Suppression of Neuroinflammation, Compositions and Methods therefor

Abstract

Provided are a pharmaceutical composition for suppressing neuroinflammation, a pharmaceutical composition for assisting in implantation of neural progenitor cells, pluripotent stem cells, and / or neurons, a pharmaceutical composition for promoting stabilization of Nrf2 protein in glial cells, and a pharmaceutical composition for protecting neural cells from neuroinflammation. The pharmaceutical compositions contain, as an active ingredient, a compound capable of inhibiting the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof.

Description

Technical Field

[0001] The present disclosure relates to the suppression of neuroinflammation, compositions and methods therefor. The present disclosure relates, in one or more aspects, to the assistance of transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons, compositions and methods therefor. The present disclosure relates, in one or more aspects, to the promotion of stabilization of Nrf2 (NF-E2-related factor 2) protein in glial cells, compositions and methods therefor. The present disclosure relates, in one or more aspects, to the protection of nerve cells from neuroinflammation, compositions and methods therefor.

Background Art

[0002] Parkinson's disease is a progressive neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantia nigra striatum. From previous clinical studies, improvement of motor symptoms in Parkinson's disease patients has been confirmed by transplantation of fetal midbrain cells. Based on such facts, cell replacement therapy is considered as a treatment method for Parkinson's disease. Pluripotent stem cells, particularly induced pluripotent stem cells (iPS cells or iPSCs), have the potential to supply a large amount of dopaminergic neurons. Therefore, pluripotent stem cells are considered as a new donor cell source. However, neural progenitor cells and dopaminergic neurons differentiated from stem cells such as iPS cells have an extremely low survival rate after transplantation into the brain (Patent Document 1).

[0003] Microglia are a type of glial cell present in the central nervous system (brain and spinal cord). Microglia are also called microglial cells or Hortega cells. Microglia are considered to be immune cells like macrophages. Microglia have various functions and roles such as antigen presentation that serves as the starting point of the immune response, innate immune action against foreign substances, phagocytosis against foreign substances and waste products, assistance in the formation of neural circuits, and production of various substances that affect surrounding cells. Microglia become activated by external stimuli, stress, etc., and produce useful substances such as antioxidants and trophic factors. However, microglia, upon pathological activation, secrete inflammatory cytokines, chemokines, nucleic acids, excitatory amino acids such as glutamate, reactive oxygen species, proteases, etc., and damage surrounding cells, thereby becoming the starting point of neuroinflammation. For this reason, microglia can also cause neurodegeneration in the central nervous system (Patent Document 2).

[0004] Compounds having an inhibitory ability against the phosphorylation activity of DYRK1A protein, which is a protein phosphorylation enzyme, have been reported to be effective for neurogenesis or neuronal proliferation (Patent Documents 3 and 4). DYRK is a general term for enzymes meaning dual specificity tyrosine-phosphorylation-regulated kinase.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Summary of the Invention

Problems to be Solved by the Invention

[0006] In one aspect, the present disclosure provides a pharmaceutical composition for suppressing neuroinflammation. In one aspect, the present disclosure provides a pharmaceutical composition for assisting the transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons. In one aspect, the present disclosure provides a pharmaceutical composition for promoting the stabilization of Nrf2 protein in glial cells. The present disclosure provides, in one aspect, a pharmaceutical composition for protecting nerve cells from neuroinflammation. **Means for Solving the Problems**

[0007] The present disclosure relates, in one aspect, to a pharmaceutical composition for suppressing neuroinflammation, which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof.

[0008] The present disclosure relates, in other aspects, to a pharmaceutical composition for assisting the transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons, which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof.

[0009] The present disclosure relates, in other aspects, to a pharmaceutical composition for promoting the stabilization of Nrf2 protein in glial cells, which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof.

[0010] The present disclosure relates, in other aspects, to a pharmaceutical composition for protecting nerve cells from neuroinflammation, which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof.

[0011] The present disclosure relates, in other aspects, to a method for suppressing neuroinflammation, which includes administering an effective amount of the pharmaceutical composition according to the present disclosure to a subject. The present disclosure relates, in other aspects, to a method for improving the survival rate of the transplanted cells, which includes administering an effective amount of the pharmaceutical composition according to the present disclosure to a recipient before, simultaneously with, or after the transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons. The present disclosure relates, in other aspects, to a method for promoting the stabilization of Nrf2 protein in glial cells and protecting nerve cells from neuroinflammation, which includes administering an effective amount of the pharmaceutical composition according to the present disclosure to a subject. In other aspects, the present disclosure relates to a method for improving, suppressing progression, and / or treating a disease associated with neuroinflammation selected from the group consisting of frontotemporal dementia, amyotrophic lateral sclerosis, and multiple sclerosis, which comprises administering an effective amount of the pharmaceutical composition according to the present disclosure to a subject.

Advantages of the Invention

[0012] According to the pharmaceutical composition of the present disclosure, in one or more embodiments, neuroinflammation can be suppressed. According to the pharmaceutical composition of the present disclosure, in one or more embodiments, the survival rate after transplantation of neural progenitor cells, stem cells, and / or neurons to be transplanted can be improved. According to the pharmaceutical composition of the present disclosure, in one or more embodiments, the stabilization of Nrf2 protein in glial cells can be promoted and improved. According to the pharmaceutical composition of the present disclosure, in one or more embodiments, neurons can be protected from neuroinflammation.

Brief Description of the Drawings

[0013]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

[0014] The present disclosure is based on the discovery that compounds capable of inhibiting the phosphorylation activity of DYRK1A protein can suppress neuroinflammation. The present disclosure is also based on the finding that compounds capable of inhibiting the phosphorylation activity of DYRK1A protein can improve the attachment rate (survival rate, survival rate) of neurons transplanted into the brain. The present disclosure is also based on the finding that compounds capable of inhibiting the phosphorylation activity of DYRK1A protein can promote the stabilization of Nrf2 protein in glial cells and protect nerve cells from neuroinflammation.

