A thiophene-pyrimidine compound, a pharmaceutical composition comprising the same, and its use

Abstract

This article discloses a thienopyrimidine compound, pharmaceutical compositions comprising the same, and their applications. The thienopyrimidine compound has the structure shown in Formula I. This compound interferes with the interaction between menin protein and MLL1, MLL2, or MLL-fusion oncoproteins, and holds promise as a treatment for tumors, diabetes, and other diseases dependent on the activity of MLL1, MLL2, MLL fusion proteins, and / or menin proteins.

Description

Technical Field

[0001] This application belongs to the field of medicinal chemistry, specifically relating to a thiophene-pyrimidine compound, a pharmaceutical composition containing the same, and its application. Background Technology

[0002] Mixed-lineage leukemia (MLL) protein is a histone methyltransferase that plays a crucial role in gene transcriptional regulation. Most acute leukemias, including acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), and mixed-lineage leukemia, have been found to involve the MLL gene, located at the q23 band on chromosome 11, which frequently translocates to form an MLL fusion (MLL-r) protein with one of approximately 80 proteins (such as AF4, AF9, ENL, AF10, ELL, AF6, AF1p, GAS7, etc.). The MLL-r protein retains approximately 1400 amino acids of the N-terminal MLL protein but lacks the C-terminal methyltransferase activity region. It abnormally regulates the transcription of various oncogenes, including HOX and MEIS1, promoting cell proliferation and ultimately leading to cancer. Leukemia patients with MLL gene chromosomal translocations generally have a poor prognosis, with a 5-year survival rate of less than 40% (Slany, Haematologica, 2009, 94, 984-993).

[0003] Menin protein, encoded by the Multiple Endocrine Neoplasia (MEN) gene, is a widely expressed nuclear protein that interacts with DNA replication and repair proteins, chromatin modification proteins, and various transcription factors (Agarwal et al., Horm Metab Res, 2005, 37, 369-374). Menin protein can bind to the N-terminus of MLL proteins, including MLL1, MLL2, and MLL-r proteins, and this binding is essential for the oncogenic activity of MLL proteins (Yokoyama et al., Cell, 2005, 123, 207-218; Cierpicki and Grembecka, FutureMed. Chem., 2014, 6, 447-462). Interfering with the interaction between menin and MLL-r protein can selectively inhibit the proliferation of MLL-r leukemia cells in vitro and in vivo (Grembecka et al., Nat. Chem. Biol., 2012, 8, 277-284; Borkin et al., Cancer cell, 2015, 27, 589-602).

[0004] Certain hematologic malignancies exhibit specific gene abnormalities or mutations, such as nucleoporin 98 (NUP98) gene fusion, nucleophosphorus protein (NPM1) gene mutation, DNA methyltransferase 3A (DNMT3A) mutation, and MLL gene amplification. These abnormalities or mutations are often accompanied by high levels of HOX gene expression. In Ewing's sarcoma, retrograde HOXD genes, especially HOXD13, are abnormally overexpressed, accompanied by high levels of menin and MLL1 proteins. HOXD13 is a downstream gene regulated by menin and MLL1. Therefore, interfering with the interaction between menin and MLL proteins, especially through covalent binding, is a promising strategy for treating tumors.

[0005] Therefore, there is an urgent need in this field to develop effective drugs that can interfere with the interaction between menin and MLL proteins. Summary of the Invention

[0006] The following is an overview of the subject matter described in detail herein. This overview is not intended to limit the scope of the claims.

[0007] In view of the shortcomings of the prior art, the purpose of this application is to provide a thienopyrimidine compound, a pharmaceutical composition containing the same, and the application thereof, wherein the thienopyrimidine compound and the pharmaceutical composition containing the same are capable of interfering with the interaction between menin and MLL proteins.

[0008] To achieve this objective, the following technical solution is adopted in this application:

[0009] In a first aspect, this application provides a thienopyrimidine compound, the structural formula of which is shown in Formula I below:

[0010]

[0011] in,

[0012] R 1 Selected from H, halogens, substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C3-C8 cycloalkyl groups, substituted or unsubstituted 4-8 membered heterocyclic groups, substituted or unsubstituted C1-C6 alkyl carbonyl groups, substituted or unsubstituted... Substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted C1-C6 alkyl sulfone, substituted or unsubstituted C1-C6 alkyl sulfoxide, Among them, R a R b Each is independently selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclic group, or R. a With R b Substituted or unsubstituted 4-8 membered heterocycles formed by bonding with N, wherein R a With R b The heterocycle formed by bonding with N contains 1-3 heteroatoms selected from N, O, S, and P;

[0013] R 2 Selected from H, halogens, substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C3-C8 cycloalkyl groups, substituted or unsubstituted C6-C16 aryl groups, substituted or unsubstituted 5-16-membered heteroaryl groups, substituted or unsubstituted 4-8-membered heterocyclic groups, substituted or unsubstituted C1-C6 alkyl carbonyl groups, substituted or unsubstituted... Substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted C1-C6 alkyl sulfone, substituted or unsubstituted C1-C6 alkyl sulfoxide, Among them, R c R d Each is independently selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclic group, or R. c With R d Substituted or unsubstituted 4-8 membered heterocycles formed by bonding with N, wherein R c With R dThe heterocycle formed by the bond with N contains 1-3 heteroatoms selected from N, O, S, and P;

[0014] R 3 Each of the following is independently selected from H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, -NH2, -NO2, -COOH, -CN, -OH, substituted or unsubstituted C1-C6 alkylsulfonyl, substituted or unsubstituted C1-C6 alkylsulfoxide, substituted or unsubstituted C1-C6 alkylthio. Among them, R 3 Selected from H, methyl, or fluorine; R 3 "Selected from chlorine or bromine, Indicates the connection position of the group;

[0015] Y and Z are independently selected from N or CH, and at least one of Y and Z is N;

[0016] W is selected from N or C;

[0017] U 1 U 2 U 3 U 4 U 5 U 6 U 7 U 8 Selected independently And U 1 U 2 U 3 U 4 At most one of them is U 5 U 6 At most one of them is U 7 U 8 At most one of them is in, Indicates the connection position of the group.

