Acrylamide Compounds

Abstract

The present invention provides a novel acrylamide compound represented by the following general formula [I] or a salt thereof, which is useful for promoting platelet production from platelet precursor cells such as megakaryocytes in vitro: JPEG2023505366000081.jpg4981 (wherein each symbol has the same meaning as in the specification)

Description

[Technical field]

[0001] The present invention relates to an acrylamide compound, and more particularly to an acrylamide compound that promotes in vitro production of platelets from platelet precursor cells such as megakaryocytes. [Background technology]

[0002] Platelet preparations are administered to patients who have experienced heavy bleeding during surgery or injury, or who have a tendency to bleed due to thrombocytopenia following anti-cancer drug treatment, for the purpose of treating the symptoms and preventing unexpected bleeding. Currently, platelet preparations are dependent on blood donations, but the shelf life of platelet preparations is very short, at around four days. Furthermore, if we continue to rely solely on blood donations due to the declining number of blood donors, it is predicted that there will be a shortage of platelet preparations in the near future. To meet these needs, methods for producing platelets in vitro are being investigated. As an in vitro platelet production method, a method has been developed in which various stem cells are differentiated to obtain megakaryocytes, which are then cultured to release platelets. For example, Takayama et al. have succeeded in inducing the differentiation of human ES cells into megakaryocytes and platelets (Non-Patent Document 1). Furthermore, as a method for producing platelets from hematopoietic progenitor cells in vitro, a method has been proposed in which hematopoietic progenitor cells are cultured in the presence of an aromatic hydrocarbon receptor antagonist and a thrombopoietin (TPO) or ROCK (Rho-associated coiled-coil forming kinase) inhibitor (Patent Documents 1, 2, and 3; Non-Patent Documents 2, 3, and 4). Indolyl acrylamide compounds have been disclosed as transcription factor inhibitors (Patent Document 4, Non-Patent Document 5). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] WO2014 / 138485 publication [Patent Document 2] WO2016 / 204256 publication [Patent Document 3] WO2010 / 059401 publication [Patent Document 4] WO2019 / 167973 publication [Non-patent literature]

[0004] [Non-Patent Document 1] Takayama et al., Blood, 111, 5298 (2008) [Non-Patent Document 2] Boitano et al., Science, 329, 1345 (2010) [Non-Patent Document 3] Strassel et al., Blood, 127, 2231 (2016) [Non-Patent Document 4] Ito et al., Cell, 174, 636 (2018) [Non-Patent Document 5] Perron et al., J. Biol. Chem., 293, 8285 (2018) Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a novel acrylamide compound or a salt thereof which is useful for promoting in vitro platelet production from platelet precursor cells such as megakaryocytes. Another object of the present invention is to provide a platelet production promoter that is useful for promoting in vitro platelet production from platelet precursor cells such as megakaryocytes. [Means for solving the problem]

[0006] As a result of extensive investigations aimed at solving the above problems, the present inventors have found that an acrylamide compound represented by the following formula [I] or [I'] or a salt thereof has the effect of promoting platelet production, and have thus completed the present invention.

[0007] That is, the present invention includes the following aspects. [1-1] General formula [I]: JPEG0007657800000001.jpg4982[In the formula, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 Alkyl; R 2 is a hydrogen atom or -C 1-6 is alkyl, R 3 is halogen, -Q k -(C 1-6 Alkyl) m -Q p -R 31 , optionally substituted phenyl or optionally substituted heteroaryl selected from the group consisting of furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and pyrimidyl; R 31 -C 1-6 Alkyl or -C 3-8 is cycloalkyl, Q are independently the same or different and each is an oxygen atom, a sulfur atom, -C(=O)-O-, or -NH-; k, m, and p are 0 or 1; n is 0, 1, or 2. When n is 2, R 3 are each independently the same or different substituents. W is a carbon atom or a nitrogen atom; X is a carbon atom, a nitrogen atom, or NR 12 and Y is a carbon atom or a nitrogen atom; Z's are independently the same or different and each is a nitrogen atom or CH; However, X and Y are not carbon atoms at the same time. R 12 is a hydrogen atom, -C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)-C 1-6 Alkyl, -C(=O)-aryl or -C(=O)-OC 1-6 is alkyl, Ring A is aryl or heteroaryl; --- is a single or double bond, However, when X is NH, W and Y are carbon atoms, and all Z are CH, ring A is 2-(-OC 1-6 alkyl)phenyl or 2,5-di(-OC 1-6 (alkyl)phenyl] A compound represented by the formula: [1-2] In general formula [I], JPEG0007657800000002.jpg32150[where, R 11 , W, X, Y, Z and --- is as defined above] The compound or a salt thereof according to [1-1], [1-3] In general formula [I], JPEG0007657800000003.jpg18150[R 3 and n is as defined above] The compound or a salt thereof according to [1-1], [1-4] In the general formula [I], the heteroaryl in ring A is Selected from the group consisting of furan, thiophene, pyridine and quinoline; The compound or a salt thereof according to [1-1]. [1-5] In general formula [I], JPEG0007657800000004.jpg32150 (wherein V is the same or different and each V is independently a nitrogen atom or CH; R 4 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC1-6 is alkyl. The compound or salt thereof according to [1-1], [1-6] General formula [Ia] JPEG0007657800000005.jpg3458[where, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl, R 12 is a hydrogen atom or -C(=O)-OC 1-6 is alkyl, JPEG0007657800000006.jpg1421 is pyridylbenzene, (halogen, -C 1-6 Alkyl or -OC 1-6 pyrimidyl)benzene, phenylthiophene, pyridylthiophene or pyrimidylthiophene, which may be substituted with alkyl. The compound according to [1-1], or a salt thereof. [1-7] The compound according to [1-1], selected from the group consisting of the following compounds, or a salt thereof: JPEG0007657800000007.jpg95150[2-1] General formula [I']: JPEG0007657800000008.jpg4982[In the formula, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 Alkyl; R 2 is a hydrogen atom or -C 1-6 is alkyl, R 3 is halogen, -Q k -(C 1-6 Alkyl) m -Q p -R 31 , optionally substituted phenyl or optionally substituted heteroaryl selected from the group consisting of furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and pyrimidyl; R31 -C 1-6 Alkyl or -C 3-8 is cycloalkyl, Q are independently the same or different and each is an oxygen atom, a sulfur atom, -C(=O)-O-, or -NH-; k, m, and p are 0 or 1; n is 0, 1, or 2. When n is 2, R 3 are each independently the same or different substituents. W is a carbon atom or a nitrogen atom; X is a carbon atom, a nitrogen atom, or NR 12 and Y is a carbon atom or a nitrogen atom; Z's are independently the same or different and each is a nitrogen atom or CH; However, X and Y are not carbon atoms at the same time. R 12 is a hydrogen atom, -C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)-C 1-6 Alkyl, -C(=O)-aryl or -C(=O)-OC 1-6 is alkyl, Ring A is aryl or heteroaryl; --- is a single or double bond. A platelet production promoter comprising a compound represented by the formula: [2-2] In the general formula [I'], JPEG0007657800000009.jpg32150[where, R 11 , W, X, Y, Z and --- is as defined above] The platelet production promoter according to [2-1], comprising a compound represented by the formula: [2-3] In the general formula [I'], JPEG0007657800000010.jpg19150[R 3 and n is as defined above] The platelet production promoter according to [2-1], comprising a compound represented by the formula: [2-4] The platelet production promoter according to [2-1], wherein in the general formula [I'], the heteroaryl in ring A includes a compound selected from the group consisting of furan, thiophene, pyridine and quinoline, or a salt thereof. [2-5] In the general formula [I'], JPEG0007657800000011.jpg32150 (wherein V is the same or different and each V is independently a nitrogen atom or CH; R 4 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl. The platelet production promoter according to [2-1], comprising a compound represented by the formula: [2-6] General formula [Ia] JPEG0007657800000012.jpg3459[where, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl, R 12 is a hydrogen atom or -C(=O)-OC 1-6 is alkyl, JPEG0007657800000013.jpg1421 is pyridylbenzene, (halogen, -C 1-6 Alkyl or -OC 1-6 pyrimidyl)benzene, phenylthiophene, pyridylthiophene or pyrimidylthiophene, which may be substituted with alkyl. The platelet production promoter according to [2-1], comprising a compound represented by the following formula: [2-7] The platelet production promoter according to [2-1], comprising a compound selected from the group consisting of the following compounds or a salt thereof: JPEG0007657800000014.jpg95150[2-8] General formula [Ia'] JPEG0007657800000015.jpg4270[In the formula, R 3a -OC 1-6Alkyl; R 3b is a hydrogen atom or -OC 1-6 Alkyl; R 11 -C 1-6 Alkyl or -OC 1-6 Alkyl; R 12 is a hydrogen atom or -C 1-6 indicates alkyl] A platelet production promoter comprising a compound represented by the formula: [2-9] In the general formula [Ia'], R 3a is -O-methyl or -O-ethyl; R 3b is a hydrogen atom or -O-methyl; R 11 is methyl or -O-methyl; R 12 is a hydrogen atom or a methyl The platelet production promoter according to [2-8], comprising a compound represented by the formula: [2-10] The platelet production promoter according to [2-8], comprising a compound selected from the group consisting of the following compounds or a salt thereof: TIFF0007657800000016.tif80160[2-11] A platelet production promoter described in any of [2-1] to "2-10" for use in combination with an aromatic hydrocarbon receptor inhibitor. [2-12] The platelet production promoter according to [2-11], wherein the aromatic hydrocarbon receptor inhibitor is selected from the group consisting of the following: JPEG0007657800000017.jpg106150 [3-1] Use of a compound or a salt thereof according to any one of [2-1] to [2-10] for promoting platelet production. [3-2] The use described in [3-1], used together with an aryl hydrocarbon receptor inhibitor. [3-3] The use according to [3-2], wherein the aryl hydrocarbon receptor inhibitor is selected from the following: JPEG0007657800000018.jpg106150 [4-1] A compound or a salt thereof according to any one of [2-1] to [2-10] for use in promoting platelet production. [4-2] The compound or salt thereof according to [4-1], used in combination with an aromatic hydrocarbon receptor inhibitor. [4-3] The compound or salt thereof according to [4-2], wherein the aromatic hydrocarbon receptor inhibitor is selected from the group consisting of: JPEG0007657800000019.jpg106150 [5-1] A method for promoting platelet production, comprising culturing platelet precursor cells in the presence of a compound or a salt thereof according to any one of [2-1] to [2-10]. [5-2] The method according to [5-1], wherein the compound or a salt thereof is used together with an aromatic hydrocarbon receptor inhibitor. [5-3] The method according to [5-2], wherein the aromatic hydrocarbon receptor inhibitor is selected from the group consisting of the following: JPEG0007657800000020.jpg106150[6-1] A method for producing platelets, comprising culturing platelet precursor cells in the presence of a compound or a salt thereof according to any one of [2-1] to [2-10]. [6-2] The method according to [6-1], further comprising culturing platelet precursor cells in the presence of an aryl hydrocarbon receptor inhibitor. [6-3] The method according to [6-2], wherein the aromatic hydrocarbon receptor inhibitor is selected from the group consisting of the following: JPEG0007657800000021.jpg106150[7-1] A method for culturing platelet precursor cells to promote platelet production, comprising culturing platelet precursor cells in the presence of a compound or a salt thereof according to any one of [2-1] to [2-10]. [7-2] The method described in [7-1], further comprising culturing platelet precursor cells in the presence of an aryl hydrocarbon receptor inhibitor. [7-3] The method according to [7-2], wherein the aromatic hydrocarbon receptor inhibitor is selected from the group consisting of the following: JPEG0007657800000022.jpg106150 Effect of the Invention

[0008] The compound according to the present invention or a salt thereof has excellent efficacy in promoting the production of platelets from platelet precursor cells in vitro. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The words and terms used in this specification are explained in detail below.