[0015] The mechanism by which compounds capable of inhibiting the phosphorylation activity of DYRK1A protein exert their anti-neuroinflammation effect has not been elucidated in detail, but is presumed to be as follows. Normally, microglia, which play a role in supporting neurons, become activated when subjected to stress such as injury or neuroinflammation, releasing inflammatory cytokines and reactive oxygen species and causing neuronal degeneration. At this time, when DYRK1A is inhibited, cyclin D1 and p21 are stabilized, the degradation of Nrf2 stops, and Nrf2 is stabilized. This Nrf2 suppresses the expression of genes that produce pro-inflammatory cytokines, and the excessive activation of microglia is suppressed. By suppressing the activation of microglia, which is the starting point of neuroinflammation, it is considered that neuroinflammation is suppressed and the effect of improving transplantation efficiency and / or protecting nerve cells is exerted. However, the present disclosure may not be construed as being limited to these mechanisms.

[0016] [Neuroinflammation inhibitor] The present disclosure relates to a pharmaceutical composition for suppressing neuroinflammation (hereinafter also referred to as the neuroinflammation inhibitor according to the present disclosure), which contains, as an active ingredient, a compound having an inhibitory ability against the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof. In one or more embodiments, the neuroinflammation inhibitor according to the present disclosure can suppress neuroinflammation by suppressing the excessive activation of microglia. In one or more embodiments, the neuroinflammation inhibitor according to the present disclosure can suppress the excessive activation of microglia and suppress neuroinflammation by promoting the stabilization of Nrf2 protein in glial cells. In the present disclosure, glial cells refer to non-neuronal cells that support neurons (nerve cells). In one or more embodiments, examples include microglia, astrocytes, and oligodendrocytes, and preferably microglia. Nrf2 is a transcription factor important for maintaining the homeostasis of the living body. In the present disclosure, the stabilization of Nrf2 protein means, in one or more embodiments, that the degradation of Nrf2 protein is suppressed and the intracellular amount of Nrf2 protein increases. The stabilization of Nrf2 can be confirmed with reference to the examples in one or more embodiments.

[0017] The neuroinflammation inhibitor according to the present disclosure can be formulated into a dosage form suitable for the administration form by applying well-known formulation techniques in one or more embodiments. Examples of the administration form include, but are not limited to, oral administration in dosage forms such as tablets, capsules, granules, powders, pills, troches, syrups, solutions, etc. Alternatively, parenteral administration in dosage forms such as injections, solutions, aerosols, suppositories, patches, poultices, lotions, liniments, ointments, eye drops, etc. can be mentioned. These formulations can be manufactured by well-known methods using additives such as excipients, lubricants, binders, disintegrants, stabilizers, flavoring and odor-correcting agents, and diluents, among others. The pharmaceutical composition of the present disclosure may further contain, in one or more embodiments, a pharmaceutically acceptable carrier, preservative, surfactant, pH adjuster, diluent, the above additives, or other pharmaceutically acceptable components.

[0018] Examples of the excipient include, but are not limited to, starches such as starch, potato starch, and corn starch, lactose, crystalline cellulose, calcium hydrogen phosphate, and the like. Examples of the lubricant include, but are not limited to, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, shellac, talc, carnauba wax, paraffin, and the like. Examples of the binder include, but are not limited to, polyvinylpyrrolidone, macrogol, and compounds similar to the excipient. Examples of the disintegrant include, but are not limited to, compounds similar to the excipient and chemically modified starches and celluloses such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone. Examples of the stabilizer include, but are not limited to, paraoxybenzoic acid esters such as methyl paraben and propyl paraben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenols such as phenol and cresol; thimerosal; dehydroacetic acid; and sorbic acid. Examples of the flavoring and odor-correcting agent include, but are not limited to, commonly used sweeteners, acidulants, fragrances, and the like.

[0019] In the production of the liquid preparation, solvents that can be used include, but are not limited to, ethanol, phenol, chlorocresol, purified water, distilled water, etc., and surfactants or emulsifiers can also be used as necessary. Examples of the surfactant or emulsifier include, but are not limited to, polysorbate 80, polyoxyl 40 stearate, lauromacrogol, and the like.

[0020] The neuroinflammation inhibitor according to the present disclosure can be administered to a subject suffering from a disease accompanied by neuroinflammation. Examples of the disease accompanied by neuroinflammation include, in one or more embodiments, frontotemporal lobe degeneration, amyotrophic lateral sclerosis, and multiple sclerosis. The subjects include humans and non-human animals. In one or more embodiments, examples of the animals include mammals such as mice, rats, guinea pigs, hamsters, rabbits, cats, dogs, sheep, pigs, cows, horses, goats, monkeys, etc.

[0021] The dosage of the neuroinflammation inhibitor according to the present disclosure may vary depending on symptoms, age, administration method, etc. The usage method is not limited to these, but it can be administered orally, transdermally, submucosally, subcutaneously, intramuscularly, intravascularly, intracranially, or intraperitoneally intermittently or continuously so that the in-vivo concentration of the compound of the active ingredient is between 100 nM and 1 mM. As a non-limiting embodiment, in the case of oral administration, for a subject (an adult if human), per day, in terms of the compound of the active ingredient, the lower limit is 0.01 mg (preferably 0.1 mg), and the upper limit is 2000 mg (preferably 500 mg, more preferably 100 mg), which can be administered once or divided into several times according to the symptoms. As a non-limiting embodiment, in the case of intravenous administration, for a subject (an adult if human), per day, the lower limit is 0.001 mg (preferably 0.01 mg), and the upper limit is 500 mg (preferably 50 mg), which can be administered once or divided into several times according to the symptoms.

[0022] Therefore, in other aspects, the present disclosure relates to a method for suppressing neuroinflammation, which includes administering an effective amount of the neuroinflammation inhibitor according to the present disclosure to a subject. In addition, in other aspects, the present disclosure relates to a method for suppressing excessive activation of microglia and suppressing neuroinflammation, which includes administering an effective amount of the neuroinflammation inhibitor according to the present disclosure to a subject. In addition, in other aspects, the present disclosure relates to a method for suppressing excessive activation of microglia and suppressing neuroinflammation by promoting the stabilization of Nrf2 protein, which includes administering an effective amount of the neuroinflammation inhibitor according to the present disclosure to a subject. In addition, in other aspects, the present disclosure relates to a method for improving, suppressing the progression of, and / or treating a disease accompanied by neuroinflammation, which includes administering an effective amount of the neuroinflammation inhibitor according to the present disclosure to a subject.

