Compound or salt thereof, method for producing same, pharmaceutical composition containing compound or salt thereof, and method for producing same

Abstract

A compound or a salt thereof, the compound or the salt thereof being represented by formula (I), in the formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a substituted or unsubstituted linear or branched alkylene group having 1 to 13 carbon atoms, X is a group represented by-(CH2CH2O) s-,-(CH2CH2CH2O) s-,-(CH (CH3) CH2O) s-, or-(CH2) tO-, s is a number from 1 to 8, and t is a number from 2 to 10.

Description

Technical Field

[0001] This disclosure relates to compounds or salts thereof and methods for manufacturing them, and pharmaceutical compositions comprising compounds or salts thereof and methods for manufacturing them.

[0002] This application claims priority based on Japanese Patent Application No. 2023-143583, filed in Japan on September 5, 2023, the contents of which are incorporated herein by reference. Background Technology

[0003] Epigenetic modifications such as methylation and acetylation of histone proteins control gene expression and regulate various life phenomena. Abnormalities in these modifications can lead to cancer, neurological disorders, and other diseases. To combat these abnormalities or the diseases they cause, low-molecular-weight epigenetic modifications have emerged as a promising therapeutic strategy.

[0004] Lysine demethylase 5 (KDM5) family proteins (KDM5A-D) are iron(II) / α-ketoglutarate-dependent oxidases that oxidatively remove the methyl group from trimethyl or dimethyl lysine 4 of histone H3 (H3K4me3 / 2). In addition to their enzymatic activity, KDM5 functions as a scaffold protein, recruiting repressive transcription factors such as histone deacetylase 1 (HDAC1) and HDAC2. In other words, KDM5 does not work alone but in collaboration with other proteins to epigenetically control gene expression. Furthermore, this suggests that KDM5 is associated with the pathogenesis of various cancers, drug resistance, and neurodegenerative diseases such as Alzheimer's disease.

[0005] To date, several KDM5 inhibitors have been identified.

[0006] In Non-Patent Literature 1, a compound (KDM5-C49) with the following formula is described as a KDM5 inhibitor.

[0007] In Non-Patent Literature 2, a compound (CPI-455) with the following formula is described as a KDM5 inhibitor.

[0008] In Non-Patent Literature 3, a compound (S1) with the following formula is described as a KDM5 inhibitor.

[0009] In Non-Patent Document 4, a compound (S2) with the following formula is described as a KDM5 inhibitor.

[0010] In Non-Patent Literature 5, a compound (S3) with the following formula is described as a KDM5 inhibitor.

[0011] In Non-Patent Literature 6, a compound (TK-129) with the following formula is described as a KDM5 inhibitor.

[0012] [Chemistry 1]

[0013] However, these KDM5 inhibitors did not produce significant pharmacological effects such as anti-cancer activity in cell and animal experiments.

[0014] Existing technical documents Non-patent literature Non-patent document 1: JohanssonC.,VelupillaiS.,TumberA.,SzykowskaA.,HookwayE.S.,NowakR.P.,Strain-DamerellC.,GileadiC.,PhilpottM.,Burgess-BrownN.,WuN.,KopecJ.,Nuzzi A.,SteuberH.,EgnerU.,BadockV.,MunroS.,LaThangueN.B.,WestawayS.,BrownJ.,AthanasouN.,PrinjhaR.,BrennanP.E.,OppermannU.,Nat.Chem.Biol.,12,539-545(2016). Non-patent document 2: VinogradovaM.,GehlingV.S.,GustafsonA.,AroraS.,TindellC.A.,WilsonC.,WilliamsonK.E.,GulerG.D .,GangurdeP.,ManieriW.,BusbyJ.,FlynnE.M.,LanF.,KimH.J.,OdateS.,CochranA.G.,LiuY.,WongchenkoM.,Yan gY.,CheungT.K.,MaileT.M.,LauT.,CostaM.,HegdeG.V.,JacksonE.,PittiR.,ArnottD.,BaileyC.,BellonS.,Cum mingsR.T.,AlbrechtB.K.,HarmangeJ.C.,KieferJ.R.,TrojerP.,ClassonM.,Nat.Chem.Biol.12,531-538(2016). Non-patent document 3: Vazquez-RodriguezS.,WrightM.,RogersC.M.,CribbsA.P.,VelupillaiS.,PhilpottM.,LeeH.,DunfordJ.E.,HuberK.VM,RobersM.B.,VastaJ.D.,ThezenasM.L.,BonhamS.,K esslerB.,BennettJ.,FedorovO.,RaynaudF.,DonovanA.,BlaggJ.,BavetsiasV.,OppermannU.,BountraC.,KawamuraA.,BrennanP.E.,Angew.Chem.Int.Ed.Engl.,58,515-519(2019). Non-patent literature 4: Miyake Y., Itoh Y., Suzuma Y., Kodama H., Uchida S., Suzuki T., ACS Catal., 10, 5383-5392 (2020). Non-patent literature 5: Zhao B., Liang Q., Ren H., Zhang X., Wu Y., Zhang K., Ma L.Y., Zheng Y.C., Liu H.M., Eur. J Med. Chem., 192, 112161 (2020). Non-patent literature 6: TangK., JiaoL.M., QiY.R., WangT.C., LiY.L., XuJ.L., WangZ.W., YuB., LiuH.M., ZhaoW., J.Med.Chem., 65, 12979-13000(2022). Summary of the Invention The problem that the invention aims to solve This disclosure was made to solve the above-mentioned problems, and its purpose is to provide compounds or salts thereof that produce significant pharmacological effects such as anticancer effects in cell and animal experiments.

[0015] means for solving problems To address the aforementioned problems, the inventors of this invention conducted in-depth research, focusing on "proteolysis-targeting chimeras (hereinafter also referred to as PROTACs)" that degrade target proteins and inhibit their overall function in order to identify compounds exhibiting stronger pharmacological activity. PROTACs are small molecules composed of two ligands. Based on KDM5 inhibitors identified to date, novel KDM5PROTAC candidate compounds were designed and synthesized. Cellular analysis results showed that the disclosed compounds or their salts exhibited significant synaptic growth-promoting activity in neuroblastoma neuropathy 2a cells via KDM5A degradation. These results suggest that KDM5PROTACs are promising drug candidates for the treatment of neurological diseases. The inventors of this invention discovered that the aforementioned problems could be solved by using the disclosed compounds or their salts, thus completing this disclosure.

[0016] That is, the purpose of this disclosure is as follows [1] to

[10] .

[0017] [1] A compound or a salt thereof, wherein the compound or a salt thereof is represented by the following formula (I). [Chemistry 2]

[0018] In formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a straight-chain or branched alkylene group with 1 to 13 substituted or unsubstituted carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 8, and t is a number from 2 to 10.

[0019] [2] According to the compound or salt thereof as described in [1], wherein in the formula (I), A is a single bond or phenylene, L is a straight-chain alkylene with 3 to 10 carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

[0020] [3] The compound or its salt according to [1] or [2], wherein in the formula (I), A is a single bond, p-phenylene or m-phenylene, L is a straight-chain alkylene with 3 to 10 carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

[0021] [4] A compound or a salt thereof according to any one of [1] to [3], wherein the compound is a compound represented by formula (I-1), a compound represented by formula (I-2), or a compound represented by formula (I-3). [Chemistry 3]

[0022] In equation (I-1), L 1 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 1 For -(CH2CH2O) s - The group shown, s is a number from 1 to 5, in formula (I-2), L 2 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 2 For -(CH2CH2O) s -or- (CH2) t The O- group represents a number from 1 to 5, and t represents a number from 2 to 10. In formula (I-3), L 3 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 3 For -(CH2CH2O) s -or- (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

[0023] [5] The compound or salt thereof according to any one of [1] to [4], wherein the compound is a compound of the following composition: In the above formula (I-1), L 1 It is a straight-chain alkylene group with 5 carbon atoms, X 1 For -(CH2CH2O) s - The group shown is 3; In the above formula (I-1), L 1 It is a straight-chain alkylene group with 8 carbon atoms, X 1 For -(CH2CH2O) s - The group shown is 3; In the above formula (I-2), L 2It is a straight-chain alkylene group with 5 carbon atoms, X 2 For -(CH2CH2O) s - The group shown, s is 3; In the above formula (I-2), L 2 It is a straight-chain alkylene group with 5 carbon atoms, X 2 For -(CH2) t The group represented by O-, and the number t is 6; In the aforementioned formula (I-3), L 3 It is a straight-chain alkylene group with 5 carbon atoms, X 3 For -(CH2CH2O) s - The group shown, s is 3; or In the aforementioned formula (I-3), L 3 X is a straight-chain alkylene group with 5 carbon atoms. 3 For -(CH2) t The O- group represents a number t of 6.

[0024] [6] A pharmaceutical composition comprising any one of [1] to [5] a compound or a salt thereof.

[0025] [7] The pharmaceutical composition according to [6], wherein the pharmaceutical composition is a KDM5 inhibitor and a KDM5 degrader.

[0026] [8] The pharmaceutical composition according to [6] or [7], wherein the pharmaceutical composition is a preventive and / or therapeutic agent for KDM5-related diseases.

[0027] [9] The compound or salt thereof according to any one of [1] to [5], wherein the compound or salt thereof is effective for the prevention and / or treatment of KDM5-related diseases by KDM5 inhibition and KDM5 degradation.

[0028]

[10] The pharmaceutical composition according to any one of [6] to [8], wherein the pharmaceutical composition is used for the prevention and / or treatment of KDM5-related diseases that are effective in inhibiting and degrading KDM5.

[0029]

[11] A method for preventing and / or treating KDM5-related diseases that are effective in inhibiting and degrading KDM5, wherein the method comprises administering to a patient a compound or a salt thereof as described in any one of [1] to [5] and [9].

[0030]

[12] A method for preventing and / or treating KDM5-related diseases that are effective in inhibiting and degrading KDM5, wherein the method comprises administering to a patient the pharmaceutical composition described in any one of [6] to [8].

[0031]

[13] Use of a compound or a salt thereof, wherein the compound or a salt thereof is any one of [1] to [5] and [9], and the use is in the preparation of a pharmaceutical composition effective for the prevention and / or treatment of KDM5-related diseases with KDM5 inhibition and KDM5 degradation.

[0032]

[14] A preventive and / or therapeutic agent for KDM5-related diseases, wherein the preventive and / or therapeutic agent for KDM5-related diseases comprises any one of [1] to [5] and [9] or a salt thereof as an active ingredient.

[0033] Invention Effects According to this disclosure, compounds or salts thereof can provide significant pharmacological effects, such as anticancer effects, in cell and animal experiments. Attached Figure Description

[0034] Figure 1 It represents compound 14a. 1 The figure of H-NMR spectrum.

[0035] Figure 2 It represents compound 14a. 13 A C-NMR spectrum.

[0036] Figure 3 This is a graph showing the UV absorption spectrum of compound 14a by HPLC.

[0037] Figure 4 It represents compound 14b. 1 The figure of H-NMR spectrum.

[0038] Figure 5 It represents compound 14b. 13 A C-NMR spectrum.

[0039] Figure 6 This is a graph showing the UV absorption spectrum of compound 14b by HPLC.

[0040] Figure 7 It represents compound 20a. 1 The figure of H-NMR spectrum.

[0041] Figure 8 It represents compound 20a. 13 A C-NMR spectrum.

[0042] Figure 9 This is a graph showing the UV absorption spectrum of compound 20a by HPLC.