[0010] In the present specification, "halogen" refers to fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine.

[0011] In this specification, "C 1-6 Alkyl" is a group with 1 to 6 carbon atoms (C 1-6 ) straight-chain or branched-chain alkyl, specific examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 3-methylpentyl, and the like. Also, "C 1-6 "Alkyl" refers to a C alkyl group in which 1 to 7 hydrogen atoms have been replaced with deuterium atoms. 1-6 Alkyl is also included.

[0012] In this specification, "C 3-8 Cycloalkyl" is a group with 3 to 8 carbon atoms (C 3-8 ) is a cycloalkyl, specific examples of which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

[0013] In the present specification, "aryl" refers to a monocyclic or polycyclic aromatic ring. Specific examples of "aryl" include benzene, naphthalene, anthracene, and the like.

[0014] In the present specification, "heteroaryl" refers to a heteroaromatic ring containing 1 to 3 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur as ring-constituting elements. Specific examples of "heteroaryl" include furan, thiophene, oxazole, thiazole, pyrazole, pyridine, pyrimidine, pyridazine, pyrazine, quinoline, isoquinoline, quinazoline, etc.

[0015] In the present specification, the term "phenyl which may have a substituent" refers to unsubstituted phenyl or phenyl substituted with 1 to 3 substituents. The substituents include halogen, -C 1-6 Alkyl, -OC 1-6 Specific examples include phenyl, fluorophenyl, chlorophenyl, bromophenyl, iodophenyl, etc.

[0016] In the present specification, "heteroaryl which may have a substituent selected from furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and pyrimidyl" refers to furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl or pyrimidyl which is unsubstituted or substituted with 1 to 3 substituents. The substituents include halogen, -C 1-6 Alkyl, -OC 1-6Specific examples include furyl, fluorofuryl, chlorofuryl, bromofuryl, iodofuryl, methylfuryl, ethylfuryl, methoxyfuryl, ethoxyfuryl, thienyl, fluorothienyl, chlorothienyl, bromothienyl, iodothienyl, methylthienyl, ethylthienyl, methoxythienyl, ethoxythienyl, oxazolyl, fluorooxazolyl, chlorooxazolyl, bromooxazolyl, iodooxazolyl, methyloxazolyl, ethyloxazolyl, methoxyoxazolyl, ethoxyoxazolyl, thiazolyl, fluorothiazolyl, chlorothiazolyl, bromothiazolyl, iodothiazolyl, methylthiazolyl, ethylthiazolyl, methoxythiazolyl, ethoxythiazolyl, pyrazolyl, fluoropyrazolyl, chloropyrazolyl, bromopyrazolyl, iodopyrazolyl, Examples include pyridyl, methylpyrazolyl, ethylpyrazolyl, methoxypyrazolyl, ethoxypyrazolyl, pyridyl, fluoropyridyl, chloropyridyl, bromopyridyl, iodopyridyl, methylpyridyl, ethylpyridyl, methoxypyridyl, ethoxypyridyl, pyrazyl, fluoropyrazyl, chloropyrazyl, bromopyrazyl, iodopyrazyl, methylpyrazyl, ethylpyrazyl, methoxypyrazyl, ethoxypyrazyl, pyridazyl, fluoropyridazyl, chloropyridazyl, bromopyridazyl, iodopyridazyl, methylpyridazyl, ethylpyrazyl, methoxypyridazyl, ethoxypyridazyl, pyrimidyl, fluoropyrimidyl, chloropyrimidyl, bromopyridazyl, iodopyridazyl, methylpyridazyl, ethylpyridazyl, methoxypyridazyl, ethoxypyridazyl, pyrimidyl, fluoropyrimidyl, chloropyrimidyl, bromopyrimidyl, iodopyrimidyl, methylpyrimidyl, ethylpyrimidyl, methoxypyrimidyl, ethoxypyrimidyl, and the like.

[0017] In the present specification, the term "pyrimidyl which may have a substituent" refers to unsubstituted pyrimidyl or pyrimidyl substituted with 1 to 3 substituents. The substituents include halogen, -C 1-6 Alkyl, -OC 1-6 Specific examples include pyrimidyl, fluoropyrimidyl, chloropyrimidyl, bromopyrimidyl, iodopyrimidyl, methylpyrimidyl, ethylpyrimidyl, methoxypyrimidyl, ethoxypyrimidyl, and the like.

[0018] In the present specification, examples of the "alkyl halide" include iodomethane, iodoethane, 1-iodopropane, 2-iodopropane, 1-iodobutane, 2-iodobutane, 1-iodo-2-methylpropane, tert-butyl iodide, 1-iodopentane, 2-iodopentane, 1-iodo-2,2-dimethylpropane, 1-iodohexane, 2-iodohexane, and 3-iodomethylpentane.

[0019] In this specification, examples of the "acid anhydride" include acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride, pivalic anhydride, n-hexanoic anhydride, heptanoic anhydride, and benzoic anhydride.

[0020] In the present specification, examples of the "acid halide" include benzoyl chloride, acetyl chloride, acetyl bromide, propionyl chloride, n-butyryl chloride, isobutyryl chloride, pentanoyl chloride, isopentanoyl chloride, DL-2-methylbutyryl chloride, pivaloyl chloride, n-hexanoyl chloride, 4-methylpentanoyl chloride, and heptanoyl chloride.

[0021] In the present specification, examples of the "halocarboxylic acid ester" include methyl chloroformate, ethyl chloroformate, propyl chloroformate, isopropyl chloroformate, butyl chloroformate, sec-butyl chloroformate, isobutyl chloroformate, pentyl chloroformate, neopentyl chloroformate, and n-hexyl chloroformate.

[0022] In the present specification, the "condensing agent" is not particularly limited, and specific examples thereof include 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (WSC·HCl), N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), N,N'-carbonyldiimidazole (CDI), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM), benzotriazol-1-yloxytris(dimethylamino)ethyl, and the like. (7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU), (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylaminomorpholinocarbenium hexafluorophosphate (COMU), and the like, and preferably WSC·HCl, HATU, or COMU.

[0023] In the present specification, the "additive" is not particularly limited, but includes 1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azabenzotriazole (HOAt), N-hydroxysuccinimide (HOSu), ethyl (hydroxyimino)cyanoacetate (Oxyma), 4-dimethylaminopyridine (DMAP), triethylamine (TEA), diisopropylethylamine (DIPEA), N-methylmorpholine, and the like, and is preferably HOBt, TEA, or DIPEA.

[0024] In the present specification, the term "leaving group" specifically refers to halogen, C 1-18 Alkanesulfonyl, lower alkanesulfonyloxy, arylsulfonyloxy, aralkylsulfonyloxy, perhaloalkanesulfonyloxy, sulfonio, toluenesulfoxy, etc. Preferred leaving groups include halogens.

[0025] The above "halogen" is fluorine, chlorine, bromine or iodine.

[0026] The above "C 1-18 Examples of "alkanesulfonyl" include linear or branched alkanesulfonyl having 1 to 18 carbon atoms, and specific examples thereof are methanesulfonyl, 1-propanesulfonyl, 2-propanesulfonyl, butanesulfonyl, cyclohexanesulfonyl, dodecanesulfonyl, octadecanesulfonyl, and the like.

[0027] Examples of the above-mentioned "lower alkanesulfonyloxy" include linear or branched alkanesulfonyloxy having 1 to 6 carbon atoms, and specific examples thereof are methanesulfonyloxy, ethanesulfonyloxy, 1-propanesulfonyloxy, 2-propanesulfonyloxy, 1-butanesulfonyloxy, 3-butanesulfonyloxy, 1-pentanesulfonyloxy, 1-hexanesulfonyloxy, and the like.

[0028] Examples of the above "arylsulfonyloxy" include phenylsulfonyloxy and naphthylsulfonyloxy which may have 1 to 3 groups selected from the group consisting of linear or branched alkyl having 1 to 6 carbon atoms, linear or branched alkoxy having 1 to 6 carbon atoms, nitro, and halogen as substituents on the phenyl ring. Specific examples of the above "phenylsulfonyloxy which may have a substituent" are phenylsulfonyloxy, 4-methylphenylsulfonyloxy, 2-methylphenylsulfonyloxy, 4-nitrophenylsulfonyloxy, 4-methoxyphenylsulfonyloxy, 2-nitrophenylsulfonyloxy, 3-chlorophenylsulfonyloxy, etc. Specific examples of the above "naphthylsulfonyloxy" are α-naphthylsulfonyloxy, β-naphthylsulfonyloxy, etc.

[0029] Examples of the above "aralkylsulfonyloxy" include linear or branched alkanesulfonyloxy having 1 to 6 carbon atoms substituted with phenyl, which may have 1 to 3 groups selected from the group consisting of linear or branched alkyl having 1 to 6 carbon atoms, linear or branched alkoxy having 1 to 6 carbon atoms, nitro, and halogen as a substituent on the phenyl ring, and linear or branched alkanesulfonyloxy having 1 to 6 carbon atoms substituted with naphthyl. Specific examples of the above "alkanesulfonyloxy substituted with phenyl" are benzylsulfonyloxy, 2-phenylethylsulfonyloxy, 4-phenylbutylsulfonyloxy, 4-methylbenzylsulfonyloxy, 2-methylbenzylsulfonyloxy, 4-nitrobenzylsulfonyloxy, 4-methoxybenzylsulfonyloxy, 3-chlorobenzylsulfonyloxy, etc. Specific examples of the above "alkanesulfonyloxy substituted with naphthyl" are α-naphthylmethylsulfonyloxy, β-naphthylmethylsulfonyloxy, etc.

[0030] Specific examples of the above-mentioned "perhaloalkanesulfonyloxy" include trifluoromethanesulfonyloxy.

[0031] Specific examples of the above-mentioned "sulfonio" include dimethylsulfonio, diethylsulfonio, dipropylsulfonio, di(2-cyanoethyl)sulfonio, di(2-nitroethyl)sulfonio, di-(aminoethyl)sulfonio, di(2-methylaminoethyl)sulfonio, di-(2-dimethylaminoethyl)sulfonio, di-(2-hydroxyethyl)sulfonio, di-(3-hydroxypropyl)sulfonio, di-(2-methoxyethyl)sulfonio, di-(2-carbamoylethyl)sulfonio, di-(2-carbamoylethyl)sulfonio, di-(2-carboxyethyl)sulfonio, di-(2-methoxycarbonylethyl)sulfonio, diphenylsulfonio, and the like.

[0032] In the present specification, the term "palladium compound" is not particularly limited, but examples thereof include tetravalent palladium catalysts such as sodium hexachloropalladate(IV) tetrahydrate and potassium hexachloropalladate(IV); [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane adduct (Pd(dppf)Cl2·CH2Cl2), (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate (XPhos Pd G3), palladium(II) chloride, palladium(II) bromide, palladium(II) acetate, palladium acetylacetonate(II), dichlorobis(benzonitrile)palladium(II), dichlorobis(acetonitrile)palladium(II), dichlorobis(triphenylphosphine)palladium(II), dichlorotetraamminepalladium(II), dichloro(cycloocta-1,5-diene)palladium(II), palladium trifluoroacetate(II), 1,1'-bis(diphenylphosphino)ferrocenedichloropalladium(II)-dichloromethane complex and other divalent palladium catalysts; bis(tri-t-butylphosphine)palladium(0) Examples of such palladium catalysts include zero-valent palladium catalysts such as Pd(tBu3P)2, tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3), tris(dibenzylideneacetone)dipalladium chloroform complex(0), and tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4). These palladium compounds are used alone or in combination of two or more.

