] The maturation promoter according to [6], wherein the subject has a neurological disorder. [8] The maturation promoter according to [7], wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or bone marrow damage. [9] The maturation promoter according to any one of [1] to [8], wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells, preferably induced pluripotent stem cells (iPS).

[10] An agent for inhibiting the proliferation of stem cell-derived non-mature neurons, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

[11] A method for promoting the maturation of stem cell-derived non-mature neurons, comprising the step of coexisting stem cell-derived non-mature neurons with a maturation promoter in vitro or ex vivo, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

[12] The method for promoting maturation described in

[11] , wherein the stem cell-derived non-mature nerve cells are immature nerve cells or progenitor nerve cells.

[13] The method for promoting maturation according to

[11] or

[12] , wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells, preferably from induced pluripotent stem cells (iPS).

[14] The method for promoting maturation according to any one of

[11] to

[13] , wherein the stem cell-derived non-mature neurons are located in a cerebral organoid.

[15] The method for promoting maturation according to any one of

[11] to

[13] , wherein the stem cell-derived non-mature neurons are located in a tissue or tissue fragment.

[16] A method for inhibiting the proliferation of stem cell-derived non-mature neurons, comprising a step of coexisting the stem cell-derived non-mature neurons with a proliferation inhibitor in vitro or ex vivo, wherein the proliferation inhibitor comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

[17] A method for producing a stem cell-derived neural cell population, comprising culturing a cell population containing stem cell-derived immature neural cells in the presence of a maturation promoter to mature the immature neural cells in the cell population, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further comprise inducing differentiation of stem cells (preferably pluripotent stem cells, more preferably iPS cells) into neural cells.

[18] A method for producing a cerebral organoid, comprising culturing a three-dimensional cell mass containing stem cell-derived immature neural cells in the presence of a maturation promoter to mature the immature neural cells in the cell mass to obtain a cerebral organoid, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further comprise the step of culturing stem cells (preferably pluripotent stem cells, more preferably iPS cells) in a suspended aggregate form to obtain a three-dimensional cell mass.

[19] A pharmaceutical composition for assisting transplantation, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, which is administered to a subject who will receive or has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue fragment containing stem cell-derived non-mature neurons (e.g., organoids, preferably cerebral organoids).

[20] A pharmaceutical composition for treating neurological disorders, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, which is administered to a subject who will receive or has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue fragment containing stem cell-derived non-mature neurons (e.g., organoids, preferably cerebral organoids).

[21] The maturation promoter according to any of [1] to [9], the proliferation inhibitor according to

[10] , or the pharmaceutical composition according to

[19] or

[20] , which are administered orally or parenterally.

[22] The maturation promoter, proliferation inhibitor, or pharmaceutical composition according to

[21] , wherein the parenteral administration is intravenous administration, intraperitoneal administration, or intrathecal administration. The limitations of [1] to [9] above also apply to

[19] and

[20] .

[0007] In a second aspect, the present invention provides the following [1] to

[15] . [1] [1a], [1b], or [1c] below: [1a] at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof, for use in promoting the maturation of stem cell-derived non-mature neurons, which is administered to a subject who will receive or has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons. [1b] at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof, for use in treating a neurological disorder, which is administered to a subject who will receive or has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons (or used in combination with a transplant of stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons). [1c] At least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, for use in assisting transplantation, wherein the compound is administered to a subject receiving or who has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons. [2] The compound for use according to [1], wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons. [3] The compound for use according to [1] or [2], wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells, preferably induced pluripotent stem cells (iPS cells). [4] The compound for use according to any of [1] to [3], wherein the compound is asenapine or a salt thereof, preferably the maleate or hydrochloride salt, more preferably the maleate salt. [5] The compound for use according to any of [1] to [4], wherein the compound is entacapone or a salt thereof, preferably entacapone.[6] The compound for use according to any one of [1] to [5], wherein the compound promotes maturation of the transplanted stem cell-derived immature nerve cells in the subject and / or suppresses proliferation of the transplanted stem cell-derived immature nerve cells. [7] The compound for use according to any one of [1] to [6], wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or a bone marrow injury. [8] The compound for use according to any one of [1] to [7], wherein the tissue is a cerebral organoid. [9] The compound for use according to any one of [1] to [8], wherein the compound is administered orally or parenterally, preferably orally, intravenously, intraperitoneally, or intrathecally.

[10] [10a] A method for promoting the maturation of stem cell-derived non-mature neurons, the method comprising the step of coexisting the stem cell-derived non-mature neurons in vitro or ex vivo with at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof. [10b] A method for inhibiting the proliferation of stem cell-derived non-mature neurons, the method comprising the step of coexisting the stem cell-derived non-mature neurons in vitro or ex vivo with at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof. [10c] A method for producing a stem cell-derived neuronal cell population (enriched with mature neurons), comprising the step of culturing a cell population containing stem cell-derived non-mature neurons in the presence of a maturation promoter to mature the non-mature neurons in the cell population, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further comprise the step of inducing differentiation of the stem cells into neurons.[10d] A method for producing brain organoids, comprising culturing a three-dimensional cell mass containing stem cell-derived immature neurons in the presence of a maturation promoter to mature the immature neurons in the cell mass and obtain brain organoids, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further comprise culturing the stem cells in a suspension aggregate to obtain a three-dimensional cell mass.

[11] The method according to

[10] , wherein the stem cell-derived immature neurons are immature neurons or progenitor neurons.

[12] The method according to

[10] or

[11] , wherein the stem cell-derived immature neurons are derived from pluripotent stem cells, preferably induced pluripotent stem cells (iPS cells).

[13] The method according to any one of

[10] to

[12] , wherein the compound is asenapine or a salt thereof, preferably the maleate or hydrochloride salt, more preferably the maleate salt.

[14] The method according to any one of

[10] to

[13] , wherein the compound is entacapone or a salt thereof, preferably entacapone.

[15] The method according to any one of

[10] to

[14] , wherein the stem cell-derived non-mature neural cells are present in a tissue or tissue fragment, preferably in a cerebral organoid.

[0008] In a third aspect, the present invention provides the following [1] to

[35] . [1] A method for promoting the maturation of stem cell-derived non-mature neurons, comprising coexisting (contacting) the stem cell-derived non-mature neurons with at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. [2] The method according to [1], wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons. [3] The method according to [1], wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells. [4] The method according to [1], wherein the stem cell-derived non-mature neurons are derived from induced pluripotent stem cells (iPS cells). [5] The method according to [1], wherein the compound is asenapine or a salt thereof. [6] The method according to [1], wherein the compound is entacapone or a salt thereof. [7] The method according to [1], comprising administering the compound to a subject in need thereof. [8] The method of [1], which comprises administering the compound to a subject who will receive or has received a transplant of stem cell-derived non-mature nerve cells, or a tissue or tissue graft containing stem cell-derived non-mature nerve cells. [9] The method of [8], wherein the subject has a neurological disorder.

[10] The method of [8], wherein the tissue is a cerebral organoid.

[11] The method of [1], wherein the method is an in vitro or ex vivo method.

[13] The following [13a] or [13b] [13a] A method for treating a neurological disorder, which comprises administering to a subject at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, wherein the subject is a subject who will receive or has received a transplant of stem cell-derived non-mature nerve cells, or a tissue or tissue graft containing stem cell-derived non-mature nerve cells.[13b] A method for treating a neurological disorder, comprising transplanting stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons, into a subject in need thereof, and administering to the subject at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

[14] The method of

[13] , wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons.

[15] The method of

[13] , wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells.

[16] The method of

[13] , wherein the stem cell-derived non-mature neurons are derived from induced pluripotent stem cells (iPS cells).

[17] The method of

[13] , wherein the compound is asenapine or a salt thereof.

[18] The method of

[13] , wherein the compound is entacapone or a salt thereof.

[19] The method of

[13] , wherein the tissue is a cerebral organoid.

[20] A method for assisting transplantation, comprising administering to a subject at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, wherein the subject is a recipient or has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons.

