Nitrogen-containing oxaspirocyclic compounds and uses thereof

Abstract

The application relates to a nitrogen-containing oxaspirocyclic compound and application thereof, in particular to a compound of formula (I) or a pharmaceutically acceptable salt thereof and application of the compound as an LSD1 inhibitor in treating blood tumors, small cell lung cancer, squamous non-small cell lung cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, glioma or Ewing's sarcoma and the like.

Description

[0001] This application claims the following priority:

[0002] CN202110694019.0, June 22, 2021. Technical Field

[0003] This invention relates to nitrogen-containing oxospirocyclic compounds and their applications, specifically to compounds of formula (I) or pharmaceutically acceptable salts thereof. Background Technology

[0004] Histone post-translational modifications, including methylation, acetylation, phosphorylation, and ubiquitination, are important regulatory mechanisms in epigenetics, influencing gene expression by altering chromatin structure [Xueshun Wang, Boshi Huang, Takayoshi Suzuki et al., Epigenomics, 2015, 1379-1396]. Although these modifications do not change the basic DNA sequence, these epigenetic changes may persist throughout the cell life cycle or during cell iteration through cell division [Adrian Bird, Nature, 2007, 396-398]. Therefore, epigenetic dysfunction is closely related to the pathological processes of various diseases [James T Lynch, William J Harris & Tim CP Somervaille, Expert Opin. Ther. Targets, 2012, 1239-1249], such as various solid tumors, hematologic malignancies, viral infections, and neurological disorders. Consequently, epigenetics has become a research hotspot in drug development. Histone methylation is regulated by both histone methyltransferases and histone demethylases. Lysine-specific demethylase 1 (LSD1, also known as KDM1A) was the first reported histone lysine demethylase. By regulating the methylation status of histone lysine, it participates extensively in transcriptional regulation, influencing numerous physiological processes such as cell proliferation and differentiation, and embryonic stem cell pluripotency. [Yujiang Shi, Fei Lan, Caitlin Matson et al., Cell, 2004, 941–953][Daniel P. Mould, Alison E. McGonagle, Daniel H. Wiseman et al., Medicinal Research Reviews, 2015, 35, 586–618]. The LSD1 structure consists of three main parts: an N-terminal SWIRM domain, a C-terminal aminooxidase domain (AOL), and a central Tower domain. [Ruchi Anand, Ronen Marmorstein, Journal of Biological Chemistry, 2007, 35425–35429]. The C-terminal aminooxidase domain includes two active pockets: one is the FAD-binding site, and the other is the site for recognizing and binding to the substrate [Pete Stavropoulos, Günter Blobel, André Hoelz, Nature Structral & Molecular Biology, 2006, 626-632].The function of the SWIRM domain is not yet fully understood. It does not directly participate in the binding of FADs or substrates, but mutations or removals in this region reduce LSD1 activity. Therefore, it is speculated that this region may affect the function of the active domain by adjusting its conformation [Yong Chen, Yuting Yang, Feng Wang et al., Biochemistry, 2006, 13956–13961]. The Tower domain is the binding domain for LSD1 to other protein factors. After binding to different protein factors, LSD1 acts on different substrates, thereby playing different regulatory roles on histones and gene expression. For example, when LSD1 binds to CoREST, it preferentially acts on histone H3K4, removing activation-related histone markers and inhibiting gene transcription through demethylation. When it binds to androgen receptor proteins, recombinant LSD1 preferentially acts on H3K9, activating androgen receptor-related gene transcription through demethylation [Ruchi Anand, Ronen Marmorstein, Journal of Biological Chemistry, 2007, 35425–35429; Eric Metzger, Melanie Wissmann, Na Yin et al., Nature, 2005, 436-439.]. Furthermore, LSD1 also regulates the methylation status of some non-histone substrates, including the tumor suppressor gene p53 and DNA methyltransferase 1 (DNMT1) [Yi Chao Zheng, Jinlian Ma, Zhiru Wang, Medicinal Research Reviews, 2015, 1032–1071].

[0005] LSD1 is a FAD-dependent amino oxidase, with proton transfer considered its most likely oxidation mechanism [Zheng YC, Yu B, Chen ZS, et al. Epigenomics, 2016, 8, 651-666.]. First, through proton transfer, the N-CH3 bond of the substrate is converted to an imine bond. This imine ion intermediate undergoes hydrolysis, producing a demethylated amine on one side and formaldehyde on the other. In this catalytic cycle, FAD is reduced to FADH2, which is then oxidized back to FAD by a molecule of oxygen, simultaneously generating a molecule of H2O2 [Yujiang Shi, Fei Lan, Caitlin Matson, Cell, 2004, 941–953].

[0006] LSD1 is aberrantly expressed in various types of tumors. It is highly expressed in acute myeloid leukemia (AML) subtypes and is an important factor in maintaining the potential of leukemia stem cells (LSCs). LSD1 is also highly expressed in various solid tumors such as lung cancer, breast cancer, prostate cancer, liver cancer, and pancreatic cancer, and is closely associated with poor tumor prognosis. LSD1 inhibits cadherin expression, which is closely related to tumor invasion and epithelial-mesenchymal transition (EMT) [Hosseini A, Minucci S. Epigenomics, 2017, 9, 1123-1142.].