[0033] In the present specification, the term "base" is not particularly limited, but examples thereof include inorganic bases and organic bases. Examples of the "inorganic base" include alkali metal hydroxides (e.g., lithium hydroxide, sodium hydroxide, potassium hydroxide), alkaline earth metal hydroxides (e.g., magnesium hydroxide, calcium hydroxide, barium hydroxide), alkali metal carbonates (e.g., sodium carbonate, potassium carbonate, cesium carbonate), alkaline earth metal carbonates (e.g., magnesium carbonate, calcium carbonate, barium carbonate), alkali metal bicarbonates (e.g., sodium bicarbonate, potassium bicarbonate), alkali metal phosphates (e.g., sodium phosphate, potassium phosphate, cesium phosphate), alkaline earth metal phosphates (e.g., magnesium phosphate, calcium phosphate), alkali metal alkoxides (e.g., sodium methoxide, sodium ethoxide, sodium tert-butoxide, potassium tert-butoxide), alkali metal hydrides (e.g., sodium hydride, potassium hydride), and the like. Examples of the "organic base" include trialkylamines (e.g., trimethylamine, triethylamine, N,N-diisopropylethylamine (DIPEA)), dialkylamines (e.g., diethylamine, diisopropylamine), 4-dimethylaminopyridine (DMAP), N-methylmorpholine, picoline, 1,5-diazabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane, and 1,8-diazabicyclo[5.4.0]-7-undecene (DBU). Preferred are DMAP and TEA. These bases can be used alone or in combination.

[0034] In the present specification, the "solvent" used in the reaction may be any solvent inert to the reaction, and examples thereof include water, ethers (e.g., dioxane, tetrahydrofuran, diethyl ether, 1,2-dimethoxyethane, diethylene glycol dimethyl ether, ethylene glycol dimethyl ether), halohydrocarbons (e.g., methylene chloride, chloroform, 1,2-dichloroethane, carbon tetrachloride), aromatic hydrocarbons (e.g., benzene, toluene, xylene), lower alcohols (e.g., methanol, ethanol, isopropanol), polar solvents (e.g., N,N-dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylsulfoxide (DMSO), hexamethylphosphoric acid triamide, acetonitrile). These solvents may be used alone or in combination of two or more.

[0035] Each of the substituents in the compound represented by the general formula [I] or [I'] in this specification (hereinafter referred to as "compound [I]") is explained below.

[0036] R in compound [I] 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 It is alkyl, preferably a hydrogen atom, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 3-methylpentyl, -O-methyl, -O-ethyl, -On-propyl, -O-isopropyl, -On-butyl, -O-isobutyl, -O-sec-butyl, -O-tert-butyl, -On-pentyl, -O-isopentyl, -O-neopentyl, -On-hexyl, -O-isohexyl or -O-3-methylpentyl, more preferably a hydrogen atom, chlorine, methyl or -O-methyl.

[0037] R in compound [I] 12 is a hydrogen atom, -C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6Alkyl, -C(=O)-C 1-6 Alkyl, -C(=O)-aryl or -C(=O)-OC 1-6 It is an alkyl group, preferably a hydrogen atom, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 3-methylpentyl, -methyl-O-methyl, -methyl-O-ethyl, -methyl-O-propyl, -ethyl-O-methyl, -ethyl-O-ethyl, -ethyl-O-propyl, -propyl-O-methyl, -propyl-O-ethyl, -propyl-O-propyl, -C(=O)-methyl, -C(=O)-ethyl, -C(=O)-n-propyl, -C(=O)-isopropyl, -C(=O)-n-butyl, -C(=O)-isobutyl, -C(=O)-sec-butyl, -C(=O)-tert-butyl, -C(=O)-n-pentyl, -C(=O)-isopentyl, -C(=O)-neopentyl, -C(=O)-n-hexyl, -C(=O)-isohexyl, -C(=O)-3-methylpentyl, -C(=O)-phenyl, -C(=O)-naphthyl, -C(=O)-O-methyl, -C(=O)-O-ethyl, -C(=O)-On-propyl, -C(=O)-O-isopropyl, -C(=O)-On-butyl, -C(=O)-O-isobutyl, -C(=O)-O-sec-butyl, -C(=O)- O-tert-butyl, -C(=O)-On-pentyl, -C(=O)-O-isopentyl, -C(=O)-O-neopentyl, -C(=O)-On-hexyl, -C(=O)-O-isohexyl or -C(=O)-O-3-methylpentyl, more preferably a hydrogen atom, methyl, -ethyl-O-methyl, -C(=O)-methyl, -C(=O)-phenyl or -C(=O)-O-methyl.

[0038] R in compound [I] 2 is a hydrogen atom or -C 1-6It is alkyl, preferably a hydrogen atom, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl or 3-methylpentyl, more preferably a hydrogen atom or methyl.

[0039] R in compound [I] 3 is halogen, -Q k -(C 1-6 Alkyl) m -Q p -R 31 and heteroaryl consisting of phenyl or furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and pyrimidyl, which may have a substituent, and which is preferably a halogen, -Q k -(C 1-6 Alkyl) m -Q p -R 31, optionally substituted phenyl, furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and optionally substituted pyrimidyl, more preferably fluorine, chlorine, bromine, iodine, -O-methyl, -O-ethyl, -O-propyl, -O-butyl, -O-methyl-O-methyl, -O-ethyl-O-methyl, -O-ethyl-O-ethyl, -O-methyl-cyclopropyl, -O-methyl-cyclobutyl, -O-methyl-cyclopentyl, -O-ethyl ... -propyl, -O-ethyl-cyclobutyl, -O-ethyl-cyclopentyl, -S-methyl, -S-ethyl, -S-propyl, -methyl-S-methyl, -methyl-S-ethyl, -ethyl-S-ethyl, -NH-methyl, -NH-ethyl, -C(=O)-O-methyl, -C(=O)-O-ethyl, -C(=O)-On-propyl, -C(=O)-O-isopropyl, -C(=O)-On-butyl, -C(=O)-O-isobutyl, -C(=O)-O-sec-butyl, -C(=O)-O-tert-butyl, -C (=O)-On-pentyl, -C(=O)-O-isopentyl, -C(=O)-O-neopentyl, -C(=O)-On-hexyl, -C(=O)-O-isohexyl, -C(=O)-O-3-methylpentyl, phenyl, fluorophenyl, chlorophenyl, bromophenyl, iodophenyl, furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyrimidyl, fluoropyrimidyl, chloropyrimidyl, bromopyrimidyl, iodopyrimidyl, methylpyrimidyl, ethylpyrimidyl diethyl, methoxypyrimidyl, ethoxypyrimidyl or pyridazyl, more preferably fluorine, methyl, -O-methyl, -O-ethyl, -O-ethyl-O-methyl, -O-methyl-cyclopropyl, -S-ethyl, -methyl-S-methyl, -NH-ethyl, -C(=O)-O-methyl, phenyl, fluorophenyl, furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyrimidyl, fluoropyrimidyl, methylpyrimidyl, methoxypyrimidyl or pyridazyl.

[0040] R in compound [I] 3a -OC 1-6It is alkyl, preferably -O-methyl or -O-ethyl.

[0041] R in compound [I] 3b is a hydrogen atom or -OC 1-6 It is alkyl, preferably a hydrogen atom or -O-methyl.

[0042] R in compound [I] 31 -C 1-6 Alkyl or -C 3-8 Cycloalkyl, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 3-methylpentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, more preferably methyl or cyclopropyl.

[0043] R in compound [I] 4 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 It is alkyl, preferably a hydrogen atom, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, 3-methylpentyl, -O-methyl, -O-ethyl, -O-propyl or -O-butyl, and more preferably a hydrogen atom, fluorine, methyl or -O-methyl.

[0044] In compound [I], each Q is independently the same or different and is an oxygen atom, a sulfur atom, -C(=O)-O- or -NH-.

[0045] In compound [I], k, m and p are the same or different and each independently represents 0 or 1.

[0046] In compound [I], n is 0, 1 or 2. When n is 2, R 3 are each independently the same or different substituents, preferably 1 or 2.

[0047] In compound [I], V may be the same or different and each independently represents a nitrogen atom or CH.

[0048] W in the compound [I] is a carbon atom or a nitrogen atom, and is preferably a carbon atom.

[0049] X in compound [I] is a carbon atom, a nitrogen atom or NR 12 It is.

[0050] In the compound [I], Y is a carbon atom or a nitrogen atom.

[0051] In compound [I], each Z is independently the same or different and is a nitrogen atom or CH.

[0052] Ring A in compound [I] is aryl or heteroaryl. The aryl can be, for example, benzene, naphthalene, anthracene, and preferably benzene. The heteroaryl can be, for example, furan, thiophene, oxazole, thiazole, pyrazole, pyridine, pyrimidine, pyridazine, pyrazine, quinoline, isoquinoline, quinazoline, and the like, and preferably furan, thiophene, pyridine, and quinoline.

[0053] In addition, in compound [I] The image is JPEG0007657800000023.jpg32150.

[0054] In compound [I] Specific examples of JPEG0007657800000024.jpg1421 include ethoxybenzene, methoxyethoxybenzene, cyclopropylmethoxybenzene, ethylsulfanylbenzene, methylsulfanylmethylbenzene, ethylaminobenzene, methyl benzoate, biphenyl, fluorobiphenyl, methoxybiphenyl, pyridylbenzene, pyrimidylbenzene, (fluoropyrimidyl)benzene, (methylpyrimidyl)benzene, (methoxypyrimidyl)benzene, pyrazylbenzene, pyridazylbenzene, furylbenzene, thienylbenzene, oxazolylbenzene, thiazolylbenzene, pyrazolylbenzene, phenylfuran, ethoxythiophene, phenylthiophene, furi. Examples of the pyridylthiophene include thiophene, thienylthiophene, pyridylthiophene, pyrimidylthiophene, methylquinoline, methoxyquinoline, and ethoxypyridine, and are preferably pyridylbenzene, pyrimidylbenzene, (fluoropyrimidyl)benzene, (methylpyrimidyl)benzene, (methoxypyrimidyl)benzene, phenylthiophene, pyridylthiophene, and pyrimidylthiophene, and are more preferably 2-pyridylbenzene, 2-pyrimidylbenzene, 2-(5-fluoropyrimidyl)benzene, 2-(5-methylpyrimidyl)benzene, 2-(5-methoxypyrimidyl)benzene, 3-phenylthiophene, 3-(2-pyridyl)thiophene, and 3-(2-pyrimidyl)thiophene.

[0055] In addition, in compound [I] The image is JPEG0007657800000025.jpg19150.

[0056] In addition, in compound [I] The image is JPEG0007657800000026.jpg32150.

[0057] In compound [I] --- is a single bond or a double bond.

[0058] Preferable compound [I] is, for example, a compound represented by the general formula [I]: R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl, R 2 is a hydrogen atom, R 3 is phenyl, pyridyl, (halogen, -C 1-6 Alkyl or -OC 1-6 pyrimidyl (optionally substituted with alkyl); X is NH; W and Y are carbon atoms; Z's are independently the same or different and each is a nitrogen atom or CH; Examples of the compound include compounds in which ring A is benzene or thiophene.