[21] The method according to

[20] , wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons.

[22] The method according to

[20] , wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells.

[23] The method according to

[20] , wherein the stem cell-derived non-mature neurons are derived from induced pluripotent stem cells (iPS cells).

[24] The method according to

[20] , wherein the compound is asenapine or a salt thereof.

[25] The method according to

[20] , wherein the compound is entacapone or a salt thereof.

[26] The method according to

[20] , wherein the subject has a neurological disorder.

[27] The method according to

[20] , wherein the tissue is a cerebral organoid.

[28] The method according to [9], wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or a bone marrow injury.

[29] The method according to

[13] , wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or bone marrow damage.

[30] The method according to

[26] , wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or bone marrow damage.

[31] The method according to [1], wherein the compound is administered orally, intravenously, intraperitoneally, or intrathecally.

[32] The method according to

[13] , wherein the compound is administered orally, intravenously, intraperitoneally, or intrathecally.

[33] The method according to

[20] , wherein the compound is administered orally, intravenously, intraperitoneally, or intrathecally.

[34] A method for producing a stem cell-derived neuronal population, comprising the step of culturing a cell population containing stem cell-derived non-mature neurons in the presence of a maturation promoter to mature the non-mature neurons in the cell population to obtain a neuronal population, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine, a salt thereof, a solvate thereof, or a derivative thereof, and entacapone, a salt thereof, a solvate thereof, or a derivative thereof.

[35] A method for producing a cerebral organoid, comprising the steps of culturing a three-dimensional cell mass containing stem cell-derived immature neurons in the presence of a maturation promoter to mature the immature neurons in the cell mass and obtain a cerebral organoid, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof.

[0009] In a fourth aspect, the present invention provides the following [1] to

[25] . [1] Use of at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, in the manufacture of a medicament for promoting the maturation of stem cell-derived non-mature neurons. [2] The use according to [1], wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons. [3] The use according to [1] or [2], wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells, preferably induced pluripotent stem cells (iPS cells). [4] The use according to any of [1] to [3], wherein the compound is asenapine or a salt thereof, preferably the maleate or hydrochloride salt, more preferably the maleate salt. [5] The use according to any of [1] to [4], wherein the compound is entacapone or a salt thereof, preferably entacapone. [6] The use according to any one of [1] to [5], wherein the medicament is for a subject receiving or who has received a transplant of stem cell-derived immature neurons, or a tissue or tissue fragment containing stem cell-derived immature neurons. [7] The use according to [6], wherein the compound promotes maturation of the transplanted stem cell-derived immature neurons in the subject and / or suppresses proliferation of the transplanted stem cell-derived immature neurons. [8] The use according to [6] or [7], wherein the subject has a neurological disorder (preferably, a brain disorder, a neurodegenerative disease, or a bone marrow injury). [9] The use according to any one of [6] to [8], wherein the tissue is a cerebral organoid.

[10] The use according to any one of [1] to [9], wherein the medicament is a formulation for oral or parenteral administration, preferably a formulation for oral, intravenous, intraperitoneal, or spinal administration.

[11] [11a] or [11b] [11a] Use of at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, in the manufacture of a medicament for treating a neurological disorder, wherein the medicament is for a subject who will receive or has received a transplant of stem cell-derived immature nerve cells, or a tissue or tissue graft containing stem cell-derived immature nerve cells.[11b] Use of at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, in the manufacture of a medicament for assisting transplantation, wherein the medicament is for a subject receiving or who has received a transplant of stem cell-derived non-mature neurons, or a tissue or tissue graft containing stem cell-derived non-mature neurons.

[12] The compound for use according to

[11] , wherein the stem cell-derived non-mature neurons are immature neurons or progenitor neurons.

[13] The compound for use according to

[11] or

[12] , wherein the stem cell-derived non-mature neurons are derived from pluripotent stem cells, preferably induced pluripotent stem cells (iPS cells).

[14] The compound for use according to any of

[11] to

[13] , wherein the compound is asenapine or a salt thereof, preferably maleate or hydrochloride, more preferably maleate.

[15] The use according to any of

[11] to

[14] , wherein the compound includes entacapone or a salt thereof, preferably entacapone.

[16] The use according to any one of

[11] to

[15] , wherein the compound promotes maturation of the transplanted stem cell-derived immature neurons in the subject and / or suppresses proliferation of the transplanted stem cell-derived immature neurons.

[17] The use according to any one of

[11] to

[16] , wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or bone marrow injury.

[18] The use according to any one of

[11] to

[17] , wherein the tissue is a cerebral organoid.

[19] The use according to any one of

[11] to

[18] , wherein the pharmaceutical agent is a formulation for oral or parenteral administration, preferably a formulation for oral, intravenous, intraperitoneal, or spinal administration.

[20] [20a], [20b], [20c], or [20d] below. [20a] A method for promoting maturation of stem cell-derived immature neurons, the method comprising the step of coexisting the stem cell-derived immature neurons in vitro or ex vivo with at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof.[20b] A method for inhibiting the proliferation of stem cell-derived non-mature neurons, the method comprising a step of coexisting the stem cell-derived non-mature neurons in vitro or ex vivo with at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. [20c] A method for producing a stem cell-derived neuronal population, the method comprising a step of culturing a cell population containing stem cell-derived non-mature neurons in the presence of a maturation promoter to mature the non-mature neurons in the cell population, the maturation promoter comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further comprise a step of inducing differentiation of the stem cells into neurons. [20d] A method for producing brain organoids, comprising culturing a three-dimensional cell mass containing stem cell-derived immature neurons in the presence of a maturation promoter to mature the immature neurons in the cell mass and obtain brain organoids, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further comprise culturing the stem cells in a suspension aggregate to obtain a three-dimensional cell mass.

[21] The method of

[20] , wherein the stem cell-derived immature neurons are immature neurons or progenitor neurons.

[22] The method of

[20] or

[21] , wherein the stem cell-derived immature neurons are derived from pluripotent stem cells, preferably induced pluripotent stem cells (iPS cells).

[23] The method of any of

[20] to

[23] , wherein the compound is asenapine or a salt thereof, preferably the maleate or hydrochloride salt, more preferably the maleate salt.

[24] The method according to any one of

[20] to

[23] , wherein the compound is entacapone or a salt thereof, preferably entacapone.

[25] The method according to any one of

[20] to

[24] , wherein the stem cell-derived non-mature neural cells are present in a tissue or tissue fragment, preferably in a cerebral organoid.

[0010] According to the present invention, the maturation of the stem cell-derived immature nerve cells can be promoted. Therefore, for example, in a treatment method in which stem cell-derived immature nerve cells such as cerebral organoids are transplanted into a subject with brain damage, by applying the present invention, the maturation of the stem cell-derived immature nerve cells can be promoted, thereby suppressing the proliferation of the stem cell-derived immature nerve cells. Therefore, the present invention can be said to be a useful technology in the treatment of neurological disorders such as brain damage.

[0011] Figure 1 is a graph showing the expression ratio of marker proteins in organoids after drug treatment in Example 1, with the left graph showing the results for CTIP2 protein, a marker for mature neurons, and the right graph showing the results for PAX6 protein, a marker for immature neurons. Figure 2 shows the serum and brain penetration of entacapone and asenapine (hydrochloride) in Example 2. The graph shows the brain concentration of asenapine hydrochloride measured by LLOQ chromatography. The table shows the concentrations of entacapone and asenapine in serum and brain samples.

[0012] Unless otherwise specified, terms used in this specification can be used in the sense commonly used in the art.

[0013] As used herein, unless otherwise specified, the term "cell" includes at least one cell, and can refer to, for example, either a single cell or a cell population.

[0014] As used herein, unless otherwise specified, the term "cell population" includes at least one type of cell, and is not limited to, for example, one type of cell, but can also be used to refer to two or more types of cells. A cell population is also referred to as, for example, a cell group.