[0043] Figure 10 It represents compound 20b. 1 The figure of H-NMR spectrum.

[0044] Figure 11 It represents compound 20b. 13 A C-NMR spectrum.

[0045] Figure 12 This is a graph showing the UV absorption spectrum of compound 20b by HPLC.

[0046] Figure 13 It represents compound 23a. 1 The figure of H-NMR spectrum.

[0047] Figure 14 It represents compound 23a. 13 A C-NMR spectrum.

[0048] Figure 15 This is a graph showing the UV absorption spectrum of compound 23a by HPLC.

[0049] Figure 16 It represents compound 23b. 1 The figure of H-NMR spectrum.

[0050] Figure 17 It represents compound 23b. 13 A C-NMR spectrum.

[0051] Figure 18 This is a graph showing the UV absorption spectrum of compound 23b by HPLC.

[0052] Figure 19 (A) in the diagram represents the structures of KDM5 inhibitor 24 and vorinostat (25). Figure 19 (B) in the figure represents the effects of individual treatments based on compounds 24 and 25 on N2a differentiation after 24 hours and 48 hours of treatment, as well as the effects of co-treatment with compounds 24 and 25.

[0053] Figure 20 These are representative images of N2a cells after 24 hours and 48 hours of treatment with either compound 24 or compound 25 alone, and after treatment with both compounds 24 and 25 together. Scale bar is 100 μm.

[0054] Figure 21(A) in the text represents the design of KDM5PROTAC. Figure 21 In the diagram, (B) represents the effect of 0.2 μM of 14a, 14b, 20a, 20b, 23a, and 23b on N2a differentiation after 48 hours of treatment. Figure 21 In the figure, (C) represents the dose-dependent N2a cell synaptic growth activity of compounds 20b, 23b, and 26 after 48 hours of treatment. Figure 21 (D) in the figure represents the time-dependent N2a cell synaptic growth activity at 0.02 μM for 20b and 23b. The bar chart represents the mean ± SD of three independent experiments.

[0055] Figure 22 This is a representative image of N2a cells after treatment with compounds 14a, 14b, 20a, 20b, 23a, and 23b for 48 hours. The scale bar is 100 μm.

[0056] Figure 23 This is a representative image of N2a cells after treatment with compounds 20b, 23b, and 26 for 48 hours. The scale bar is 100 μm.

[0057] Figure 24 These are representative images of N2a cells treated with compounds 20b and 23b for 24, 48, and 72 hours. Scale bar: 100 μm.

[0058] Figure 25 This is a Western blot diagram showing the KDM5A level in N2a cells. Figure 25 (A) in the text indicates that after treatment with compounds 20b and 23b for 24 hours, Figure 25 (B) indicates treatment with compound 23b for 24 hours, and treatment with compounds 24 and 26 together for 24 hours. Figure 25 (C) indicates that after treatment with compound 20b and the proteasome inhibitor MG-132 for 24 hours, Figure 25 (D) in the text indicates treatment with the proteasome inhibitor MG-132 and compound 23b for 24 hours.

[0059] Figure 26 This is a Western blot diagram showing the levels of H3K4me3 and H3K27Ac in N2a cells after treatment with compounds 20b and 23b for 24 hours.

[0060] Figure 27This is a graph showing the KDM5A inhibitory activity and HDAC1 inhibitory activity of compounds 20b and 23b. Detailed Implementation

[0061] The following is a more detailed explanation of this disclosure.

[0062] This disclosure, in addition to the compounds shown in formula (I) and their salts, also includes their N-oxides, solvates, isomers, mixtures of multiple isomers, single crystal forms, mixtures of multiple crystal forms, eutectics, and isotopically labeled compounds. Hereinafter, they are also collectively referred to as "the compounds".

[0063] Unless otherwise specified, "alkyl" includes straight-chain, branched, and cyclic monovalent saturated hydrocarbon groups. The same applies to alkyl groups in alkoxy groups.

[0064] Unless otherwise specified, "alkylene" includes straight-chain, branched, and cyclic divalent saturated hydrocarbon groups.

[0065] Unless otherwise specified, "alkenyl" includes linear, branched, and cyclic divalent unsaturated hydrocarbon groups.

[0066] Halogen atoms can be listed as fluorine atoms, chlorine atoms, bromine atoms, and iodine atoms.

[0067] When described as "substituted or unsubstituted", it includes both cases where a hydrogen atom (-H) is substituted with a monovalent substituent and cases where a methylene (-CH2-) or methine (>CH- or =CH-) is replaced with a divalent substituent.

[0068] Monovalent substituents can be listed as alkyl, alkoxy, halogen atom, haloalkyl, hydroxy, carboxyl, amino, nitro, etc.

[0069] The alkyl group used as a substituent is preferably an alkyl group having 1 to 5 carbon atoms, and most preferably methyl, ethyl, propyl, n-butyl, or tert-butyl.

[0070] The alkoxy group used as a substituent is preferably an alkoxy group having 1 to 5 carbon atoms, more preferably methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, or tert-butoxy, and most preferably methoxy or ethoxy.

[0071] The halogen atom used as a substituent is preferably a fluorine atom or an iodine atom.

[0072] Haloalkyl groups that are substituents can be exemplified as alkyl groups having 1 to 5 carbon atoms, such as methyl, ethyl, propyl, n-butyl, tert-butyl, etc., in which some or all of the hydrogen atoms are replaced by the halogen atom.

[0073] "Divalent substituents" can be listed as -O-, -C(=O)-O-, -OC(=O)-, -C(=O)-, -OC(=O)-O-, -C(=O)-NH-, -C(=O)-NH-C(=O)-, -N=, -NH-, -NH-C(=NH)- (H can be replaced by hydrocarbon, acyl, alkoxyalkyl, etc. substituents), -S-, -S(=O)2-, -S(=O)2-O-, and general formula -Y. 21 -OY 22 -、-Y 21 -O-、-Y 21 -C(=O)-O-、-C(=O)-OY 21 -、-[Y 21 -C (=O) -O m” -Y 22 -、-Y 21 -OC (=O) -Y 22 -or-Y 21 -S(=O)2-OY 22 - The group shown in the formula [where Y] 21 and Y 22 [Each is an independently substituted or unsubstituted divalent hydrocarbon group, O is an oxygen atom, and m" is an integer from 0 to 3] etc.

[0074] This compound In the compounds or salts thereof disclosed herein, the compounds are those represented by formula (I).

[0075] [Chemistry 4]

[0076] In formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a straight-chain or branched alkylene group with 1 to 13 substituted or unsubstituted carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 8, and t is a number from 2 to 10.

[0077] As A, it is preferably a single bond, phenylene, or naphthylene; more preferably a single bond, 1,2-phenylene, 1,3-phenylene (m-phenylene), 1,4-phenylene (p-phenylene), 1,2-naphthylene, 1,4-naphthylene, 1,5-naphthylene, 2,3-naphthylene, or 2,6-naphthylene; and even more preferably a single bond, 1,3-phenylene (m-phenylene), or 1,4-phenylene (p-phenylene).

[0078] The L is preferably a straight-chain alkylene group having 3 to 10 carbon atoms, more preferably a straight-chain alkylene group having 4 to 9 carbon atoms, and even more preferably a straight-chain alkylene group having 5 to 8 carbon atoms.

[0079] As X, it is preferably -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t The group indicated by O-; more preferably -(CH2CH2O). s -、-(CH2CH2CH2O) s - or - (CH2) t The group represented by O- is further preferably -(CH2CH2O). s - or - (CH2) t The group indicated by O-.

[0080] The number s is preferably 1 to 5, more preferably 2 to 4, further preferably 2 or 3, and particularly preferably 3.

[0081] The number t is preferably 2 to 10, more preferably 3 to 8, even more preferably 4 to 7, and particularly preferably 6.

[0082] In the formula (I), A is preferably a single bond or a phenylene group, L is a straight-chain alkylene group with 3 to 10 carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

[0083] In the aforementioned formula (I), it is more preferable that A is a single bond, p-phenylene, or m-phenylene, L is a straight-chain alkylene group with 3 to 10 carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -or- (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

[0084] The compound is further preferably a compound represented by formula (I-1), a compound represented by formula (I-2), or a compound represented by formula (I-3).

[0085] [Chemistry 5]

[0086] In equation (I-1), L 1 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 1 For -(CH2CH2O) s - The group shown, s is a number from 1 to 5, in formula (I-2), L 2 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 2 For -(CH2CH2O) s -or- (CH2) t The O- group represents a number from 1 to 5, and t represents a number from 2 to 10. In formula (I-3), L 3 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 3 For -(CH2CH2O) s -or- (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

[0087] The compound is particularly preferably at least one selected from the group consisting of compounds of the following: in formula (I-1), L 1 It is a straight-chain alkylene group with 5 carbon atoms, X 1 For -(CH2CH2O) s - The group shown, the compound with s = 3 (14a); in the formula (I-1), L 1 It is a straight-chain alkylene group with 8 carbon atoms, X 1 For -(CH2CH2O) s - The group shown, the compound with s = 3 (14b); in the formula (I-2), L 2 It is a straight-chain alkylene group with 5 carbon atoms, X 2 For -(CH2CH2O) s - The group shown, the compound with s being 3 (20a); in the formula (I-2), L 2 It is a straight-chain alkylene group with 5 carbon atoms, X 2 For -(CH2) t Compounds with the O- group and t = 6 (23a); in formula (I-3), L 3 It is a straight-chain alkylene group with 5 carbon atoms, X 3 For -(CH2CH2O) s - The group shown, the compound with s = 3 (20b); and in the formula (I-3), L 3 It is a straight-chain alkylene group with 5 carbon atoms, X 3 For -(CH2)t Compounds with the O- group and a t number of 6 (23b).

[0088] Salts of the compounds include all pharmacologically acceptable salts. Preferably, all pharmacologically acceptable salts are water-soluble salts with low toxicity. Examples of suitable salts include acid addition salts (e.g., inorganic acid salts [e.g., hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, nitrate, etc.], organic acid salts [e.g., acetate, trifluoroacetate, lactate, tartrate, oxalate, fumarate, maleate, benzoate, citrate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, hydroxyethanesulfonate, glucuronide, gluconate, etc.], and salts of acidic natural amino acids [e.g., aspartic acid, glutamic acid, etc.]).

[0089] This compound can be converted into N-oxides and solvates.

[0090] N-oxides refer to substances formed by the oxidation of nitrogen atoms in this compound. This compound can be converted into N-oxides by known methods.

[0091] A solvate is a solvated form in which the compound is reacted with pharmacologically acceptable solvents such as water and ethanol. This compound can be converted into a solvate by known methods.

[0092] This compound can form a cocrystal with a suitable cocrystal forger. The cocrystal is preferably a pharmaceutically acceptable cocrystal formed from a pharmaceutically acceptable cocrystal forger. A cocrystal is generally defined as a crystal formed by two or more molecules through nonionic intermolecular interactions. The cocrystal can be a complex of a neutral molecule and a salt. The cocrystal can be prepared by known methods such as melt crystallization, recrystallization from a solvent, or physical pulverization of the constituent components.