[0007] Currently, no LSD1 inhibitors have been approved for marketing, but eight drugs are in clinical trials, primarily for the treatment of hematological malignancies, small cell lung cancer, and Ewing's sarcoma. However, given the significant unmet market demand, this field still needs candidate compounds with better activity and pharmacokinetic parameters to advance into clinical trials to meet therapeutic needs. Summary of the Invention

[0008] This invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0009]

[0010] in,

[0011] m is 1 or 2;

[0012] n is 0, 1, or 2;

[0013] g can be 0, 1, or 2.

[0014] In some embodiments of the present invention, n is 0 or 1, and other variables are as defined in the present invention.

[0015] In some embodiments of the present invention, g is 0 or 1, and other variables are as defined in the present invention.

[0016] In some embodiments of the present invention, when m is 1, n is 1, g is 1, and other variables are as defined in the present invention.

[0017] In some embodiments of the present invention, when m is 2, n is 1, g is 1, and other variables are as defined in the present invention.

[0018] In some embodiments of the present invention, when m is 1, n is 0, g is 0, and other variables are as defined in the present invention.

[0019] In some embodiments of the present invention, the above-mentioned structural unit for Other variables are as defined in this invention.

[0020] Some solutions in this invention are derived from arbitrary combinations of the above-mentioned variables.

[0021] In some embodiments of the present invention, the above-mentioned compound is

[0022]

[0023] in,

[0024] m, n, and g are as defined in this invention.

[0025] The present invention also provides compounds of the following formula or pharmaceutically acceptable salts thereof.

[0026]

[0027] In some embodiments of the present invention, the above-mentioned compound is,

[0028]

[0029] In some embodiments of the present invention, the salts described above are selected from hydrochloride and p-benzenesulfonate.

[0030] The present invention also provides the use of the above-mentioned compounds or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating LSD1-related diseases.

[0031] In some embodiments of the present invention, the aforementioned diseases are hematologic malignancies, small cell lung cancer, squamous non-small cell lung cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, glioma, or Ewing's sarcoma.

[0032] In some embodiments of the present invention, the aforementioned hematologic malignancy is preferably human acute myeloid leukemia.

[0033] The present invention also provides a method for treating LSD1-related diseases in subjects in need, the method comprising providing the subject with an effective dose of a compound defined by any of the above-described technical solutions or a pharmaceutically acceptable salt thereof.

[0034] The present invention also provides a method for treating hematologic malignancies, small cell lung cancer, squamous non-small cell lung cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, glioma, or Ewing's sarcoma in a subject in need, the method comprising providing the subject with an effective dose of any of the above-described technical solutions of the compound or a pharmaceutically acceptable salt thereof.

[0035] The present invention also provides a method for treating hematologic malignancies, wherein the hematologic malignancies are human acute myeloid leukemia.

[0036] Technical effect

[0037] As a novel LSD1 inhibitor, the compound of this invention exhibits significant in vitro activity and good pharmacokinetic properties.

[0038] Definitions and Explanations

[0039] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0040] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0041] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0042] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0043] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0044] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0045] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0046] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0047] Unless otherwise stated, "(+)" indicates right-handed rotation, "(-)" indicates left-handed rotation, and "(±)" indicates racemic rotation.

[0048] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and straight dashed key

[0049] Unless otherwise stated, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom in the double bond is attached to two different substituents (in a double bond containing a nitrogen atom, the lone pair of electrons on the nitrogen atom is considered as one of the substituents it is attached to), if the atoms in the double bond and their substituents in the compound are separated by a wavy line... The ligature indicates that the compound exists as a (Z) type isomer, an (E) type isomer, or a mixture of both isomers. For example, formula (A) indicates that the compound exists as a single isomer of formula (A-1) or formula (A-2) or as a mixture of the two isomers of formula (A-1) and (A-2); formula (B) indicates that the compound exists as a single isomer of formula (B-1) or formula (B-2) or as a mixture of the two isomers of formula (B-1) and (B-2); formula (C) indicates that the compound exists as a single isomer of formula (C-1) or formula (C-2) or as a mixture of the two isomers of formula (C-1) and (C-2).

[0050]

[0051]

[0052] Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers with different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. Tautomers can be chemically equilibrated if possible (e.g., in solution). For example, proton tautomers (also called prototropic tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions involving the rearrangement of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the tautomers pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0053] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0054] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0055] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated, and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0056] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0057] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0058] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0059] When a substituent is vacant, it means that the substituent does not exist. For example, if X is vacant in AX, it means that the structure is actually A.

[0060] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (such as a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonates; chlorine, bromine, and iodine; sulfonate groups, such as methanesulfonates, toluenesulfonates, p-bromobenzenesulfonates, p-toluenesulfonates, etc.; acyloxy groups, such as acetoxy groups, trifluoroacetoxy groups, etc.

[0061] The term "protecting group" includes, but is not limited to, "amino protecting group," "hydroxy protecting group," or "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the nitrogen position of an amino group. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenemethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), triphenylmethyl (Tr), 1,1-di-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxyl protecting group" refers to a protecting group suitable for preventing hydroxyl side reactions. Representative hydroxyl protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanolyl groups (e.g., acetyl); arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (diphenylmethyl, DPM); silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc.