[0015] In the present invention, as terms indicating the differentiation stage of a cell, "mature" means a state in which the final stage of differentiation has been reached, and "non-mature" means a state in which the final stage of differentiation has not been reached. In the present invention, "non-mature" includes "precursor" and "immature." "Precursor" means a state in which a certain degree of differentiation has progressed even in the non-mature stage, and the cell that will ultimately become has been determined, and "immature" means a state in which the precursor stage has not been reached.

[0016] <Maturation promoter for stem cell-derived non-mature neurons> The maturation promoter for stem cell-derived non-mature neurons of the present invention is characterized by comprising at least one compound selected from the group consisting of asenapine, a salt, solvate, or derivative thereof, and entacapone, a salt, solvate, or derivative thereof. Hereinafter, the stem cell-derived non-mature neurons are also simply referred to as non-mature neurons.

[0017] As described above, the maturation promoter of the present invention can promote the maturation of the stem cell-derived immature neurons. Therefore, the maturation promoter of the present invention can be used, for example, in regenerative medicine, in which the stem cell-derived immature neurons are transplanted. In the present invention, promoting the maturation of the immature neurons means promoting the induction of differentiation from the immature neurons into neurons. The maturation promoter of the present invention can also be referred to as, for example, a differentiation inducer.

[0018] Generally, immature cells, which are non-mature cells, have the ability to proliferate and proliferate as multiple species; progenitor cells, which are non-mature cells, have the ability to proliferate and proliferate as a single species; and mature cells are a single species and do not have the ability to proliferate. When a graft of the non-mature neurons is used in regenerative medicine, for example, some of the non-mature neurons in the graft may differentiate into mature neurons, while some of the non-mature neurons may continue to proliferate without being induced to differentiate. In contrast, in the presence of the maturation promoter of the present invention, for example, maturation of non-mature neurons with the ability to proliferate into mature cells without the ability to proliferate can be promoted in a cell population of the graft. This maturation promotion can increase the proportion of mature cells without the ability to proliferate in the cell population and reduce the number of non-mature neurons with the ability to proliferate, thereby suppressing the increase in the number of proliferating cells from the non-mature neurons. Therefore, the maturation-promoting agent of the present invention can inhibit the proliferation of the immature neurons without being induced to differentiate, thereby increasing the proportion of mature neurons differentiated from the immature neurons in the cell group and reducing the proportion of the immature neurons.

[0019] In the present invention, an increase in the proportion of mature neurons and a decrease in the proportion of non-mature neurons can have the following meaning: when a cell group containing the non-mature neurons is treated (e.g., incubated for a certain period of time) under the same conditions except for the presence and absence of an active ingredient, the proportion of the mature neurons in the cell group after treatment increases and the proportion of the non-mature neurons decreases when treated in the presence of the active ingredient compared to when treated in the absence of the active ingredient.

[0020] As described above, the active ingredient in the maturation-accelerating agent of the present invention is one or more of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. Hereinafter, asenapine and its salts, solvates, or derivatives are also referred to as asenapine-class compounds. Hereinafter, entacapone and its salts, solvates, or derivatives are also referred to as entacapone-class compounds. Hereinafter, the asenapine-class compounds and the entacapone-class compounds are also referred to as maturation-accelerating compounds of the present invention.

[0021] Asenapine is a compound (C) represented by the following formula (1-1): １７ H １６ ClNO), i.e., trans-5-chloro-2-methyl-2,3,3a,12b-tetrahydro-1H-dibenzo[2,3:6,7]oxepino[4,5-c]pyrrole. Asenapine exists in multiple optical isomers, but as long as the object of the present invention is not impaired, asenapine may be an optically active form or a racemic form. Asenapine of the following formula (1-1) will hereinafter also be referred to as free asenapine.

[0022]

[0023] The type of asenapine salt is not particularly limited and includes, for example, alkali metal salts, alkaline earth metal salts, amphoteric element salts, amine salts, inorganic acid salts, organic acid salts, etc. Examples of inorganic acid salts include hydrochloride, hydrobromide, sulfate, phosphate, etc. Examples of organic acid salts include fumarate, succinate, oxalate, lactate, maleate, etc.

[0024] A specific example of the asenapine salt is asenapine maleate, which is represented by the following formula (1-2): １７ H １６ ClNO.C ４ H ４ O ４ ).

[0025]

[0026] The type of the asenapine solvate is not particularly limited. The asenapine solvate may be an asenapine hydrate or an asenapine organic solvate. Examples of solvents that can form the hydrate include water, and examples of solvents that can form the organic solvate include alcohols such as methanol, ethanol, and isopropyl alcohol, and organic solvents such as acetone, ethyl acetate, and methylene chloride.

[0027] The asenapine derivative is not particularly limited as long as it has the same function as asenapine. Examples of the asenapine derivative include those in which a part of the chemical active site is modified, prodrugs, etc.

[0028] Entacapone is a compound represented by the following formula (2) (C １４ H １５ N ３ O ５ ), i.e., (E)-2-cyano-3-(3,4-dihydroxy-5-nitrophenyl)-N,N-diethyl-2-propenamide.

[0029]

[0030] The type of the entacapone salt is not particularly limited, and examples thereof include alkali metal salts, alkaline earth metal salts, amphoteric element salts, amine salts, inorganic acid salts, and organic acid salts.

[0031] The type of the entacapone solvate is not particularly limited. The entacapone solvate may be, for example, an entacapone hydrate or an entacapone organic solvate. Examples of solvents that can form the hydrate include water, and examples of solvents that can form the organic solvate include alcohols such as methanol, ethanol, and isopropyl alcohol, and organic solvents such as acetone, ethyl acetate, and methylene chloride.

[0032] The entacapone derivative is not particularly limited as long as it has the same function as entacapone. Examples of the entacapone derivative include those in which a part of the chemical active site is modified, prodrugs, etc.

[0033] The maturation accelerator of the present invention may contain any one of the maturation-promoting compounds of the present invention as an active ingredient, and other components are not particularly limited. In the maturation accelerator of the present invention, the maturation-promoting compound may be, for example, one, two, three, or more types. The combination of the maturation-promoting compounds is not particularly limited, and examples thereof include a combination of the asenapine-class compound and the entacapone-class compound. Specific examples include a combination of at least one of asenapine and an asenapine salt with entacapone, and more specifically, a combination of asenapine maleate with entacapone. The maturation accelerator of the present invention may further contain, as the active ingredient, other substances involved in maturation promotion in addition to the maturation-promoting compound.

[0034] The maturation accelerator of the present invention may contain, for example, only the active ingredient, or may further contain other additives. The additives are preferably pharmaceutically acceptable additives. The additives are not particularly limited and can be appropriately selected depending on the form of use (e.g., dosage form, etc.). Examples of the additives include solvents, carriers, excipients, stabilizers, lubricants, sweeteners, preservatives, suspending agents, dispersants, thickeners, pH adjusters, antifoaming agents, and flavorings.

[0035] Examples of the solvent include aqueous solvents such as water, physiological saline, isotonic solutions, and buffer solutions; oily solvents such as soybean oil; and emulsified solvents which are mixtures of the aqueous solvents and the oily solvents.

[0036] The form of the maturation accelerator of the present invention is not particularly limited and can be selected depending on, for example, the method of use (e.g., administration method, etc.). Examples of the form include liquids, gels, creams, and solids such as powders. For example, the maturation accelerator of the present invention may be the same or different before use (e.g., at the time of distribution) and at the time of use. When used in vitro or ex vivo, the maturation accelerator of the present invention is preferably in a liquid form when added to a subject (i.e., at the time of use) so that it can coexist with the immature neurons. Before use, the maturation accelerator may be in a form that can be prepared into a liquid (e.g., a concentrated liquid, a powder, etc.).

[0037] In the maturation-accelerating agent of the present invention, the content of the maturation-accelerating compound is not particularly limited and can be appropriately determined depending on the form of use.

[0038] As described above, the maturation promoter of the present invention can be used for the stem cell-derived non-mature nerve cells. The non-mature nerve cells are derived from stem cells, and specifically, preferably from pluripotent stem cells.