[0059] More preferred compounds [I] include those represented by the general formula [Ia] JPEG0007657800000027.jpg3459[where, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl, R 12 is a hydrogen atom or -C(=O)-OC 1-6 is alkyl, JPEG0007657800000028.jpg1421 is pyridylbenzene, (halogen, -C 1-6 Alkyl or -OC 1-6 and the like is a pyrimidyl benzene, phenylthiophene, pyridylthiophene or pyrimidylthiophene, each of which may be substituted with an alkyl group; In particular, in the general formula [Ia], R 11 is a hydrogen atom, methyl or -O-methyl, R 12 is a hydrogen atom or -C(=O)-O-methyl, JPEG0007657800000029.jpg1421 includes compounds that are pyridylbenzene, pyrimidylbenzene, (fluoropyrimidyl)benzene, (methylpyrimidyl)benzene, (methoxypyrimidyl)benzene, phenylthiophene, pyridylthiophene, or pyrimidylthiophene.

[0060] More preferred examples of the compound [I] include compounds selected from the group consisting of the following: JPEG0007657800000030.jpg95150

[0061] Preferred examples of the compound [I] include compounds represented by the general formula [Ia'] JPEG0007657800000031.jpg4370[In the formula, R 3a -OC 1-6 Alkyl; R 3b is a hydrogen atom or -OC 1-6 Alkyl; R 11 -C 1-6 Alkyl or -OC 1-6 Alkyl; R 12 is a hydrogen atom or -C 1-6 indicates alkyl] In particular, the compound represented by the general formula [Ia'] is R 3a is -O-methyl or -O-ethyl, R 3b is a hydrogen atom or -O-methyl, R 11 is methyl or -O-methyl, R 12 is a hydrogen atom or methyl Compounds include:

[0062] More preferred examples of the compound [Ia'] include compounds selected from the group consisting of the following: JPEG0007657800000032.jpg75150

[0063] Compound [I] or a salt thereof is useful as a platelet production promoter. Accordingly, one aspect of the present invention is a platelet production promoter comprising compound [I] or a salt thereof. This embodiment also includes a platelet production promoter for use in combination with an aromatic hydrocarbon receptor inhibitor.

[0064] One aspect of the present invention is the use of compound [I] or a salt thereof for promoting platelet production. This aspect also includes use in conjunction with an aryl hydrocarbon receptor inhibitor.

[0065] One aspect of the present invention is compound [I] or a salt thereof for use in promoting platelet production. This embodiment also includes compound [I] or a salt thereof, which is used in combination with an aryl hydrocarbon receptor inhibitor.

[0066] One embodiment of the present invention is a method for promoting platelet production, which comprises culturing platelet precursor cells in the presence of compound [I] or a salt thereof. This embodiment also includes a method further comprising culturing the platelet precursor cells in the presence of an aryl hydrocarbon receptor inhibitor.

[0067] One aspect of the present invention is a method for producing platelets, which comprises culturing platelet precursor cells in the presence of compound [I] or a salt thereof. This embodiment also includes a method further comprising culturing the platelet precursor cells in the presence of an aryl hydrocarbon receptor inhibitor.

[0068] One aspect of the present invention is a method for culturing platelet precursor cells to promote platelet production, which comprises culturing platelet precursor cells in the presence of compound [I] or a salt thereof. This embodiment also includes a method further comprising culturing the platelet precursor cells in the presence of an aryl hydrocarbon receptor inhibitor.

[0069] Throughout this specification, the presentation of preferred embodiments and options for different features of the compound [I] or its salts, uses, methods and compositions of the present invention also includes the presentation of combinations of preferred embodiments and options for the different features, so long as they are combinable and not inconsistent.

[0070] The manufacturing method of compound [I] will be described below. Compound [I] can be manufactured based on the manufacturing method shown below. Compound [I] can also be manufactured by the method described in WO2019 / 167973, for example. These manufacturing methods are merely examples, and the manufacturing method of compound [I] is not limited to these.

[0071] In the following reaction formulas, when an alkylation reaction, amination reaction, esterification reaction, amidation reaction, etherification reaction, nucleophilic substitution reaction, addition reaction, oxidation reaction, reduction reaction, etc. are carried out, these reactions are carried out according to a method known per se. Examples of such methods include those described in Experimental Chemistry Lectures (5th Edition, edited by the Chemical Society of Japan, Maruzen Co., Ltd.), ORGANIC FUNCTIONAL GROUP PREPARATIONS 2nd Edition, published by ACADEMIC PRESS, INC. in 1989, Comprehensive Organic Transformations, published by VCH Publishers Inc. in 1989, and "Greene's Protective Groups in Organic Synthesis" (4th Edition, 2006) by PGM Wuts and T.W. Greene.

[0072] General synthetic route for compound [I] (1) JPEG0007657800000033.jpg38150 (wherein each symbol is as defined above)

[0073] The compound [I] of the present invention can be produced by the reaction shown in the above reaction formula. Specifically, the compound [I] can be produced by condensing the compound [II] and the compound [III].

[0074] Other reaction conditions (such as reaction temperature and reaction time) can be appropriately determined based on a condensation reaction known per se.

[0075] General synthetic route for compound [I] (2) JPEG0007657800000034.jpg43150 (in the formula, R 12a -C 1-6 alkyl, and each of the other symbols is as defined above.

[0076] The compound [Ic] of the present invention can be produced by the reaction shown in the above reaction formula. Specifically, the compound [Ic] can be produced by reacting the compound [Ib] with an alkyl halide.

[0077] Other reaction conditions (such as reaction temperature and reaction time) can be appropriately determined based on a condensation reaction known per se.

[0078] General synthetic route for compound [I] (3) JPEG0007657800000035.jpg42150 (in the formula, R 12b is -C(=O)-C 1-6 Alkyl, -C(=O)-aryl or -C(=O)-OC 1-6 alkyl, and each of the other symbols is as defined above.

[0079] The compound [Id] of the present invention can be produced by the reaction shown in the above reaction formula. Specifically, the compound [Ib] can be produced by reacting the compound [Id] with an acid anhydride, an acid halide or a halocarboxylate.

[0080] Other reaction conditions (such as reaction temperature and reaction time) can be appropriately determined based on a condensation reaction known per se.

[0081] General synthetic route for compound [I] (4) JPEG0007657800000036.jpg36150 (wherein ring B is an optionally substituted benzene or thiophene, U is a leaving group, and the other symbols are as defined above.)

[0082] The compound [Ie] of the present invention can be produced by the reaction shown in the above reaction formula. Specifically, the compound [Ie] can be produced by coupling the compound [IV] having a leaving group (U) with the compound [V] in the presence of a palladium compound.

[0083] The "boronic acid" or "boronic acid ester" (compound [V] in the formula) used in this reaction may be produced separately and isolated and purified before use. Alternatively, for example, a precursor halogenated compound or the like may be reacted with bis-pinacoldiborane in the presence of a palladium compound and used in the coupling reaction without isolation and purification.

[0084] Other reaction conditions (such as reaction temperature and reaction time) can be appropriately determined based on known coupling reactions.

[0085] In each reaction in the above reaction scheme, the product can be used in the next reaction as the reaction solution or as a crude product, or it can be isolated from the reaction mixture in a conventional manner or easily purified by conventional separation means such as recrystallization, distillation, and chromatography.

[0086] The starting compounds, intermediate compounds, and target compounds in each step, as well as the compounds of the present invention or their salts, include geometric isomers, stereoisomers, optical isomers, and tautomers. Various isomers can be separated by a general optical resolution method. They can also be produced from suitable optically active starting compounds.

[0087] The compound of the present invention or a salt thereof can be produced by the synthesis methods shown in the above reaction schemes or methods analogous thereto.

[0088] In the production of the compound of the present invention or a salt thereof, a starting compound may be a commercially available compound, or a compound produced according to a known method or a method similar thereto, unless a specific production method is described.

[0089] The starting compounds and the target compounds in each step above can be used in the form of suitable salts. Examples of such salts include the same salts as those exemplified below as salts of the compound [I] of the present invention.

[0090] The compound or salt of the present invention includes the salt form, and may form an acid addition salt or a salt with a base depending on the type of the substituent. Examples of such "acids" include inorganic acids (e.g., hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc.); and organic acids (e.g., methanesulfonic acid, p-toluenesulfonic acid, acetic acid, citric acid, tartaric acid, maleic acid, fumaric acid, malic acid, lactic acid, etc.). Examples of such "bases" include inorganic bases (e.g., sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, etc.); and organic bases (e.g., methylamine, diethylamine, trimethylamine, triethylamine, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, tris(hydroxymethyl)methylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, guanidine, pyridine, picoline, choline, etc.); and ammonium salts. In addition, it may form a salt with an amino acid such as lysine, arginine, aspartic acid, or glutamic acid.

[0091] The compounds of the present invention or salts thereof include compounds in which one or more atoms are replaced with one or more isotope atoms. Examples of isotope atoms include deuterium ( 2 H), tritium ( 3 H), 13 C.15 N, 18 Examples include O.

[0092] The compound of the present invention or a salt thereof has the function of promoting the production of platelets from platelet precursor cells in vitro.

[0093] Hereinafter, a method for producing platelets from platelet precursor cells using the compound of the present invention or a salt thereof will be described.

[0094] Platelets can be produced by culturing platelet precursor cells (e.g., megakaryocytes or their precursor cells) in the presence of one or more of the compounds of the present invention or salts thereof. The concentration of the compounds of the present invention or salts thereof in the medium is not particularly limited and can be appropriately determined by those skilled in the art depending on the platelet production promoter, and is, for example, 1 nM to 100 μM, preferably 10 nM to 100 μM, and more preferably 100 nM to 10 μM, but may be outside this range as long as the desired effect is achieved.

[0095] The compound of the present invention or a salt thereof can increase the amount of platelet production from megakaryocytes. The compound of the present invention or a salt thereof can increase the platelet count by, for example, but not limited to, 200% or more, preferably 300% or more, and more preferably 400% or more compared to the control.

[0096] The timing of adding (allowing to be present) the compound of the present invention or its salt to the medium is not particularly limited as long as the desired effect is achieved. For example, the compound of the present invention or its salt is added to megakaryocyte cells or their precursor cells. The megakaryocyte cells may be megakaryocyte cells before multinucleation or may be multinucleated megakaryocyte cells, and the multinucleated megakaryocyte cells may be megakaryocyte cells in the process of platelet production. As described below, in a cell less differentiated from megakaryocytes, at least one gene selected from the group consisting of oncogenes, polycomb genes, and apoptosis inhibitor genes is forcibly expressed to prepare immortalized megakaryocyte cells, and then the forced expression is released to promote the multinucleation of the immortalized megakaryocyte cells, it is preferable to add the compound [I] of the present invention or its salt to the medium after the forced expression is released. The compound of the present invention or its salt may be added to the medium at the same time as the start of the culture for platelet production, or 1 day, 2 days, 3 days, 4 days, 5 days, or 6 days after the start of the culture.

[0097] Known megakaryocyte cells can be used as megakaryocyte cells that can be used in the present invention. For example, immortalized megakaryocyte cells can be prepared using the method disclosed in WO2016 / 204256.

[0098] The origin of megakaryocytes or their precursor cells is not particularly limited as long as they have the ability to produce platelets, and can be, for example, pluripotent stem cells, particularly induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). The origin of iPS cells and ES cells is not particularly limited, and can be, for example, human-derived cells.