[0062] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0063] The structures of the compounds of this invention can be confirmed using conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) can be used. The cultured single crystal is analyzed using a Bruker D8venture diffractometer to collect diffraction intensity data. The light source is CuKα radiation, and the scanning mode is φ / scan. After collecting the relevant data, the crystal structure can be further analyzed using the direct method (Shelxs 97) to confirm the absolute configuration. Detailed Implementation

[0064] The present invention will be described in detail below with reference to embodiments, but this does not imply any adverse limitation on the invention. The present invention has been described in detail, and specific embodiments thereof have been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope thereof.

[0065] Example 1

[0066]

[0067] Synthesis route:

[0068]

[0069] first step

[0070] At 0–10 °C, 1M sodium hydroxide (10.5 L), compound 1-1 (2502.41 g, 8.77 mol), and dichloromethane (7.5 L) were added sequentially to a reaction vessel. The reaction vessel temperature was adjusted to 20–30 °C, and the mixture was stirred for 1–2 hours. The pH of the aqueous phase was monitored; when the pH was alkaline and the reaction solution was clear, the reaction was stopped. The reaction solution was transferred to a separatory funnel, allowed to stand for separation, and the aqueous phase was extracted with dichloromethane (4.0 L × 2). The combined organic phases were washed with water (6.3 L × 1), then with saturated brine (6.3 L × 1), allowed to stand for separation, and the organic phase was dried with anhydrous sodium sulfate (629.31 g). The mixture was filtered, and the filtrate was concentrated under reduced pressure at an external temperature of 40–50 °C to obtain compound 1-2. 1 H NMR (400MHz, CD3OD) δ7.23-7.17(m,2H),7.13-7.07(m,1H),7.04-6.97(m,2H),2.43-2.41(m,1H),1.85-1.84(m,1H),1.03-0.93(m,2H).

[0071] Step 2

[0072] Anhydrous dichloromethane (19.0 L) and compound 1-2 (1087.12 g, 8.16 mol) were dissolved in dichloromethane (3.0 L) and added sequentially to the reactor at 15–25 °C. Compound 1-3 (2349.70 g, 9.20 mol) was added to the reactor in portions at 0–10 °C, followed by sodium borohydride acetate (4382.43 g, 20.68 mol) in portions. The reactor temperature was adjusted to 15–25 °C, and the mixture was stirred for 2–3 hours. The reaction solution was slowly added to another reaction vessel containing saturated sodium carbonate aqueous solution (32.0 L) for quenching. The temperature of the reaction vessel was controlled at 15-25℃. The mixture was introduced into a separatory funnel, allowed to stand for separation, and the aqueous phase was extracted with dichloromethane (12.0 L × 2). The organic phases were combined, washed once with saturated sodium chloride aqueous solution (6.0 L × 1), allowed to stand for separation, and the organic phase was dried with anhydrous sodium sulfate (8048.34 g). The mixture was filtered, and the filtrate was concentrated under reduced pressure at an external temperature of 40-50℃ to obtain compounds 1-4. 1 ¹H NMR (400MHz, CD₃OD) δ 7.24–7.21 (m, 2H), 7.14–7.10 (m, 1H), 7.05–7.03 (m, 2H), 4.00–3.97 (m, 1H), 3.69–3.55 (m, 4H), 3.35–3.32 (m, 2H), 2.30–2.27 (m, 1H), 2.13–2.07 (m, 1H), 1.94–1.86 (m, 1H), 1.71–1.58 (m, 4H), 1.53–1.49 (m, 1H), 1.46 (s, 9H), 1.10–0.99 (m, 2H). MS-ESI calculated value [M+H]+373, measured value 373.

[0073] Step 3

[0074] Compounds 1-4 (3390.58 g, 9.10 mol) were dissolved in acetonitrile (56.0 L × 2). Compound (-)-O,O-di-p-toluamide-L-tartaric acid (3366.99 g, 8.72 mol) was added to the reaction solution. The reaction solution was stirred at 55 °C for 2 hours, then cooled to 25 °C and stirred for 12 hours. The reaction solution was filtered, and the solid was washed with acetonitrile (8.0 L × 2) and dried under vacuum at 40 °C to obtain a solid. The solid was dispersed in acetonitrile (35.0 L), and the reaction solution was stirred at 55 °C for 1.5 hours, then at 35 °C for 1.5 hours, and then at 25 °C for 12 hours. The reaction solution was filtered, and the solid was washed with acetonitrile (4.0 L × 1) and dried under vacuum to obtain a solid. The mother liquor was concentrated to obtain compounds 1-6. The solid was added to acetonitrile (23.0 L) and water (6.0 L). The reaction solution was heated to 55 °C and stirred until dissolved. The reaction solution was then slowly cooled to 25 °C and stirred for 12 hours. The reaction solution was filtered, and the solid was washed with acetonitrile (4.0 L × 1) and dried under vacuum to obtain compounds 1-5. Compounds 1-5 and 1-6 were then isolated and analyzed by SFC (chiral column: Chiralpak AD-3 50 × 4.6 mm ID, 3 μm; mobile phase: [A: CO2, B: 0.05% diethylamine methanol solution]; gradient: from 5% to 40% 2 min, 40% 1.2 min, 5% 0.8 min (B: 0.05% diethylamine methanol solution), 4 mL / min). The retention time of compound 1-5 was 1.550 min; the retention time of compound 1-6 was 1.121 min.