[0018] Each R' is independently selected from: H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

[0019] Each "R" is independently selected from: H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

[0020] Each R”' is independently selected from: H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

[0021] Each R is independently selected from: H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

[0022] A is selected from substituted or unsubstituted C6-C16 aryl groups, substituted or unsubstituted 5-16 heteroaryl groups, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O, S, and P;

[0023] L 1 For non-existent in, Indicates the connection position of the group, R L1' R L1” Each is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, or R. L1' With R L1” It forms substituted or unsubstituted C3-C8 cycloalkyl groups or substituted or unsubstituted 4-8 membered heterocycles with the attached carbon atom, wherein R L1' With R L1” The heterocycle formed with the attached carbon atom contains 1-3 heteroatoms selected from N, O, S, and P;

[0024] L 2 Selected from in, Indicates the connection position of the group;

[0025] X is selected from carbon atoms or

[0026] R 4 Selected from Wherein, R4' is selected from fluorine or chlorine, R4" is selected from H, methyl or fluorine, and R4"' is selected from H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, substituted or unsubstituted (C1-C4 alkyl)2amino, substituted or unsubstituted C1-C4 alkylthio, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclic group, or substituted or unsubstituted C2-C4 acyl group, wherein the heterocyclic group contains 1-3 heteroatoms selected from N, O, S, and P. Indicates the connection position of the group.

[0027] Preferably, the R 1The heterocyclic group is selected from H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclic group, wherein the heterocyclic group contains 1-3 heteroatoms selected from N, O, S, P.

[0028] Preferably, the R 1 Selected from substituted or unsubstituted C1-C6 alkyl groups, more preferably halogen-substituted C1-C6 alkyl groups, and even more preferably trifluoroethyl.

[0029] Preferably, the R 2 The group is selected from H, halogens, substituted or unsubstituted C1-C6 alkyl groups, substituted or unsubstituted C3-C8 cycloalkyl groups, substituted or unsubstituted C6-C16 aryl groups, substituted or unsubstituted 5-16-membered heteroaryl groups, and substituted or unsubstituted 4-8-membered heterocyclic groups, wherein the heteroaryl or heterocyclic group contains 1-3 heteroatoms selected from N, O, S, and P. More preferably, the R... 2 For H.

[0030] Preferably, the R 3 It can be H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, -NH2 or -CN.

[0031] Preferably, Y and Z are each selected from N.

[0032] Preferably, W is C.

[0033] Preferably, the U 1 U 2 U 3 U 4 U 5 U 6 U 7 U 8 Selected independently And U 1 U 2 U 3 U 4 At most one of them is U 5 U 6 At most one of them is U 7 U 8 At most one of them is in, Indicates the connection position of the group.

[0034] Each R' is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, and cyano;

[0035] Each "R" is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, and cyano;

[0036] Each R”' is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

[0037] Each R is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, and cyano.

[0038] Preferably, A is selected from substituted or unsubstituted C6-C10 aryl groups or substituted or unsubstituted 5-12 heteroaryl groups, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O, S, and P.

[0039] Preferably, A is a substituted or unsubstituted benzene ring, a substituted or unsubstituted pyridine ring, a substituted or unsubstituted pyridazine ring, a substituted or unsubstituted pyrimidine ring, a substituted or unsubstituted triazine ring, a substituted or unsubstituted thiophene ring, a substituted or unsubstituted thiazole ring, a substituted or unsubstituted imidazole ring, a substituted or unsubstituted pyrrole ring, a substituted or unsubstituted pyrazole ring, a substituted or unsubstituted oxazole ring, a substituted or unsubstituted isoxazole ring, or a substituted or unsubstituted triazine ring.

[0040] Preferably, the L 1 It is either absent or -CH2-, preferably -CH2-.

[0041] Preferably, the L 2 Selected from Preferred

[0042] Preferably, X is selected from carbon atoms.

[0043] Preferably, the R 4 Selected from Wherein, R4' is selected from fluorine or chlorine, R4” is selected from H, methyl or fluorine, and R4”' is selected from H, substituted or unsubstituted C1-C4 alkyl. Indicates the connection position of the group.

[0044] Preferably, the spirocyclic portion of the compound represented by Formula I has the following structural formula: in The position of the group is indicated by selecting any one of the following groups:

[0045]

[0046] Preferably, the spirocyclic portion of the compound represented by Formula I has the following structural formula: in The position of the group is indicated by selecting any one of the following groups:

[0047]

[0048] Preferably, the structural formula in Formula I is: The loop portion shown, wherein The position of the group is indicated by selecting any one of the following groups:

[0049]

[0050]

[0051] Among them, R e R f Each group is independently selected from H, methyl, trifluoromethyl, difluoromethyl, methoxy, ethoxy, trifluoromethoxy, halogen, hydroxyl, amino, cyano, methylamino, dimethylamino, ethylamino, methylethylamino, diethylamino, trifluoroethylamino, carboxyl, methoxycarbonyl, ethoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, methylethylcarbamoyl, or diethylcarbamoyl.

[0052] Preferably, the structural formula in Formula I is: The annular portion shown, in which The position of the group is indicated by selecting any one of the following groups:

[0053] Preferably, the R 4 Selected from -CH2F, -CH2Cl,

[0054] Preferably, the compound represented by Formula I is selected from any one of the following compounds:

[0055]

[0056]

[0057]

[0058]

[0059] Preferably, the thienopyrimidine compound further includes any one of the following: a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, polymorph, or deuterated derivative of the compound represented by Formula I.

[0060] In a second aspect, this application provides a pharmaceutical composition comprising a thiophene-pyrimidine compound as described in the first aspect and a pharmaceutically acceptable carrier.

[0061] Preferably, the pharmaceutical composition further comprises other pharmaceutically acceptable therapeutic agents, particularly other antitumor drugs. These therapeutic agents include, but are not limited to: antitumor drugs that act on the chemical structure of DNA, such as cisplatin; antitumor drugs that affect nucleic acid synthesis, such as methotrexate (MTX) and 5-fluorouracil (5FU); antitumor drugs that affect nucleic acid transcription, such as doxorubicin, epirubicin, aclarubicin, and sclerosomycin; antitumor drugs that act on microtubule synthesis, such as paclitaxel and vinorelbine; aromatase inhibitors, such as ammoniaglutide, lantron, letrozole, and renin; and cell signaling pathway inhibitors, such as epidermal growth factor receptor inhibitors, such as imatinib, gefitinib, erlotinib, and lapatinib.

[0062] Thirdly, this application provides the use of a thiophene-pyrimidine compound as described in the first aspect or a pharmaceutical composition as described in the second aspect, wherein the use is selected from any one of (a)-(c) below:

[0063] (a) To prepare medicines for the prevention or treatment of tumors, diabetes and other diseases associated with the activity of MLL1, MLL2, MLL fusion protein and / or menin protein;

[0064] (b) To prepare inhibitors for in vitro non-therapeutic use related to the activity of MLL1, MLL2, MLL fusion protein, and / or menin protein;

[0065] (c) Preparation of proliferation inhibitors for non-therapeutic tumor cells in vitro.