[0023] [Transplantation assisting agent] In one aspect, the present disclosure relates to a pharmaceutical composition (hereinafter also referred to as the transplantation assisting agent according to the present disclosure) for assisting the transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons, which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof. In the present disclosure, in one or more embodiments, the transplantation of cells refers to transplanting cells to a specific site of a transplantation target (recipient) and making them engraft (settle) at the transplanted site and / or its surrounding site, and / or appropriately differentiating them according to the surrounding environment. In one or more embodiments, the specific site to be transplanted includes the nervous system, the central nervous system (e.g., the brain, spinal cord), the peripheral nervous system, or these tissues. In the present disclosure, "assisting transplantation" refers to, in one or more embodiments, improving the survival rate, engraftment rate, and / or residual rate of transplanted cells after transplantation. The improvement of the survival rate, engraftment rate, and / or residual rate of cells can be confirmed with reference to the examples in one or more embodiments. In the present disclosure, "transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons" may, in one or more embodiments, be transplantation of these cells themselves, or may be transplantation in a form including other cells, or may be transplantation in a form in which neural progenitor cells, stem cells, and / or neurons are included in an organ (viscus), tissue, or part of an aggregate.

[0024] In the present disclosure, "neural progenitor cells" refer to cells that can differentiate into nerve cells, and their differentiation stage is not particularly limited. The neural progenitor cells used in the present disclosure may be neural stem cells in one or more embodiments. The neural progenitor cells used in the present disclosure may be cells isolated from mammalian brain tissues such as humans in one or more embodiments. The neural progenitor cells used in the present disclosure may be cells obtained by inducing differentiation from pluripotent stem cells such as embryonic stem cells (ES cells) and human induced pluripotent stem cells (iPS cells) (in some cases, they may be referred to as ES cell-derived cells and iPS cell-derived cells, respectively). The pluripotent stem cells used in the present disclosure include, in one or more embodiments, pluripotent stem cells capable of differentiating into nerve cells and neural stem cells. Examples of pluripotent stem cells include, in one or more embodiments, ES cells, iPS cells, embryonic stem (ntES) cells derived from cloned embryos obtained by nuclear transfer, germline stem cells (GS cells), embryonic germ cells (EG cells), pluripotent stem cells derived from cultured fibroblasts and bone marrow stem cells (Muse cells), and the like. Neurons (nerve cells) are not particularly limited, but in one or more embodiments, neurons induced to differentiate from neural progenitor cells are included. The neural progenitor cells and neurons to be transplanted include, in one or more embodiments, dopamineergic neuron precursor cells induced to differentiate from pluripotent stem cells and dopamineergic neurons induced to differentiate from pluripotent stem cells.

[0025] The transplantation adjuvant according to the present disclosure can be used to be administered to a recipient before transplantation, simultaneously with transplantation, or after transplantation. Alternatively, the transplantation adjuvant may be added to the cells to be transplanted before transplantation. Recipients include humans or animals other than humans. Examples of the animals include, in one or more embodiments, mammals such as mice, rats, guinea pigs, hamsters, rabbits, cats, dogs, sheep, pigs, cows, horses, goats, and monkeys. The cells to be transplanted can also be cells of the above-mentioned humans or animals other than humans. The recipient and the type of cells to be transplanted may be the same or different.

[0026] The dosage of the transplantation adjuvant according to the present disclosure varies depending on the purpose of administration, the administration method, and the condition of the administration subject (gender, age, weight, medical condition, etc.). However, when administered to humans, in one or more embodiments, the active ingredient may be used so that 10 to 1200 mg, or 100 to 1200 mg is administered per day. Alternatively, it may be the same as the neuroinflammation inhibitor according to the present disclosure described above.

[0027] As the administration route of the transplantation adjuvant according to the present disclosure, it can be directly contacted with the transplantation site or the transplanted cells, or can be administered orally, transdermally, submucosally, subcutaneously, intramuscularly, intravascularly, intracerebrally, or intraperitoneally. Examples of commonly used administration forms include solvents, tablets, capsules, granules, fine granules, powders, sublingual tablets, syrups, suspensions, etc. The transplantation adjuvant in the form of a liquid agent may be administered parenterally as an injection. The above administration forms can be produced by formulating the active ingredient according to the present disclosure with acceptable ordinary carriers, excipients, binders, stabilizers, etc. When the transplantation adjuvant according to the present disclosure is used as an injection, acceptable buffers, solubilizing agents, isotonic agents, etc. can also be added.

[0028] When using the transplantation adjuvant according to the present disclosure, in one or more embodiments, the survival rate, engraftment rate, and / or residual rate of neurons after transplantation in the target may be improved. Therefore, the transplantation adjuvant according to the present disclosure can be used for transplantation of organs (viscera), tissues, or cells for regenerative medicine in one or more embodiments. In addition, the transplantation adjuvant according to the present disclosure can be used for transplantation in the surgery (treatment) of neurological diseases such as neurodegenerative diseases showing progressive nerve loss, such as cerebral infarction, spinal cord infarction, cerebral hemorrhage, spinal cord hemorrhage, facial nerve paralysis, peripheral nerve paralysis, Lewy body dementia, Down syndrome, depression, neurodegenerative diseases, Alzheimer's disease, Parkinson's disease, and Huntington's disease in one or more embodiments.

[0029] Thus, in other aspects, the present disclosure relates to a method for improving the survival rate of transplanted cells, which includes administering an effective amount of the transplantation adjuvant according to the present disclosure to a recipient before, simultaneously with, or after transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons. In addition, in other aspects, the present disclosure relates to a method for transplanting neural progenitor cells, pluripotent stem cells, and / or neurons, which includes administering an effective amount of the transplantation adjuvant according to the present disclosure to a recipient before, simultaneously with, or after transplantation.