[0093] This disclosure includes all geometric isomers, optical isomers based on asymmetric carbon, stereoisomers, tautomers, and mixtures of isomers that arise structurally from the compound, and is not limited to the formulas used for convenience. Any single isomer or mixture thereof may exist. Therefore, it is possible for optically active forms and racemic mixtures to exist within the molecule due to the presence of asymmetric carbon atoms, but this is not a limitation of this disclosure. Furthermore, polymorphs may sometimes exist, but these are also not limited; any single crystalline form or mixture thereof, or hydrates other than anhydrous substances, are all included within the scope of this disclosure.

[0094] This disclosure also includes isotopically labeled compounds of this compound, which are identical to this compound except that one or more atoms are replaced by atoms having atomic masses or mass numbers different from those normally found in nature. Isotopes that can be intercalated into this compound include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, phosphorus, sulfur, iodine, and chlorine. 2 H, 3 H, 11 C 14 C 13 N、 15 O、 18 F, 35 S, 123 I, and 125 I etc.

[0095] Compounds containing the aforementioned isotopes and / or other isotopes, and their pharmaceutically acceptable derivatives (e.g., salts), are within the scope of this disclosure. The isotope-labeled compounds of this disclosure, for example, those embedded with… 3 H and / or 14 Compounds containing radioactive isotopes such as C are useful for tissue distribution analysis of pharmaceuticals and / or matrices. It is believed that... 3 H and 14 C is useful due to the ease of their preparation and detection. It is believed that isotopes... 11 C and 18 F is useful in PET (positron emission tomography), and it is believed that isotopes... 125 I is useful in SPECT (single-photon emission computed tomography) and in all brain imaging. Based on 2 The substitution with heavier isotopes such as H produces greater metabolic stability, resulting in certain therapeutic advantages such as increased in vivo half-life or reduced dosage, and is therefore considered useful in some cases. The isotopically labeled compounds of this invention can be prepared similarly by using readily available isotopically labeled reagents instead of non-isotopically labeled reagents and performing the steps disclosed in the following diagrams and / or examples.

[0096] The prodrug of this compound refers to a compound that is transformed into this compound through a reaction with enzymes, gastric acid, etc. in a living organism. Examples of prodrugs of this compound include compounds in which the amino group is acylated, alkylated, or phosphorylated when the compound has an amino group (e.g., compounds in which the amino group is converted to eicosanoyl, alanyl, pentylaminocarbonyl, (5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methoxycarbonyl, tetrahydrofuranyl, pyrrolidinylmethyl, neopentyloxymethyl, acetoxymethyl, tert-butyl, etc.); and compounds in which the hydroxyl group is acylated, alkylated, phosphorylated, or converted to a borate (e.g., compounds in which the hydroxyl group is converted to acetyl, palmitoyl, propionyl, neopentyl, succinyl, fumaroyl, alanyl, dimethylaminomethylcarbonyl, etc., etc.). As prodrugs of this compound, examples include those converted to this compound under physiological conditions as disclosed in "Pharmaceutical Development," Volume 7, "Molecular Design," 1990, Hirokawa Shoten Co., Ltd., pp. 163-198. Prodrugs of this compound can be manufactured using methods known to them. Like the compound itself, prodrugs of this compound can, for example, form salts such as acid addition salts, or can form solvates with water or alcoholic solvents (such as ethanol).

[0097] Method for manufacturing this compound This compound can be manufactured, for example, by the method described below as shown in Scheme I. In the manufacturing method described below, the raw material compound can be a salt. As a salt, for example, it may contain a salt of a compound represented by general formula (I) as described above.

[0098] [Chemistry 6]

[0099] In Scheme I, compounds 1, 6, 8, and 11, besides being available as commercially available products, can be readily synthesized from commercially available products using known methods. In the chemical formulas, A, L, and X are the same as those described above in the description of the compounds shown in Formula (I). R 6 The alkyl group has 1 to 6 carbon atoms, preferably methyl or ethyl.

[0100] In step (a), compound 2 can be manufactured by protecting the primary amino group of compound 1 with a Boc group (tert-butoxycarbonyl).

[0101] The protection of primary amines by the Boc group is well known. For example, it can be carried out in solvents such as dichloromethane, in the presence of Boc group-introducing agents such as (Boc)₂O, and bases such as triethylamine at 0–80 °C.

[0102] In step (b), compound 3 can be manufactured by replacing the hydrogen atom on the nitrogen atom of compound 2 with a methyl group.

[0103] The reaction of replacing a hydrogen atom on a nitrogen atom with a methyl group is well known. For example, it can be carried out in solvents such as N,N-dimethylformamide (hereinafter also called DMF) in the presence of methylating agents such as MeI and bases such as NaH at 0–80 °C.

[0104] In step (c), compound 4 can be produced by a cross-coupling reaction of compound 3 and an organoboronic acid compound using a palladium catalyst.

[0105] Cross-coupling reactions using palladium catalysts are well known. For example, they can be carried out in solvents such as dimethyl sulfoxide (hereinafter also referred to as DMSO) at temperatures ranging from 40 to 190 °C in the presence of compound 3, organoboronic acid compounds such as bis(pinacol)diboron, palladium catalysts such as Pd(dppf)Cl2, and bases such as KOAc.

[0106] In step (d), compound 5 can be manufactured by deprotecting the Boc-protected amino group of compound 4.

[0107] Deprotection of Boc-protected amino groups is well known. For example, it can be carried out in solvents such as dichloromethane, in the presence of acids such as trifluoroacetic acid, at 0–80 °C.

[0108] In step (e), compound 7 can be produced by reductive amination of the secondary amino group of compound 5 with the organic aldehyde compound 6.

[0109] Reductive amination is well known. For example, it can be carried out in solvents such as 1,2-dichloroethane at temperatures ranging from 0 to 80°C in the presence of a reducing agent such as sodium triacetoxyborohydride.

[0110] In step (f), compound 9 can be produced by a cross-coupling reaction of compound 7 and organohalogen compound 8 using a palladium catalyst.

[0111] Cross-coupling reactions using palladium catalysts are well known. For example, they can be carried out in solvents such as 1,2-dimethoxyethane (hereinafter also called DME), in the presence of palladium catalysts such as compound 7, compound 8, and Pd(dppf)Cl2, and bases such as Na2CO3, at temperatures ranging from 40 to 190 °C.

[0112] In step (g), compound 10 can be manufactured by converting the ester of compound 9 to a carboxyl group through hydrolysis.

[0113] Hydrolysis reactions that convert esters to carboxyl groups are well known. For example, they can be carried out in solvents such as tetrahydrofuran (hereinafter also called THF), methanol, and water, in the presence of a base such as sodium hydroxide, at 0–80 °C.

[0114] In step (h), compound 12 can be produced by the synthesis reaction of compound 11, which has a phenolic hydroxyl group, with an aryl ether of an alcohol having an amino group protected by a Boc group.

[0115] The synthesis of aryl ethers of organic compounds with phenolic hydroxyl groups from alcohols is well known. For example, it can be carried out in solvents such as THF at 0–80 °C in the presence of compound 11, the specific alcohols mentioned above, photoresist such as di-2-methoxyethyl azodicarbonate, and organophosphorus compounds such as triphenylphosphine.

[0116] Alternatively, in step (h), compound 12 can be formed by reacting compound 11, which has a phenolic hydroxyl group, with an S-haloalkyl group having an amino group protected by a Boc group. N 2. To produce it through reaction.

[0117] Compound 11, which has a phenolic hydroxyl group, and S of a haloalkyl group N 2. The reaction is well known. For example, it can be carried out in solvents such as DMF, in the presence of compound 11, the specific haloalkyl group mentioned above, a base such as KHCO3, and an iodizing agent such as KI, at 0 to 150 °C.

[0118] In step (i), compound 13 can be manufactured by deprotecting the Boc-protected amino group of compound 12 to form an amino hydrochloride.

[0119] Deprotection of Boc-protected amino groups is well known. For example, it can be carried out in solvents such as dichloromethane, in the presence of acids such as trifluoroacetic acid, at 0–80 °C.

[0120] Reactions that convert amino groups into salts are well known. For example, after deprotection, the reaction can be carried out in ethyl acetate (hereinafter also referred to as AcOEt) in the presence of an acid such as hydrochloric acid at 0–80 °C.

[0121] In step (j), compound 14, which is to be prepared prior to the hydrochloride salt, can be produced by the condensation reaction of the carboxyl group of compound 10 with the secondary amino group of compound 13.

[0122] The condensation reaction of carboxyl and amino groups is well known. For example, it can be carried out in DMF in the presence of condensing agents such as compound 10, compound 13, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (hereinafter also known as HATU), and bases such as diisopropylethylamine (hereinafter also known as i-Pr2NEt) at 0 to 80 °C.

[0123] In step (k), the hydrochloride salt of compound 14 can be manufactured by forming the tertiary amino group in compound 14 into a hydrochloride salt.

[0124] Reactions that form tertiary amino groups into salts are well known. For example, they can be carried out in AcOEt at 0–80 °C in the presence of acids such as hydrochloric acid.

[0125] Based on this, compound 6 can also be manufactured by the method described below as shown in Scheme II.

[0126] [Chemistry 7]

[0127] In Scheme II, compounds 6a and 6b are not only available as commercially available products, but can also be easily synthesized from commercially available products using known methods.

[0128] In the chemical formula, L is the same as the L mentioned above in the description of the compound shown in formula (I).

[0129] A is a substituted or unsubstituted divalent aromatic hydrocarbon group; preferably phenylene, naphthylene, etc.; more preferably 1,2-phenylene, 1,3-phenylene (m-phenylene), 1,4-phenylene (p-phenylene), 1,2-naphthylene, 1,4-naphthylene, 1,5-naphthylene, 2,3-naphthylene, and 2,6-naphthylene, etc.; even more preferably 1,3-phenylene (m-phenylene) and 1,4-phenylene (p-phenylene).

[0130] L' is a straight-chain or branched alkenyl group with 2 to 13 carbon atoms, either substituted or unsubstituted, preferably a straight-chain alkenyl group with 3 to 10 carbon atoms, more preferably a straight-chain alkenyl group with 4 to 9 carbon atoms, and even more preferably a straight-chain alkenyl group with 5 to 8 carbon atoms.

[0131] Hal represents a halogen atom, preferably a chlorine atom, bromine atom, or iodine atom.

[0132] R 6 The alkyl group has 1 to 6 carbon atoms, preferably methyl or ethyl.

[0133] In step (l), compound 6 can be produced by a cross-coupling reaction of unsaturated alcohol 6a with haloaryl 6b using a palladium catalyst.

[0134] Cross-coupling reactions using palladium catalysts are well known. For example, they can be carried out in solvents such as DMF, in the presence of palladium catalysts such as compound 6a, compound 6b, and Pd(OAc)2, bases such as LiOAc and tetrabutylammonium chloride (hereinafter also referred to as n-Bu4NCl), and reaction promoters such as LiCl at 40–190 °C.

[0135] <Application> This compound possesses both KDM5 inhibitory and KDM5 degrading activities, and therefore can be used as a KDM5 inhibitor and a KDM5 degrader, and can be used as a preventive and / or therapeutic agent for KDM5-related diseases in mammals, especially humans.

[0136] This compound can be a chemical probe for targeting proteins that capture physiologically active low-molecular-weight compounds. Specifically, this compound can be converted into an affinity chromatography probe, photoaffinity probe, etc., by introducing a labeling group, linker, etc., into a part different from the structural part necessary for the active expression of the compound, using methods described in J. Mass Spectrum. Soc. Jpn. Vol. 51, No. 5, 2003, pp. 492-498 or WO2007 / 139149, etc.