[0039] The pluripotent stem cells can be induced from, for example, fertilized eggs, cloned embryos, germline stem cells, tissue stem cells, somatic cells, etc. Examples of the pluripotent stem cells include embryonic stem cells (ES cells), embryonic germ cells (EG cells), and induced pluripotent stem cells (iPS cells). Furthermore, the pluripotent stem cells may be, for example, Muse cells (Multi-lineage Differentiating Stress Enduring cells) obtained from mesenchymal stem cells (MSCs), and GS cells prepared from germ cells (e.g., testes), etc. When the pluripotent stem cells are, for example, embryonic stem cells or any cells derived from a human embryo, they may be cells produced by destroying an embryo or cells produced without destroying an embryo, but from an ethical standpoint, cells (cell lines) produced without destroying an embryo are preferred. Furthermore, human embryonic stem cells may be established from a human embryo within 14 days of fertilization, for example. Preferably, the pluripotent stem cells are induced pluripotent stem cells (iPS cells).

[0040] The stem cell-derived non-mature neural cells are cells that can differentiate into neural cells. They may be precursor cells (progenitor neural cells) whose differentiation direction has been determined to a certain extent, or immature cells (immature neural cells) that have not yet reached the precursor cell stage, with the former precursor cells being preferred. In the treatment of neurological disorders such as brain damage, it is preferable to efficiently regenerate neural cells at the target site. Therefore, when the goal is to regenerate neural cells (e.g., brain cells) in vivo, the non-mature neural cells are preferably precursor cells whose differentiation direction has been determined to a certain extent. Specifically, for example, the differentiation direction is preferably toward neurons rather than glial cells, and neural precursor cells at a certain differentiation stage are preferred. Since the maturation-promoting agent of the present invention promotes the maturation of such neural precursor cells into neural cells, for example, proliferation at the precursor stage can be suppressed in vivo, and further differentiation into neural cells can be effectively achieved.

[0041] Whether the stem cell-derived non-mature neurons are at the precursor cell stage or the earlier immature cell stage can be determined, for example, by detecting a marker corresponding to the differentiation stage of the cells. Examples of immature cell markers include the neural stem cell marker PAX6, and examples of neural progenitor cell markers include LMX1A (dopamine). The maturation of non-mature neurons can be determined by the disappearance of the immature cell marker or neural progenitor cell marker and the expression of mature neuron markers. Examples of mature neuron markers include TUJ1 and CTIP2 (cerebral nerve).

[0042] The non-mature nerve cells to which the maturation promoter of the present invention is applied may be, for example, a single cell, but preferably a cell population. The form of the cell population is not particularly limited, and may be, for example, a state in which each of multiple cells is free, a structure in which multiple cells form a two-dimensional structure (two-dimensional structure), or a structure in which multiple cells form a three-dimensional structure (three-dimensional structure). The maturation promoter of the present invention is preferably a three-dimensional structure, for example, because it is suitable for regenerative medicine as described above. The three-dimensional structure is also known as, for example, an organoid.

[0043] The type of organoid of the immature nerve cells is not particularly limited, and for example, brain organoids such as cerebral organoids, cerebellar organoids, brainstem organoids can be mentioned. In addition, the brain organoids can be, for example, cortical organoids or medulla organoids. As mentioned above, the organoid of the immature nerve cells can be used for the treatment of nerve disorders, so for example, when used for therapeutic purposes, the type of the brain organoid can be selected according to the brain site to be transplanted. Specific examples of the brain organoids include, for example, cerebral organoids, cerebral cortex organoids, etc.

[0044] As described above, the cell population (e.g., the organoid) contains the stem cell-derived non-mature nerve cells. The cell population may contain other cells in addition to the stem cell-derived non-mature nerve cells, but preferably the majority of the cells are the stem cell-derived non-mature nerve cells, and the proportion of the non-mature nerve cells is, for example, 10% or more, 20% or more, or 30% or more.

[0045] The non-mature neural cells in the cell population may be, for example, progenitor cells, immature cells that have not yet reached the progenitor stage, or both. From the viewpoint of therapeutic efficiency, the cell population preferably contains progenitor cells whose differentiation destination has been determined to some extent, and more preferably contains neural progenitor cells. Specifically, the cell population preferably contains a relatively large number of progenitor cells as the non-mature neural cells, for example, compared to the immature cells, and more preferably, progenitor cells make up the majority. The proportion of progenitor cells in the non-mature neural cells is preferably, for example, 10% or more, 25% or more, or 50% or more.

[0046] The maturation-promoting agent of the present invention may be used in vivo, ex vivo, or in vitro as long as it is coexistent with the immature neurons (so as to be in contact with them). The coexistence of the maturation-promoting agent of the present invention with the immature neurons can also be referred to as, for example, contact between the maturation-promoting compound in the maturation-promoting agent of the present invention and the immature neurons.

[0047] First, as an example of a method for using the maturation promoter of the present invention, in vivo use is exemplified below. The method for using the maturation promoter of the present invention is not limited to the following example. In addition, although organoids are sometimes cited as an example of the subject to which the maturation promoter of the present invention is applied, as mentioned above, this is not limited thereto, and can be interpreted as, for example, the above-mentioned free immature nerve cells, two-dimensional structures of immature nerve cells, etc.

[0048] (1) In vivo Use The maturation promoter of the present invention can be used in vivo. For example, by coexisting the maturation promoter of the present invention with the immature neurons or organoids containing immature neurons in vivo, the maturation of the immature neurons transplanted into a subject can be promoted, and the proliferation of the immature neurons can also be suppressed.

[0049] The type of subject is not particularly limited, and examples thereof include humans and non-human animals. Examples of the non-human animals include non-human mammals such as mice, rats, cats, dogs, horses, cows, and camels.

[0050] The subject is, for example, a subject to which the non-mature neural cells are to be transplanted or have been transplanted, specifically, a subject to which the non-mature neural cells are to be transplanted or have been transplanted for the treatment of a neurological disorder.

[0051] The nerve disorder is not particularly limited and is a disorder (disease) caused by nerve degeneration, nerve damage, etc., and examples thereof include disorders caused by degeneration or damage to brain nerves (brain disorders), disorders caused by bone marrow damage, etc. The type of the nerve disorder is not particularly limited and examples thereof include neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS), cerebrovascular disorders such as cerebral infarction, bone marrow damage, etc.

[0052] Asenapine and entacapone are used to improve neurological disorders such as Parkinson's disease, but they are not known to promote the maturation of stem cell-derived immature neurons when applied to subjects into which stem cell-derived immature neurons have been transplanted or administered.

[0053] The transplantation site of the immature neurons in the subject is not particularly limited, and may be, for example, a site where neural degeneration or damage has occurred, specifically the brain or spinal cord. The immature neurons may also be transplanted intravenously, for example. When the transplantation site is the brain, specific examples include the forebrain, including the cerebrum and diencephalon, the brainstem, including the midbrain and rhombencephalon, and the like, preferably the cerebrum. The brain region may be, for example, the cortex (gray matter) or the medulla (white matter), preferably the cerebral cortex. The immature neurons transplanted into the subject may be, for example, free immature neurons, the two-dimensional structure, or the three-dimensional structure (organoid). When directly transplanted into the transplantation site, the immature neurons are preferably transplanted into the subject as, for example, a tissue fragment (graft) of the organoid.

[0054] As specific examples of transplantation, in the case of neurodegenerative diseases such as Parkinson's disease, for example, cell transplantation into the brain is preferred; in the case of cerebral infarction, for example, cell transplantation into a vein is preferred; and in the case of bone marrow injury, for example, cell transplantation into the bone marrow is preferred.

[0055] The maturation promoter of the present invention can be administered to the subject, for example, to allow it to coexist (so that it can come into contact with) the non-mature neurons. The administration method is not particularly limited, and can be, for example, any method that delivers the maturation-promoting compound of the maturation promoter to the non-mature neurons (e.g., tissue fragments of the organoid) transplanted into the subject. The administration method can be, for example, parenteral administration or oral administration. The administration method can be appropriately set, for example, depending on the transplant site of the non-mature neurons in the subject.