[0075] Step 4

[0076] At 0–10 °C, saturated sodium bicarbonate (13.0 L), compounds 1-5 (2546.03 g, 3.36 mol), and dichloromethane (13.0 L) were sequentially added to a reaction vessel. The reaction vessel temperature was adjusted to 20–30 °C, and the mixture was stirred for 1–2 hours. The reaction solution was transferred to a separatory funnel, allowed to stand for separation, and the aqueous phase was extracted with dichloromethane (6.0 L × 2). The combined organic phases were washed with saturated brine (6.4 L × 1), allowed to stand for separation, and the organic phase was dried with anhydrous sodium sulfate (467.51 g). The mixture was filtered, and the filtrate was concentrated under reduced pressure at 40–50 °C to obtain compounds 1-7. 1¹H NMR (400MHz, CD₃OD) 7.28–7.19 (m, 2H), 7.16–7.09 (m, 1H), 7.08–7.00 (m, 2H), 4.04–3.96 (m, 1H), 3.67–3.52 (m, 4H), 3.39–3.31 (m, 2H), 2.33–2.25 (m, 1H), 2.15–2.07 (m, 1H), 1.97–1.88 (m, 1H), 1.73–1.57 (m, 4H), 1.56–1.48 (m, 1H), 1.45 (s, 9H), 1.09–0.98 (m, 2H). MS-ESI calculated values ​​[M+H] + The measured value is 373.

[0077] Step 5

[0078] Compounds 1-7 (7.00 g, 18.79 mmol) were dissolved in acetonitrile (70 mL), and p-toluenesulfonic acid monohydrate (9.71 g, 56.38 mmol) was added to the reaction solution. The reaction solution was stirred at 25 °C for 12 hours. The reaction solution was filtered, and the filter cake was dried under vacuum to give di-p-toluenesulfonate of compound 1. 1 H NMR (400MHz, CD3OD) 7.70 (d, J=8.4Hz, 4H), 7.31-7.27 (m, 2H), 7.23-7.21 (m, 5H), 7.15-7.13 (m, 2H), 4.19-4.05 (m, 3H), 3.27-3.15 (m, 4H), 3.00-2.97 (m, 1H), 2.52-2.48 (m, 1H), 2.43-2.38 (m, 1H), 2.35 (s, 6H), 2.04-1.95 (m, 4H), 1.81-1.73 (m, 1H), 1.55-1.50 (m, 1H), 1.40-1.35 (m, 1H). MS-ESI calculated value [M+H]+273, measured value 273.

[0079] Example 2

[0080]

[0081] Synthesis route:

[0082]

[0083] first step

[0084] Saturated sodium bicarbonate (0.44 mL), compounds 1-6 (0.6 g, 0.718 mmol), and dichloromethane (10 mL) were stirred and mixed. The reaction vessel temperature was 25 °C, and the mixture was stirred for 1 hour. The reaction solution was allowed to stand and separate into layers. The aqueous phase was extracted with dichloromethane (5.0 mL × 2). The organic phases were combined, washed with saturated brine (5.0 mL × 1), allowed to stand and separate into layers, and the organic phase was dried with anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-1. 1 ¹H NMR (400MHz, CD₃OD) 7.26–7.22 (m, 2H), 7.15–7.11 (m, 1H), 7.07–7.05 (m, 2H), 4.02–3.98 (m, 1H), 3.65–3.50 (m, 4H), 3.37–3.29 (m, 2H), 2.30–2.22 (m, 1H), 2.13–2.04 (m, 1H), 1.95–1.86 (m, 1H), 1.69–1.56 (m, 4H), 1.51–1.48 (m, 1H), 1.46 (s, 9H), 1.09–1.03 (m, 2H). MS-ESI calculated values ​​[M+H] + The measured value is 373.

[0085] Step 2

[0086] Compound 2-1 (250 mg, 0.603 mmol) was dissolved in acetonitrile (10 mL), and p-toluenesulfonic acid monohydrate (311.56 mg, 1.81 mmol) was added to the reaction solution. The reaction solution was stirred at 25 °C for 12 hours. The reaction solution was filtered, and the filter cake was dried under vacuum to obtain di-p-toluenesulfonate of compound 2. 1 H NMR(400MHz, CD3OD)7.71(d,J=8.4Hz,4H),7.35-7.27(m,2H),7.24-7.21(m,5H),7. 16-7.15(m,2H),4.24-4.14(m,1H),4.12-4.09(m,2H),3.28-3.17(m,4H),3.05-2.9 7(m,1H), 2.56-2.52(m,1H), 2.55-2.47(m,1H), 2.37(s,6H), 2.03-1.98(m,4H), 1.84-1.74(m,1H), 1.60-1.51(m,1H), 1.42-1.35(m,1H). MS-ESI calculated value [M+H]+273, measured value 273.