[0137] Labeling groups, linkers, etc. used for chemical probes can be listed, for example, as groups represented by the following groups (1) to (5).

[0138] (1) Photoaffinity labeling groups (e.g., benzoyl, benzophenone, azide, carbonyl azide, diaziridinyl, enone, diazo, and nitro, etc.) and chemophilic groups (e.g., ketones, carbamoyl, esters, alkylthiols, α, β-unsaturated ketones, esters, Michael acceptors, and ethylene oxide, etc., where the α carbon atom is replaced by a halogen atom) and other protein labeling groups; (2) Linkers that can be cleaved, such as -SS-, -O-Si-O-, monosaccharides (glucosyl, galactosyl, etc.) or disaccharides (lactose, etc.), as well as oligopeptide linkers that can be cleaved by enzyme reactions; (3) Biotin, 3-(4,4-difluoro-5,7-dimethyl-4H-3a,4a-diaza-4-boraza-s-indarsen-3-yl)propionyl and other fishing tag groups; (4) 125 I, 32 P, 3 H, 14Radioactive labeling groups such as C; fluorescent labeling groups such as fluorescein, rhodamine, dansyl, umbelliferone, 7-nitrofuran, and 3-(4,4-difluoro-5,7-dimethyl-4H-3a,4a-diaza-4-borza-s-indarin-3-yl)propionyl; chemiluminescent groups such as fluorescein and luminol; detectable labels such as lanthanide metal ions and radium ions; or (5) Groups that are combined with solid carriers such as glass beads, glass column beds, microporous titration plates, agarose beads, agarose column beds, polystyrene beads, polystyrene column beds, nylon beads, and nylon column beds.

[0139] The probe prepared by introducing the marker group selected from the group consisting of (1) to (5) above into the compound according to the methods described in the above literature can be used as a chemical probe for identifying useful marker proteins for the exploration of novel drug development targets.

[0140] Pharmaceutical Compositions The pharmaceutical compositions disclosed herein (hereinafter collectively referred to as "the pharmaceutical compositions") comprise the compound.

[0141] When this compound is used for pharmaceutical purposes, it may of course be used as a single agent, for example, to (1) assist and / or enhance the effect of the compound for the prevention, treatment and / or improvement of symptoms, (2) improve pharmacokinetics and absorption, reduce the dosage of the compound, and / or (3) reduce the side effects of the compound, or as a compounding agent in combination with other pharmaceutical agents.

[0142] The pharmaceutical composition disclosed herein may contain the present compound in an amount of 0.000001 to 99.5% by weight, preferably 0.000001 to 90% by weight, relative to the total mass of the pharmaceutical composition.

[0143] In combination with other pharmaceutical agents, this compound and other agents can be administered as a combination drug containing both components in a single dose, or the respective formulations can be administered via the same or different routes of administration. Simultaneous administration of different formulations is not required; they can be administered sequentially with a time difference. In the case of sequential administration, the order or method of administration is not particularly limited and can be appropriately adjusted to obtain the desired efficacy.

[0144] The dosage of other agents used in combination with this compound may be appropriately increased or decreased based on their clinical dosage or similar agents. Furthermore, the ratio of this compound to other agents may be appropriately adjusted considering the subject's age, weight, method of administration, time of administration, disease, and condition. Generally, other agents may be combined in the range of 0.01 to 100 parts by weight relative to 1 part by weight of this compound. Multiple other agents may also be used. These other agents may be based on the aforementioned agents and may also have the same mechanism of action as the aforementioned agents. These other agents include not only known agents but also agents to be discovered in the future.

[0145] The dosage of this compound varies depending on age, weight, disease condition, treatment effect, method of administration, and duration of administration. This compound can be administered orally to adults once or several times a day in the range of 0.1 mg to 300 mg, or non-oral or intravenously continuously for 1 hour to 24 hours a day in the range of 0.1 mg to 150 mg.

[0146] Pharmaceutical compositions containing this compound can be formulated into various dosage forms. Examples of dosage forms include, for instance, oral administration preparations (e.g., tablets, capsules, granules, powders, oral liquids, syrups, oral gels, etc.), oral preparations (e.g., oral tablets, oral sprays, oral semi-solid preparations, oral mouthwashes, etc.), injectable preparations (e.g., injections, etc.), dialysis preparations (e.g., dialysis preparations, etc.), inhalation preparations (e.g., inhalers, etc.), ophthalmic preparations (e.g., eye drops, ointments, etc.), otolaryngological preparations (e.g., ear drops, etc.), nasal preparations (e.g., nasal drops, etc.), rectal preparations (e.g., suppositories, rectal semi-solid preparations, enemas, etc.), vaginal preparations (e.g., vaginal tablets, vaginal suppositories, etc.), and skin preparations (e.g., solid topical preparations, liquid topical preparations, sprays, ointments, creams, gels, patches, etc.).

[0147] In the manufacture of oral solid dosage forms, excipients, binders, disintegrants, lubricants, colorants, etc., can be added to this compound as needed to manufacture tablets, granules, powders, and capsules using conventional methods. Furthermore, tablets, granules, powders, and capsules can be coated as required.

[0148] Examples of excipients include lactose, corn starch, and crystalline cellulose; examples of binders include hydroxypropyl cellulose and hydroxypropyl methylcellulose; examples of disintegrants include calcium carboxymethyl cellulose and sodium croscarmellose; examples of lubricants include magnesium stearate and calcium stearate; examples of colorants include titanium dioxide; and examples of coating agents include hydroxypropyl cellulose, hydroxypropyl methylcellulose, and methylcellulose, but these are not the only examples.

[0149] These solid dosage forms, such as tablets, capsules, granules, and powders, typically contain 0.001 to 99.5% by weight, preferably 0.001 to 90% by weight, of this compound relative to the total mass of the solid dosage form.

[0150] In the manufacture of injectable preparations (for intravenous, intramuscular, subcutaneous, and intraperitoneal administration, etc.), pH adjusters, buffers, suspending agents, solubilizers, antioxidants, preservatives, isotonic agents, etc., can be added to this compound as needed, and the injectable preparations can be manufactured using conventional methods. Alternatively, freeze-drying can be performed to prepare a freeze-dried formulation that dissolves upon use.

[0151] Examples of pH adjusters and buffers include organic or inorganic acids and / or their salts; examples of suspending agents include methylcellulose, polysorbate 80, and sodium carboxymethylcellulose; examples of solubilizers include polysorbate 80 and polyoxyethylene sorbitan monolaurate; examples of antioxidants include α-tocopherol; examples of preservatives include methylparaben and ethylparaben; and examples of isotonic agents include glucose, sodium chloride, and mannitol, but these are not the only examples.

[0152] These injections typically contain 0.000001 to 99.5% by weight, preferably 0.000001 to 90% by weight, of this compound relative to the total mass of the injection.

[0153] In the manufacture of topical preparations, base ingredients can be added to this compound, and as needed, preservatives, stabilizers, pH adjusters, antioxidants, colorants, etc., can be added to manufacture, for example, transdermal preparations (ointments, patches, etc.), eye drops, nasal drops, suppositories, etc., by conventional methods.

[0154] As raw materials for the base agent used, various raw materials commonly used in pharmaceuticals, quasi-pharmaceuticals, cosmetics, etc., can be used. Specifically, examples include animal and vegetable oils, mineral oils, ester oils, waxes, emulsifiers, higher alcohols, fatty acids, silicone oils, surfactants, phospholipids, alcohols, polyols, water-soluble polymers, clay minerals, purified water, etc.

[0155] These topical agents typically contain 0.000001 to 99.5% by weight, preferably 0.000001 to 90% by weight, of this compound relative to the total mass of the topical agent.

[0156] The dosage of this compound varies depending on the severity of symptoms, age, sex, weight, form of administration, type of salt, and specific type of disease. Generally, in adults, the oral dosage is about 30 μg to 10 g, preferably 100 μg to 5 g, more preferably 100 μg to 1 g, and the injectable dosage is about 30 μg to 1 g, preferably 100 μg to 500 mg, more preferably 100 μg to 300 mg, administered once or in several divided doses.

[0157] <Application> This compound and the pharmaceutical composition have both KDM5 inhibitory activity and KDM5 degrading activity, and therefore can be used as a KDM5 inhibitor and a KDM5 degrader, and can be used as a preventive and / or therapeutic agent for KDM5-related diseases (mammals, especially humans).

[0158] Examples of such diseases include proliferative disorders, cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, multiple sclerosis, Huntington's disease, Alzheimer's disease, cystic fibrosis, viral diseases, autoimmune diseases, atherosclerosis, restenosis, psoriasis, rheumatoid arthritis, inflammatory bowel disease, asthma, allergic diseases, inflammation, neurological disorders, hormone-related disorders, symptoms associated with organ transplantation, immunodeficiency diseases, destructive bone disorders, proliferative diseases, infectious diseases, symptoms associated with cell death, and thrombin. Induced platelet aggregation, liver disease, pathological immune status associated with T cell activation, central nervous system disorders, myeloproliferative disorders, Parkinson's disease, Lewy body disease, frontotemporal degeneration, mild cognitive impairment, dementia, cerebrovascular disease, schizophrenia, depression, anxiety disorders, bipolar disorder, autism spectrum disorder, attention deficit / hyperactivity disorder, learning disabilities, motor disorders, obsessive-compulsive disorder, personality disorders, sleep disorders, delirium, amyotrophic lateral sclerosis, developmental disorders, intellectual disabilities, post-traumatic stress disorder, hepatitis, etc.

[0159] In particular, this compound and the pharmaceutical composition are useful for the prevention and / or treatment of cancer, Huntington's disease, Alzheimer's disease, Parkinson's disease, Lewy body disease, frontotemporal degeneration, mild cognitive impairment, dementia, cerebrovascular disease, schizophrenia, depression, anxiety disorders, bipolar disorder, autism spectrum disorder, attention deficit / hyperactivity disorder, learning disabilities, motor disorders, obsessive-compulsive disorder, personality disorders, sleep disorders, delirium, amyotrophic lateral sclerosis, developmental disorders, intellectual disabilities, post-traumatic stress disorder, or hepatitis. This compound or the pharmaceutical composition is particularly suitable for the prevention and / or treatment of cancer and Alzheimer's disease.

[0160] When this pharmaceutical composition is used as a single agent or in combination with other pharmaceutical agents for the prevention and / or treatment of the aforementioned diseases, the active ingredient is typically formulated using pharmaceutically acceptable carriers such as various additives or solvents. The resulting formulation is administered systemically or locally, orally or non-orally. Here, a pharmaceutically acceptable carrier refers to a substance other than the active ingredient commonly used in pharmaceutical formulations. Preferably, a pharmaceutically acceptable carrier does not exhibit pharmacological activity at the dosage of the formulation, is harmless, and does not interfere with the therapeutic effect of the active ingredient. Pharmaceutically acceptable carriers can be used to improve the usability of the active ingredient and the formulation, facilitate formulation, stabilize quality, and improve usability. Specifically, substances listed in the 2000 "Dictionary of Pharmaceutical Additives" and the Pharmaceutical Affairs Daily (IPEC JAPAN edition) can be appropriately selected as needed.

[0161] Methods for Manufacturing Pharmaceutical Compositions Pharmaceutical compositions containing this compound can be manufactured by mixing other pharmaceutical agents, carriers, excipients, and other additives into this compound as needed.

[0162] Example The present disclosure will be specifically described below through embodiments, but the present disclosure is not limited to these embodiments.