[0030] [Nrf2 stabilizer] In one aspect, the present disclosure relates to a pharmaceutical composition for promoting the stabilization of Nrf2 protein in glial cells (hereinafter also referred to as the Nrf2 stabilizer according to the present disclosure), which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof. The Nrf2 stabilizer according to the present disclosure can promote the stabilization of Nrf2 protein in glial cells in one or more embodiments. By promoting the stabilization of Nrf2 protein in glial cells, the Nrf2 stabilizer according to the present disclosure can suppress the excessive activation of glial cells in one or more embodiments. Since the activation of microglia is the starting point of neuroinflammation, the Nrf2 stabilizer according to the present disclosure can suppress the activation of glial cells and neuroinflammation in one or more embodiments.

[0031] The administration form, dosage form, dosage, etc. of the Nrf2 stabilizer according to the present disclosure can be the same as those of the neuroinflammation inhibitor according to the present disclosure.

[0032] The Nrf2 stabilizer according to the present disclosure can be administered to a subject suffering from a disease accompanied by neuroinflammation. In one or more embodiments, diseases accompanied by neuroinflammation include frontotemporal lobar degeneration, amyotrophic lateral sclerosis, and multiple sclerosis. Subjects include humans and non-human animals.

[0033] Therefore, in other aspects, the present disclosure relates to a method for promoting the stabilization of Nrf2 protein in glial cells, which includes administering an effective amount of the Nrf2 stabilizer according to the present disclosure to a subject. In addition, in other aspects, the present disclosure relates to a method for promoting the stabilization of Nrf2 protein in glial cells and suppressing neuroinflammation, which includes administering an effective amount of the Nrf2 stabilizer according to the present disclosure to a subject. In addition, in other aspects, the present disclosure relates to a method for improving, suppressing the progression of, and / or treating a disease accompanied by neuroinflammation, which includes administering an effective amount of the Nrf2 stabilizer according to the present disclosure to a subject.

[0034] [Neuroprotective agent] In one aspect, the present disclosure relates to a pharmaceutical composition for protecting neurons from neuroinflammation (hereinafter also referred to as the neuroprotective agent according to the present disclosure), which contains, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof. In one or more embodiments, the neuroprotective agent according to the present disclosure can suppress the excessive activation of glial cells by promoting the stabilization of Nrf2 protein in glial cells. Since the activation of microglia is the starting point of neuroinflammation, neuroinflammation can be suppressed by suppressing the activation of glial cells, and the protection of neurons can be achieved.

[0035] The dosage form, formulation type, dosage, etc. of the neuroprotective agent according to the present disclosure can be the same as those of the neuroinflammation inhibitor according to the present disclosure.

[0036] The neuroprotective agent according to the present disclosure can be administered to a subject suffering from a disease accompanied by neuroinflammation. In one or more embodiments, the diseases accompanied by neuroinflammation include frontotemporal lobe degeneration, amyotrophic lateral sclerosis, and multiple sclerosis. Subjects include humans and non-human animals.

[0037] Thus, in other aspects, the present disclosure relates to a method for protecting neurons from neuroinflammation, which includes administering an effective amount of the neuroprotective agent according to the present disclosure to a subject. Also, in other aspects, the present disclosure relates to a method for protecting neurons by suppressing neuroinflammation, which includes administering an effective amount of the neuroprotective agent according to the present disclosure to a subject. Also, in other aspects, the present disclosure relates to a method for protecting neurons by suppressing the activation of glial cells to suppress neuroinflammation, which includes administering an effective amount of the neuroprotective agent according to the present disclosure to a subject. Also, in other aspects, the present disclosure relates to a method for improving, suppressing the progression of, and / or treating a disease accompanied by neuroinflammation, which includes administering an effective amount of the neuroprotective agent according to the present disclosure to a subject.

[0038] [Active ingredient] The active ingredient of the pharmaceutical composition according to the present disclosure (neuroinflammation inhibitor, transplantation adjuvant, Nrf2 stabilizer, and neuroprotective agent) (also referred to as the active ingredient according to the present disclosure) is a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof. In one or more embodiments, compounds having an ability to inhibit the phosphorylation activity of DYRK1A protein that are disclosed in WO2018 / 043674 and WO2015 / 107945 can be used. The contents of these documents are incorporated herein by reference as part of the present disclosure. DYRK1A promotes the degradation of cyclin D1 and p21 by phosphorylating cyclin D1.

[0039] In one or more embodiments, the compound having an ability to inhibit the phosphorylation activity of DYRK1A protein in the present disclosure includes at least one selected from the group consisting of compounds represented by the following formulas (I) to (III). [Chemical formula] In formula (I), R 1 and R 2is, independently of each other, a hydrogen atom or a hydrocarbon chain having 1 to 6 carbon atoms, R 3 is -CH2-CH2- or -CH=CH-, R 4 is a hydrogen atom, a halogen atom or an alkyl group having 1 to 6 carbon atoms, In formulas (II) and (III), R 5 , R 6 , R 7 and R 8 are, independently of each other, a hydrogen atom, a halogen atom, a carboxyl group, an amino group, a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, or an alkyl group having 1 to 4 carbon atoms substituted with a halogen atom.

[0040] In general formula (I), R 1 is, in one or more embodiments, an alkyl group having 1 to 6 carbon atoms, and in further one or more embodiments, is a methyl group, an ethyl group, or a propyl group. In general formula (I), R 2 is, in one or more embodiments, an alkyl group having 1 to 6 carbon atoms, and in further one or more embodiments, is a methyl group. In general formula (I), R 4 is, in one or more embodiments, a hydrogen atom.

[0041] The compound represented by the general formula (I) is, in one or more embodiments,

Chemical formula

[0042] In general formulas (II) and (III), R 5 , R 6 , R 7 and R 8 are, in one or more embodiments, hydrogen atoms.

[0043] The compounds represented by the general formulas (II) and (III) are, in one or more embodiments,

Chemical formula

[0044] In the present disclosure, "pharmaceutically acceptable salts" include salts that are pharmacologically and / or pharmaceutically acceptable, and examples thereof include inorganic acid salts, organic acid salts, inorganic base salts, organic base salts, acidic or basic amino acid salts, and the like.