[0066] In a preferred embodiment, the tumor associated with the activity of MLL1, MLL2, MLL fusion protein, and / or menin protein is selected from the group consisting of: leukemia, Ewing's sarcoma, breast cancer, prostate cancer, T-cell lymphoma, B-cell lymphoma, malignant rhabdomyosarcoma, synovial sarcoma, colorectal cancer, endometrioma, gastric cancer, liver cancer, kidney cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, glioma, cholangiocarcinoma, nasopharyngeal carcinoma, cervical cancer, head and neck cancer, esophageal cancer, thyroid cancer, and bladder cancer.

[0067] "Other diseases" include, but are not limited to, autoimmune diseases, non-alcoholic hepatitis, etc.

[0068] Terminology Explanation

[0069] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0070] As used herein, when referring to a specific enumerated value, the term “about” means that the value can vary by no more than 1% from the enumerated values. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0071] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0072] In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0073] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionates, hexanoates, octanoates, decanoates, undecenoates, glycolates, gluconates, lactates, sebates, adipates, glutarate, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartate, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbic acid salts, salicylates, 4-aminosalicylic acid salts, and naphthalene disulfonates. These salts can be prepared using methods known in this field.

[0074] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

[0075] In this application, "pharmaceutical composition" refers to a formulation of the compound of this application with a medium generally accepted in the art for delivering a bioactive compound to a mammal (e.g., a human). This medium includes pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate administration to the organism, thereby promoting the absorption of the active ingredient and the exertion of its bioactivity.

[0076] As used herein, the term "pharmaceutically acceptable" means a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compound of this application and is relatively non-toxic, i.e., that the substance can be administered to an individual without causing an adverse biological reaction or interacting adversely with any component contained in the composition.

[0077] In this article, "pharmaceuticalally acceptable excipients" include, but are not limited to, any adjuvants, carriers, excipients, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that are permitted by the relevant government regulatory authorities to be acceptable for human or livestock use.

[0078] The term "tumor" as used in this application includes, but is not limited to, glioma, sarcoma, melanoma, arthroma, chondroma, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, colorectal cancer, nasopharyngeal carcinoma, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, and other diseases.

[0079] The terms “preventive,” “prevention,” and “avoidance” used in this article include reducing the likelihood of a patient developing or worsening a disease or condition.

[0080] The term "treatment" and other similar synonyms used in this article include the following meanings:

[0081] (i) To prevent the occurrence of disease or condition in mammals, especially when such mammals are susceptible to the disease or condition but have not yet been diagnosed with it;

[0082] (ii) To suppress a disease or symptom, that is, to curb its development;

[0083] (iii) To alleviate a disease or symptom, that is, to cause the condition of the disease or symptom to subside; or

[0084] (iv) To alleviate the symptoms caused by the disease or condition.

[0085] As used herein, the terms "effective amount," "therapeutic effective amount," or "pharmaceutical effective amount" refer to an amount of at least one drug or compound that, when taken, is sufficient to alleviate, to some extent, one or more symptoms of the disease or condition being treated. The result may be a reduction and / or relief of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition containing the compounds disclosed herein that is clinically necessary to provide significant symptom relief. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation testing.

[0086] As used herein, the terms “administration,” “application,” “dosage,” etc., refer to methods that deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral administration, duodenal administration, parenteral administration (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injection or infusion), local administration, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used with the compounds and methods described herein, such as those discussed in Goodman and Gilman, *The Pharmacological Basis of Therapeutics*, current ed.; Pergamon; and Remington's, *Pharmaceutical Sciences* (current edition), Mack Publishing Co., Easton, Pa. In a preferred embodiment, the compounds and compositions discussed herein are administered orally.

[0087] As used herein, the terms “drug combination,” “drug co-administration,” “combination therapy,” “administration of other treatments,” and “administration of other therapeutic agents” refer to drug therapy obtained by mixing or combining more than one active ingredient, including fixed and non-fixed combinations of active ingredients. The term “fixed combination” refers to the simultaneous administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity or single dosage form. The term “non-fixed combination” refers to the simultaneous, combined, or sequential administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity at variable intervals. These also apply to cocktail therapies, such as the administration of three or more active ingredients.

[0088] Group definition

[0089] Definitions of standard chemical terms can be found in the references (including Carey and Sundberg, "Advanced Organic Chemistry 4th Edition," Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise stated, conventional methods within the scope of the art, such as mass spectrometry, NMR, IR, UV / VIS spectroscopy, and pharmacological methods, are used. Unless specifically defined, the terminology used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, reactions and purifications can be carried out using the manufacturer's instructions for use of kits, or in accordance with methods known in the art or the descriptions in this application. The techniques and methods described herein are generally carried out according to conventional methods well known in the art, based on the descriptions in the various summary and more specific references cited and discussed in this specification. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural moieties and compounds.

[0090] When a substituent is described using a conventional chemical formula written from left to right, it also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0091] The chapter headings used in this document are for organizational purposes only and should not be construed as limiting the subject matter. All references or portions thereof cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and papers, are incorporated herein by reference in their entirety.

[0092] Certain chemical groups defined herein are preceded by simplified symbols to indicate the total number of carbon atoms present in the group. For example, C1-C6 alkyl refers to alkyl groups having a total of 1 to 6 carbon atoms as defined below. The total number of carbon atoms in the simplified symbols does not include carbons that may be present in substituents of the group.

[0093] Except as otherwise specified, when used in the specification and claims of this application, the following terms shall have the following meanings.

[0094] In this article, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0095] "Hydroxy group" refers to the -OH group.

[0096] "Hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group (-OH) as defined below.

[0097] "Carbonyl" refers to the -C(=O)- group.

[0098] "Nitro" refers to -NO2.

[0099] "Cyano" refers to -CN.

[0100] "Amino" refers to -NH2.

[0101] "Substituted amino" refers to an amino group that is substituted by one or two alkyl, alkylcarbonyl, aralkyl, or heteroaralkyl groups as defined below, such as monoalkylamino, dialkylamino, alkylamide, aralkylamino, or heteroaralkylamino.

[0102] The "carboxyl group" refers to -COOH.

[0103] As used herein, or as part of other groups (e.g., in halogen-substituted alkyl groups), the term "alkyl" refers to a fully saturated straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, having, for example, 1 to 12 (preferably 1 to 8, more preferably 1 to 6) carbon atoms, and connected to the rest of the molecule by single bonds, such as, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl, and decyl. For the purposes of this application, the term "alkyl" refers to an alkyl group containing 1 to 6 carbon atoms.