[0163] The chemical reagents and solvents used in this embodiment are commercially available products of the highest purity. The reagents and solvents were purchased from Sigma Aldrich, Fujifilm and Photochemical Industries, Ltd., TCI Tokyo Chemical Industries, Ltd., NacalaiTesque Ltd., and Kanto Chemical Co., Ltd., and were used without purification.

[0164] Reactions susceptible to air and moisture were conducted entirely in dry glassware under an argon (Ar) atmosphere. NMR spectroscopy was performed at 400 MHz. 1 The JEOLECS 400 spectrometer operates at 700 MHz (H) and at 700 MHz (H) 1 H) or 175MHz 13C) Record the data using a Bruker AVANCE III 700 spectrometer that is operating under these conditions. 1 HNMR and 13 CNMR chemical shift values, relative to solvent peaks or tetramethylsilane (TMS) (DMSO-d6: 1 HNMR 2.50, 13 The δ (ppm) of CNMR39.52 is reported, and the coupling constant is given in Hz.

[0165] Using a Shimadzu UFLC (SPD-M20A UV detector, DGU-20A3R degassing unit, LC-20AD solvent delivery unit, CBM-20A system) and a COSMOSIL packed column (5C18-AR-II, 4.6ID×150mm, NacalaiTesque INC.), the purity of all test compounds was determined by HPLC at a flow rate of 1 mL / min and UV detection (λ=254nm). The results showed that the purity of all test compounds was above 95%.

[0166] HPLC conditions: Eluent A: H2O containing 0.1% TFA; Eluent B: Acetonitrile containing 0.1% TFA.

[0167] Elution buffer B: 0–20 min, 10–90%; 20–30 min, 90%; 30–40 min, 90–10%. Positive / negative LRMS ion mass spectrometry was recorded using a Bruker HCT-Plus. High-resolution mass spectrometry (HRMS) was recorded using an LTQ Orbitrap XL (THERMO) or Shimadzu LCMS-IT-TOF mass spectrometer.

[0168] abbreviation AcOEt: Ethyl acetate; Boc2O = di-tert-butyl dicarbonate; n-Bu4NCl: Tetrabutylammonium chloride; DME: 1,2-Dimethoxyethane; DMEAD = di-2-methoxyethyl azodicarbonate; DMF = N,N-dimethylformamide; DMSO = dimethyl sulfoxide; dppf=1,1'-ferrocenebis(diphenylphosphine); HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; MeCN: Acetonitrile; MeI: Iodomethane; MeOH: Methanol; i-Pr2NEt: diisopropylethylamine; TFA = trifluoroacetic acid; THF = Tetrahydrofuran.

[0169] (1) Synthesis of compounds Example 1: Synthesis of 7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy}ethyl}heptamide hydrochloride (14a-hydrochloride) Compound 14a was synthesized according to the following reaction scheme.

[0170] [Chemistry 8]

[0171] Synthesis of <7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}heptanoic acid (10a)> Step 1: Synthesis of tert-butyl (4-bromophenylethyl) carbamate (2) A solution of 2-(4-bromophenyl)ethylamine (compound 1, 5.00 g, 25.0 mmol) and triethylamine (4.20 mL, 30.1 mmol) in CH₂Cl₂ (50 mL) was cooled in an ice bath while Boc₂O (5.72 g, 26.2 mmol) was added. The resulting mixture was stirred at room temperature for 5 hours. The reaction was then quenched with 10% citric acid and extracted with AcOEt. The organic layer was separated, washed with brine, and dried over MgSO₄. The mixture was filtered, concentrated under reduced pressure, and purified by rapid column chromatography (silica gel, hexane / AcOEt = 9 / 1–4 / 1) to give compound 2 as a colorless solid (5.83 g, 78%).

[0172] 1 HNMR(DMSO-d6,400MHz,δppm)7.46(2H,d,J=8.3Hz),7.15(2H,d,J=8.3Hz),6.86( 1H,t,J=5.2Hz),3.12(2H,td,J=6.8,6.8Hz),2.66(2H,t,J=7.0Hz),1.35(9H,s). Step 2: Synthesis of tert-butyl(4-bromophenylethyl)(methyl)carbamate (3) A DMF (50 mL) solution of compound 2 (5.83 g, 19.4 mmol) was added to an oil suspension of 60% NaH (0.820 g, 20.5 mmol) at 0 °C. After 15 minutes, MeI (1.40 mL, 22.5 mmol) was added dropwise, and the resulting mixture was stirred at 0 °C for 5 hours.

[0173] The reaction mixture was injected into water and extracted with AcOEt. The organic layer was washed with brine and dried over MgSO4. After filtration, concentration under reduced pressure, and purification by rapid column chromatography (silica gel, n-hexane / AcOEt = 9 / 1), compound 3 was given as a colorless oil (6.08 g, 99%).

[0174] 1 HNMR(DMSO-d6,400MHz,δppm)7.47(2H,d,J=8.1Hz),7.15(2H,d,J=8.1Hz),3.36(2H,t,J=6.5Hz),3.32(3H,s),2.72(2H,t,J=6.8Hz),1.25(9H,s). Step 3: Synthesis of tert-butylmethyl [4-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)phenethyl]carbamate (4) A solution of compound 3 (6.08 g, 19.3 mmol), bis(pinacol)diboron (5.70 g, 22.4 mmol), KOAc (7.30 g, 74.4 mmol), and Pd(dppf)Cl2 (670 mg, 0.916 mmol) in DMSO (60 mL) was heated at 80 °C for 7 hours. The reaction mixture was filtered, the filtrate was extracted with AcOEt, and washed with brine.

[0175] The organic layer was separated and dried over MgSO4. After filtration, concentration under reduced pressure, and purification by rapid column chromatography (silica gel, n-hexane / AcOEt=4 / 1), compound 4 was obtained as a colorless solid (5.76 g, 82%).

[0176] 1 HNMR(DMSO-d6,400MHz,δppm),7.59(2H,d,J=7.9Hz),7.21(2H,d,J=7.4Hz),3.36 (2H,t,J=7.2Hz),2.76(2H,t,J=7.2Hz),2.73(3H,s),1.36(9H,s),1.28(12H,s). Step 4: Synthesis of N-methyl-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)phenyl]ethane-1-amine (5) Trifluoroacetic acid (TFA) (16 mL) was added to a CH₂Cl₂ (35 mL) solution of compound 4 (5.76 g, 15.9 mmol), and the mixture was cooled in an ice bath. The resulting mixture was stirred at 0 °C for 2 hours. The solvent was removed under reduced pressure, and the residue was separated by adding a saturated aqueous solution of NaHCO₃ and AcOEt.

[0177] The organic layer was washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by rapid chromatography (silica gel, n-hexane / AcOEt=4 / 1) to give compound 5 as a colorless solid (3.50 g, 84%).

[0178] 1 HNMR(DMSO-d6,400MHz,δppm),8.45(1H,brs),7.65(2H,d,J=8.1Hz),7.28(2H,d ,J=8.3Hz),3.15(2H,t,J=9.2Hz),2.94-2.89(2H,m),2.59(3H,s),1.28(12H,s). Step 5: Synthesis of methyl 7-{methyl[4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)phenethyl]amino} heptanoate (7a) A solution of methyl 7-oxoheptanoate (compound 6a, 401 mg, 2.53 mmol), compound 5 (632 mg, 2.42 mmol), and NaBH(OAc)3 (820 mg, 3.87 mmol) in ClCH2CH2Cl (10 mL) was stirred overnight at room temperature. The reaction was quenched with water and extracted with AcOEt. The organic layer was separated, washed with brine, and dried on MgSO4.

[0179] After filtration, concentration under reduced pressure, and purification by rapid column chromatography (silica gel, CHCl3 / AcOEt=4 / 1 to CHCl3 / MeOH=9 / 1), compound 7a was obtained as a colorless oil (253 mg, 26%).

[0180] 1 HNMR(DMSO-d6,400MHz,δppm),7.57(2H,d,J=8.8Hz),7.22(2H,d,J=8.8Hz),3.57(3H,s),2.74-2.64(2H,m) ,2.35-2.23(4H,m),2.17(3H,brs),1.55-1.43(2H,m),1.41-1.32(2H,m),1.28(12H,s),1.25-1.14(6H,m). Step 6: Synthesis of methyl 7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}heptanoic acid (9a) A solution of compound 7a (147 mg, 0.364 mmol), compound 8 (78.3 mg, 0.331 mmol), Na₂CO₃ (70.2 mg, 0.662 mmol), and Pd(dppf)Cl₂ (24.2 mg, 0.0331 mmol) in DME / H₂O (1.5 mL / 0.5 mL) was heated at 110 °C for 3 hours. The reaction mixture was filtered, the filtrate was extracted with AcOEt, and washed with brine.

[0181] The organic layer was separated and dried over MgSO4. After filtration and concentration under reduced pressure, the mixture was purified by rapid column chromatography (silica gel, CHCl3 / AcOEt=4 / 1 to CHCl3 / MeOH=9 / 1) to give compound 9a as a colorless solid (23.5 mg, 15%).

[0182] 1 HNMR(DMSO-d6,400MHz,δppm),8.03(1H,brs),7.39(2H,d,J=7.9Hz),7.32(2H,d,J=7.9Hz),3.57(3H,s),2.88-2.76(4H,m),2.60 -2.53(3H,m),2.38(3H,s),2.28(2H,t,J=7.2Hz),1.54-1.42(4H,m),1.30-1.23(4H,m),1.18(6H,d,J=8.2Hz);MS(ESI)m / z478(MH + ). Step 7: Synthesis of 7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}heptanoic acid (10a) An aqueous solution of NaOH (4N, 0.200 mL, 0.791 mmol) was added to a MeOH solution (1.2 mL) of compound 9a (126 mg, 0.264 mmol), and the mixture was stirred overnight at room temperature. A portion of a 10% citric acid aqueous solution was added, and the resulting mixture was stirred at room temperature. The insoluble matter was collected by filtration and washed with water to give compound 10a as a colorless solid (66.0 mg, 54%).

[0183] 1HNMR(DMSO-d6,400MHz,δppm),11.9(1H,brs),9.24(1H,brs),8.02(1H,s),7 .33(4H,d,J=9.6Hz),3.11-2.92(2H,m),2.90-2.78(2H,m),2.75-2.68(1H,m) ,2.52-2.50(2H,m),2.51(3H,s),2.21(2H,t,J=7.3Hz),1.68-1.60(2H,m),1. 54-1.48(2H,m),1.36-1.27(4H,m),1.23(6H,d,J=6.9Hz);MS(ESI)m / z464(MH + ). Synthesis of <7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy}ethyl}heptamide hydrochloride (14a-hydrochloride)> Step 1: Synthesis of {2-[2-(2-{[2-(2,6-dioxopiridine-3-yl)-1,3-dioxoindololin-4-yl]oxy}ethoxy)ethoxy]ethyl}carbamate (12) To a solution of 2-(2,6-dioxadiazine-3-yl)-4-hydroxyisoindoline-1,3-dione (compound 11, 300 mg, 1.09 mmol) and PPh3 (430 mg, 1.64 mmol) in THF (3 mL), 2-[2-(2-tert-butoxyaminoethoxy)ethoxy]ethanol (300 mg, 1.20 mmol) and DMEAD (383 mg, 1.64 mmol) were added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with water and extracted with AcOEt.