[0056] Examples of parenteral administration include intravenous administration, intracerebral administration, intramedullary administration, nasal administration, subcutaneous administration, intradermal administration, transdermal administration, and administration to the affected area (direct administration to the transplant site). The site of intravenous administration is not particularly limited, and examples thereof include the central vein, internal jugular vein, subclavian vein, femoral vein, etc. The method of parenteral administration is not particularly limited, and examples thereof include administration by injection, administration by infusion, etc.

[0057] The active ingredients, the asenapine-class compounds and the entacapone-class compounds, are known to have blood-brain barrier permeability. Therefore, the antiproliferative agent of the present invention can effectively deliver the active ingredient to the affected area of ​​the brain, for example, by administration into the blood. From this perspective, the antiproliferative agent of the present invention is preferably administered into the blood, specifically, intravenously as described above. The site of intravenous administration is not particularly limited, and examples include the central vein, internal jugular vein, subclavian vein, femoral vein, etc.

[0058] Furthermore, it is presumed that the asenapine-class compound, which is an active ingredient, when administered intravenously, has a maturation-promoting effect on the immature neurons at the transplant site (e.g., in the brain). Furthermore, it is presumed that the entacapone-class compound, which is an active ingredient, when administered into the blood, does not migrate into the brain but activates specific cells such as macrophages and glial cells outside the blood-brain barrier, and the activated cells send signals into the brain, thereby suppressing the proliferation of the immature neurons transplanted into the brain.

[0059] In the case of intracerebral administration, for example, administration to the transplant site of the immature nerve cells in the brain (administration to the affected area) can be mentioned.

[0060] Furthermore, the maturation-promoting agent of the present invention may further comprise, for example, a DDS component that delivers the maturation-promoting compound to the transplant site (affected site).

[0061] As described above, the maturation promoter of the present invention is administered to a subject to which the immature neurons are to be or have been transplanted. The start of administration is not particularly limited, and may be initiated, for example, before transplantation, simultaneously with transplantation, or after transplantation. Regardless of the start time of administration, the maturation promoter of the present invention is preferably administered, for example, after the immature neurons have been transplanted. The immature neurons to be transplanted into a subject may be treated with the maturation promoter of the present invention, for example, prior to transplantation, and the in vitro description described below can be used.

[0062] The administration conditions of the maturation promoter of the present invention are not particularly limited and can be appropriately determined depending on, for example, the type, age, weight, sex, type of neuropathy, degree of progression of neuropathy, the amount of the immature neurons to be transplanted into the subject, the volume of the organoid, etc. Furthermore, the content of the maturation promoter compound in the maturation promoter of the present invention is not particularly limited and can be appropriately determined depending on, for example, the administration conditions.

[0063] (2) Ex vivo Use The maturation promoter of the present invention can be used ex vivo. For example, the maturation promoter of the present invention can be coexisted with the immature neurons (e.g., the organoid) in an isolated organ (e.g., the brain) or its tissue (e.g., brain tissue). This can promote the maturation of the transplanted immature neurons in the isolated organ or tissue, thereby suppressing the proliferation of the immature neurons.

[0064] Organs and tissues can be, for example, those isolated from a subject. The subject is not particularly limited, and as in (1), can be exemplified as a human or non-human animal. The non-mature nerve cells to be transplanted are not particularly limited, and as in (1), can be, for example, free non-mature nerve cells, the two-dimensional structure, or the three-dimensional structure (organoid), and it is preferable to transplant the organoid into an organ or tissue as a tissue fragment (graft).

[0065] The maturation promoter of the present invention is applied to, for example, an organ or tissue (e.g., brain or brain tissue) into which the immature neural cells are transplanted. The maturation promoter of the present invention may be directly injected into the brain or brain tissue, or may be added to a culture medium and allowed to coexist with the brain or brain tissue in the culture medium. The type of the culture medium is not particularly limited.

[0066] The conditions for allowing the maturation-promoting agent of the present invention to coexist with an organ or tissue (e.g., brain or brain tissue) are not particularly limited. The conditions for treatment with the maturation-promoting agent of the present invention are not particularly limited, and are, for example, 35 to 38°C and 1 to 4 days. The amount of the maturation-promoting compound added to the organ or tissue is not particularly limited, and can be determined appropriately depending on, for example, the volume of the organ or tissue and the amount of the immature nerve cells to be transplanted or transplanted (e.g., the volume of the organoid).

[0067] When the maturation-promoting agent of the present invention is used ex vivo, for example, the tissue (e.g., brain tissue) into which the immature neurons have been transplanted and treated with the maturation-promoting agent may be transplanted into a corresponding organ (e.g., brain) of a subject. When the treated tissue is transplanted into the subject, for example, it is preferable to further administer the maturation-promoting agent of the present invention to the subject after transplantation into the subject.

[0068] (3) In Vitro Use The maturation-promoting agent of the present invention can be used in vitro. For example, by coexisting the maturation-promoting agent of the present invention with the immature neurons, the maturation of the immature neurons can be promoted, thereby suppressing the proliferation of the immature neurons.

[0069] The maturation promoter of the present invention is preferably added to a culture medium, for example, and allowed to coexist with the immature neurons in the culture medium. The type of the culture medium is not particularly limited, and examples thereof include DMEM, DMEM / F12, GMEM, and NeuroBasal. The immature neurons are not particularly limited, and as in (1), may be, for example, free immature neurons, the two-dimensional structure, or the three-dimensional structure (organoid), and among these, tissue fragments of the organoid are preferred.

[0070] The conditions for allowing the maturation-promoting agent of the present invention and the immature neurons to coexist in the medium are not particularly limited. The conditions for treatment with the maturation-promoting agent of the present invention are not particularly limited, and are, for example, 35 to 38°C and 1 to 4 days. The amount of the maturation-promoting compound added to the immature neurons is also not particularly limited, and is, for example, 10 to 100 mg per immature neuron ... mm2 of the organoid. ３ The average daily dose is 15 ng to 15 μg.

[0071] When the maturation-promoting agent of the present invention is used in vitro, for example, the immature neurons may be treated with the maturation-promoting agent and then transplanted into the brain of a subject. When the immature neurons are transplanted into the subject after the treatment, it is preferable to further administer the maturation-promoting agent of the present invention to the subject after transplantation into the subject.

[0072] <Method for Promoting Maturation of Stem Cell-Derived Immature Neurons> As described above, the method for promoting maturation of stem cell-derived immature neurons of the present invention comprises a treatment step of coexisting stem cell-derived immature neurons with a maturation promoter, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. In the present invention, the maturation promoter is the maturation promoter of the present invention described above. The maturation promotion method of the present invention is characterized by using the maturation promoter of the present invention, and other steps and conditions are not particularly limited. Furthermore, unless otherwise specified, the description of the maturation promoter of the present invention can be used for the maturation promotion method of the present invention.

[0073] The method for promoting maturation of the present invention may be carried out in vivo, ex vivo, or in vitro.

[0074] The first maturation promoting method of the present invention is, for example, a method of allowing the immature neurons and the maturation promoting agent to coexist in vivo. That is, in the first maturation promoting method of the present invention, for example, the treatment step is a step of administering the maturation promoting agent to a subject whose brain has been or will be transplanted with the immature neurons. For example, the description regarding the in vivo use of the maturation promoting agent of the present invention can be used for the first maturation promoting method.

[0075] The type of the subject is not particularly limited, and as described above, can be, for example, a human or a non-human animal. The immature nerve cells to be transplanted are not particularly limited, and as described above, can be, for example, free immature nerve cells, the two-dimensional structure, or the three-dimensional structure (organoid), and among these, it is preferable to transplant them as a tissue fragment (graft) of the organoid.

[0076] The immature neurons can be used, for example, to treat neurological disorders. Therefore, in the maturation promoting method of the present invention, the subject is preferably, for example, a subject with a neurological disorder, specifically, for example, a subject with a brain disorder into which the immature neurons are to be transplanted. The method and conditions for administering the maturation promoting agent are not particularly limited and are as described above.