[0087] Example 3

[0088]

[0089] Synthesis route:

[0090]

[0091] first step

[0092] Compound 3-1 (50.0 g, 0.251 mol) was dissolved in anhydrous tetrahydrofuran (150 mL) and water (150 mL). Ammonium chloride (49.9 g, 0.934 mol) and zinc powder (49.2 g, 0.753 mol) were added to the reaction solution at 0 °C. Compound 3-2 (91.1 g, 0.753 mol) was slowly added dropwise at 0 °C. The reaction solution was stirred at 20 °C for 12 hours. The mixture was filtered, and the filtrate was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and then separated by silica gel column chromatography (3:1 petroleum ether / ethyl acetate, R...). f =0.47) yielded compound 3-3. 1 H NMR(400MHz,CD3OD)δ5.88-5.80(m,1H),5.18-5.10(m,2H),3.95-3.64(m,2H) ,3.28-3.02(m,2H),2.23-2.21(m,2H),1.58-1.48(m,4H),1.45-1.44(m,9H).

[0093] Step 2

[0094] Compound 3-3 (2.00 g, 8.29 mmol) was dissolved in N,N-dimethylformamide (20 mL). Sodium hydroxide (60%, 0.994 mg, 24.9 mmol) was added to the reaction solution under nitrogen protection at 0 °C. Compound 3-2 (3.01 g, 24.9 mmol) was then added to the reaction solution under nitrogen protection. The reaction solution was stirred at 25 °C for 2 hours. Saturated ammonium chloride (200 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and then separated by silica gel column chromatography (10:1 petroleum ether / ethyl acetate, R...). f =0.60) yielded compounds 3-4.

[0095] Step 3

[0096] Compounds 3-4 (2.15 g, 7.64 mmol) were dissolved in dichloromethane (20 mL). (1,3-dimethylimidazolidine-2-ylene(2-isopropoxybenzyl)ruthenium chloride (VI)) (0.479 g, 0.764 mmol) was added to the reaction solution. The reaction solution was stirred at 25 °C for 3 hours. The reaction was quenched with water (100 mL), extracted with ethyl acetate (100 mL × 3), and the organic phases were combined. The mixture was washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (10:1 petroleum ether / ethyl acetate, R...). f =0.51) yielded compounds 3-5. 1 H NMR (400MHz, CD3OD) δ5.68-5.61(m,2H),4.04-4.03(m,2H),3.71-3.60(m,2H),3.12-3.06(m, 2H),1.93-1.90(m,2H),1.78-1.68(m,2H),1.62-1.61(m,1H),1.39(s,9H),1.37-1.36(m,1H).

[0097] Step 4

[0098] Compounds 3-5 (1.87 g, 7.38 mmol) were dissolved in anhydrous tetrahydrofuran (10 mL). Borane tetrahydrofuran (1 M, 22.1 mL) was added to the reaction solution under nitrogen protection at 0 °C. The reaction solution was stirred at 30 °C for 7 hours. Sodium hydroxide (3.54 g, 88.6 mmol), water (10 mL), and hydrogen peroxide (27.1 g, 0.295 mol) were added to the reaction solution at 0 °C. The reaction solution was stirred at 30 °C for 1 hour. The reaction solution was quenched with water (100 mL), extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (2:1 petroleum ether / ethyl acetate, Rf = 0.20) to give compounds 3-6. 1 H NMR (400MHz, CD3OD) δ3.79-3.65(m,4H), 3.15-2.94(m,2H), 1.82-1.74(m,3H), 1.67-1.53(m,6H), 1.38(s,9H).

[0099] Step 5

[0100] Compounds 3-6 (1.65 g, 6.08 mmol) were dissolved in anhydrous dichloromethane (20 mL). Pyridinium dichromate salt (4.58 g, 12.2 mmol) was added to the reaction solution under nitrogen protection at 0 °C. The reaction solution was stirred at 30 °C for 12 hours. The mixture was filtered, and the filtrate was extracted with dichloromethane (80 mL × 1). The organic phases were combined and washed with hydrochloric acid (1 mol / L, 50 mL) and saturated sodium chloride (100 mL × 1). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and separated by silica gel column chromatography (2:1 petroleum ether / ethyl acetate, Rf = 0.59) to give compounds 3-7. 1 ¹H NMR (400MHz, CD₃OD) δ 4.03–4.00 (m, 2H), 3.83–3.81 (m, 2H), 3.22–3.16 (m, 2H), 2.53–2.48 (m, 2H), 1.93–1.77 (m, 4H), 1.58–1.52 (m, 2H), 1.48–1.47 (m, 9H). MS-ESI calculated values ​​[M-Boc+H] + 170, [M-56+H] + 214, measured value 170, 214.

[0101] Step 6

[0102] Compounds 3-7 (240 mg, 0.891 mmol) and 1-2 (142 mg, 1.07 mmol) were dissolved in dichloromethane (3 mL). Glacial acetic acid (53.5 mg, 0.891 mol) was added to the reaction solution, and the mixture was stirred at 25 °C for 10 hours. Sodium triacetoxyborohydride (378 mg, 1.78 mmol) was added to the reaction solution, and the reaction was continued for 2 hours. The reaction was quenched by adding saturated sodium bicarbonate solution (20 mL) at 25 °C. The mixture was extracted with dichloromethane (20 mL × 3), washed with saturated sodium chloride solution (30 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by thin-layer chromatography (1:1 petroleum ether / ethyl acetate, Rf = 0.3) to obtain compounds 3-8. MS-ESI calculated value [M+H] + 387, measured value 387.