[0045] Preferable examples of the inorganic acid salts include, for example, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, and the like. Preferable examples of the organic acid salts include, for example, acetate, succinate, fumarate, maleate, tartrate, citrate, lactate, stearate, benzoate, methanesulfonate, p-toluenesulfonate, and the like.

[0046] Preferable examples of the inorganic base salts include, for example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, aluminum salt, ammonium salt, and the like. Preferable examples of the organic base salts include, for example, diethylamine salt, diethanolamine salt, meglumine salt, N,N'-dibenzylethylenediamine salt, and the like.

[0047] Preferable examples of the acidic amino acid salts include, for example, aspartate, glutamate, and the like. Preferable examples of the basic amino acid salts include, for example, arginine salt, lysine salt, ornithine salt, and the like.

[0048] In the present disclosure, "salts of the compound" may include hydrates that can be formed by the compound absorbing moisture when left in the air. Further, in the present disclosure, "salts of the compound" may also include solvates that can be formed by the compound absorbing certain other solvents.

[0049] In the present disclosure, the alkyl group includes, in one or more embodiments, a linear, branched, or cyclic alkyl group. In the present disclosure, the "alkyl group having 1 to 4 carbon atoms" refers to, in one or more embodiments, a linear, branched alkyl group having 1, 2, 3, or 4 carbon atoms, or a cyclic alkyl group having 3 or 4 carbon atoms. Examples of the linear or branched alkyl group having 1, 2, 3, or 4 carbon atoms include, in one or more embodiments, a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, and a tert-butyl group. Examples of the cyclic alkyl group having 3 or 4 carbon atoms include, in one or more embodiments, a cyclopropyl group and a cyclobutyl group.

[0050] In the present disclosure, the "hydrocarbon chain having 1 to 6 carbon atoms" refers to a monovalent group derived by removing one arbitrary hydrogen atom from an aliphatic hydrocarbon having 1, 2, 3, 4, 5, or 6 carbon atoms. The hydrocarbon chain may be, in one or more embodiments, a linear structure, a branched structure, or a cyclic structure, and examples include an alkyl group, an alkenyl group, a phenyl group, or a cycloalkyl group. In the present disclosure, the "alkyl group having 1 to 6 carbon atoms" includes, in one or more embodiments, a methyl group, an ethyl group, a 1-propyl group, a 2-propyl group, a 2-methyl-1-propyl group, a 2-methyl-2-propyl group, a 1-butyl group, a 2-butyl group, a 1-pentyl group, a 2-pentyl group, a 3-pentyl group, a 2-methyl-1-butyl group, a 3-methyl-1-butyl group, a 2-methyl-2-butyl group, a 3-methyl-2-butyl group, a 2,2-dimethyl-1-propyl group, a 1-hexyl group, a 2-hexyl group, a 3-hexyl group, a 2-methyl-1-pentyl group, a 3-methyl-1-pentyl group, a 4-methyl-1-pentyl group, a 2-methyl-2-pentyl group, a 3-methyl-2-pentyl group, a 4-methyl-2-pentyl group, a 2-methyl-3-pentyl group, a 3-methyl-3-pentyl group, a 2,3-dimethyl-1-butyl group, a 3,3-dimethyl-1-butyl group, a 2,2-dimethyl-1-butyl group, a 2-ethyl-1-butyl group, a 3,3-dimethyl-2-butyl group, or a 2,3-dimethyl-2-butyl group. In the present disclosure, the halogen atom includes, in one or more embodiments, a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

[0051] The present disclosure may relate to one or more of the following embodiments; [1] A pharmaceutical composition for suppressing neuroinflammation, comprising, as an active ingredient, a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof. [2] The pharmaceutical composition according to [1], for assisting the transplantation of neural progenitor cells, pluripotent stem cells, and / or neurons. [3] The pharmaceutical composition according to [1], for promoting the stabilization of Nrf2 protein in glial cells. [4] The pharmaceutical composition according to [1], for protecting neurons from neuroinflammation. [5] The pharmaceutical composition according to [1], for improving, suppressing the progression of, and / or treating a disease associated with neuroinflammation selected from the group consisting of frontotemporal dementia, amyotrophic lateral sclerosis, and multiple sclerosis. [6] The compound having an ability to inhibit the phosphorylation activity of DYRK1A protein is at least one selected from the group consisting of compounds represented by the following formulas (I) to (III),

Chemical formula

[10] A method for protecting neurons from neuroinflammation, comprising administering to a subject an effective amount of the pharmaceutical composition according to any one of [1] to [6].

[11] A method for improving, suppressing the progression of, and / or treating a disease associated with neuroinflammation selected from the group consisting of frontotemporal dementia, amyotrophic lateral sclerosis, and multiple sclerosis, comprising administering to a subject an effective amount of the pharmaceutical composition according to any one of [1] to [6].

Examples

[0052] Hereinafter, the present disclosure will be described in more detail by way of examples, which are illustrative only and the present disclosure is not limited to these examples. All the documents cited in the present disclosure are incorporated herein by reference as part of the present disclosure.

[0053] Compound 1 The following Compound 1 was synthesized by the method disclosed in WO2018 / 043674. The IC 50 of Compound 1 in the in vitro kinase activity against DYRK1A is 76.95 nM.

Chemical formula

[0054] Compound 2 Compound 2 below was synthesized by the method disclosed in WO2015 / 083750. The IC 50 of compound 2 in the in vitro kinase activity against DYRK1A

Chemical formula

[0055] Statistics Results obtained from more than 3 experiments were expressed as mean ± SEM. Statistically significant differences were determined using the Tukey-Kramer post hoc test following two-tailed unpaired Student's t-test or one-way ANOVA. A P-value of less than 0.05 was considered significant and indicated by a single asterisk (*), and a P-value of less than 0.01 was indicated by a double asterisk (**).

[0056] Image analysis Cells were seeded into 96-well PureCoat amine-coated plates and processed as required for each specific experiment. After immunolabeling was completed, images were automatically acquired (20x magnification of the objective lens, 2x2 CCD binning, 25 fields per well) and analyzed using Opera Phenix (Perkin Elmer) equipped with Harmony software. A fluorescence microscope (BZ-9000, Keyence) or a confocal microscope (SP-8, Leica) was also used for image acquisition.