[0077] The second maturation promotion method of the present invention involves coexisting stem cell-derived non-mature neurons with the maturation promoter ex vivo. The stem cell-derived non-mature neurons are, for example, located in an isolated organ or its tissue. That is, in the second maturation promotion method of the present invention, the treatment step involves coexisting the maturation promoter of the present invention with the non-mature neurons in, for example, an isolated organ or its tissue (e.g., brain or brain tissue). For the second maturation promotion method, for example, the description regarding the ex vivo use of the maturation promoter of the present invention can be used. The method and conditions for coexistence are not particularly limited and are as described above.

[0078] A third maturation promotion method of the present invention is a method of coexisting stem cell-derived non-mature neurons with the maturation promoter in vitro. The stem cell-derived non-mature neurons are, for example, a cell population containing the stem cell-derived non-mature neurons. That is, in the third maturation promotion method of the present invention, the treatment step is, for example, a step of coexisting the non-mature neurons with the maturation promoter in a culture medium. For example, the description regarding the in vitro use of the maturation promoter of the present invention can be used for the third maturation promotion method. The method and conditions for coexistence are not particularly limited and are as described above.

[0079] <Proliferation inhibitor of stem cell-derived non-mature nerve cells> The proliferation inhibitor of stem cell-derived non-mature nerve cells of the present invention is characterized by comprising at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof.

[0080] The proliferation inhibitor of the present invention can inhibit the proliferation of immature neurons. Therefore, the proliferation inhibitor of the present invention can be used, for example, in regenerative medicine, in which stem cell-derived immature neurons are transplanted into a subject. In other words, in the absence of the proliferation inhibitor of the present invention, for example, in a graft of immature neurons, some immature neurons differentiate into mature neurons, while some immature neurons may continue to proliferate without being induced to differentiate. In contrast, in the presence of the proliferation inhibitor of the present invention, for example, maturation (differentiation induction) of immature neurons is promoted in the graft. Therefore, for example, in the graft, the proliferation of immature neurons is inhibited, and further, differentiation of the immature neurons into mature neurons is effectively induced. As a result, a decrease in the proportion of differentiated mature neurons in the graft due to proliferation of the immature neurons can be prevented, and the proportion of mature neurons differentiated from immature neurons can be increased in the graft.

[0081] As described above, the active ingredients in the growth inhibitor of the present invention are asenapine-class compounds and entacapone-class compounds, which are also referred to as growth inhibitors in the present invention. The growth inhibitors in the present invention are the same as the maturation-promoting compounds in the present invention. Therefore, the growth inhibitors of the present invention can be incorporated with reference to the description of the maturation promoter of the present invention. Specifically, the growth inhibitors of the present invention can be incorporated with reference to the description of the maturation promoter of the present invention, with "maturation promotion" replaced with "growth inhibition."

[0082] The method of use of the proliferation inhibitor of the present invention is not particularly limited, and can be used, for example, for immature nerve cells whose proliferation is to be inhibited. As mentioned above, the proliferation inhibitor of the present invention is the same as the maturation promoter of the present invention, and its target is also the stem cell-derived immature nerve cells, the same as the maturation promoter of the present invention. Therefore, the description of the maturation inhibitor of the present invention can also be used for its method of use. Specifically, the method of use of the proliferation inhibitor of the present invention can be used in its entirety by substituting "maturation promotion" for "proliferation inhibition" in the description of the method of use of the maturation promoter of the present invention.

[0083] The growth inhibitor of the present invention may be used in vivo, in vitro or ex vivo.

[0084] (1) In vivo Use The proliferation inhibitor of the present invention can be used in vivo. For example, by allowing it to coexist with immature neurons in vivo, it can promote the maturation of the immature neurons transplanted into a subject and inhibit the proliferation of the immature neurons.

[0085] (2) Ex vivo Use The growth inhibitor of the present invention can be used ex vivo. For example, the growth inhibitor of the present invention can be coexisted with the immature neurons in an isolated organ or tissue thereof (e.g., the brain or brain tissue). This can promote the maturation of the immature neurons transplanted into the isolated organ or tissue, and inhibit the proliferation of the immature neurons.

[0086] (3) In Vitro Use The growth inhibitor of the present invention can be used in vitro. For example, by coexisting the growth inhibitor of the present invention with the immature neurons, maturation of the immature neurons can be promoted and the proliferation of the immature neurons can be inhibited.

[0087] <Method for inhibiting proliferation of stem cell-derived non-mature neurons> As described above, the method for inhibiting proliferation of stem cell-derived non-mature neurons of the present invention comprises a treatment step of coexisting non-mature neurons with a proliferation inhibitor, wherein the proliferation inhibitor comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. In the present invention, the proliferation inhibitor is the proliferation inhibitor of the present invention described above. The proliferation inhibitor of the present invention is characterized by using the proliferation inhibitor of the present invention, and other steps and conditions are not particularly limited.

[0088] In the proliferation inhibitory method of the present invention, the proliferation inhibitor is the same as the maturation promoter of the present invention, as described above. Therefore, the proliferation inhibitory method of the present invention can be applied to the description of the maturation promoter and maturation promotion method of the present invention. Specifically, the proliferation inhibitory method of the present invention can be applied to the description of the maturation promoter and maturation promotion method of the present invention by replacing "maturation promotion" with "proliferation inhibition."

[0089] The proliferation-inhibiting method of the present invention may be any of in vivo, ex vivo, and in vitro methods.

[0090] The first proliferation-inhibiting method of the present invention is a method in which the immature neurons and the proliferation-inhibiting agent coexist in vivo. That is, in the first proliferation-inhibiting method of the present invention, for example, the treatment step is a step of administering the proliferation-inhibiting agent to a subject into whose brain the immature neurons are to be or have been transplanted.

[0091] The second proliferation-inhibiting method of the present invention is a method in which the non-mature nerve cells and the proliferation-inhibiting agent are allowed to coexist ex vivo. That is, in the second proliferation-inhibiting method of the present invention, the treatment step is, for example, a step in which the proliferation-inhibiting agent of the present invention and the non-mature nerve cells are allowed to coexist in an isolated organ or tissue (e.g., brain or brain tissue).

[0092] A third proliferation-inhibiting method of the present invention is a method in which the non-mature nerve cells and the proliferation inhibitor are allowed to coexist in vitro. That is, in the third proliferation-inhibiting method of the present invention, the treatment step is, for example, a step of allowing the non-mature nerve cells and the proliferation inhibitor to coexist in a culture medium.

[0093] <Pharmaceutical composition for neurological disorders and method for treating neurological disorders> The pharmaceutical composition for neurological disorders of the present invention comprises, as an active ingredient, at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof, and is characterized in that it is administered to a subject who will receive or has received a transplant of stem cell-derived immature neurons, or a tissue or tissue graft containing stem cell-derived immature neurons.

[0094] The active ingredients in the pharmaceutical composition of the present invention are the same as those in the maturation promoter and proliferation inhibitor of the present invention. Therefore, the same descriptions as those for the maturation promoter and proliferation inhibitor of the present invention can be applied to the pharmaceutical composition of the present invention.

[0095] The pharmaceutical composition of the present invention can, for example, promote the maturation (differentiation induction) of stem cell-derived immature neurons transplanted into a subject and inhibit the proliferation of the stem cell-derived immature neurons. Therefore, the pharmaceutical composition for neurological disorders of the present invention can be used, for example, for subjects into which the immature neurons are or have been transplanted. The subject is not particularly limited and may be a human or a non-human animal.

[0096] Furthermore, the pharmaceutical composition of the present invention may be, for example, a pharmaceutical kit containing the stem cell-derived non-mature nerve cells and the active ingredient.