[0103] Step 7

[0104] Compound 3-8 (100 mg, 0.259 mmol) was dissolved in ethyl acetate (5 mL), and ethyl hydrochloric acid solution (4 M, 10 mL, 40.0 mmol) was added and stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure and purified by high performance liquid chromatography (acidic, hydrochloric acid system) to obtain the hydrochloride salt of compound 3. 1¹H NMR (400MHz, CD₃OD) δ 7.35–7.28 (m, 2H), 7.27–7.16 (m, 3H), 3.95–3.91 (m, 1H), 3.86–3.78 (m, 1H), 3.73–3.66 (m, 1H), 3.29–3.21 (m, 3H), 3.11–2.97 (m, 2H), 2.59–2.52 (m, 2H), 2.18–2.13 (m, 2H), 1.98–1.89 (m, 1H), 1.84–1.68 (m, 3H), 1.65–1.56 (m, 2H), 1.47–1.41 (m, 1H). MS-ESI calculated values ​​[M+H] + 287, measured value 287.

[0105] Example 4

[0106]

[0107] Synthesis route:

[0108]

[0109] first step

[0110] Compound 4-1 (194 mg, 0.856 mmol) and compound 1-2 (114 mg, 0.856 mmol) were dissolved in dichloromethane-free solution (1 mL). Glacial acetic acid (154 mg, 2.57 mmol) was added to the reaction solution. The reaction solution was stirred at 26 °C for 2 hours, and sodium borohydride acetate (544 mg, 2.57 mmol) was added. The mixture was stirred at 26 °C for another 10 hours. Saturated sodium bicarbonate (30 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and separated by thin-layer chromatography (2:1 petroleum ether / ethyl acetate, Rf = 0.26) to obtain compound 4-2. MS-ESI calculated value [M+H] + 345, measured value 345.

[0111] Step 2

[0112] Compound 4-2 (70.0 mg, 0.203 mmol) was dissolved in ethyl acetate (1 mL), and ethyl acetate hydrochloride (4 M, 0.406 mL) was added dropwise at 0 °C. The reaction mixture was stirred at 20 °C for 1 hour, and the solvent was removed by concentration under reduced pressure. The crude product was then subjected to high performance liquid chromatography (acidic, hydrochloric acid system) to prepare the hydrochloride salt of compound 4. 1¹H NMR (400MHz, CD₃OD) δ 7.35–7.32 (m, 2H), 7.27–7.20 (m, 3H), 4.41–4.26 (m, 1H), 4.29–4.07 (m, 6H), 3.05–3.03 (m, 1H), 2.91–2.84 (m, 1H), 2.68–2.55 (m, 2H), 1.66–1.60 (m, 1H), 1.50–1.41 (m, 1H). MS-ESI calculated values ​​[M+H] + 245, measured value 245.

[0113] Example 5

[0114]

[0115] Synthesis route:

[0116]

[0117] Compounds 1-4 (250 mg, 0.671 mmol) were dissolved in ethyl acetate (5 mL), and ethyl acetate hydrochloride solution (4 M, 5 mL) was added at 15 °C. The reaction mixture was stirred at 15 °C for 10 hours, and the solvent was removed by concentration under reduced pressure. The crude product was then subjected to high performance liquid chromatography (acidic, hydrochloric acid system) to prepare the hydrochloride salt of compound 5. 1 ¹H NMR (400MHz, CD₃OD) δ 7.35–7.31 (m, 2H), 7.26–7.22 (m, 3H), 4.23–4.21 (m, 1H), 4.19–4.13 (m, 2H), 3.33–3.20 (m, 4H), 3.05–3.03 (m, 1H), 2.71–2.67 (m, 1H), 2.49–2.44 (m, 1H), 2.17–2.11 (m, 3H), 2.03–2.01 (m, 1H), 1.93–1.89 (m, 1H), 1.70–1.66 (m, 1H), 1.44–1.40 (m, 1H). MS-ESI calculated values ​​[M+H] + 273, measured value 273.

[0118] Biochemical detection:

[0119] Experiment 1: Enzyme Activity Evaluation

[0120] The purpose of this experiment was to detect the in vitro inhibitory activity of the compound against LSD1. The enzyme used was human LSD1, and the standard substrate was histone H3K4me peptide (20 μM). The activity of the compound was determined using an enzyme-linked fluorescence spectrophotometer, which detected the H2O2 generated after the reaction with horseradish peroxidase (HRP) and the fluorescent reagent Amplex Red. The compound was diluted 3-fold starting from 10 μM, and its IC50 was measured at 10 different concentrations. 50 Values. The enzyme and substrate were co-incubated for 30 minutes before the reaction was initiated. Fluorescence detector: EnVision, excitation wavelength: Ex / Em = 530 / 590 nM.

[0121] The inhibitory activity of the compounds against LSD1 was tested, and the results are shown in Table 1.

[0122] Table 1: Results of in vitro enzyme activity screening tests of the compounds of the present invention

[0123] Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> di-p-toluenesulfonate of compound 1 28.05 Hydrochloride salt of compound 4 72.92 Hydrochloride salt of compound 3 17.43 Hydrochloride salt of compound 5 4.66

[0124] Conclusion: The compounds of this invention exhibit significant inhibitory activity against LSD1.