[0104] As used herein, or as part of other groups, the term "alkenyl" refers to a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms connected to the rest of the molecule by single bonds, such as, but not limited to, vinyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl, etc.

[0105] As used herein, or as part of other groups, the term "alkynyl" refers to a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms connected to the rest of the molecule by single bonds, such as, but not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octyynyl, etc.

[0106] As used herein, or as part of other groups, the term "cyclic hydrocarbon group" refers to a stable, non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms. This group may include fused ring systems, bridged ring systems, or spirocyclic systems, having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, and may be saturated or unsaturated and may be connected to the rest of the molecule via single bonds through any suitable carbon atom. Unless otherwise specifically indicated in this specification, the carbon atoms in the cyclic hydrocarbon group may optionally be oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2,3-dihydroindenyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, and 5,6,7,8,9,10-hexahydro-benzocycloheptenyl. Cyclooctenyl, fluorenyl, bicyclo[2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantyl, octahydro-4,7-methylene-1H-indenyl and octahydro-2,5-methylene-cyclopentadienyl, etc.

[0107] As used herein, or as part of other groups, the term "heterocyclic group" refers to a stable 3- to 20-membered non-aromatic cyclic group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen, and sulfur. Unless otherwise specified in this specification, a heterocyclic group can be a monocyclic, bicyclic, tricyclic, or more ring system, which may include fused ring systems, bridged ring systems, or spirocyclic systems; the nitrogen, carbon, or sulfur atoms in the heterocyclic group may optionally be oxidized; the nitrogen atom may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be connected to the remainder of the molecule via a carbon atom or heteroatom and by a single bond. In heterocyclic groups containing fused rings, one or more rings may be aryl or heteroaryl as defined below, provided that the connection point with the remainder of the molecule is a non-aromatic ring atom. For the purposes of this application, the heterocyclic group is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged, or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur, and more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged, or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heterocyclic groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]nonane-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2.1]heptane-2-yl, aziridine, pyranyl, tetrahydropyranyl, thiaranyl, tetrahydrofuranyl, oxazinyl, dioxocyclopentyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazoalkyl, quinazinyl, thiazoalkyl, isothiazyl, isoxazylalkyl, dihydroindolyl, octahydroindolyl, octahydroisoindolyl, pyrazolinyl, pyrazolyl, phthalimide, etc.

[0108] As used herein, the term "aryl" as a group or part of another group refers to a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms (preferably 6 to 10 carbon atoms). For the purposes of this application, an aryl group can be a monocyclic, bicyclic, tricyclic, or more cyclic system, and can be fused with cycloalkyl or heterocyclic groups as defined above, provided that the aryl group is connected to the rest of the molecule via single bonds through atoms on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthraceneyl, phenanthrene, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1,4-benzoxazine-3(4H)-one-7-yl, etc.

[0109] In this article, the term "arylalkyl" refers to an alkyl group as defined above that has been replaced by an aryl group as defined above.

[0110] As used herein, the term "heteroaryl" as a group or part of other groups refers to a 5- to 16-membered conjugated cyclic group having 1 to 15 carbon atoms (preferably 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically indicated in this specification, a heteroaryl group may be a monocyclic, bicyclic, tricyclic, or more cyclic system, and may be fused with cycloalkyl or heterocyclic groups as defined above, provided that the heteroaryl group is connected to the remainder of the molecule via single bonds through atoms on the aromatic ring. The nitrogen, carbon, or sulfur atom in the heteroaryl group may optionally be oxidized; the nitrogen atom may optionally be quaternized. For the purposes of this application, the heteroaryl group is preferably a stable 5- to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen, and sulfur, more preferably a stable 5- to 10-membered aromatic group containing 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, but are not limited to, thiophene, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridinyl, pyrazinyl, pyrazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furanyl, pyrrolithyl, triazolyl, tetrazolyl, triazinyl, inazinyl, isoindolyl, indazolyl, isoindazolyl, purinyl, quinolinyl, isoquinolinyl, diazonyl, naphthidyl, quinoxolinyl, pteridyl, carbazolyl, carbazolyl, phenanthridine, phenanthroxolinyl, acridineyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothiophene, oxazolyl, etc. Triazolyl, cyclolinyl, quinazolinyl, --nitrogenindyl, o-diazaphenyl, isoxazolyl, phenoxazinyl, phenthiazinyl, 4,5,6,7-tetrahydrobenzo[b]thiophenyl, naphthopyridyl, [1,2,4]triazolo[4,3-b]pyridazine, [1,2,4]triazolo[4,3-a]pyrazine, [1,2,4]triazolo[4,3-c]pyrimidine, [1,2,4]triazolo[4,3-a]pyridine, imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine, imidazo[1,2-a]pyrazine, etc.

[0111] In this article, the term "heteroarylalkyl" refers to an alkyl group as defined above that has been replaced by a heteroaryl group as defined above.

[0112] In this document, the term "substituted or unsubstituted" means that the hydrogen in the structure is substituted by the substituent or that the hydrogen is not substituted by the substituent. Unless otherwise stated, the optionally substituted group may have a substituent at every substituted position of the group, or more than one position in the structure (up to the point where the substituted position is filled) may be substituted.