[0184] The organic layer was separated and dried on MgSO4. After filtration, concentration under reduced pressure, and purification by rapid column chromatography (silica gel, CHCl3 / AcOEt=4 / 1~1 / 1), compound 12 was obtained as a colorless oil (226 mg, 41%).

[0185] 1HNMR(DMSO-d6,400MHz,δppm),11.10(1H,brs),7.85-7.78(1H,m),7.54(1H,d,J=7.8Hz) ,7.46(1H,d,J=7.4Hz),6.74(1H,brs),5.08(1H,dd,J=12.8,5.4Hz),4.35(2H,t,J=4.5H z),3.80(2H,t,J=4.5Hz),3.64(2H,t,J=4.7Hz),3.53-3.47(2H,m),3.40-3.36(2H,m),3 .08-3.01(2H,m),2.94-2.84(1H,m),2.62-2.51(2H,m),2.05-2.01(1H,m),1.36(9H,s). Step 2: Synthesis of 4-(2-(2-aminoethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione hydrochloride (13-hydrochloride) Trifluoroacetic acid (TFA) (0.34 mL) was added to a CH₂Cl₂ (2 mL) solution of compound 12 (226 mg, 0.448 mmol) while cooling in an ice bath. After stirring at 0 °C for 3 hours, the reaction mixture was concentrated under reduced pressure. 4N hydrochloric acid from AcOEt was added to the residue, and the solvent was removed by distillation under reduced pressure to give the crude product, compound 13-hydrochloride (239 mg), which was used in the next reaction without further purification.

[0186] Step 3: Synthesis of 7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy}ethyl}heptamide (14a) HATU (20.0 mg, 0.0526 mmol) was added to a DMF (0.2 mL) solution of compound 10a (19.9 mg, 0.0429 mmol), i-Pr2NEt (0.0180 mL, 0.104 mmol), and the crude product of compound 13-hydrochloride (19.0 mg) at room temperature. The reaction mixture was stirred overnight at room temperature. Water was added to the reaction mixture, followed by extraction twice with AcOEt. The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The filtrate was concentrated and purified by reversed-phase rapid chromatography (MeCN / 0.1% TFA) to give compound 14a, which was converted to hydrochloride in the next step.

[0187] Step 4: Synthesis of 7-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy}ethyl}heptamide hydrochloride (14a-hydrochloride) Compound 14a was treated with 4N hydrochloric acid in AcOEt, and the solvent was removed by distillation under reduced pressure to obtain compound 14a-hydrochloride (5.8 mg, 15% of compound 10a), which was a colorless solid.

[0188] 1 HNMR(DMSO-d6,700MHz,δppm),δ13.44(1H,s),11.13(1H,s),8.41(1H,s),7.88(1H,brs),7.81(1H,t,J=8.4Hz),7.54(1H,d,J=8.6Hz) ,7.50-7.48(4H,m),7.47(1H,d,J=7.0Hz),5.09(1H,dd,J=12.9,5.6Hz),4.34(2H,t,J=4.3Hz),3.81-3.80(2H,m),3.66-3.64(2H,m), 3.53-3.52(2H,m),3.41-3.39(2H,m),3.32-3.25(1H,m),3.19-2.99(6H,m),2.91-2.80(1H,m),2.82(3H,s),2.60-2.58(2H,m),2.55- 2.49(2H,m),2.08-2.06(2H,m),2.04-2.00(1H,m),1.71-1.63(2H,m),1.51-1.46(2H,m),1.31-1.25(4H,m),1.23(6H,d,J=7.0Hz,6H); 13CNMR(DMSO-d6,175MHz,δppm),172.22,171.48,169.37,166.20,164.69,155.18,154.40,147 .27,144.73,142.85,138.43,136.42,132.62,130.91,128.34,128.25,119.38,115.65,114. 81,113.38,112.16,73.37,69.48,68.99,68.56,68.23,68.06,54.83,54.27,48.11,37.80,3 4.47,30.33,28.60,27.75,27.53,25.20,24.35,22.47,21.37,20.47,19.34;HRMScalcdforC 45 H 55 N8O9 + 851.4092, found 851.4077; HPLCR t 12.27 min, 97.84% purity. Example 2: Synthesis of 10-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy}ethyl}decanoamide hydrochloride (14b-hydrochloride) For compound 14b-hydrochloride, methyl 10-oxodecanoate (6b) was used instead of methyl 7-oxoheptanoate (6a), and the synthesis was carried out in the same manner as described in the synthesis of compound 10a and compound 14a-hydrochloride; yellow solid (5.9 mg, 16% of compound 10b).

[0189] 1HNMR(DMSO-d6,700MHz,δppm),13.43(1H,s),11.14(1H,s),8.42(1H,s),7.84-7.81(2H,m),7.54(1H,d,J=8.6Hz),7.50(4H,s),7.47(1H,d,J=7.3Hz),5.10(1H,dd,J=12.9,5.2Hz),4.35(2H,t,J=4.3Hz),3.81(2H,t,J=4.5Hz),3.65(2H,t,J=4.7Hz),3.53(2H,t,J=4.7Hz),3.50-3.35(2H,m),3.32-3.29(1H,m),3.22-3.04(6H,m),2.94-2.86(1H,m),2.85(3H,s),2.61-2.57(2H,m),2.55-2.48(2H,m),2.06-2.02(1H,m),2.04(2H,t,J=7.0Hz),1.72-1.60(2H,m),1.48-1.44(2H,m),1.29-1.23(10H,m),1.23(6H,d,J=7.0Hz); 13 CNMR(DMSO-d6,175MHz,δppm),172.21,171.54,169.35,166.19,164.67,157.35,157.16,155.18,154.39,147.23,144.72,142.84,138.27,136.41,132.62,130.94,128.34,128.25,119.36,115.64,114.80,113.36,112.16,73.36,69.47,68.99,68.56,68.22,68.06,54.87,54.42,52.88,48.10,41.15,37.79,34.63,30.32,28.64,28.11,28.07,28.02,27.88,27.75,25.35,24.60,22.67,21.36,20.45,19.34,17.43,16.09,11.84;HRMScalcdforC 48 H 61 N8O9 + 893.4562,found893.4545;HPLCR t 13.26min,95.59%purity. Example 3: Synthesis of 4-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy)ethoxy]ethyl}benzamide (20a) Compound 20a was synthesized according to the following reaction scheme.

[0190] [Chemistry 9]

[0191] Step 1: Synthesis of ethyl 4-(6-oxohexyl)benzoate (16a) A 10 mL solution of compound 15a (2.42 g, 8.77 mmol), 5-hexen-1-ol (1.05 g, 10.5 mmol), LiOAc (1.44 g, 21.8 mmol), LiCl (371 mg, 8.75 mmol), tetrabutylammonium chloride (1.22 g, 4.39 mmol), and Pd(OAc)₂ (195 mg, 0.869 mmol) in DMF was heated at 70 °C for 6.5 h. The reaction mixture was filtered, the filtrate was extracted with AcOEt, and washed with brine.

[0192] The organic layer was separated and dried over MgSO4. After filtration, concentration under reduced pressure, and purification by rapid column chromatography (silica gel, n-hexane / AcOEt=4 / 1), compound 16a was obtained as a colorless oil (1.96 g, 90%).

[0193] 1 HNMR(DMSO-d6,400MHz,δppm),9.65(1H,t,J=1.5Hz),7.87(2H,d,J=8.4Hz),7.34(2H,d,J=8.4Hz),4.29(2H,q,J=7.1 Hz),2.64(2H,t,J=7.6Hz),2.41(2H,td,J=7.0,2.0Hz),1.63-1.51(4H,m),1.33-1.26(2H,m),1.31(3H,t,J=7.2Hz). Steps 2-4: Synthesis of 4-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)benzoic acid (19a) For compound 19a, it was synthesized from compound 8 and compound 16a using the same steps as described in the synthesis of compound 10a (steps 5-7); yellow solid (25.0 mg, 7% of compound 16a).

[0194] 1 HNMR(DMSO-d6,400MHz,δppm),12.8(1H,brs),8.01(1H,brs),7.86(2H,d,J=7.8Hz),7.33-7.31(6H,m),3.33-3.24(1H,m),3.06 -2.93(4H,m),2.80(3H,brs),2.68-2.64(4H,m),1.67-1.56(4H,m),1.37-1.29(4H,m),1.24(6H,d,J=9.8Hz);MS(ESI)m / z540(MH + ). Step 5: Synthesis of 4-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy)ethoxy]ethyl}benzamide (20a) Compound 20a was synthesized from compound 13-hydrochloride and compound 19a using the same steps as described in the synthesis of 14a-hydrochloride (step 3); yellow solid (9.1 mg, 21%).

[0195] 1 HNMR(DMSO-d6,700MHz,δppm),11.14(1H,s),8.44(1H,t,J=5.6Hz),8.04(1H,s),7.82-7.76(3H,m),7.52(1H,d,J =7.0Hz),7.47(1H,d,J=7.0Hz),7.32-7.26(6H,m),5.10(1H,dd,J=12.9,5.2Hz),4.33-4.23(2H,m),3.84-3.80(2 H,m),3.66(2H,t,J=4.7Hz),3.57-3.53(4H,m),3.43-3.30(6H,m),2.92-2.87(2H,m),2.74(1Ht,J=6.5Hz),2.65- 2.55(5H,m),2.54-2.48(2H,m),2.04-2.00(1H,m),1.63-1.46(4H,m),1.35-1.28(4H,m),1.30(6H,d,J=18.1Hz);13 CNMR(DMSO-d6,175MHz,δppm),172.70,169.85,166.69,166.00,165.16,156.73,155.67,145.54 ,143.81,136.87,133.10,131.75,128.00,127.94,127.16,127.07,126.61,124.80,119.83,116 .14,115.26,110.56,73.97,69.95,69.55,68.81,68.68,68.54,59.65,48.59,34.72,34.27,30. 85,30.82,30.47,30.28,29.11,28.72,28.22,21.96,21.86,20.66,20.44,13.87;HRMScalcdforC 51 H 59 N8O9 + 927.4405, found 927.4390; HPLCR t 13.73 min, 95.89% purity. Example 4: Synthesis of 4-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)-N-{2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}ethoxy)ethoxy]ethyl}benzamide hydrochloride (20b-hydrochloride) For compound 20b-hydrochloride, ethyl 3-(6-oxohexyl)benzoate (16b) was used instead of ethyl 4-(6-oxohexyl)benzoate (16a), and the synthesis was carried out using the same steps as described in the synthesis of compound 20a (steps 1-5) and compound 14a-hydrochloride (step 4); yellow solid (16.6 mg, 17% of compound 8). Furthermore, for compound 16b, compound 15b was used instead of compound 15a, and the synthesis was carried out using the same steps as described in the synthesis of compound 16a (step 1).