[0125] Experiment 2: Evaluation of cell viability

[0126] A. kasumi-1, MV-4-11 cell viability assay

[0127] Experimental objective: To analyze the inhibitory effect of the compound on the proliferation of kasumi-1 and MV4-11 cells.

[0128] Experimental methods and procedures: Cell culture medium: Add 20% or 10% fetal bovine serum and 1% penicillin / streptomycin solution to RPMI 1640 cell culture medium and store at 4℃ for later use.

[0129] 1) Mix kasumi-1 / MV-411 cells and count them, then prepare 1×10⁻⁶ cells / mL of culture medium. 4 Kasumi-1 cell suspension at 1 cell / mL and 3×10 4 Add 80 μl of MV-411 cell suspension (1 cell / mL) to a 96-well cell culture plate.

[0130] 2) Dilute the compound to 50 μM using cell culture medium and place it in column 1 of the compound plate (1.5 μL 10 mM stock solution + 300 μL cell culture medium). Add 80 μL of cell culture medium to the wells in columns 2 to 8. Take 20 μL of the compound from column 1 and add it to column 2, mix well, then take 20 μL of the compound from column 2 and add it to column 3, mix well, and repeat this step up to column 8.

[0131] 3) Take 20 μL of the serially diluted compound from each well of the compound plate and place it into the corresponding position of the cell culture plate. At this point, the final concentration of the compound is 10 μM to 0.128 nM. Place the cell plate back into a carbon dioxide incubator containing 5% CO2 and continue incubation for 6 days.

[0132] 4) After 6 days of cell culture, remove the 96-well cell culture plate, add 50 μl of CTG reagent per well, mix well, centrifuge, and incubate at room temperature for 15 minutes. Read the values ​​using an Envision multilabel analyzer.

[0133] 5) Inhibition rate calculation:

[0134] Inhibition rate (%) = (RFU sample - RFU negative control) / (RFU positive control - RFU negative control) × 100%

[0135] 6) IC 50 calculate:

[0136] The inhibition rate was measured using the Prism 8 software for IC testing. 50 The calculation is performed using the following formula:

[0137] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope))

[0138] The experimental results are shown in Table 2:

[0139] Table 2: Results of in vitro cell activity tests of the compounds of the present invention

[0140]

[0141] Conclusion: The compounds of this invention exhibit significant inhibitory activity against the proliferation of kasumi-1 / MV4-11 cells.

[0142] B.KG-1 cell viability evaluation

[0143] Experimental objective: To analyze the inhibitory effect of the compound on the proliferation of human acute myeloid leukemia KG-1 cells.

[0144] Experimental materials: Fetal bovine serum (FBS) was purchased from Ecosai Biotechnology Co., Ltd.; IMDM medium was purchased from the American Type Culture Collection (ATCC); penicillin / streptomycin antibiotics were purchased from HyClone; CellTiter-Glo Luminescent Cell Viability Assay reagents were purchased from Promega; KG-1 cells were purchased from the European Collection of Authenticated Cell Cultures (ECACC); Nivo multilabel analyzer (PerkinElmer).

[0145] Experimental methods:

[0146] 1) Seed KG-1 cells in 96-well plates, 100 μL of cell suspension per well, including 1×10⁶ cells. 4 Cells / mL. The cell plate was incubated in a CO2 incubator.

[0147] 2) Transfer the prepared compound to the corresponding wells of the cell plate (start with a final concentration of 10 μM, decrease by 5X, for a total of 9 concentrations). Incubate the cell plates at 37°C for 6 days in a cell culture incubator containing 5% CO2.

[0148] 3) On day 6, remove the 96-well cell culture plate, add 50 μL of CellTiter-Glo Luminescent CellViability Assay reagent per well, mix well and shake for 10 minutes, then incubate at room temperature for 5 minutes. Read the fluorescence values ​​of the cell plate using an Envision Multilabel Plate Reader.

[0149] Data Analysis:

[0150] 1) Inhibition rate calculation:

[0151] Inhibition rate (%) = (RFU sample - RFU negative control) / (RFU positive control - RFU negative control) × 100%

[0152] 2) IC 50 Calculation: The inhibition rate was calculated using the software GraphPad Prism 9 for IC calculation. 50 The calculation is performed using the following formula, and the results are shown in Table 3.

[0153] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope))

[0154] Table 3: Inhibitory effects of compounds on cell proliferation

[0155]

[0156] Conclusion: The compounds of this invention exhibited significant antiproliferative activity in KG-1 cells.

[0157] Experiment 3: Inhibition of hERG potassium ion channels

[0158] Experimental objective: To investigate the effect of compound 1's di-p-toluenesulfonate on the current of the human Ether-a-go-go Related Gene potassium channel (hERG) using the electrophysiological manual patch-clamp method.