[0057] Animal model 8- to 9-week-old C57black / 6J mice were intraperitoneally injected with LPS (O55:B5) (Sigma, L2880) at a dose of 1 mg / kg once a day for 4 days. hiPSC-derived dopamine (DA) progenitor cells were transplanted into the striatum of 4-week-old SCID mice. Drug treatment was performed 1 hour before LPS or iPSC transplantation.

[0058] Reagents Lipopolysaccharide (LPS) was obtained from Sigma-Aldrich. Small molecule compounds were dissolved in dimethyl sulfoxide (DMSO, manufactured by Nacalai Tesque) to prepare a 50 mM stock solution for in vitro assays. siRNA was purchased from Ambion or Dharmacon. Rabbit polyclonal anti-Nrf2 (MBL) for Western blot, rabbit polyclonal anti-Nrf2 (abcam) for immunocytochemistry, mouse monoclonal anti-p21 (abcam), rabbit polyclonal anti-cyclin D1 (Cell Signaling), rat anti-Cd11b (abcam), chicken polyclonal anti-TH (abcam), rabbit polyclonal anti-Iba1 (Wako), goat polyclonal anti-GFAP (Millipore), rat anti-Nurr1 (provided by KAN Research Institute), mouse monoclonal anti-hNuclei (abcam), and rabbit polyclonal HRP-conjugated anti-GAPDH (MBL) were used.

[0059] Cell culture The microglial cell line BV-2 was maintained in high glucose Dulbecco's modified Eagle's medium (DMEM) (Nacalai Tesque) supplemented with 10% fetal bovine serum (Nichirei Bioscience), 100 U / mL penicillin, and 100 μg / mL streptomycin. Primary hippocampal and cortical neuron cultures were prepared from embryonic day 18 mice and maintained in Neurobasal medium (Life Technologies) supplemented with 2% B27 supplement, 100 U / mL penicillin, 100 μg / mL streptomycin, and 0.5 mM L-glutamine. Dopaminergic neuron cultures were prepared from the ventral midbrain of embryonic day 13 mice and maintained in Neurobasal medium (Life Technologies) supplemented with 10% fetal bovine serum, 2% B27 supplement, 10 ng / mL GDNF, 100 U / mL penicillin, and 100 μg / mL streptomycin. For glial cell removal, 5 μM cytosine β-D-arabinofuranoside, 10 μM 5-fluorouracil, and 10 μM uridine (all from Sigma-Aldrich) were added on day 2 of culture. Mixed glial cultures were obtained from neonatal mouse pups (P1–P4) and maintained in T75 or T175 flasks. The medium was changed every 3–4 days until the cells became confluent. Microglia were obtained either by vigorous shaking or by Cd11b positive selection (Invitrogen). For co-culture experiments, mouse glial cell cultures were prepared from the medial ganglionic eminence (MGE) of embryonic day 13 and cultured in PLL-coated dishes. The cultures were passaged twice with trypsin and used for co-culture assays with iPSC-derived neurons. 1039A1 iPS cells were established and maintained as previously described (Nakagawa et al., Sci Rep 4, 3594, 2014). Induction of dopaminergic progenitor cells was performed as previously described (Doi et al., Stem Cell Reports 2, 337–350, 2014, Kikuchi et al., Nature 548, 592–596, 2017).Briefly, hiPS cells 1039A1 were seeded on Laminin511 and differentiated in GMEM medium containing 8% KSR, Y27632 (Wako), A-83-01 (Wako), and LDN193178 (STEMGENT). Purmorphamine (Wako), FGF8 (Wako, from day 1 to day 7), and CHIR99021 (Wako, from day 3 to day 12) were added. After cell sorting for Corin+ cells, the cells were re-seeded into low-adhesion 96-well plates for neurosphere culture and maintained for an additional 2 weeks in neurobasal medium supplemented with B27 supplement, 2 mM L-glutamine (Invitrogen), 10 ng / mL GDNF, 200 μM ascorbic acid, 20 ng / mL BDNF (all from Wako), and 400 μM dbcAMP (Sigma-Aldrich). To avoid apoptosis, 30 μM of Y27632 (Wako) was added at the first plating.

[0060] Immunocytochemistry Cells were fixed with 4% paraformaldehyde for 10 minutes and then permeabilized with 0.2% Triton X-100 for 10 minutes. After washing with PBS, the cells were blocked with 5% normal donkey serum (Jackson ImmunoResearch Laboratories) / 1% BSA (SIGMA A7906) / PBS and labeled with each primary antibody. After washing with PBS, the primary antibody was labeled with the corresponding fluorescently labeled secondary antibody. Nuclei were detected using Hoechst33342.

[0061] RNA extraction and quantitative RT-PCR Total RNA was extracted using the RNeasy kit (Qiagen), and then cDNA was synthesized using iScript (Bio-Rad). Quantitative PCR was performed using SYBR green Ex Taq (TaKaRa). Gene primers were designed using PrimerBank (Wang et al., 2012).

[0062] Immunoblotting Total protein was extracted from cell culture samples using RIPA buffer (Wako) containing protease inhibitor cocktail (Nacalai Tesque) and phosphatase inhibitor cocktail (Nacalai Tesque). After centrifugation at 15,000 rpm for 15 minutes at 4 °C, the supernatant was collected and the protein concentration was measured using the Pierce 660 nm Protein Assay Kit (Thermo Scientific). Next, the proteins were separated on a 5–20% gradient SDS / PAGE gel (ATTO) and transferred to a polyvinylidene fluoride membrane (Millipore) by electroblotting. The membrane was blocked with Blocking One (Nacalai Tesque) and then probed with the indicated antibodies. Detection was performed using Immunostar chemiluminescence (Wako) and a ChemiDoc imaging system (Bio-Rad).

[0063] In vitro enzyme activity assay The in vitro kinase activity assay was performed as previously described (Ogawa et al., Nat Commun 1, 86, 2010).