[0196] 1HNMR(DMSO-d6,700MHz,δppm),13.44(1H,s),11.15(1H,d,J=5.2Hz,1H),8.51(1H,t,J=5.6Hz),8.43(1H,s),7.82(2H,t,J=8.0Hz),7.74-7.67(2H,m),7.53-7.47(6H,m),7.39-7.36(2H,m),5.10(1H,dd,J=12.9,5.2Hz),4.33(2H,t,J=4.5Hz),3.81(2H,t,J=4.5Hz),3.67(2H,t,J=4.7Hz),3.58(2H,t,J=4.7Hz),3.53(2H,t,J=4.7Hz),3.52-3.37(6H,m),3.35-3.27(1H,m),3.22-3.00(4H,m),2.92-2.87(1H,m),2.86(3H,s),2.65-2.59(4H,m),2.55-2.48(2H,m),2.04-2.01(1H,m),1.70-1.66(2H,m),1.66-1.63(2H,m),1.40-1.32(4H,m),1.25(6H,d,J=7.0Hz); 13 CNMR(DMSO-d6,175MHz,δppm),172.71,171.93,169.86,166.69,166.20,165.17,155.66,154.89,147.74,145.22,143.34,142.15,138.82,136.88,134.24,133.11,131.42,130.98,128.83,128.74,128.04,127.05,124.40,119.84,116.14,115.28,113.86,112.66,73.86,69.96,69.53,68.78,68.69,68.54,55.33,54.83,48.59,34.77,30.82,30.49,29.12,28.25,28.00,25.71,23.07,22.02,21.85,20.96,19.84;HRMScalcdforC 51 H 59 N8O9 + 927.4405,found927.4388;HPLCR t 13.80min,97.80%purity. Example 5: Synthesis of 4-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)-N-(9-{[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}nonyl)benzamide hydrochloride (23a-hydrochloride)] The following reaction scheme was used to synthesize compound 23a-hydrochloride.

[0197] [Chemistry 10]

[0198] Step 1: Synthesis of tert-butyl(6-{[2-(2,6-dioxoperidin-3-yl)-1,3-dioxoisoindoline-4-yl]oxy}hexyl)carbamate (21) A 3 mL solution of compound 11 (274 mg, 1.00 mmol), tert-butyl (6-bromohexyl) carbamate (280 mg, 1.00 mmol), KHCO3 (150 mg, 1.50 mmol), and KI (17.0 mg, 0.102 mmol) in DMF was heated at 80 °C for 14 hours.

[0199] After cooling to room temperature, water and AcOEt were added for separation. The organic layer was washed with brine and dried on MgSO4. After filtration, concentration under reduced pressure, and purification by rapid column chromatography (CHCl3 / AcOEt=10 / 1→1 / 1), compound 21 was obtained as a colorless oil (373 mg, 79%).

[0200] 1 HNMR(DMSO-d6,400MHz,δppm),11.08(1H,brs),7.82-7.78(1H,m),7.51(1H, d,J=8.2Hz),7.44(1H,d,J=7.6Hz),6.75(1H,brs),5.07(1H,dd,J=12.8,5.4H z),4.20(2H,t,J=6.5Hz),2.93-2.90(1H,m),2.93-2.83(2H,m),2.61-2.53( 2H,m),2.04-2.00(1H,m),1.78-1.71(2H,m),1.49-1.26(6H,m),1.36(9H,s). Step 2: Synthesis of 4-[(6-aminohexyl)oxy]-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione hydrochloride (22-hydrochloride) Compound 22-hydrochloride (404 mg) was synthesized from the crude product following the same steps as described in the synthesis of compound 14a-hydrochloride (step 2). It was not further purified and was used directly in the next reaction.

[0201] Step 3: Synthesis of 4-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)-N-(6-{[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}hexyl)benzamide hydrochloride (23a-hydrochloride) For compound 23a-hydrochloride, it was synthesized from compound 19a and compound 22-hydrochloride using the same steps as described in the synthesis of compound 14a-hydrochloride (steps 3 and 4); yellow solid (26.0 mg, 52%).

[0202] 1 HNMR(DMSO-d6,700MHz,δppm),13.43(1H,s),11.12(1H,s),8.42(1H,s),8. 41-8.38(1H,m),7.81(1H,dd,J=7.0,7.0Hz),7.76(2H,d,J=8.2Hz),7.52(1 H,d,J=8.6Hz),7.50-7.48(4H,m),7.44(1H,d,J=6.9Hz),7.27(2H,d,J=8.2 Hz),5.08(1H,dd,J=12.9,5.6Hz),4.21(2H,t,J=6.5Hz),3.32-3.28(1H,m), 3.27-3.24(2H,m),3.20-2.99(4H,m),2.91-2.86(1H,m),2.83(3H,s),2.65 -2.61(2H,m),2.60-2.56(2H,m),2.53-2.47(2H,m),2.03-2.00(1H,m),1.79 -1.75(2H,m),1.73-1.65(2H,m),1.65-1.57(2H,m),1.57-1.52(2H,m),1.5 2-1.46(2H,m),1.41-1.36(2H,m),1.36-1.31(4H,m),1.24(6H,d,J=7.0Hz); 13CNMR(DMSO-d6,175MHz,δppm),172.20,169.37,166.23,165.28,164.70,155.35,154.38,147.23, 144.76,144.71,142.83,136.42,132.60,131.60,130.92,128.32,128.23,127.47,126.55,126.0 8,119.12,115.54,114.51,113.35,112.14,73.35,68.08,54.82,54.35,48.06,34.14,30.31,29. 83,28.62,28.50,27.74,27.48,25.53,25.19,24.45,22.58,21.35,20.45,19.33;HRMScalcdforC 51 H 59 N8O7 + 895.4507, found 895.4490; HPLCR t 15.16 min, 95.44% purity. Example 6: Synthesis of 3-(6-{[4-(3-cyano-6-isopropyl-7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidin-5-yl)phenethyl](methyl)amino}hexyl)-N-(6-{[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindoline-4-yl]oxy}hexyl)benzamide hydrochloride (23b-hydrochloride)) For compound 23b-hydrochloride, the same steps as described in the synthesis of compound 23a-hydrochloride (steps 1-3) were used for synthesis; yellow solid (10.4 mg, 11%).

[0203] 1HNMR(DMSO-d6,700MHz,δppm),13.43(1H,s),11.12(1H,s),8.47-8.41(2H,m),7.80(1H,dd,J=8.0,8.0Hz),7.71-7.64(2H,m),7.52-7. 43(6H,m),7.36-7.34(2H,m),5.08(1H,dd,J=12.9,5.6Hz),4.20(2H,t,J=6.5Hz),3.33-3.30(1H,m),3.28-3.24(2H,m),3.17-3.00(4H . 73-1.66(2H,m),1.65-1.59(2H,m),1.57-1.52(2H,m),1.52-1.47(2H,m),1.41-1.37(2H,m),1.37-1.31(4H,m),1.23(6H,d,J=7.0Hz); 13 CNMR(DMSO-d6,175MHz,δppm),172.20,169.37,166.22,165.51,164.71,155.35,141.62,13 6.42,134.06,132.60,130.31,128.30,128.20,127.49,126.50,123.87,119.12,115.54,114 .52,73.35,68.08,54.80,54.28,48.06,34.28,30.31,30.00,28.59,28.47,27.74,27.50,2 5.55,25.21,24.45,23.62,22.52,21.52,21.45,21.35,20.45,19.34,13.36;HRMScalcdforC 51 H 59 N8O7 + 895.4507, found 895.4494; HPLCR t 15.26 min, 95.66% purity. (2) Analysis of neural synapse growth Mouse Neuro-2a (N2a) cell lines were obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank. N2a cells were cultured in Dulbecco modified Eagle medium (hereinafter also referred to as DMEM) containing high glucose, 10% FBS, 100 U / mL penicillin, and 100 μg / mL streptomycin at a concentration of 1 × 10⁻⁶ cells / mL. 4 The concentration of cells / mL was determined, and the plates were plated at 37°C under a humidified atmosphere of 5% CO2.

[0204] In the neural synapse growth analysis, the culture medium was changed to DMEM supplemented with 2% FBS. After incubation with the test compounds, cell morphology was observed using a microscope (Olympus CKX41).

[0205] Differentiated cells were defined as those with at least one neural synapse having a length more than twice the diameter of the cell body. Results were expressed as the proportion of differentiated cells in the counted cells. These experiments were repeated three times.

[0206] (3) Protein blotting N2a cells (5 × 10) 5Cells (2 mL / culture dish) were treated with the indicated concentration of the test compound for 24 hours in cell culture medium supplemented with 10% FBS. Cells were then recovered and extracted with SDS sample buffer. The protein concentration of the lysates was determined using BCA protein assay. Equal amounts of protein were separated from each lysate using a 5%–20% SDS-polyacrylamide gel, and the bands were transcribed onto PVDF membranes (Millipore, β-actin detection #IPVH00010, H3, H3K4me3, H3K27Ac detection #ISEQ10100). After blocking with TBS-T containing 5% skim milk, the transferred membranes were probed with a single antibody. The following antibodies were used as the primary antibodies: rabbit monoclonal KDM5A antibody (CST#3876, 1:1000 dilution); mouse monoclonal β-actin antibody (SantaCruz, #sc-47778, 1:2000 dilution); rabbit polyclonal H3K4me3 antibody (Abcam, #ab8580, 1:5000 dilution); rabbit polyclonal H3K27Ac antibody (Abcam, #ab4729, 1:1000 dilution); or rabbit polyclonal histone H3 antibody (Abcam, #ab1791, 1:200000 dilution). The membrane was washed three times with TBS-T and incubated with ECL rabbit IgGHRP-binding whole antibody (GE HealthcareLifeScience, #NA934, 1:2500 dilution) or ECL mouse IgGHRP-binding whole antibody (GE HealthcareLifeScience, #NA931, 1:2500 dilution), followed by three more washes with TBS-T. Immunoblots were visualized by using enhanced chemiluminescence with a chemiluminescent HRP substrate (Millipore, #P90718).

[0207] (4) Results and Investigation Prior to the study of KDM5PROTAC, it was suggested that compounds inhibiting KDM5A could be therapeutic candidates for neurodegenerative diseases, including Alzheimer's disease. Therefore, synaptic growth analysis, frequently used in the early screening of drugs for neurodegenerative diseases, was employed to investigate compound 24 (…). Figure 19 Experiment (A) was conducted. Figure 19As shown in (B), even culturing mouse neuroblastoma N2a cells with 0.2 μM compound 24 for 24 or 48 hours failed to strongly induce synaptic growth. This indicates that previous KDM5 inhibitors that only inhibit the catalytic function of KDM5A do not exhibit strong synaptic growth activity. Based on the hypothesis that the overall function of KDM5A in synaptic growth is important, as it involves a scaffold that controls histone deacetylation by interacting with HDAC1 and HDAC2, the synaptic growth-promoting activity of combining a KDM5 inhibitor with the HDAC inhibitor vorinostat (25) was investigated. Figure 19 (A)). It is worth noting that this combination significantly induced synaptic growth compared to compound 24 or compound 25 alone. Figure 19 (B) This indicates that inducing histone methylation and acetylation through the inhibition of KDM5A and HDAC, respectively, is an effective means of promoting synaptic growth. Based on these results, it is believed that TPD-based degradation of KDM5A disrupts both its catalytic and scaffold functions, thus strongly inducing synaptic growth in N2a cells. This idea led to the synthesis of a KDM5PROTAC candidate exhibiting potent synaptic growth activity.

[0208] also, Figure 19 (A) represents the structures of KDM5 inhibitor 24 and vorinostat (25). Figure 19 (B) in the figure represents the effects of individual treatments with compounds 24 and 25, and the effects of co-treatment with compounds 24 and 25, on N2a differentiation after 24 and 48 hours of treatment. The bar charts represent the mean ± SD of the three independent experiments. p-values ​​were determined using Tukey's multiple comparison test, *p < 0.05. Representative images of N2a cells treated with compounds 24 and 25 alone, and N2a cells treated with both compounds 24 and 25 are shown below. Figure 20 .