[0113] For example, in the absence of an explicit list of substituents, the terms “substituted” or “replaced by” as used herein mean that one or more hydrogen atoms on a given atom or group are independently substituted by one or more, for example, 1, 2, 3, or 4 substituents, which are independently selected from: deuterium (D), halogen, -OH, mercapto, cyano, -CD3, -C1-C6 alkyl (preferably -C 1-3 Alkyl), C2-C6 alkenyl, C2-C6 alkynyl, cycloalkyl (preferably 3-8 membered cycloalkyl), aryl, heterocyclic (preferably 3-8 membered heterocyclic), heteroaryl, aryl-C1-C6 alkyl-, heteroaryl-C1-C6 alkyl-, C1-C6 haloalkyl-, -OC1-C6 alkyl (preferably -OC1-C3 alkyl), -OC2-C6 alkenyl, -OC1-C6 alkylphenyl, -C1-C6 alkyl-OH (preferably -C1-C4 alkyl-OH), -C1-C6 alkyl-SH, -C1-C6 alkyl-O-C1-C6 alkyl, -OC1-C 6-Halogenated alkyl, -NH2, -C1-C6 alkyl-NH2 (preferably -C1-C3 alkyl-NH2), -N(C1-C6 alkyl)2 (preferably -N(C1-C3 alkyl)2), -NH(C1-C6 alkyl) (preferably -NH(C1-C3 alkyl)), -N(C1-C6 alkyl)(C1-C6 alkylphenyl), -NH(C1-C6 alkylphenyl), nitro, -C(O)-OH, -C(O)OC1-C6 alkyl (preferably -C(O)OC1-C3 alkyl), -CONRiRii (where Ri and Ri are H, D and C respectively) 1-6 Alkyl, preferably C 1-3Alkyl), -NHC(O) (C1-C6 alkyl), -NHC(O) (phenyl), -N(C1-C6 alkyl)C(O) (C1-C6 alkyl), -N(C1-C6 alkyl)C(O) (phenyl), -C(O)C1-C6 alkyl, -C(O) heteroaryl (preferably -C(O)-5-7 heteroaryl), -C(O)C1-C6 alkylphenyl, -C(O)C1-C6 haloalkyl, -OC(O)C1-C6 alkyl (preferably -OC(O)C1-C3 alkyl) -S(O)2-C1-C6 alkyl, -S(O)-C1-C6 alkyl, -S(O)2-phenyl, -S(O)2-C1-C6 haloalkyl, -S(O)2NH2, -S(O)2NH (C1-C6 alkyl), -S(O)2NH (phenyl), -NHS(O)2 (C1-C6 alkyl), -NHS(O)2 (phenyl) and -NHS(O)2 (C1-C6 haloalkyl), wherein the alkyl, cycloalkyl, phenyl, aryl, heterocyclic and heteroaryl groups are used. Each of the following may optionally be further substituted by one or more substituents selected from: halogen, -OH, -NH2, cycloalkyl, 3-8 membered heterocyclic, C1-C4 alkyl, C1-C4 haloalkyl-, -OC1-C4 alkyl, -C1-C4 alkyl-OH, -C1-C4 alkyl-O-C1-C4 alkyl, -OC1-C4 haloalkyl, cyano, nitro, -C(O)-OH, -C(O)OC1-C6 alkyl, -CON(C1-C6 alkyl)2, -CONH(C1 -C6 alkyl), -CONH2, -NHC(O) (C1-C6 alkyl), -NH(C1-C6 alkyl)C(O) (C1-C6 alkyl), -SO2 (C1-C6 alkyl), -SO2 (phenyl), -SO2 (C1-C6 haloalkyl), -SO2NH2, -SO2NH (C1-C6 alkyl), -SO2NH (phenyl), -NHSO2 (C1-C6 alkyl), -NHSO2 (phenyl), and -NHSO2 (C1-C6 haloalkyl). When an atom or group is substituted by multiple substituents, the substituents may be the same or different. The terms "part," "structural part," "chemical part," "group," and "chemical group" as used herein refer to a specific segment or functional group in a molecule. A chemical part is generally considered to be a chemical entity embedded in or attached to a molecule.

[0114] "Non-existent" means that the two sides of the group defined above are directly connected by chemical bonds. For example, "B is non-existent in ABC" means "AC".

[0115] "Stereoisomers" are compounds composed of identical atoms bonded by the same bonds, but with different three-dimensional structures. This application will cover various stereoisomers and mixtures thereof.

[0116] When the compounds of this application contain alkene double bonds, unless otherwise stated, the compounds of this application are intended to contain E- and Z-geometric isomers.

[0117] "Tautomer" refers to an isomer formed when a proton is transferred from one atom of a molecule to another atom of the same molecule. All tautomer forms of the compounds in this application will also be included within the scope of this application.

[0118] The compounds of this application, or their pharmaceutically acceptable salts, may contain one or more chiral carbon atoms, and thus may produce enantiomers, diastereomers, and other stereoisomers. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. This application aims to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of this application may select racemic, diastereomer, or enantiomers as starting materials or intermediates. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0119] In this application, Ester groups representing C1-C6 alkyl substituted groups, for example, could be

[0120] In this application, (C1-C4 alkyl)2amino represents an amine substituted with two C1-C4 alkyl groups, such as... wait.

[0121] In this application, “each R’”, “each R””, “each R””” and “each R””” refer to “each R’”, “each R””, “each R”” and “each R””” in the thiophenepyrimidine compounds shown in Formula I.

[0122] Conventional techniques for preparing / separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemic mixtures (or racemic mixtures of salts or derivatives) using, for example, chiral high-performance liquid chromatography, as seen in Gerald Gübitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; AMStalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3: 341-63, 2010; Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup. TH ED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; Heller, Acc. Chem. Res. 1990, 23, 128.

[0123] Those skilled in the art will also understand that, in the methods described below, the functional groups of the intermediate compounds may require protection by appropriate protecting groups. Such functional groups include hydroxyl, amino, mercapto, and carboxylic acids. Suitable hydroxyl protecting groups include trialkylsilyl or diarylalkylsilyl (e.g., tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, etc. Suitable amino, amidine, and guanidine protecting groups include tert-butoxycarbonyl, benzyloxycarbonyl, etc. Suitable mercapto protecting groups include -C(O)-R" (where R is alkyl, aryl, or aralkyl), p-methoxybenzyl, triphenylmethyl, etc. Suitable carboxyl protecting groups include alkyl, aryl, or aralkyl esters.

[0124] Protecting groups can be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting groups is detailed in Greene, TW & PGMUTS, Protective Groups in OrganiSynthesis, (1999), 4th Ed., Wiley. Protecting groups can also be polymer resins.

[0125] Compared with the prior art, this application has the following advantages:

[0126] (1) This application provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof.

[0127] (2) This application provides a compound of Formula I for preparing a pharmaceutical composition for preventing and treating diseases related to the activity of MLL1, MLL2, MLL fusion protein, and / or menin protein.

[0128] After reading and understanding the detailed description, other aspects can be understood. Detailed Implementation

[0129] The technical solution of this application will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely to help understand this application and should not be regarded as specific limitations on this application.

[0130] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. Percentages and parts are weight percentages and parts by weight.

[0131] Unless otherwise specified, all experimental materials and reagents used in the following examples are available from commercially available sources.

[0132] In each embodiment, 1 ¹H NMR was recorded using a BRUKER AVANCE NEO 400MHz NMR spectrometer, and chemical shifts are expressed as δ (ppm). Liquid chromatography-mass spectrometry (LC-MS) was performed using a Shimadzu LC-20AD, SIL-20A, CTO-20AC, SPD-M20A, CBM-20A, or LCMS-2020 mass spectrometer. Preparative HPLC separations were performed using a Gilson-281 liquid chromatograph.