[0097] The method for treating a neurological disorder of the present invention is characterized by comprising an administration step of administering the pharmaceutical composition of the present invention, i.e., at least one compound selected from the group consisting of asenapine-class compounds and entacapone-class compounds, to a subject suffering from a neurological disorder into which the stem cell-derived non-mature neurons have been transplanted. The method for treating a neurological disorder of the present invention is characterized by the use of the pharmaceutical composition of the present invention, and other configurations and conditions are not particularly limited. The description of the in vivo method for promoting maturation and the method for inhibiting proliferation of the present invention can be used for the method for treating a neurological disorder of the present invention. The type of neurological disorder is not particularly limited, and the above examples can be used for the treatment.

[0098] The therapeutic method of the present invention includes, for example, a transplantation step of transplanting the stem cell-derived immature neural cells into a subject with a neurological disorder, and an administration step of administering the pharmaceutical composition. In the therapeutic method of the present invention, the order of the transplantation step and the administration step is not particularly limited, and the administration step may be performed before the transplantation step, after the transplantation step, before or after the transplantation step, or simultaneously with the transplantation step.

[0099] In the treatment method of the present invention, the administration step is preferably carried out before the transplantation step. By administering the pharmaceutical composition prior to transplantation in this manner, it becomes possible, for example, to allow the transplanted cells to engraft early after transplantation and further to start promoting maturation early.

[0100] <Pharmaceutical composition for supporting transplantation and method for supporting transplantation> The pharmaceutical composition for supporting transplantation of the present invention contains, as an active ingredient, at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof, and is characterized in that it is administered to a subject who will receive or has received a transplant of stem cell-derived immature nerve cells, or a tissue or tissue fragment containing stem cell-derived immature nerve cells.

[0101] The method for assisting transplantation of the present invention comprises administering at least one compound selected from the group consisting of asenapine and a salt, solvate, or derivative thereof, and entacapone and a salt, solvate, or derivative thereof to a subject, wherein the subject is a recipient or has been a recipient of a transplant of stem cell-derived immature nerve cells, or a tissue or tissue graft containing stem cell-derived immature nerve cells.

[0102] <Method for producing a stem cell-derived neural cell population> The present specification also provides a method for producing a neural cell population, including the above-mentioned method for promoting the maturation of stem cell-derived non-mature neural cells. The method includes a step of culturing a cell population containing stem cell-derived non-mature neural cells in the presence of a maturation promoter to mature the non-mature neural cells in the cell population, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives. The method may further include a step of inducing differentiation of stem cells (preferably pluripotent stem cells, more preferably iPS cells) into neural cells to obtain a cell population containing stem cell-derived non-mature neural cells.

[0103] For example, the method of the present invention for producing a neural cell population derived from stem cells comprises: 1) a step of inducing differentiation of pluripotent stem cells into neural cells; and 2) a step of culturing the cell population after differentiation induction in the presence of a maturation promoter to mature immature neural cells in the cell population and obtain a neural cell population.

[0104] Methods for inducing differentiation of pluripotent stem cells into a neural cell population are known in the art, and step 1 can be performed according to such known methods or using a commercially available neural cell differentiation induction kit (e.g., the STEMdiff SMADi Neural Induction Kit). Generally, pluripotent stem cells are subjected to cell aggregation based on suspension culture, and then neural differentiation is induced in that state by adjusting the amount of added factors. The cell population obtained in step 1 contains a significant amount of immature neural cells. In step 2, the cell population after differentiation induction is cultured in the presence of a maturation promoter to mature the immature neural cells in the cell population. This results in a neural cell population enriched for mature neural cells compared to conventional methods that do not include a maturation step. Mature neural cells can be characterized by the expression of markers (e.g., CTIP2 and / or TUJ1) as described above.

[0105] <Method for producing brain organoids> This specification also provides a method for producing brain organoids, including the above-mentioned method for promoting the maturation of stem cell-derived immature neurons.The method comprises the step of culturing a three-dimensional cell mass comprising stem cell-derived immature neurons in the presence of a maturation promoter, and maturing the immature neurons in the cell mass to obtain brain organoids, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates or derivatives, and entacapone and its salts, solvates or derivatives.The method may further comprise the step of obtaining a three-dimensional cell mass from stem cells (preferably pluripotent stem cells, more preferably iPS cells).

[0106] For example, the method for producing cerebral organoids of the present invention includes the steps of: 1) culturing pluripotent stem cells as floating aggregates to obtain a three-dimensional cell mass containing immature neurons; and 2) culturing the three-dimensional cell mass in the presence of a maturation promoter to mature the immature neurons in the cell mass to obtain a cerebral organoid.

[0107] Methods for obtaining brain organoids from pluripotent stem cells are known in the art, and include, for example, the SFEB method, the SFEBq method, and modifications thereof. Step 1 can be carried out according to such known methods. The cell population obtained in step 1 contains a considerable amount of immature neurons. In step 2, the cell population after differentiation induction is cultured in the presence of a maturation promoter to mature the immature neurons in the cell population. This results in a neuronal population enriched in mature neurons compared to conventional methods that do not include a maturation step.

[0108] <Uses> The present invention relates to at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, for use in promoting the maturation of stem cell-derived immature neurons. The present invention also relates to use of at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, in the manufacture of an agent for promoting the maturation of stem cell-derived immature neurons.

[0109] The present invention relates to at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, for use in inhibiting the proliferation of stem cell-derived non-mature nerve cells. The present invention also relates to use of at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, in the manufacture of an agent for inhibiting the proliferation of stem cell-derived non-mature nerve cells.

[0110] The present invention relates to at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, for use in treating neurological disorders in subjects into which stem cell-derived non-mature neural cells have been transplanted. The present invention also relates to the use of at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, in the manufacture of a pharmaceutical composition for neurological disorders.

[0111] In the uses of the present invention, the asenapine class compound and the entacapone class compound, and methods for using them, may refer to the descriptions of the maturation promoter and maturation promoting method of the present invention, the proliferation inhibitor and proliferation inhibiting method of the present invention, and the pharmaceutical composition and method for treating neurological disorders of the present invention.

[0112] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these.

[0113] Example 1 The inhibition of proliferation and promotion of differentiation induction of stem cell-derived immature nerve cells by asenapine and entacapone were confirmed in vitro.

[0114] (1) Preparation of organoids Using undifferentiated iPS cells (cell line 201B7) as human-derived induced pluripotent stem cells, iPS-derived cerebral organoids were prepared as follows. The prepared organoids are hereinafter referred to as "untreated organoids."

[0115] (First half differentiation medium) GMEM + NEAA + SodiumPyruvate + 2ME +KSR + SB +IWR (second half differentiation medium) DMEM / F12 + GlutaMAX + CDLipid + P / S + AmphotericinB + N2 (Culture conditions) 37°C, 5% CO ２

[0116] (First half of differentiation) The induced pluripotent stem cells were cultured in an undifferentiated state using StemFit (AK02) medium. The proliferated induced pluripotent stem cells were detached from the culture vessel by enzyme treatment (Triple Select) and cultured at a density of 15,000 cells / cm. ２ At the time of cell seeding, iMatrix (1.5 μg / cm) was added to maintain the undifferentiated state of the induced pluripotent stem cells. ２ ) and a ROCK inhibitor, Y-27632 (10 μmol / L), were added. The induced pluripotent stem cells adhered to the TS-02 substrate were then cultured for proliferation for 5 to 7 days, and when they reached confluence, they were switched to a differentiation-inducing medium.

[0117] The differentiation-inducing medium (medium for the first half of differentiation) was a conditioned medium based on GMEM medium, supplemented with NEAA (1%), sodium pyruvate (1%), and 2-mercaptoethanol (2ME, 1%), and further supplemented with KSR (20%), SB (5 μmol / L), and IWR (3 μmol / L) to promote neural differentiation. Differentiation induction was performed using this medium for 16 to 20 days. Subsequently, the cells (cell populations containing immature neurons) adhered to the TS-02 substrate were naturally detached using PBS or medium (medium for the second half of differentiation) chilled at 4°C.