[0209] To design KDM5PROTAC, compound 24 was chosen as the KDM5 ligand. Simulations of compound 24 binding to KDM5A revealed that the hexyl chain oriented towards the protein surface. Based on this simulation, a linker was introduced at the end of the hexyl group.

[0210] Since linker structure largely affects degradation activity, various linkers were explored, including amide-bonded polyethylene glycol linkers (14) and linkers containing benzene rings (20 and 23). Figure 21 (A)). Furthermore, thalidomide (26), which is most widely used in the PROTAC study, was selected as the E3 ligand.

[0211] Next, compounds 14a, 14b, 20a, 20b, 23a, and 23b were synthesized as novel KDM5PROTAC candidates. Figure 21 In (A) (referring to the chemical formula above), synthetic compounds are screened through neural synaptic growth activity analysis. For example... Figure 21 As shown in (B), treatment of N2a cells with compounds 14a, 14b, 20a, 20b, 23a, and 23b promoted synaptic growth. Compound 14b exhibited a stronger synaptic growth-promoting effect than compound 14a. This suggests that the introduction of a spacer between the hexyl group of the KDM5 inhibitor and the amide group of the linker is important for synaptic growth-promoting activity. Furthermore, by substituting the spacer with a benzene ring, it is expected to contribute to the rigidity of the linker orientation, maintaining or increasing this effect. Importantly, the meta-substituted compounds 20b and 23b showed better activity than the para-substituted compounds 20a and 23a. Further, the carbon linker compound 23b showed slightly better activity than the polyethylene linker compound 20b. Through preliminary structure-activity correlation studies, it was clarified that compounds with linkers containing meta-substituted benzene rings exhibit strong synaptic growth-promoting activity. As a result, compounds 20b and 23b showed significant synaptic growth-promoting activity.

[0212] In addition, experiments were conducted on compounds 20b and 23b at different concentrations and incubation times. Figure 21 (C) and Figure 21 (D)). Both compounds promoted synaptic growth in the range of 0.02-2 μM. Figure 21 In (C) of the study, when applied at 0.02 μM, the number of neural synaptic cells increased in a time-dependent manner. Figure 21 (D) Furthermore, the effect of compound 26, as an E3 ligand, on synaptic growth activity was investigated (D). Figure 21 (C)). Importantly, compound 26 did not show strong synaptic growth activity compared to compounds 20b and 23b. These results suggest that strong synaptic growth is mediated by the degradation of KDM5 in PROTAC compounds.

[0213] also, Figure 21 (A) in the text represents the design of KDM5PROTAC. Figure 21In the diagram, (B) represents the effect of 0.2 μM compounds 14a, 14b, 20a, 20b, 23a, and 23b on N2a differentiation after 48 hours of treatment. Figure 21 In the figure, (C) represents the dose-dependent N2a cell synaptic growth activity of compounds 20b, 23b, and 26 after 48 hours of treatment. Figure 21 In the figure, (D) represents the time-dependent synaptic growth activity of N2a cells at 0.02 μM for compounds 20b and 23b. The bar chart represents the mean ± SD of three independent experiments. Representative images of N2a cells treated with the test compounds are shown below. Figures 22-24 .

[0214] Next, Western blot analysis was used to investigate the KDM5A degradation activity of compounds 20b and 23b, confirming their role as KDM5PROTACs. Treatment of N2a cells with compounds 20b and 23b resulted in a dose-dependent decrease in KDM5A levels. Figure 25 (A)). The combination of compounds 24 and 26, which are the parent compounds of compounds 20b and 23b, did not reduce the KDM5A level ( Figure 25 (B)). Furthermore, the reduction in KDM5A levels induced by compounds 20b and 23b was hindered by the proteasome inhibitor MG-132. Figure 25 (C) and Figure 25 (D)). These results suggest that compounds 20b and 23b act as PROTACs, reducing KDM5A levels in N2a cells.

[0215] also, Figure 25 Western blot analysis of KDM5A levels in N2a cells was shown. Figure 25 (A) in the text indicates that after treatment with compounds 20b and 23b for 24 hours, Figure 25 (B) indicates treatment with compound 23b for 24 hours, and treatment with compounds 24 and 26 together for 24 hours. Figure 25 (C) indicates that after treatment with compound 20b and the proteasome inhibitor MG-132 for 24 hours, Figure 25 (D) in the text indicates treatment with the proteasome inhibitor MG-132 and compound 23b for 24 hours.

[0216] Finally, the effects of PROTAC compounds 20b and 23b on histone methylation and acetylation were investigated. Since H3K4me3 is a substrate of KDM5s, the methylation level of H3K4 in N2a cells treated with KDM5PROTAC was analyzed. Figure 26 As shown, H3K4me3 levels accumulated in a dose-dependent manner when treated with 20b and 23b. This suggests that 20b and 23b inhibit the catalytic function of KDM5s in N2a cells. Subsequently, it was investigated whether compounds 20b and 23b affected the level of acetylated lysine 27 (H3K27Ac) of histone H3 through interactions with HDAC1 and HDAC2, which are direct substrates of HDAC1 and HDAC2 and indirect substrates of KDM5s. As expected, 20b and 23b also increased H3K27Ac levels. Figure 26 Furthermore, compounds 20b and 23b strongly inhibited KDM5A in in vitro analyses without affecting HDAC1. Figure 27 Here, in evaluating the inhibitory activity against HDAC1, vorinostat (25), an HDAC inhibitor, was used as a positive control, exhibiting 99% inhibition at 1 μM. These results indicate that degradation of KDM5A based on 20b and 23b simultaneously inhibits both the catalytic function of KDM5s controlling H3K4me3 and the scaffold function of KDM5s that impedes the deacetylation activity of HDAC1 and HDAC2 in N2a cells.

[0217] also, Figure 26 Western blot analysis of H3K4me3 and H3K27Ac levels in N2a cells after treatment with compounds 20b and 23b for 24 hours.

[0218] (5) Conclusion While the potential effectiveness of KDM5s inhibition in neurodegenerative diseases has been reported, in this study, the existing KDM5 inhibitor 24 failed to induce strong synaptic growth activity in N2a cells. On the other hand, the synergistic effect of compound 24 with the HDAC inhibitor vorinostat (25) significantly induced synaptic growth in N2a cells, suggesting that both the inhibition of KDM5's catalytic activity and its binding as a scaffold for HDAC influence synaptic growth. Therefore, focusing on KDM5PROTACs capable of inhibiting both of these functions, compounds 14a, 14b, 20a, 20b, 23a, and 23b were synthesized as KDM5PROTAC candidates. Among these, compounds 20b and 23b significantly promoted synaptic growth in N2a cells. Furthermore, biological analysis revealed that when N2a cells were treated with compounds 20b and 23b, KDM5A was degraded via proteasome degradation mediated by PROTAC. This degradation, in contrast to previous KDM5 inhibitors, affects the overall function of KDM5, thus suggesting that KDM5PROTAC holds promise as a treatment for neurological diseases.

[0219] Industrial applicability This compound has shown significant pharmacological effects, including anticancer activity, in cell and animal experiments.

Claims

1. A compound or a salt thereof, wherein, The compound or a salt thereof is represented by the following formula (I). [Chemistry 1] In formula (I), A is a single bond or a substituted or unsubstituted divalent aromatic hydrocarbon group, L is a straight-chain or branched alkylene group with 1 to 13 substituted or unsubstituted carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 8, and t is a number from 2 to 10.

2. The compound or a salt thereof according to claim 1, wherein, In formula (I), A is a single bond or a phenylene group, L is a straight-chain alkylene group with 3 to 10 carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s -、-(CH(CH3)CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

3. The compound or a salt thereof according to claim 1, wherein, In formula (I), A is a single bond, p-phenylene, or m-phenylene, L is a straight-chain alkylene group with 3 to 10 carbon atoms, and X is -(CH2CH2O). s -、-(CH2CH2CH2O) s - or - (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

4. The compound or a salt thereof according to claim 1, wherein, The compound is a compound represented by formula (I-1), a compound represented by formula (I-2), or a compound represented by formula (I-3). [Chemistry 2] In equation (I-1), L 1 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 1 For -(CH2CH2O) s - The group shown, s is a number from 1 to 5, in formula (I-2), L 2 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 2 For -(CH2CH2O) s -or- (CH2) t The O- group represents a number from 1 to 5, and t represents a number from 2 to 10. In formula (I-3), L 3 X is a straight-chain alkylene group with 3 to 10 carbon atoms. 3 For -(CH2CH2O) s -or- (CH2) t For the O- group, s is a number from 1 to 5, and t is a number from 2 to 10.

5. The compound or a salt thereof according to claim 4, wherein, The compound is the following compound: In the above formula (I-1), L 1 It is a straight-chain alkylene group with 5 carbon atoms, X 1 For -(CH2CH2O) s - The group shown is 3; In the above formula (I-1), L 1 It is a straight-chain alkylene group with 8 carbon atoms, X 1 For -(CH2CH2O) s - The group shown is 3; In the above formula (I-2), L 2 It is a straight-chain alkylene group with 5 carbon atoms, X 2 For -(CH2CH2O) s - The group shown, s is 3; In the above formula (I-2), L 2 It is a straight-chain alkylene group with 5 carbon atoms, X 2 For -(CH2) t The group represented by O-, and the number t is 6; In the aforementioned formula (I-3), L 3 It is a straight-chain alkylene group with 5 carbon atoms, X 3 For -(CH2CH2O) s - The group shown, s is 3; or In the aforementioned formula (I-3), L 3 X is a straight-chain alkylene group with 5 carbon atoms. 3 For -(CH2) t The O- group represents a number t of 6.

6. A pharmaceutical composition, wherein, The pharmaceutical composition comprises any one of the compounds or salts thereof according to claims 1 to 5.

7. The pharmaceutical composition according to claim 6, wherein, The pharmaceutical composition is a KDM5 inhibitor and a KDM5 degrader.

8. The pharmaceutical composition according to claim 6, wherein, The pharmaceutical composition is a preventive and / or therapeutic agent for KDM5-related diseases.

9. The compound or a salt thereof according to any one of claims 1 to 5, wherein, The compound or its salts are effective for the prevention and / or treatment of KDM5-related diseases by inhibiting and degrading KDM5.

10. The pharmaceutical composition according to claim 6, wherein, The pharmaceutical composition is used for the prevention and / or treatment of KDM5-related diseases with effective KDM5 inhibition and KDM5 degradation.

11. A method for preventing and / or treating KDM5-related diseases that are effective against KDM5 inhibition and KDM5 degradation, wherein, The method comprises administering to a patient the compound or a salt thereof as described in any one of claims 1 to 5.

12. A method for preventing and / or treating KDM5-related diseases that are effective against KDM5 inhibition and KDM5 degradation, wherein, The method comprises administering the pharmaceutical composition of claim 6 to a patient.

13. The use of a compound or a salt thereof in the preparation of a pharmaceutical composition, wherein, The compound or its salt is the compound or its salt according to any one of claims 1 to 5, and the pharmaceutical composition is used to prevent and / or treat KDM5-related diseases that are effective in inhibiting and degrading KDM5.

14. A preventive and / or therapeutic agent for KDM5-related diseases, wherein, The preventive and / or therapeutic agent for KDM5-related diseases comprises the compound or salt thereof as any one of claims 1 to 5 as an active ingredient.

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