[0133] Preparation of intermediates

[0134] 1. Preparation of intermediate A

[0135]

[0136] The synthetic route for intermediate A is shown below:

[0137]

[0138] (1) Triethylamine (5.84 g, 57.7 mmol) and compound A-2 (7.27 g, 63.5 mmol) were added to a solution of compound A-1 (5.0 g, 28.9 mmol) in dichloromethane (25.0 mL). The reaction mixture was stirred at 0 °C for 1 hour. The reaction was quenched by adding saturated sodium bicarbonate aqueous solution (40.0 mL). The mixture was extracted with dichloromethane (40.0 mL × 3). The combined organic phases were washed with saturated brine (30.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound A-3.

[0139] 1 H NMR (400MHz, CDCl3) δ5.24-5.15(m,1H),4.31-4.23(m,2H),4.13-4.06(m,2H),3.09-3.04(m,3H),1.46-1.42(m,9H).

[0140] (2) Compound A-4 (6.54 g, 57.3 mmol) was added to a solution of compound A-3 (7.2 g, 28.6 mmol) in N,N-dimethylformamide (70.0 mL). The reaction mixture was stirred at 85 °C for 12 hours. The reaction was quenched with water (50.0 mL), extracted with ethyl acetate (50.0 mL × 3), and the combined organic phases were washed with saturated brine (40.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 100:1 to 1:1) to obtain compound A-5.

[0141] 1 H NMR (400MHz, MeOD) δ4.41-4.32(m,2H), 4.21-4.12(m,1H), 3.83-3.72(m,2H), 2.34-2.31(m,3H), 1.45-1.42(m,9H).

[0142] (3) Compound A-5 (5.0 g, 28.9 mmol) was dissolved in acetic acid (20.0 mL) and water (2.0 mL), and N-chlorosuccinimide (865.9 mg, 6.48 mmol) was added. The reaction solution was stirred at 25 °C for 0.5 hours. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL × 2), and the combined organic phases were washed with saturated brine (10.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain intermediate A;

[0143] 1 H NMR (400MHz, CDCl3) δ4.56-4.47(m,1H), 4.42-4.30(m,4H), 1.48-1.44(m,9H).

[0144] 2. Preparation of intermediate B

[0145]

[0146] The synthetic route for intermediate B is shown below:

[0147]

[0148] (1) Dimethyl sulfoxide (6.10 g, 78.1 mmol) was slowly added dropwise to a solution of oxalyl chloride (5.45 g, 42.9 mmol) in dichloromethane (50.0 mL) over 30 minutes, and the reaction mixture was maintained at -78 °C. Subsequently, a solution of compound B-1 (5.0 g, 39.0 mmol) in dichloromethane (50.0 mL) was added, and the reaction mixture was stirred at -78 °C for 1 hour. Triethylamine (19.8 g, 195 mmol) was added, and the mixture was stirred at -78 °C for 0.5 hours. The temperature was then raised to 0 °C and the reaction was carried out for 1 hour. The reaction was quenched with water (40.0 mL), extracted with dichloromethane (40.0 mL × 2), and the combined organic phases were washed with saturated brine (40.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure to obtain compound B-2.

[0149] 1 H NMR (400MHz, CDCl3) δ9.72 (s, 1H), 2.67-2.71 (m, 2H), 2.29-2.41 (m, 2H).

[0150] (2) Compound B-3 (3.27 g, 38.9 mmol), triethylamine (7.87 g, 77.7 mmol), and sulfur (1.31 g, 40.8 mmol) were added to a solution of compound B-2 (4.9 g, 38.8 mmol) in N,N-dimethylformamide (80.0 mL). The reaction mixture was stirred at 25 °C for 12 hours. The reaction was quenched with water (50.0 mL), extracted with ethyl acetate (50.0 mL × 2), and the combined organic phases were washed with saturated brine (50.0 mL × 4), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound B-4.

[0151] MS-ESI[M+H] + Calculated value 225, measured value 225.

[0152] (3) Compound B-5 (2.67 g, 18.0 mmol) was added to a solution of compound B-4 (1.0 g, 4.46 mmol) in acetic acid (4.0 mL). The reaction mixture was stirred at 120 °C for 12 hours. The reaction was quenched with water (20.0 mL), extracted with ethyl acetate (20.0 mL × 3), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound B-6.

[0153] MS-ESI[M+H] + Calculated value 235, measured value 235.

[0154] (4) Compound B-6 (800 mg, 3.42 mmol) was dissolved in phosphorus oxychloride (3.0 mL), and the reaction solution was stirred at 115 °C for 3 hours. The reaction solution was directly concentrated under reduced pressure to remove phosphorus oxychloride, and the pH was adjusted to greater than 7 by adding ice and saturated sodium bicarbonate solution. The solution was extracted with ethyl acetate (20.0 mL × 2), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain intermediate B;

[0155] 1 H NMR (400MHz, CDCl3) δ8.85(s,1H),7.35(s,1H),3.69-3.78(m,2H).

[0156] 3. Preparation of intermediate C

[0157]

[0158] The synthetic route for intermediate C is shown below:

[0159]

[0160] (1) Compound C-1 (5.0 g, 18.6 mmol), compound C-2 (3.74 g, 27.9 mmol), cesium carbonate (12.1 g, 37.2 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (1.36 g, 1.86 mmol) were dissolved in dioxane (40.0 mL) and water (40.0 mL). The reaction mixture was stirred at 110 °C for 1 hour under nitrogen protection. 100 mL of aqueous solution was added, and the mixture was extracted with dichloromethane (100 mL × 2). The combined organic phases were washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound C-3.

[0161] MS-ESI[M+H] + The calculated value is 261, and the measured value is 261.

[0162] 1H NMR (400MHz, CDCl3) δ7.36-7.42(m,1H),7.22-7.26(m,1H),6.77-6.86(m,1H),6.11-6.19(m,1H) ,5.44-5.51(m,1H),4.55-4.63(m,2H),3.71-3.79(m,2H),2.97-3.05(m,2H),1.50-1.51(m,9H).

[0163] (2) Compound C-3 (1.6 g, 6.15 mmol), sodium periodate (3.94 g, 18.4 mmol), and potassium osmium tetroxide (452 ​​mg, 1.23 mmol) were dissolved in tetrahydrofuran (15.0 mL) and water (24.0 mL). The reaction mixture was stirred at 25 °C for 1 hour under nitrogen protection. The reaction was quenched with water (100 mL), extracted with dichloromethane (100 mL × 2), and the combined organic phases were washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound C.