[0099] The compound of the present invention or a salt thereof can be used as a platelet production promoter in combination with one or more aromatic hydrocarbon receptor inhibitors (AhR antagonists), one or more thrombopoietin (TPO) or TPO receptor agonists, one or more Rho-associated coiled-coil forming kinase (ROCK) inhibitors, and / or one or more ADAM (a disintegrin and metalloprotease) inhibitors, etc.

[0100] The compound of the present invention or a salt thereof exhibits a more excellent platelet production promoting effect when platelet precursor cells are cultured in the coexistence of an aromatic hydrocarbon receptor inhibitor.

[0101] The aromatic hydrocarbon receptor inhibitor that can be used together with the compound of the present invention or a salt thereof is not particularly limited as long as it has a platelet production promoting effect, and examples thereof include the compounds disclosed in WO2020 / 050409. Specific examples thereof include the following compounds. 4-[2-[[2-benzo[b]thien-3-yl-9-(1-methylethyl)-9H-purin-6-yl]amino]ethyl]phenol (Compound A1) JPEG0007657800000037.jpg3641·N-[2-(1H-indol-3-yl)ethyl]-9-(1-methylethyl)-2-(5-methyl-3-pyridinyl)-9H-Purin-6-amine (Compound A2) JPEG0007657800000038.jpg3442·4-(2-Methyl-4-pyridinyl)-N-[4-(3-pyridinyl)phenyl]-benzeneacetamide (Compound A3) JPEG0007657800000039.jpg3050·1-Methyl-N-[2-methyl-4-[2-(2-methylphenyl)diazenyl]phenyl]-1H-pyrazole-5-carboxamide (Compound A4) JPEG0007657800000040.jpg2147·3-[5-[2-[[2-(5-Fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl]amino]ethyl]-2-hydroxyphenyl]benzonitrile (Compound A5) JPEG0007657800000041.jpg3156·2-(2-fluorophenyl)-4-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl]amino]ethyl]phenol (Compound A6) JPEG0007657800000042.jpg3354·2-(5-fluoropyridin-3-yl)-4-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl]amino]ethyl]phenol (Compound A7) JPEG0007657800000043.jpg3251·2-(2-fluorophenyl)-4-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]thiazol-4-yl]amino]ethyl]phenol (Compound A8) JPEG0007657800000044.jpg3250

[0102] The concentration of aromatic hydrocarbon receptor inhibitor is not particularly limited, and can be appropriately determined by those skilled in the art according to the compound.The concentration of aromatic hydrocarbon receptor inhibitor can be, for example, 1.0 nM to 1000 μM, 10 nM to 100 μM, 100 nM to 100 μM, or 100 nM to 10 μM, but may be outside this range as long as it has the desired effect.

[0103] Examples of ROCK inhibitors include, but are not limited to, Y27632, Y39983, Fasudil hydrochloride, Ripasudil, SLX-2119, RKI-1447, Azaindole 1, SR-3677, Staurosporine, H1152 dihydrochloride, AR-12286, INS-117548, etc. The concentration of the ROCK inhibitor is not particularly limited and can be appropriately determined by those skilled in the art depending on the compound. The concentration of the ROCK inhibitor can be, for example, 1.0 nM to 1.0 mM, 10 nM to 0.1 mM, 100 nM to 0.1 mM, or 100 nM to 0.01 mM, but may be outside this range as long as the desired effect is achieved.

[0104] Thrombopoietin includes thrombopoietin (TPO) and human recombinant thrombopoietin. In addition, examples of TPO receptor agonists include, but are not limited to, TA-316. The concentrations of TPO and human recombinant TPO are not particularly limited and can be appropriately determined by those skilled in the art. The concentrations of TPO and human recombinant TPO can be, for example, 0.5 ng / mL to 5 μg / mL, preferably 5 to 500 ng / mL, and more preferably 50 ng / mL, but may be outside this range as long as the desired effect is achieved. The concentration of the TPO receptor agonist is not particularly limited, and can be appropriately determined by those skilled in the art depending on the compound. The concentration of the TPO receptor agonist can be, for example, 0.1 ng / mL to 1 mg / mL, preferably 1 ng / mL to 100 μg / mL, more preferably 10 ng / mL to 10 μg / mL, but may be outside this range as long as it has the desired effect.

[0105] Examples of ADAM inhibitors include, but are not limited to, KP-457. The concentration of the ADAM inhibitor is not particularly limited, and can be appropriately determined by those skilled in the art according to the compound. The concentration of the ADAM inhibitor can be, for example, 1.0 nM to 1.0 mM, preferably 10 nM to 0.1 mM, more preferably 100 nM to 0.1 mM, but may be outside this range as long as it has the desired effect.

[0106] The compound of the present invention or a salt thereof can be made into a kit in combination with one or more aromatic hydrocarbon receptor antagonists, one or more TPO or TPO receptor agonists, one or more ROCK inhibitors, and / or one or more ADAM inhibitors, etc.

[0107] The timing of adding the compound to be used in combination (coexisting with the compound of the present invention or its salt in the medium) is not particularly limited as long as it has the desired effect. The compound to be used in combination can be added to the medium before, after, or simultaneously with the addition of the compound of the present invention or its salt to the medium. In cells less differentiated than megakaryocytes, at least one gene selected from the group consisting of oncogenes, polycomb genes, and apoptosis suppressor genes is forced to be expressed to prepare immortalized megakaryocyte cells, and then forced expression is released to promote the multinucleation of immortalized megakaryocyte cells, so it is preferable to add the compound to the medium after (including simultaneously with) the release of forced expression.

[0108] The period of forced expression is not particularly limited and can be appropriately determined by those skilled in the art. After forced expression, the cells may be subcultured, and the period from the last subculture to the day when forced expression is released is not particularly limited, but may be, for example, 1 day, 2 days, or 3 days or more.

[0109] When the compound of the present invention or a salt thereof is added to the medium after the above-mentioned forced expression is released, the period from the release of the forced expression to the addition of the compound of the present invention or a salt thereof to the medium is not particularly limited, but for example, the culture may be started within 1, 2, 3, 4, 5, or 6 days. The period for culturing the cells in the presence of the compound of the present invention or a salt thereof is also not particularly limited. Usually, functional platelets are gradually released from about the first day after the compound of the present invention or a salt thereof is added to the medium, and the number increases with the number of days of culture. The period for culturing the cells in the presence of the compound of the present invention or a salt thereof is, for example, 5 to 10 days, but the number of days of culture may be shorter or longer. The compound of the present invention or a salt thereof may be added to the medium once or more during the culture period.

[0110] The cell culture conditions can be normal conditions. For example, the temperature can be about 35°C to about 42°C, preferably about 36°C to about 40°C, more preferably about 37°C to about 39°C, and may be 5% CO2 and / or 20% O2. Static culture or shaking culture may be used. The shaking speed in the case of shaking culture is not particularly limited, and can be, for example, 10 rpm to 200 rpm, preferably 30 rpm to 150 rpm, etc.

[0111] When megakaryocytes and / or their precursor cells are contacted with the compound of the present invention or a salt thereof and cultured, mature megakaryocytes are obtained, and platelets are produced from the cytoplasm. Here, the maturation of megakaryocytes means that megakaryocytes become multinucleated and are able to release platelets.

[0112] The medium for culturing megakaryocytes is not particularly limited, and any known medium suitable for producing platelets from megakaryocytes or a medium similar thereto can be used as appropriate. For example, a medium used for culturing animal cells can be prepared as a basal medium. Examples of basal medium include IMDM medium, Medium 199 medium, Eagle's Minimum Essential Medium (EMEM) medium, αMEM medium, Dulbecco's modified Eagle's Medium (DMEM) medium, Ham's F12 medium, RPMI1640 medium, Fischer's medium, Neurobasal Medium (Life Technologies), and mixed media thereof.

[0113] The medium may contain serum or plasma, or may be serum-free. When serum is used, fetal bovine serum (FBS) or human serum can be used. If necessary, the medium may also contain one or more substances such as albumin, insulin, transferrin, selenium, fatty acids, trace elements, 2-mercaptoethanol, thiolglycerol, monothioglycerol (MTG), lipids, amino acids (e.g., L-glutamine), ascorbic acid, heparin, non-essential amino acids, vitamins, growth factors, low molecular weight compounds, antibiotics, antioxidants, pyruvic acid, buffers, inorganic salts, and cytokines. Cytokines are proteins that promote blood cell differentiation, and examples of such substances include VEGF, TPO, TPO receptor agonists, SCF, ITS (insulin-transferrin-selenite) supplements, and ADAM inhibitors.

[0114] The agents and amounts used, the timing of their addition to the medium, the platelet precursor cells, the culture method and culture conditions, and the like, which have been explained regarding the platelet production promoter and the method for producing platelets, can be similarly applied to other aspects of the present invention (agents, uses, methods, etc.).

[0115] The disclosures of all patent and non-patent publications cited herein are hereby incorporated by reference in their entirety. EXAMPLES

[0116] The present invention will be further explained in detail by the following Test Examples, Reference Examples and Examples, but these do not limit the present invention and may be modified without departing from the scope of the present invention. The following abbreviations may be used in this specification:

[0117] JPEG0007657800000045.jpg207141 JPEG0007657800000046.jpg231141 JPEG0007657800000047.jpg104141

[0118] In the following examples, "room temperature" generally refers to about 10° C. to about 35° C. Ratios shown in mixed solvents are volume ratios unless otherwise specified. % refers to weight % unless otherwise specified. 1 HNMR (proton nuclear magnetic resonance spectrum) was measured by Fourier transform NMR (either Bruker AVANCE III 400 (400 MHz) or Bruker AVANCE III HD (500 MHz)). Mass spectra were measured by LC / MS (ACQUITY UPLC H-Class). The ionization method used was ESI, and the data was the actual value (found). Usually, the molecular ion peak ([M+H] + , [MH] - In the case of salts, the free molecular ion peak or fragment ion peak is usually observed. In silica gel column chromatography, aminopropylsilane-bonded silica gel was used when basic was mentioned. The absolute configuration of the compound was determined by a known X-ray crystal structure analysis method (for example, Shigeru Ohba and Shigenobu Yano, "Basic Lectures for Chemists 12: X-ray Crystal Structure Analysis" (1st edition, 1999)) or estimated from the empirical rule of asymmetric epoxidation (Waldemar Adam, Rainer T. Fell, Chantu R. Saha-Moller and Cong-Gui Zhao: Tetrahedron: Asymmetry 1998, 9, 397-401. Yuanming Zhu, Yong Tu, Hongwu Yu, Yian Shi: Tetrahedron Lett. 1988, 29, 2437-2440).

[0119] [Reference example] Reference example 1 Preparation of (E)-N-[2-(2-bromophenyl)ethyl]-3-(7-methoxy-1H-indol-3-yl)prop-2-enamide (E)-3-(7-methoxy-1H-indol-3-yl)prop-2-enoic acid (25.0 mg), 2-bromophenethylamine (19.8 μl) in DCM (2 ml) was added with DIPEA (40.2 μl) and COMU (59.1 mg) and stirred overnight at room temperature. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt) to obtain the target product (28 mg).

[0120] Reference example 2 Preparation of 2-(2-aminoethyl)-N-ethylaniline dihydrochloride To a solution of tert-butyl N-[2-(2-aminoethyl)phenyl]-N-ethylcarbamate (180 mg) in EtOH (2 ml), 4N HCl / AcOEt (1 ml) was added and stirred for 1.5 hours at 50° C. After concentrating the reaction solution, the residue was dispersed and washed with AcOEt to obtain the target product (170 mg).