[0064] Drug treatment study Compound 1 was first dissolved in DMSO at a concentration of 100 mg / mL, diluted to the desired concentration with 10% Tween 80 (Polysorbate 80 (HX2), manufactured by HOF Corporation) in physiological saline, and delivered subcutaneously in an amount of 0.05 mL / kg. Compound 2 was suspended in 0.5% carboxymethyl cellulose (Nacalai Tesque) and orally administered in an amount of 0.1 mL / kg at the desired concentration. The drug-treated animals were anesthetized with isoflurane for blood sampling and then perfused with physiological saline. Brain homogenates were prepared using a Beads Crusher μT-12 system (TAITEC) with 5 volumes of physiological saline. The concentrations of the target compound, dopamine, in serum and brain homogenates were analyzed by LC / MS using an Agilent 6420 Q-TOF mass spectrometer equipped with an Agilent 1290 nanoflow HPLC system (Agilent Technologies). For the measurement of dopamine, 50 mg / mL ascorbic acid was added to prevent oxidation.

[0065] Immunohistochemistry The brains of adult mice were perfused with PBS, fixed with 4% paraformaldehyde, equilibrated with 30% sucrose / PBS, and then cut into 40 μm sections with a vibratome (Leica) or 20 μm sections with a cryostat (Leica). After antigen retrieval using HistoOne (Nacalai Tesque), the tissues were stained with the designated antibodies.

[0066] [Experimental Example 1: Induction of p21 and Nrf2] It was confirmed that Compounds 1 and 2 induced p21 and Nrf2 in glial cells. Figure 1A shows the results of Western blot analysis when microglial cell line BV-2 was treated with Compound 1. BV-2 cells were treated with Compound 1 at the designated concentration and for the designated period. The treated samples were subjected to Western blot analysis using the indicated antibodies. As shown in Figure 4A, in BV-2 cells, cyclin D1, p21, and Nrf2 were upregulated by Compound 1 in a time-dependent and dose-dependent manner. Figure 1B is a representative image of glial cells treated with compounds 1 and 2. Cells were visualized with anti-Nrf2 (pseudo-colored), Cd11b (magenta, microglia) and GFAP (green, astrocytes) antibodies. Scale bar = 25 μm. Figure 1C is an example of quantification of the signal intensity of Nrf2 in the nuclei of Cd11b-positive cells. Glial cells were treated with the indicated concentrations of compounds 1 and 2 for the indicated periods. From Figures 1B and C, it was confirmed that treatment with compound 1 or 2 induced the expression of Nrf2 in Cd11b-positive microglial cells. Furthermore, Nrf2 expression did not increase upon treatment with compound 1 or 2 in cells treated with p21 siRNA, but increased upon treatment with compound 1 or 2 in cells treated with control siRNA (data not shown). From these results, it is considered that treatment with compound 1 or 2 mediates the induction of Nrf2 by stabilizing the cyclin D1 / p21 complex.

[0067] [Experimental Example 2: Compounds 1 and 2 suppress neuroinflammation through the stabilization of Nrf2] It was investigated whether compounds 1 and 2 suppress cytokine production in microglia. Figure 2A shows the results of evaluating the production of cytokines, chemokines, and iNOS mRNA by qPCR during LPS treatment. *P < 0.05. Figure 2B shows the results of quantifying cytokines produced by LPS stimulation by ELISA. *P < 0.05. As shown herein, LPS stimulation dramatically induces inflammatory cytokine gene expression, but these upregulations were effectively suppressed by treatment with compound 1 or 2. Furthermore, the induction of some chemokine and iNOS mRNA expression was also suppressed (Figure 2A). Figure 2C shows the results of qPCR analysis of the production of the indicated cytokines in the presence or absence of Nrf2 by compound 1. Cytokine production suppression was not observed in cells treated with Nrf2 siRNA. *P < 0.05. These results indicate that the treatment with Compound 1 or 2 can effectively suppress neuroinflammation. Furthermore, these results strongly suggest that the suppression of neuroinflammation (suppression of cytokine production) by Compounds 1 and 2 is mediated via Nrf2.

[0068] [Experimental Example 3: Effect of improving the transplantation efficiency of iPSC-derived cells (in vivo)] Dopaminergic neurons (DA neurons) were generated from human iPSCs and transplanted into the mouse brain, and it was confirmed that the treatment with Compound 2 improved the efficiency of the engraftment of iPSC-derived DA neurons. The outline is shown in Fig. 3. Fig. 3A is the experimental scheme. The recipient (mouse) was administered Compound 2 one hour before the transplantation surgery of hiPSC-derived DA neurons. Drug administration was performed continuously for 4 days until 4 days after the surgery when strong glial activation was expected, and the animals were left untreated for 4 weeks. Fig. 3B is a representative image of the cells transplanted into the striatal tissue. The arrows indicate iPSC-derived cells co-labeled with TH or Nurr1 (both are dopaminergic markers). Scale bar = 50 μm. The transplanted DA neurons were identified by hNuclei staining, and the surviving iPSC-derived DA neurons were quantified by co-staining with TH and Nurr1 dopaminergic markers. Fig. 3C shows the results of the quantitative analysis of the transplanted cells. The number of double-positive cells for hNuclei and TH or Nurr1 was normalized to the number of hNuclei-positive cells. N = 7 and 5 for each condition. *P < 0.05, **P < 0.01. From the results of Fig. 3, it was shown that the treatment with Compound 2 can improve the survival rate of iPSC-derived DA neurons after 4 weeks of transplantation.