[0164] 1 H NMR (400MHz, CDCl3) δ10.02-10.05(m,1H),7.80-7.84(m,1H),7.58-7.63(m ,1H),4.68-4.72(m,2H),3.79-3.84(m,2H),3.09-3.15(m,2H),1.51(s,9H).

[0165] 4. Preparation of intermediate D

[0166]

[0167] The synthetic route for intermediate D is shown below:

[0168]

[0169] (1) To a solution of compound D-1 (390 mg, 1.40 mmol) in dichloromethane (15 mL), 1-hydroxybenzotriazole (284 mg, 2.1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (403 mg, 2.10 mmol), diisopropylethylamine (543 mg, 4.20 mmol), and compound D-2 (205 mg, 2.10 mmol) were added. The reaction mixture was stirred at 25 °C for 16 hours. Saturated sodium bicarbonate aqueous solution (20.0 mL) was added, and the mixture was extracted with dichloromethane (15.0 mL × 3). The organic phase was washed with saturated brine (15.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound D-3.

[0170] MS-ESI[M+H] + The calculated value is 322, and the measured value is 322.

[0171] 1 H NMR (400MHz, MeOD) δ 8.64 (s, 1H), 7.91 (s, 1H), 4.68 (s, 2H), 3.70-3.73 (m, 2H), 3.60 (s, 3H), 3.38 (s, 3H), 2.91-2.94 (t, J = 5.6Hz, 2H), 1.50 (s, 9H).

[0172] (2) Diisobutylaluminum hydride (1.5 mol / L, 2.49 mL, 3.74 mmol) was added to a tetrahydrofuran (15.0 mL) solution of compound D-3 (400 mg, 1.24 mmol) at -78 °C. The reaction solution was stirred at 0 °C for 0.5 hours under nitrogen protection. The reaction was quenched with hydrochloric acid (1 mol / L), and the pH was adjusted to 7. Then, water (20.0 mL) was added, and the mixture was extracted with ethyl acetate (20.0 mL × 2). The combined organic phases were washed with saturated brine (20.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain intermediate D.

[0173] 1 H NMR (400MHz, MeOD) δ10.05(s,1H),8.85(s,1H),8.09(s,1H),4.72(s,2H),3.71-3.74(t,J=5.6Hz,2H),2.95-2.98(t,J=6.0Hz,2H)1.50(s,9H).

[0174] 5. Preparation of intermediate E

[0175]

[0176] The synthetic route for intermediate E is shown below:

[0177]

[0178] Under nitrogen protection at -78°C, N,N-dimethylformamide (344 mg, 4.71 mmol) was added to a tetrahydrofuran (15.0 mL) solution of compound E-1 (1.0 g, 3.14 mmol), followed by dropwise addition of n-butyllithium (2.5 mol / L, 1.89 mL, 4.73 mmol). The reaction mixture was stirred at -78°C for 2 hours under nitrogen protection. The reaction was quenched at 0°C by adding saturated ammonium chloride aqueous solution (30.0 mL), followed by the addition of water (40.0 mL). The mixture was extracted with ethyl acetate (40.0 mL × 2), and the combined organic phases were washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain intermediate E.

[0179] 1 H NMR (400MHz, CDCl3) δ9.82-9.85(m,1H),7.46-7.50(m,1H),4.52-4.57(m,2H),3.75(br t,J=5.3Hz,2H),2.90-2.96(m,2H),1.50-1.51(m,9H).

[0180] Example 1

[0181] This embodiment provides a compound 1 represented by Formula I, the structural formula of which is shown below:

[0182]

[0183] The synthetic route for compound 1 is shown below:

[0184]

[0185] (1) Potassium carbonate (437 mg, 3.16 mmol) was added to an N,N-dimethylformamide (8.0 mL) solution of intermediate B (400 mg, 1.58 mmol) and compound 1-1 (394 mg, 1.74 mmol). The reaction solution was stirred at 50 °C for 3 hours. Ethyl acetate (20.0 mL) was added, and the mixture was washed with saturated brine (20.0 mL × 5). The mixture was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1 to 1:1) to obtain compound 1-2.

[0186] 1 H NMR(400MHz, CDCl3)δ8.43(s,1H),7.34(s,1H),3.95(br d,J=6.4Hz,2H),3.76(q,J=10.8Hz,2H),3.63(q,J=10.0Hz,2H),3.55-3.45(m,2 H),3.44-3.32(m,2H),2.15-2.02(m,2H),1.96(dt,J=6.4,12.4Hz,2H),1.46(br s,9H).

[0187] (2) A solution of dioxane (4 mol / L, 2.0 mL) containing 576 mg, 1.30 mmol of compound 1-2 was added to a solution of dichloromethane (5.0 mL), and the reaction mixture was stirred at 20 °C for 2 hours. The reaction mixture was then concentrated under reduced pressure to obtain the hydrochloride salt of compound 1-3.

[0188] 1 H NMR (400MHz, DMSO-d6) δ9.66(br s,2H),8.59(s,1H),7.90(br s,1H),3.94(brs,6H),3.38-3.14(m,4H),2.25-1.93(m,4H).

[0189] (3) Triethylamine was added to a methanol (15.0 mL) solution of the hydrochloride salts of compounds 1-3 (370 mg, 0.98 mmol) to adjust the pH to 7. Compounds 1-4 (255 mg, 0.98 mmol) were added, and the reaction mixture was stirred at 25 °C for 15 minutes. Sodium cyanoborohydride (245 mg, 3.91 mmol) was then added, and the mixture was stirred at 25 °C for 12 hours. The reaction was quenched with water (20.0 mL), extracted with ethyl acetate (10.0 mL × 1), the organic phase was washed with saturated brine (10.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 100:1 to 1:1) to obtain compounds 1-5.

[0190] MS-ESI[M+H] + Calculated value 588, measured value 588.

[0191] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compounds 1-5 (347 mg, 0.59 mmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetates of compounds 1-6. The crude product is used directly in the next reaction step;

[0192] (5) Triethylamine was added to a solution of trifluoroacetate (340 mg, 565 μmol) of compounds 1-6 in N,N-dimethylformamide (8.0 mL) to adjust the pH to 8. Then, intermediate A (289 mg, 1.13 mmol) was added, and the reaction mixture was stirred at 25 °C for 0.5 hours. The reaction was quenched with water (15.0 mL), extracted with ethyl acetate (10.0 mL × 3), and the combined organic phases were washed with saturated brine (10.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compounds 1-7 were obtained by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1).