[0159] Experimental methods

[0160] 3.1 Cell Culture

[0161] Chinese hamster ovary cells stably expressing the hERG channel were cultured in 35 mm diameter cell culture dishes at 37°C in a 5% CO2 incubator. The cells were passaged every 48 hours at a 1:5 ratio. The culture medium consisted of 90% F12 (Invitrogen), 10% fetal bovine serum (Gibco), 100 g / mL G418 (Invitrogen), and 100 g / mL Hygromycin B (Invitrogen). On the day of the experiment, the cell culture medium was aspirated, the cells were washed once with extracellular fluid, and then 0.25% Trypsin-EDTA (Invitrogen) solution was added for digestion at room temperature for 3-5 minutes. The digestion fluid was aspirated, the cells were resuspended in extracellular fluid, and then transferred to experimental dishes for electrophysiological recording.

[0162] 3.2 Compound Preparation

[0163] The compound was dissolved in DMSO to prepare a 20 mM stock solution. On the day of testing, the stock solution was serially diluted 3-fold with DMSO, i.e., 10 μL of the stock solution was added to 20 μL of DMSO to obtain intermediate concentrations of the compound obtained by serial dilution with DMSO: 20, 6.66, 2.22, 0.74, 0.25, and 0.082 mM, respectively. Then, 10 μL of the intermediate concentration was added to 4990 μL of extracellular fluid and diluted 500-fold to obtain the final concentration to be tested. The highest test concentration was 40 μM, with subsequent concentrations of 40, 13.3, 4.44, 1.48, 0.49, and 0.16 μM. Preparation of the positive control compound cisapride: A 150 μM cisapride stock solution was serially diluted 3-fold with 100% DMSO. Specifically, 10 μL of the 150 μM cisapride stock solution was added to 20 μL of DMSO to obtain five intermediate cisapride concentrations obtained through serial dilution with DMSO: 150, 50, 16.7, 5.56, and 1.85 μM. Then, 10 μL of each intermediate cisapride concentration was added to 4990 μL of extracellular fluid and diluted 500-fold to obtain the final concentration to be tested. The highest test concentration was 300 nM, with five concentrations obtained: 300, 100, 33.3, 11.1, and 3.70 nM. The DMSO content in the final test concentration did not exceed 0.2%, as this concentration of DMSO had no effect on hERG potassium channels.

[0164] 3.3 Electrophysiological Recording Process

[0165] CHO (Chinese Hamster Ovary) cells stably expressing the hERG potassium channel were used to record hERG potassium channel currents at room temperature using whole-cell patch-clamp technique. Glass microelectrodes were fabricated from glass electrode blanks (BF150-86-10, Sutter) using a stretching instrument. The tip resistance after perfusion with electrode fluid was approximately 2-5 MΩ. The glass microelectrodes were inserted into the amplifier probe to connect to an Axopatch 200B (Molecular Devices) patch-clamp amplifier. Clamping voltage and data recording were controlled and recorded using pClamp 10 software via computer, with a sampling frequency of 10 kHz and a filtering frequency of 2 kHz. After obtaining whole-cell recordings, the cells were clamped at -80 mV. The step voltage to induce hERG potassium current (IhERG) was applied from -80 mV with a 2-s depolarization voltage to +20 mV, then repolarized to -50 mV, held for 1 second, and then returned to -80 mV. This voltage stimulation is applied every 10 seconds. Once the hERG potassium current stabilizes (1 minute), the drug administration process begins. Compound concentrations are administered continuously, starting from low test concentrations, with each test concentration administered for at least 1 minute. At least 3 cells (n≥3) are tested for each compound concentration, and at least 2 cells (n≥2) are tested for each positive compound concentration.

[0166] 3.4 Data Analysis

[0167] Data analysis and processing were performed using pClamp 10, GraphPad Prism 5, and Excel software. The degree of inhibition of hERG potassium current (the peak hERG tail current induced at -50mV) by different compound concentrations was calculated using the following formula:

[0168] Inhibition%=[1–(I / Io)]×100%

[0169] Where Inhibition% represents the percentage of inhibition of hERG potassium current by the compound, and I and Io represent the amplitude of hERG potassium current before and after drug administration, respectively.

[0170] Compound IC 50 The following equations were fitted and calculated using GraphPad Prism 5 software:

[0171] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope))

[0172] Where X is the Log value of the detected concentration of the test sample, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0173] 3.5 Test Results

[0174] Example compound hERG IC 50 The results are shown in Table 4.

[0175] Table 4: Compounds in Examples, hERG IC 50 Value Result

[0176] Test sample <![CDATA[hERG IC 50 (μM)]]> <![CDATA[IC 50 (ng / mL)]]> di-p-toluenesulfonate of compound 1 >40 >24671.6

[0177] Conclusion: The compounds of this invention have no inhibitory effect on hERG potassium ion channels.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, ， in, m is 1 or 2; n is 1; g is 1.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, Structural unit for or .

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: ， in, m, n, and g are as defined in claim 1.

4. A compound of the following formula or a pharmaceutically acceptable salt thereof, or .

5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , or .

6. The compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, wherein the salt is selected from hydrochloride and p-toluenesulfonate.

7. Use of the compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating LSD1-related diseases.

8. The application according to claim 7, wherein the disease is a hematologic malignancy, small cell lung cancer, squamous non-small cell lung cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, glioma, or Ewing's sarcoma.

9. The application according to claim 8, wherein the hematologic malignancy is human acute myeloid leukemia.

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