[0069] [Experimental Example 4: Effect of suppressing neurodegeneration caused by neuroinflammation (in vivo)] It was confirmed that Compound 2 can reduce neurodegeneration caused by neuroinflammation. The outline is shown in Fig. 4. Figure 4A shows the experimental scheme. Neuroinflammation was induced by intraperitoneal administration of LPS to mice. The drug (Compound 2) was administered 1 hour before the LPS injection to the mice. The drug was orally administered once a day at the designated dose. On the first day after administration, glial activation and cytokine production were evaluated. On the 7th and 14th days, the degeneration of dopaminergic neurons was evaluated (Figures 4B and 4C). Figure 4B is a representative image of the substantia nigra of the treated animals. TH (green), Iba1 (grayscale), and GFAP (magenta) were used as markers for DA neurons, microglia, and astrocytes, respectively. Scale bar = 200 μm. Figure 4C shows the result of quantifying the number of TH-positive cells in the substantia nigra pars compacta (SNpc). N = 5 - 6 animals were analyzed for each condition. Error bars represent SEM. *P < 0.05. It was confirmed that the administration of Compound 2 suppressed glial activation (Figure 4D) and cytokine production (Figures 4E and F). Figure 4D shows the result of quantitative analysis of glial activation by qPCR of striatal tissue on the first day after the last LPS injection. Error bars represent SEM. *P < 0.05. Figures 4E and F show the results of analyzing the levels of cytokines and chemokines shown from striatal tissue on Day 1 by qPCR (E) and ELISA (F). Error bars represent SEM. *P < 0.05. Importantly, the loss of dopaminergic neurons was rescued by the administration of Compound 2 (Figures 4B and C). That is, Compound 2 was able to reduce the neurodegeneration caused by neuroinflammation.

[0070] [Experimental Example 5: Neuroprotective mechanism via glial cells (in vitro)] It was confirmed with Compound 2 that the site of action of the neuroprotective functions (improvement of the effect of iPS cell transplantation and inhibitory effect on cell degeneration due to cell inflammation) shown in Experimental Examples 1 and 2 is glial cells. To confirm the protection of neurons via glial cells, a co-culture system was set up. hiPSC-derived dopaminergic neural progenitor cells were co-cultured by mixing with glial culture solution isolated from the 13-day-old mouse brain. The mixed glial culture solution contained GFAP-positive and Iba1-positive cells, but no TH-positive cells. Oxidative stress was induced in the mixed glial culture solution by H2O2 treatment (Figure 5). The survival of neurons was evaluated by the number of human nuclei via immunostaining (Figures 5A and B). Dopaminergic markers of Nurr1 and TH were evaluated by qPCR (Figure 5C). Figure 5A is a representative image of hiPSC-derived dopaminergic neural progenitor cells. hiPSC-derived DA neurons were visualized using antibodies against anti-hNuclei (green), anti-Nurr1 (red), and anti-TH (grayscale). Scale bar = 50 μm. Figure 5B is a quantitative analysis of the number of hNuclei. The data were normalized to the control conditions without H2O2 treatment. *P<0.05. Figure 5C is a quantification by qPCR. Nurr1 and TH were used as markers for dopaminergic neurons in early differentiation and maturation, respectively. As shown in Figure 5, oxidative stress caused by H2O2 treatment reduces the survival of DA neurons, but the survival amount increases depending on the added amount of Compound 2. However, the protection of neurons by this Compound 2 does not occur in the absence of glia and occurs only during co-culture with glia. These strongly indicate that the neuroprotective function of Compound 2 is exerted via glial cells.

Claims

1. A pharmaceutical composition for suppressing neuroinflammation, comprising: a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof as an active ingredient; The pharmaceutical composition, wherein the compound is any of the following compounds. 【Chemical 1】

2. The pharmaceutical composition according to claim 1, wherein the compound is any of the following compounds. 【Chemical Formula 2】

3. The pharmaceutical composition according to claim 1 or 2, for improving, suppressing the progression of, and / or treating a disease associated with neuroinflammation.

4. The pharmaceutical composition according to any one of claims 1 to 3, for treating Alzheimer's disease.

5. The pharmaceutical composition according to any one of claims 1 to 4, for oral administration.

6. A pharmaceutical composition for promoting the stabilization of Nrf2 protein in glial cells to protect neurons, comprising: a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof as an active ingredient; The pharmaceutical composition, wherein the compound is any of the following compounds. 【Chemical 3】

7. The pharmaceutical composition according to claim 6, wherein the compound is any of the following compounds. 【Chemical Formula 4】

8. The pharmaceutical composition according to claim 6 or 7, for improving, suppressing the progression of, and / or treating a disease associated with neuroinflammation.

9. The pharmaceutical composition according to any one of claims 6 to 8, wherein the pharmaceutical composition is a pharmaceutical composition for treating Alzheimer's disease.

10. The pharmaceutical composition according to any one of claims 6 to 9, wherein the pharmaceutical composition is a pharmaceutical composition for oral administration.

11. A pharmaceutical composition for improving the engraftment rate of transplanted cells after transplantation, comprising: a compound having an ability to inhibit the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof as an active ingredient; wherein the cells are neural progenitor cells, pluripotent stem cells, and / or neurons; The pharmaceutical composition, wherein the compound is any of the following compounds. 【Chemical Formula 5】

12. The pharmaceutical composition according to claim 11, wherein the compound is any of the following compounds. [Chemical Formula 6]

13. The pharmaceutical composition according to claim 11 or 12, for improving the engraftment rate after transplantation in a surgery (treatment) for a neurological disease. The pharmaceutical composition, wherein the neurodegenerative disease is a neurodegenerative disease showing progressive neuronal loss selected from cerebral infarction, spinal cord infarction, cerebral hemorrhage, spinal cord hemorrhage, facial nerve palsy, peripheral nerve palsy, Lewy body dementia, Down syndrome, depression, neurodegenerative disease, Alzheimer's disease, Parkinson's disease and Huntington's disease.

14. A pharmaceutical composition for improving the survival rate and / or residual rate of transplanted cells after transplantation, which contains, as an active ingredient, a compound having an inhibitory ability against the phosphorylation activity of DYRK1A protein or a pharmaceutically acceptable salt thereof, wherein the cells are neural progenitor cells, pluripotent stem cells, and / or neurons, and the compound is any of the following compounds. 【Chemical Formula 7】

15. The pharmaceutical composition according to claim 14, wherein the compound is the following compound. 【Chemical 8】

16. The pharmaceutical composition according to claim 14 or 15 for improving the engraftment rate after transplantation in the surgery (treatment) of a neurodegenerative disease, wherein the neurodegenerative disease is a neurodegenerative disease showing progressive neuronal loss selected from cerebral infarction, spinal cord infarction, cerebral hemorrhage, spinal cord hemorrhage, facial nerve palsy, peripheral nerve palsy, Lewy body dementia, Down syndrome, depression, neurodegenerative disease, Alzheimer's disease, Parkinson's disease and Huntington's disease.

Images