[0118] (Late differentiation) For the suspension culture of cell fragments after detachment, differentiation induction medium (late differentiation medium) was used, based on DMEM / F12 medium, GlutaMAX, CD lipid concentrate (1%), Penicillin / Streptomycin (1%), Amphotericin B (0.1%), N2-supplement (1%) was added to the conditioned medium. For suspension culture, first, the cell fragments were transferred onto a 90 mm dish (MS-1390R (Sumitomo Bakelite Co., Ltd.)) and subjected to static culture for 4 days, then switched to rotational culture and performed for 13 days or more, allowing organoids to form. The medium was changed once every 1 to 3 days.

[0119] (2) Drug Treatment Organoids prepared in (1) above on day 35 of suspension culture (hereinafter referred to as "pre-treatment organoids") were treated with drugs as follows: Asenapine (Example 1-1, asenapine maleate) or entacapone (Example 1-2) was used as the drug in this example.

[0120] Size of one organoid on day 35: Diameter approximately 1.5 mm to 2 mm Drug concentration in medium: 50 μmol / L Medium: DMEM / F12 + GlutaMAX + CDLipid + P / S + Amphotericin B + N2

[0121] The drug was added to the medium to prepare a drug-added medium. The organoids were transferred one by one to each well of a 96-well plate, and the drug-added medium was added to 180 μL / well. The well plate was then placed in an incubator at 37°C and 5% CO ２ The organoids were cultured for 4 days in DAPT-containing medium. As a positive control, the organoids were cultured in a 96-well plate in the same manner as above, using DAPT-containing medium containing the gamma-secretase inhibitor DAPT (GSI-IX, LY-374973, Sigma-Aldrich) instead of the drug. As a control, the organoids were cultured in a 96-well plate in the same manner as above, using DMSO-containing medium containing DMSO instead of the drug.

[0122] Then, for the cultured organoids (hereinafter referred to as "treated organoids"), the expression of PAX6 (paired box 6) protein, a marker for immature neurons, and CTIP2 protein, a marker for mature neurons, was measured by protein quantification using a cell sorter.

[0123] The relative expression level of the protein for each system was evaluated as a ratio to the control measurement value, which was set to 1. These results are shown in Figure 1. Figure 1 is a graph showing the expression ratio of marker proteins, with the left graph showing the results for CTIP2 protein, a marker for mature neurons, and the right graph showing the results for PAX6 protein, a marker for immature neurons.

[0124] As shown in the left graph of Figure 1, the organoids (Examples) treated with asenapine or entacapone showed a significant increase in the expression level of mature neuronal marker (CTIP2 protein) compared to the control. In particular, asenapine showed an expression level comparable to that of DAPT (positive control). And, as shown in the right graph of Figure 1, the organoids (Examples) treated with asenapine or entacapone showed a significant decrease in the expression level of immature neuronal marker (PAX6 protein) compared to the control. In addition, the organoids (Examples) treated with asenapine or entacapone showed a lower expression level of immature neuronal marker than DAPT (positive control), and entacapone in particular showed a significant decrease. From these results, it was found that asenapine and entacapone can promote the differentiation of immature neurons into mature neurons in organoids. Furthermore, it was found that by promoting the differentiation of immature neurons into mature neurons, the proportion of immature neurons with the ability to proliferate in the organoid cell population can also be reduced.

[0125] Example 2 Entacapone and asenapine, which are effective against cultured cells, were examined for their ability to penetrate into the brain in order to anticipate their effects on intracerebral grafts.

[0126] Entacapone (100 mg / kg) and asenapine hydrochloride (100 mg / kg) were intraperitoneally administered to separate mice. Three hours after the single administration, the drug concentrations were determined using serum and brain parenchyma samples from the animals by LLOQ chromatography (outsourced to Shimadzu Techno Research Center).

[0127] The results are shown in Figure 2. Asenapine (hydrochloride) was confirmed to be efficiently delivered to the brain parenchyma by intraperitoneal administration. Entacapone was below the detection limit. Because asenapine hydrochloride penetrates the blood-brain barrier and reaches the brain parenchyma, oral or parenteral administration of asenapine or its salts to subjects receiving or who have received grafts containing stem cell-derived immature neurons is expected to inhibit the proliferation and promote the maturation of immature neurons. Regarding entacapone, it is thought that its effect on stem cell-derived immature neurons in an in vitro environment before cell transplantation or its simultaneous administration with stem cell-derived immature neurons during transplantation may be effective.

[0128] Although the present invention has been described above with reference to the embodiments, the present invention is not limited to the above-described embodiments. Various modifications that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.

[0129] According to the present invention, the maturation of the stem cell-derived immature nerve cells can be promoted. Therefore, for example, the proliferation of the stem cell-derived immature nerve cells can be inhibited. Furthermore, by applying the present invention to a treatment method in which stem cell-derived immature nerve cells such as organoids are transplanted into a subject with a nerve disorder, the proliferation of the stem cell-derived immature nerve cells can be inhibited. Therefore, the present invention can be said to be a useful technology in the treatment of nerve disorders such as brain disorders.

[0130] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A maturation promoter for stem cell-derived immature neurons, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

2. The maturation promoter according to claim 1, wherein the stem cell-derived non-mature nerve cells are immature nerve cells or progenitor nerve cells.

3. The maturation promoter according to claim 1, wherein the compound is asenapine or a salt thereof.

4. The maturation promoter according to claim 1, comprising entacapone or a salt thereof as the compound.

5. The maturation promoter according to claim 1, comprising asenapine or a salt thereof and entacapone or a salt thereof as the compounds.

6. The maturation promoter according to claim 1, which is administered to a subject who will receive or has received a transplant of stem cell-derived immature nerve cells, or a tissue or tissue fragment containing stem cell-derived immature nerve cells.

7. The maturation promoter of claim 6, wherein the subject has a neurological disorder.

8. The maturation promoter according to claim 7, wherein the neurological disorder is a brain disorder, a neurodegenerative disease, or bone marrow damage.

9. A maturation promoter according to any one of claims 1 to 8, wherein the stem cell-derived non-mature neural cells are derived from pluripotent stem cells.

10. A proliferation inhibitor of stem cell-derived non-mature nerve cells, characterized by containing at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

11. A method for promoting the maturation of stem cell-derived non-mature neurons, comprising the step of coexisting stem cell-derived non-mature neurons with a maturation promoter in vitro or ex vivo, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

12. The method for promoting maturation according to claim 11, wherein the stem cell-derived non-mature nerve cells are immature nerve cells or progenitor nerve cells.

13. The method for promoting maturation described in claim 11, wherein the stem cell-derived non-mature neural cells are derived from pluripotent stem cells.

14. A maturation promotion method described in any one of claims 11 to 13, wherein the stem cell-derived non-mature neural cells are within a cerebral organoid.

15. A method for promoting maturation described in any one of claims 11 to 13, wherein the stem cell-derived non-mature neural cells are within a tissue or tissue fragment.

16. A method for inhibiting the proliferation of stem cell-derived non-mature nerve cells, comprising a step of coexisting stem cell-derived non-mature nerve cells in vitro or ex vivo with a proliferation inhibitor, wherein the proliferation inhibitor comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

17. A method for producing a stem cell-derived neural cell population, comprising the step of culturing a cell population containing stem cell-derived immature neural cells in the presence of a maturation promoter to mature the immature neural cells in the cell population, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

18. A method for producing cerebral organoids, comprising the step of culturing a three-dimensional cell mass containing stem cell-derived immature neurons in the presence of a maturation promoter to mature the immature neurons in the cell mass and obtain cerebral organoids, wherein the maturation promoter comprises at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives.

19. A pharmaceutical composition for assisting transplantation, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, wherein the pharmaceutical composition is administered to a subject who will receive or has received a transplant of stem cell-derived immature nerve cells, or a tissue or tissue graft containing stem cell-derived immature nerve cells.

20. A pharmaceutical composition for treating a neurological disorder, comprising at least one compound selected from the group consisting of asenapine and its salts, solvates, or derivatives, and entacapone and its salts, solvates, or derivatives, wherein the pharmaceutical composition is administered to a subject who will receive or has received a transplant of stem cell-derived immature nerve cells, or a tissue or tissue graft containing stem cell-derived immature nerve cells.

Images