[0121] Reference example 3 Preparation of tert-butyl N-[2-(2-aminoethyl)phenyl]-N-ethylcarbamate To a solution of tert-butyl N-[2-(2-azidoethyl)phenyl]-N-ethylcarbamate (300 mg) in EtOH (3 ml), 10% Pd / C (50 mg) was added and stirred at room temperature under a hydrogen atmosphere for 3 hours. The solid matter was filtered through Celite, and the filtrate was concentrated to obtain the target product (208 mg).

[0122] Reference example 4 Preparation of tert-butyl N-[2-(2-azidoethyl)phenyl]-N-ethylcarbamate To a solution of tert-butyl N-[2-(2-azidoethyl)phenyl]carbamate (1.0 g) in DMF (3 ml), NaH (0.18 g) and iodoethane (0.37 ml) were added and stirred at room temperature overnight. Water was added to the reaction solution and extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. It was filtered and the filtrate was concentrated. The residue was purified by column chromatography (Hexane / AcOEt) to obtain the target product (940 mg).

[0123] Reference example 6 Preparation of 2-(3-ethoxythiophen-2-yl)ethanamine hydrochloride To a solution of tris(pentafluorophenyl)borane (14.7 mg) in DCM (2 ml), diethylsilane (310 μl) and a solution of 2-(3-ethoxythiophen-2-yl)acetonitrile (160 mg) in DCM (1 ml) were added at 0°C under a nitrogen atmosphere, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated, and 4N HCl / AcOEt (718 μl) was added to the residue. The precipitated solid was collected by filtration to obtain the target product (38 mg).

[0124] Reference example 7 Preparation of 2-(3-ethoxythiophen-2-yl)acetonitrile A solution of TosMIC (502 mg) in DME (3 ml) was added dropwise to a suspension of KOtBu (524 mg) in DME (4 ml) at -50°C under a nitrogen atmosphere, and then a solution of 3-ethoxythiophene-2-carbaldehyde (365 mg) in DME (3 ml) was added dropwise and stirred for 1 hour. The mixture was brought to room temperature, MeOH (10 ml) was added, and the mixture was stirred for 1 hour under heating and reflux. Water was added to the reaction solution, and the mixture was extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (Hexane / AcOEt) to obtain the target product (162 mg).

[0125] Reference example 8 Preparation of (E)-N-[2-(2-bromo-5-fluorophenyl)ethyl]-3-(7-methoxy-1H-indol-3-yl)prop-2-enamide (E)-3-(7-methoxy-1H-indol-3-yl)prop-2-enoic acid (25.0 mg), 2-bromo-5-fluorophenethylamine (30.1 mg) in DCM (2 ml) was added with DIPEA (40.2 μl) and HATU (52.5 mg) and stirred at room temperature for 1 hour. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt) to obtain the target product (43 mg).

[0126] Reference example 13 Preparation of 2-(2-pyrimidin-2-ylphenyl)ethanamine hydrochloride To a solution of tert-butyl N-[2-(2-bromophenyl)ethyl]carbamate (200 mg) in toluene (4 ml), 2-tributylstannylpyrimidine (232 μl) and Pd(PPh3)4 (77.0 mg) were added under an argon atmosphere, and the mixture was stirred overnight at reflux. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt). 4N HCl / AcOEt (0.5 ml) was added to a solution of the purified product in EtOH (1 ml), and the mixture was stirred at 50°C for 1.5 hours. The reaction solution was concentrated to obtain the target product (76.0 mg).

[0127] Reference example 14 Preparation of (E)-N-[2-(3-bromothiophen-2-yl)ethyl]-3-(7-methoxy-1H-indol-3-yl)prop-2-enamide A solution of 3-bromo-2-[(E)-2-nitroethenyl]thiophene (400 mg) in THF (3 ml) was added dropwise to a suspension of LAH (0.084 g) in THF (4 ml) at 0°C under a nitrogen atmosphere, and the mixture was stirred at room temperature for 2 hours. Water (0.15 ml), 15% NaOH aqueous solution (0.15 ml), and water (0.45 ml) were added, the mixture was filtered through Celite, and the filtrate was concentrated. To a solution of the residue in DCM (1 ml), (E)-3-(7-methoxy-1H-indol-3-yl)prop-2-enoic acid (40.0 mg), DIPEA (0.048 ml), and HATU (91.0 mg) were added, and the mixture was stirred at room temperature overnight. The reaction solution was purified by column chromatography (Hexane / AcOEt) to obtain the target product (0.032 g).

[0128] Reference example 15 Preparation of 2-(3-thiophen-2-ylthiophen-2-yl)ethanamine hydrochloride A mixture of tert-butyl N-[2-(3-bromothiophen-2-yl)ethyl]carbamate (57.0 mg), 2-thiopheneboronic acid (40.5 mg), PdCl2(dppf)DCM (7.6 mg), K3PO4 (79.0 mg), and 1,4-dioxane / water (4 / 1) (1 ml) was stirred at 90°C for 2 hours under a nitrogen atmosphere. The reaction solution was purified by column chromatography (Hexane / AcOEt). 4N HCl / AcOEt (0.5 ml) was added to a solution of the purified product in EtOH (0.5 ml) and stirred overnight at room temperature. The reaction solution was concentrated to obtain the target product (38.2 mg).

[0129] Reference example 19 Preparation of 2-(2-pyrimidin-4-ylphenyl)ethanamine hydrochloride A mixture of tert-butyl N-[2-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethyl]carbamate (150 mg), 4-chloropyrimidine hydrochloride (98.0 mg), PdCl2(dppf)DCM (35.3 mg), K3PO4 (183 mg), and DME / water (4 / 1) (2 ml) was stirred overnight under nitrogen atmosphere at reflux. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt). 4N HCl / AcOEt (0.5 ml) was added to a solution of the purified product in EtOH (1 ml), and the mixture was stirred at 50°C for 1.5 hours. The reaction solution was concentrated to obtain the target product (55.0 mg).

[0130] Reference example 20 Preparation of (E)-3-(7-methoxy-1H-pyrrolo[2,3-c]pyridin-3-yl)prop-2-enoic acid To a solution of 7-methoxy-1H-pyrrolo[2,3-c]pyridine (420 mg) in AcOH (3 ml), hexamethylenetetramine (265 mg) was added and stirred at 100°C for 6 hours. A saturated aqueous solution of NaHCO3 was added to the reaction solution and extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. It was filtered and the filtrate was concentrated. The residue was suspended in DCM (3 ml), DIBOC (439 μl) and DMAP (23.1 mg) were added, and the mixture was stirred for 30 minutes. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt). To a solution of diethylphosphonoethyl acetate (113 μl) in THF (3 ml), NaH (22.7 mg) was added and stirred for 30 minutes, after which a solution of the purified product (104 mg) in THF (2 ml) was added dropwise and stirred at room temperature for 1 hour. Water was added to the reaction solution and it was extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. It was filtered and the filtrate was concentrated. The residue was purified by column chromatography (Hexane / AcOEt). A 5N NaOH solution (240 μl) was added to a solution of the purified product (104 mg) in THF-MeOH-water (1:1:1) (6 ml) and the mixture was stirred overnight at reflux. After concentration, the residue was neutralized with 1N HCl solution, and the solid matter was filtered to obtain the target product (48.0 mg).

[0131] Reference example 22 Preparation of (E)-3-(4-methoxyindol-1-yl)prop-2-enoic acid Cs2CO3 (996 mg) and ethyl propiolate (248 μl) were added to a solution of 4-methoxyindole (300 mg) in DMF (3 ml), and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction solution, and the mixture was extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated. The residue was purified by column chromatography (Hexane / AcOEt). A 5N NaOH aqueous solution (636 μl) was added to a solution of the purified product in THF-tert-butanol-water (1:1:0.5) (9 ml), and the mixture was stirred for 3 hours under reflux. After concentration, a 1N HCl aqueous solution was added to the residue, and the solid matter was filtered to obtain the target product (212 mg).

[0132] Reference example 23 Preparation of (E)-3-(8-methoxyimidazo[1,2-a]pyridin-3-yl)prop-2-enoic acid To a solution of diethylphosphonoethyl acetate (378 μl) in THF (5 ml), NaH (76.0 mg) was added and stirred for 1 hour, then a solution of 8-methoxyimidazo[1,2-a]pyridine-3-carbaldehyde (280 mg) in THF (10 ml) was added dropwise and stirred at room temperature for 1 hour. Water was added to the reaction solution and extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. It was filtered and the filtrate was concentrated. The residue was washed with IPE. A 5N NaOH aqueous solution (804 μl) was added to a solution of the purified product in THF-MeOH-water (1:1:1) (6 ml) and stirred overnight under reflux. After concentration, a 5N HCl aqueous solution was added to the residue to make it weakly acidic, and the solid matter was filtered to obtain the target product (212 mg).

[0133] Reference example 26 Preparation of 2-[2-(5-fluoropyrimidin-2-yl)phenyl]ethanamine hydrochloride A mixture of tert-butyl N-[2-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethyl]carbamate (222 mg), 2-chloro-5-fluoropyrimidine (118 μl), Pd(tBu3P)2 (16.3 mg), K3PO4 (271 mg), and 1,4-dioxane / water (4 / 1) (2.5 ml) was stirred at 90°C for 7 hours under a nitrogen atmosphere. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt). 4N HCl / AcOEt (0.5 ml) was added to a solution of the purified product in EtOH (1 ml), and the mixture was stirred at 50°C for 1.5 hours. The reaction solution was concentrated to obtain the target product (128 mg).

[0134] The compounds of Reference Examples 5, 9 to 12, 16 to 18, 21, 24, 25, 27 and 28 were produced in the same manner as in Reference Examples 1 to 4, 6 to 8, 13 to 15, 19, 20, 22, 23 and 26. The structural formulas and physical property data of the compounds of Reference Examples 1 to 28 are shown in Tables 1-1 to 1-5, respectively.

[0135] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0136] [Example] Example 10 Preparation of (E)-N-[2-[2-(cyclopropylmethoxy)phenyl]ethyl]-3-(7-methoxy-1-methylindol-3-yl)-N-methylprop-2-enamide To a solution of (E)-N-[2-[2-(cyclopropylmethoxy)phenyl]ethyl]-3-(7-methoxy-1H-indol-3-yl)-N-methylprop-2-enamide (25.0 mg) and DMF (1 ml), iodomethane (5.80 μl) and Cs2CO3 (40.3 mg) were added and stirred at room temperature for 5 hours. Water was added to the reaction solution, and the mixture was extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (Hexane / AcOEt) to obtain the target product (23.0 mg).

[0137] Example 17 Preparation of (E)-3-(7-methoxy-1H-indol-3-yl)-N-[2-(2-pyrimidin-2-ylphenyl)ethyl]prop-2-enamide DIPEA (96.0 μl) and COMU (71.0 mg) were added to a solution of (E)-3-(7-methoxy-1H-indol-3-yl)prop-2-enoic acid (30.0 mg) and 2-(2-pyrimidin-2-ylphenyl)ethanamine hydrochloride (71.6 mg) in DCM (3 ml), and the mixture was stirred at room temperature for 2 hours. A saturated aqueous solution of NaHCO3 was added to the reaction solution, and the mixture was extracted with AcOEt. The organic layer was washed with saturated saline and then dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated. The residue was purified by column chromatography (Hexane / AcOEt) to obtain the target product (40 mg).

[0138] Example 19 Preparation of (E)-3-(7-methoxy-1H-indol-3-yl)-N-[2-(2-thiophen-2-ylphenyl)ethyl]prop-2-enamide A mixture of (E)-N-[2-(2-bromophenyl)ethyl]-3-(7-methoxy-1H-indol-3-yl)prop-2-enamide (30.0 mg), 2-thiopheneboronic acid (12.5 mg), PdCl2(dppf)DCM (3.1 mg), K3PO4 (31.9 mg), and 1,4-dioxane / water (4 / 1) (1 ml) was stirred at 90°C for 6 hours under a nitrogen atmosphere. The reaction solution was purified by column chromatography (Hexane / AcOEt) to obtain the target product (24.7 mg).

[0139] Example 24 Preparation of (E)-3-(1-acetyl-7-methoxyindol-3-yl)-N-[2-(2-phenylphenyl)ethyl]prop-2-enamide To a solution of (E)-3-(7-methoxy-1H-indol-3-yl)-N-[2-(2-phenylphenyl)ethyl]prop-2-enamide (25.0 mg) in DCE (0.6 ml), TEA (0.050 ml), DMAP (7.2 mg), and acetic anhydride (0.011 ml) were added and stirred at room temperature overnight. The reaction solution was purified by column chromatography (Hexane / AcOEt). To a solution of the purified product in DCE (0.6 ml), TEA (0.050 ml), DMAP (3.0 mg), and acetic anhydride (0.011 ml) were added and stirred at room temperature for 1 hour. The reaction solution was purified by column chromatography (Hexane / AcOEt) to obtain the target product (20.2 mg).

[0140] Example 25 Preparation of (E)-3-(1-benzoyl-7-methoxyindol-3-yl)-N-[2-(2-phenylphenyl)ethyl]prop-2-enamide To a solution of (E)-3-(7-methoxy-1H-indol-3-yl)-N-[2-(2-phenylphenyl)ethyl]prop-2-enamide (19.7 mg) in DCE (0.6 ml), TEA (0.039 ml), DMAP (5.7 mg), and benzoyl chloride (0.011 ml) were added and stirred at room temperature for 3 hours. The reaction solution was purified by column chromatography (Hexane / AcOEt) to obtain the target product (22.2 mg).

[0141] Example 29 Preparation of (E)-N-[2-(2-ethoxypyridin-3-yl)ethyl]-3-(7-methoxy-1H-indol-3-yl)prop-2-enamide To a mixture of 2-(2-ethoxypyridin-3-yl)acetonitrile (140 mg), NaBH4 (140 mg), and THF (3 ml), TFA (0.28 ml) was added at 0°C and stirred at room temperature for 1 hour. Water and saturated aqueous NaHCO3 were added, extracted with AcOEt, and the organic layer was concentrated. To a solution of the residue in DCM (1 ml), (E)-3-(7-methoxy-1H-indol-3-yl)prop-2-enoic acid (30.0 mg), DIPEA (0.036 ml), and HATU (68.3 mg) were added and stirred at room temperature for 1 hour. The reaction solution was purified by column chromatography (Hexane / AcOEt) to obtain the target product (10.8 mg).

[0142] Example 45 Preparation of (E)-3-(7-methoxy-1H-indol-3-yl)-N-[2-(3-pyrimidin-2-ylthiophen-2-yl)ethyl]prop-2-enamide 2-(3-pyrimidin-2-ylthiophen-2-yl)ethanamine hydrochloride (16.2 mg), DCM (0.6 ml) was mixed with DIPEA (82.0 μl), (E)-3-(7-methoxy-1H-indol-3-yl)prop-2-enoic acid (15.0 mg), and HATU (33.1 mg), and stirred overnight at room temperature. The reaction solution was purified by column chromatography (Hexane / AcOEt) to obtain the target product (11.8 mg).

[0143] Example 47 Preparation of (E)-3-(1H-indol-3-yl)-N-[2-(2-pyrimidin-2-ylphenyl)ethyl]prop-2-enamide 2-(2-pyrimidin-2-ylphenyl)ethanamine hydrochloride (45.0 mg), DCM (0.6 ml), DIPEA (128 μl), (E)-3-(1H-indol-3-yl)prop-2-enoic acid (27.5 mg), and HATU (72.6 mg) were added to the mixture and stirred at room temperature overnight. The reaction solution was purified by column chromatography (Hexane / AcOEt / MeOH) to obtain the target product (36.1 mg).

[0144] The compounds of Examples 1 to 9, 11 to 16, 18, 20 to 23, 26 to 28, 30 to 44, 46, and 48 to 59 were produced in the same manner as in Examples 10, 17, 19, 24, 25, 29, 45, and 47. The structural formulas and physical property data of the compounds of Examples 1 to 59 are shown in Tables 2-1 to 2-13, respectively.

[0145] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Table 2-11] [Table 2-12] [Table 2-13]

[0146] [Production Example] Preparation Example 1: Preparation of 3-[5-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl]amino]ethyl]-2-hydroxyphenyl]benzonitrile (Compound A5) JPEG0007657800000066.jpg105160

[0147] (1) Preparation of tert-butyl N-[2-[3-bromo-4-(methoxymethoxy)phenyl]ethyl]carbamate (Compound IM1) To a solution of tert-butyl N-[2-(3-bromo-4-hydroxyphenyl)ethyl]carbamate (9.40 g) in DCM (150 ml), DIPEA (7.79 ml) and chloromethyl methyl ether (2.94 ml) were added at 0° C. and stirred at room temperature for 3 days. After concentrating the reaction solution, the residue was purified by column chromatography (Hexane / AcOEt) to obtain compound IM1 (10.9 g). NMR2(500 MHz); 7.38 (1H, d, J=1.9 Hz), 7.11-7.03 (2H, m), 5.22 (2H, s), 4.53 (1H, s), 3.52 (3H, s), 3.37-3.30 (2H, m), 2.72 (2H, t, J=7.0 Hz), 1.44 (9H, s).

[0148] (2) Preparation of tert-butyl N-[2-[3-(3-cyanophenyl)-4-(methoxymethoxy)phenyl]ethyl]carbamate (Compound IM2) A mixture of compound IM1 (350 mg), 3-cyanophenylboronic acid (186 mg), K3PO4 (412 mg), PdCl2(dppf)DCM (39.7 mg), and 1,4-dioxane / water (4 / 1) (5 ml) was stirred under a nitrogen atmosphere at 90° C. for 4 hours. After concentrating the reaction solution, the residue was purified by column chromatography (Hexane / AcOEt) to obtain compound IM2 (366 mg). NMR2(500 MHz); 7.83 (1H, t, J=1.7 Hz), 7.74 (1H, dt, J=7.9, 1.5 Hz), 7.61 (1H, dt, J=7.7, 1.4 Hz), 7.51 (1H, t, J=7.8 Hz), 7.19-7.15 (2H, m), 7.12 (1H, s), 5.13 (2H, s), 4.57 (1H, s), 3.41-3.34 (5H, m), 2.79 (2H, t, J=7.1 Hz), 1.43 (9H, s).

[0149] (3) Preparation of 3-[5-(2-aminoethyl)-2-hydroxyphenyl]benzonitrile hydrochloride (compound IM3) To a solution of compound IM2 (364 mg) in EtOH (2 ml), 4N HCl / AcOEt (2 ml) was added and stirred at room temperature for 7 hours. The reaction solution was concentrated to give compound IM3 (242 mg). NMR1(500 MHz); 9.81 (1H, s), 7.99 (1H, t, J=1.8 Hz), 7.97-7.88 (4H, m), 7.77 (1H, dt, J=7.7, 1.4 Hz), 7.62 (1H, t, J=7.8 Hz), 7.24 (1H, d, J=2.2 Hz), 7.11 (1H, dd, J=8.3, 2.3 Hz), 6.96 (1H, d, J=8.3 Hz), 3.09-2.98 (2H, m), 2.86-2.79 (2H, m).

[0150] (4) Preparation of 2-amino-6-chloro-9-(1-hydroxy-2-methylpropan-2-yl)-7H-purin-8-one (compound IM4) A solution of 2,5-diamino-4,6-dichloropyrimidine (10.0 g) and 2-amino-2-methyl-1-propanol (11.7 ml) in NMP (10 ml) was stirred at 140° C. overnight. The reaction solution was purified by column chromatography (Hexane / AcOEt / MeOH). CDI (19.9 g) was added to a solution of the purified product in THF (150 ml) at 0° C. and stirred for 1 hour. A 50% MeOH aqueous solution (300 ml) and a 5N NaOH aqueous solution (44.7 ml) were added and stirred for 1 hour. The reaction solution was concentrated, a 5N HCl aqueous solution was added to the residue, and the precipitated solid was collected by filtration to obtain compound IM4 (10.9 g). NMR1(500 MHz); 11.16 (1H, s), 6.48 (2H, s), 4.87 (1H, t, J=6.6 Hz), 3.79 (2H, d, J=6.6 Hz), 1.60 (6H, s).

[0151] (5) Preparation of 4-chloro-2-iodo-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazole (compound IM5) To a suspension of compound IM4 (10.90 g) and triphenylphosphine (13.31 g) in THF (200 ml), diisopropyl azodicarboxylate (40% toluene solution) (26.7 ml) was added dropwise at 0°C under a nitrogen atmosphere and stirred for 2 hours. The reaction solution was concentrated, and the residue was purified by column chromatography (Hexane / AcOEt). To a solution of the purified product in THF (200 ml), copper(I) iodide (8.06 g), diiodomethane (10.24 ml), and tert-butyl nitrite (7.55 ml) were added and stirred at 60°C for 5 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated. The residue was purified by column chromatography (Hexane / AcOEt) to obtain compound IM5 (9.29 g). NMR1(500 MHz); 5.02 (2H, s), 1.68 (6H, s).

[0152] (6) Preparation of 3-[2-hydroxy-5-[2-[(2-iodo-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl)amino]ethyl]phenyl]benzonitrile (compound IM6) A suspension of compound IM5 (150 mg), compound IM3 (153 mg), and DIPEA (0.22 ml) in IPA (2 ml) was stirred overnight at 80° C. Water was added, and the precipitated solid was collected by filtration to obtain compound IM6 (211 mg). NMR1(500 MHz); 9.62 (1H, s), 7.95 (1H, s), 7.88 (1H, d, J=7.9 Hz), 7.75 (1H, d, J=7.7 Hz), 7.67 (1H, s), 7.60 (1H, t, J=7.8 Hz), 7.22 (1H, s), 7.08 (1H, dd, J=8.3, 2.2 Hz), 6.88 (1H, d, J=8.2 Hz), 4.85 (2H, s), 3.92-3.51 (2H, m), 2.80 (2H, t, J=7.3 Hz), 1.60 (6H, s).

[0153] (7) Preparation of Compound A5 A mixture of compound IM6 (244 mg), 5-fluoropyridine-3-boronic acid (93 mg), PdCl2(dppf)DCM (18.0 mg), K3PO4 (188 mg), and 1,4-dioxane / water (4 / 1) (1 ml) was stirred at 90°C for 3 hours under a nitrogen atmosphere. The reaction solution was purified by column chromatography (Hexane / AcOEt). This was washed with Hexane / AcOEt to obtain compound A5 (197 mg). NMR1(500 MHz); 9.58 (1H, s), 9.34 (1H, s), 8.61 (1H, d, J=2.9 Hz), 8.37-8.30 (1H, m), 7.90 (1H, s), 7.84 (1H, d, J=7.9 Hz), 7.73 (1H, dt, J=7.8, 1.4 Hz), 7.59-7.52 (2H, m), 7.24 (1H, s), 7.12 (1H, dd, J=8.2, 2.2 Hz), 6.87 (1H, d, J=8.2 Hz), 4.91 (2H, s), 3.79 (2H, s), 2.90 (2H, t, J=7.2 Hz), 1.71 (6H, s).

[0154] Production Example 2: Production of 2-(2-fluorophenyl)-4-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl]amino]ethyl]phenol (Compound A6) The target compound was produced in the same manner as in the production of compound IM2, except that 3-cyanophenylboronic acid was changed to 2-fluorophenylboronic acid. NMR1(500 MHz); 9.37-9.31 (2H, m), 8.62 (1H, d, J=2.8 Hz), 8.38-8.31 (1H, m), 7.53 (1H, s), 7.39-7.31 (1H, m), 7.31-7.25 (1H, m), 7.21-7.10 (3H, m), 7.10-7.03 (1H, m), 6.84 (1H, d, J=8.2 Hz), 4.91 (2H, s), 3.77 (2H, s), 2.88 (2H, t, J=7.4 Hz), 1.71 (6H, s).

[0155] Preparation Example 3: Preparation of 2-(5-fluoropyridin-3-yl)-4-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]oxazol-4-yl]amino]ethyl]phenol (Compound A7) The target compound was produced in the same manner as in the production of compound IM2, except that 3-cyanophenylboronic acid was changed to 5-fluoropyridine-3-boronic acid. NMR1(500 MHz); 9.69 (1H, s), 9.32 (1H, s), 8.61 (1H, d, J=2.9 Hz), 8.56 (1H, s), 8.47 (1H, d, J=2.8 Hz), 8.36-8.30 (1H, m), 7.80-7.74 (1H, m), 7.54 (1H, s), 7.27 (1H, s), 7.15 (1H, dd, J=8.3, 2.2 Hz), 6.88 (1H, d, J=8.2 Hz), 4.90 (2H, s), 3.80 (2H, s), 2.91 (2H, t, J=7.2 Hz), 1.71 (6H, s).

[0156] Preparation Example 4: Preparation of 2-(2-fluorophenyl)-4-[2-[[2-(5-fluoropyridin-3-yl)-8,8-dimethyl-7H-purino[8,9-b][1,3]thiazol-4-yl]amino]ethyl]phenol (Compound A8)

[0157] (1) Preparation of 4-chloro-2-iodo-8,8-dimethyl-7H-purino[8,9-b][1,3]thiazole (compound IM5') A solution of 2,5-diamino-4,6-dichloropyrimidine (10.0 g) and 2-amino-2-methyl-1-propanol (12.8 ml) was stirred at 140°C for 4 hours. Water was added at room temperature, and the precipitated solid was collected by filtration. To a solution of the filtered solid in THF (100 ml), TCDI (20.5 g) was gradually added at 0°C, and the mixture was stirred at room temperature for 1 hour. After concentrating the reaction solution, water was added at 0°C, and the precipitated solid was collected by filtration. A suspension of the filtered solid, copper(I) iodide (4.19 g), diiodomethane (7.09 ml), and tert-butyl nitrite (3.93 ml) in THF (80 ml) was stirred overnight at 60°C. The reaction solution was filtered through Celite, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (Hexane / AcOEt). This was washed with IPA to obtain the desired product (3.96 g). NMR1(500 MHz); 3.95 (2H, s), 1.72 (6H, s).

[0158] (2) Preparation of Compound A8 The target compound was produced in the same manner as in the production method for compound IM2, except that 3-cyanophenylboronic acid was changed to 2-fluorophenylboronic acid and compound IM5 was changed to compound IM5'. NMR1(500 MHz); 9.35 (1H, s), 9.33 (1H, s), 8.63 (1H, d, J=2.9 Hz), 8.38-8.32 (1H, m), 7.81 (1H, s), 7.38-7.30 (1H, m), 7.27-7.23 (1H, m), 7.21-7.10 (3H, m), 7.07 (1H, s), 6.84 (1H, d, J=8.2 Hz), 3.90 (2H, s), 3.78 (2H, s), 2.89 (2H, t, J=7.4 Hz), 1.78 (6H, s).

[0159] [Test example] Test Example 1 (Platelet Production: Shaking Culture) The immortalized megakaryocyte cell line obtained by the method described in WO 2016 / 204256 was washed twice with D-PBS(-) and cultured in a medium containing no doxycycline to disable forced expression (gene expression OFF culture). The cells were cultured in a 125 mL polycarbonate Erlenmeyer flask (Corning #431143) at a seeding density of 1 × 10 5 The cells were seeded at 25 mL / flask and cultured in the following medium with shaking at 100 rpm under the following conditions: 37°C, 5% CO2. The medium was prepared by adding the following components (concentrations are final concentrations) to IMDM as the base medium. FBS 15% L-Glutamine 2 mM ITS 100x dilution MTG 450 μM Ascorbic acid 50 μg / mL SCF 50 ng / mL TA-316 0.1 μg / mL ADAM inhibitor 15 μM ROCK inhibitor 0.5 μM At the same time as the cells were seeded, an aromatic hydrocarbon receptor inhibitor (compound A5, final concentration 0.1 μM) or DMSO (control) was added and culture was started. On the third day after the culture, the compound of the present invention (Examples 1 to 61, final concentration 10 μM) or DMSO (control) was added, and after culturing for a total of 6 days, the number of platelets was measured. The measurement method is as follows. The same operation was performed for the control. After 6 days of gene expression OFF culture, a portion of the culture supernatant was collected and suspended in the following antibodies and Flow-Count Fluorospheres (Beckman Coulter #7547053) for staining. Anti-CD41 antibody APC label (BioLegend #303710) Anti-CD42a antibody eFluor 450 label (eBioscience #48-0428-42) Anti-CD42b antibody PE-labeled (BioLegend #303906) Thirty minutes after the staining reaction, the platelet count (CD41, CD42a, CD42b positive cells) was calculated using Flow-Count Fluorospheres with a BD Japan FACSVerse. The platelet count was expressed as a ratio to the control. Table 3 shows the results of culturing in the case where DMSO was added simultaneously with cell seeding, and Table 4 shows the results of culturing in the case where an aromatic hydrocarbon receptor inhibitor was added simultaneously with cell seeding. + and ++ indicate that the amount of platelet production was increased by 1.5 to less than 6.5 times and 6.5 times or more, respectively, compared with the control.

[0160] Examples 60 and 61 are known compounds and were prepared by the method described in WO2019 / 167973. JPEG0007657800000067.jpg63126 [Table 3]

[0161] [Table 4]

[0162] Test Example 2 (Platelet Production: Shaking Culture) In the same manner as in Test Example 1, the compounds of Examples 57 to 61 were used, and the compounds A1 (final concentration 0.75 μM), A2 (final concentration 0.1 μM), A3 (final concentration 10 μM), A4 (final concentration 1 μM), and A6 to A8 (final concentration 0.1 μM) were added as aryl hydrocarbon receptor inhibitors, and the results of the culture are shown in Table 5. In addition, the results of the culture using only the aryl hydrocarbon receptor inhibitor are also shown as a comparative example.

[0163] [Table 5]

Claims

1. General formula [I]: [In the formula, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 Alkyl; R 2 is a hydrogen atom or -C 1-6 is alkyl, R 3 is halogen, -Q k -(C 1-6 Alkyl) m -Q p -R 31 , optionally substituted phenyl or optionally substituted heteroaryl selected from the group consisting of furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and pyrimidyl; R 31 -C 1-6 Alkyl or -C 3-8 is cycloalkyl, Q are independently the same or different and each is an oxygen atom, a sulfur atom, -C(=O)-O-, or -NH-; k, m, and p are 0 or 1; n is 0, 1, or 2. When n is 2, R 3 are each independently the same or different substituents. W is a carbon atom or a nitrogen atom; X is a carbon atom, a nitrogen atom, or NR 12 and Y is a carbon atom or a nitrogen atom; Z's are independently the same or different and each is a nitrogen atom or CH; However, X and Y are not carbon atoms at the same time. R 12 is a hydrogen atom, -C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)-C 1-6 Alkyl, -C(=O)-aryl or -C(=O)-OC 1-6 is alkyl, Ring A is aryl or heteroaryl; --- is a single or double bond; However, when X is NH, W and Y are carbon atoms, and all Z are CH, ring A is 2-(-OC 1-6 alkyl)phenyl or 2,5-di(-OC 1-6 (alkyl)phenyl) However, the following compounds are not included, or salts thereof: 。

2. In general formula [I], [In the formula, R 11 , W, X, Y, Z and --- are as defined above.

2. The compound according to claim 1, wherein:

3. In general formula [I], [In the formula, R 3 and n is as defined above.

2. The compound according to claim 1, wherein:

4. In the general formula [I], the heteroaryl in ring A is selected from the group consisting of furan, thiophene, pyridine and quinoline.

2. The compound according to claim 1 or a salt thereof.

5. In general formula [I], [In the formula, V is the same or different and each V is independently a nitrogen atom or CH; R 4 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl] 2. The compound according to claim 1, wherein:

6. General formula [Ia] [In the formula, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 is alkyl, R 12 is a hydrogen atom or -C(=O)-OC 1-6 is alkyl, is pyridylbenzene, (halogen, -C 1-6 Alkyl or -OC 1-6 pyrimidyl)benzene, phenylthiophene, pyridylthiophene or pyrimidylthiophene, which may be substituted with alkyl. The compound according to claim 1, wherein the compound is represented by the formula:

7. The compound according to claim 1, selected from the group consisting of the following compounds, or a salt thereof:

8. General formula [I']: [In the formula, R 11 is a hydrogen atom, halogen, -C 1-6 Alkyl or -OC 1-6 Alkyl; R 2 is a hydrogen atom or -C 1-6 is alkyl, R 3 is halogen, -Q k -(C 1-6 Alkyl) m -Q p -R 31 , optionally substituted phenyl or optionally substituted heteroaryl selected from the group consisting of furyl, thienyl, oxazolyl, thiazolyl, pyrazolyl, pyridyl, pyrazyl, pyridazyl and pyrimidyl; R 31 -C 1-6 Alkyl or -C 3-8 is cycloalkyl, Q are independently the same or different and each is an oxygen atom, a sulfur atom, -C(=O)-O-, or -NH-; k, m, and p are 0 or 1; n is 0, 1, or 2. When n is 2, R 3 are each independently the same or different substituents. W is a carbon atom or a nitrogen atom; X is a carbon atom, a nitrogen atom, or NR 12 and Y is a carbon atom or a nitrogen atom; Z's are independently the same or different and each is a nitrogen atom or CH; However, X and Y are not carbon atoms at the same time. R 12 is a hydrogen atom, -C 1-6 Alkyl, -C 1-6 Alkyl-OC 1-6 Alkyl, -C(=O)-C 1-6 Alkyl, -C(=O)-aryl or -C(=O)-OC 1-6 is alkyl, Ring A is aryl or heteroaryl; --- is a single bond or a double bond. A platelet production promoter comprising a compound represented by the formula:

9. The enhancer of claim 8 for use in combination with an aromatic hydrocarbon receptor inhibitor.

10. 9. The method of claim 8, wherein the aromatic hydrocarbon receptor inhibitor is selected from the group consisting of:

11. Use of the compound or salt thereof according to claim 8 for promoting platelet production in vitro.

12. 10. The compound or salt thereof according to claim 8 for use in promoting platelet production.

13. A method for promoting platelet production in vitro, comprising culturing platelet precursor cells in the presence of the compound or salt thereof according to claim 8.

14. 10. A method for producing platelets in vitro, comprising culturing platelet precursor cells in the presence of the compound or salt thereof according to claim 8.

15. A method for culturing platelet precursor cells in vitro to promote platelet production, comprising culturing platelet precursor cells in the presence of the compound or salt thereof according to claim 8.

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