[0193] MS-ESI[M+H] + Calculated value 707, measured value 707.

[0194] (6) Add 1.5 mL of trifluoroacetic acid to a solution of compounds 1-7 (100 mg, 0.14 mmol) in dichloromethane (4.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetates of compounds 1-8. The crude product is used directly in the next reaction step.

[0195] (6) Compounds 1-9 (34.5 mg, 208 μmol) and triethylamine were added to a solution of trifluoroacetate (100 mg, 138 μmol) of compounds 1-8 in N,N-dimethylformamide (5.0 mL) to adjust the pH to 8. Then, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (105 mg, 277 μmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL × 1), and the combined organic phases were washed with saturated brine (10.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The trifluoroacetate of compound 1 was obtained by high performance liquid chromatography (Kromasil C18, 150 mm × 30 mm 5 μm, A: water (0.2% trifluoroacetic acid); B: acetonitrile, 5%-65%: 20 min; 100%: 5 min) to prepare the crude product.

[0196] MS-ESI[M+H] + Calculated value 718, measured value 718.

[0197] 1H NMR(400MHz,MeOD)δ8.44-8.37(m,1H),7.72-7.62(m,1H),7.42-7.28(m,3H) ,6.79-6.69(m,1H),6.52-6.44(m,1H),4.85-4.81(m,1H),4.66-4.54(m,4H) ,4.45-4.35(m,3H),4.28-4.22(m,2H),4.08-3.85(m,8H),3.70-3.62(m,3H) ,3.52-3.36(m,2H),3.02-2.96(m,2H),2.92-2.87(m,6H),2.38-2.09(m,4H).

[0198] Example 2

[0199] This embodiment provides a compound 2 represented by Formula I, the structural formula of which is shown below:

[0200]

[0201] The synthetic route for compound 2 is shown below:

[0202]

[0203] Triethylamine was added to adjust the pH to 8 to a solution of trifluoroacetate (100 mg, 139 μmol) of intermediates 1-8 from Example 1 in dichloromethane (5.0 mL). Then, compound 2-1 (18.8 mg, 208 μmol, 17.0 μL) was added, and the reaction mixture was stirred at -78°C for 0.5 hours. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL × 3), and the combined organic phases were washed with saturated brine (10.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high-performance liquid chromatography (Kromasil C18, 150 mm × 30 mm 5 μm, A: water (0.2% trifluoroacetic acid); B: acetonitrile, 5%–65%: 20 min; 100%: 5 min) to obtain trifluoroacetate of compound 2.

[0204] MS-ESI[M+H] + Calculated value: 661, Measured value: 661.

[0205] 1H NMR (400MHz, MeOD) δ8.49(s,1H),7.74(s,1H),7.38(d,J=7.8Hz,1H),7.34-7.26(m,2H),6.35-6.18(m,2H),5.76(d,J=9.6Hz,1H),4.61-4. 48(m,4H),4.44-4.31(m,3H),4.20(d,J=6.8Hz,2H),4.13-3.93(m,6H),3.67(t,J=6.0Hz,4H),3.48(s,1H),3.12-2.89(m,3H),2.30(s,4H).

[0206] Example 3

[0207] This embodiment provides a compound 3 represented by Formula I, the structural formula of which is shown below:

[0208]

[0209] The synthetic route for compound 3 is shown below:

[0210]

[0211] (1) Potassium carbonate (656 mg, 4.75 mmol) was added to a solution of intermediate B (400 mg, 1.58 mmol) and compound 3-1 (358 mg, 1.58 mmol) in N,N-dimethylformamide (10.0 mL). The reaction solution was stirred at 50 °C under nitrogen protection for 12 hours. Water (50.0 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20.0 mL × 2). The mixture was washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1 to 0:1) to obtain compound 3-2.

[0212] MS-ESI[M+H] + Calculated value 443, measured value 443.

[0213] 1 H NMR (400MHz, CDCl3) δ 8.44 (s, 1H), 7.13 (s, 1H), 4.13 (br s, 4H), 3.64 (q, J = 10.0Hz, 2H), 3.45 (br s, 4H), 1.84 (br t, J = 5.6Hz, 4H), 1.48 (s, 9H).

[0214] (2) A solution of dioxane (4 mol / L, 5.0 mL) of hydrogen chloride was added to an ethyl acetate (5.0 mL) solution of compound 3-2 (270 mg, 1.30 mmol), and the reaction solution was stirred at 25 °C for 3 hours. The reaction solution was concentrated under reduced pressure to obtain the hydrochloride salt of compound 3-3;

[0215] MS-ESI[M+H] + Calculated value 343, measured value 343.

[0216] (3) Triethylamine (177 mg, 1.75 mmol), compound 3-4 (153 mg, 584 μmol), and sodium cyanoborohydride (110 mg, 1.75 mmol) were added to a methanol (10.0 mL) solution of the hydrochloride salt of compound 3-3 (200 mg, 584 μmol), and stirred at 25 °C for 12 hours. The reaction was quenched by adding ammonium chloride aqueous solution (50.0 mL), extracted with ethyl acetate (50.0 mL × 1), washed with saturated brine (50.0 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 3-5;

[0217] MS-ESI[M+H] + Calculated value 588, measured value 588.

[0218] (4) Add 3.0 mL of trifluoroacetic acid to a solution of compound 3-5 (150 mg, 584 μmol) in dichloromethane (3.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Concentrate the reaction solution under reduced pressure to obtain trifluoroacetate of compound 3-6. The crude product is used directly in the next reaction step.

[0219] (5) Triethylamine (81 mg, 800 μmol) and intermediate A (82 mg, 320 μmol) were added to a solution of trifluoroacetate (130 mg, 267 μmol) in dichloromethane (3.0 mL). The reaction mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (10.0 mL), extracted with ethyl acetate (20.0 mL × 1), and the combined organic phases were washed with saturated brine (10.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 93:7) to obtain compound 3-7.

[0220] MS-ESI[M+H] + Calculated value 707, measured value 707.

[0221] 1H NMR(400MHz, CDCl3)δ8.43(s,1H),7.08-7.18(m,3H),7.01-7.07(m,1H),4.48-4.57(m,2H),4.20-4.30(m,2H),4.08-4.12(m,4H),3.95(s, 1H),3.62-3.69(m,4H),3.43-3.51(m,2H),2.89-2.98(m,2H),2.33-2 .48(m,2H),1.82-1.98(m,4H),1.41-1.46(m,9H),1.28-1.35(m,4H).

[0222] (6) Add 2.0 mL of trifluoroacetic acid to a solution of compound 3-7 (143 mg, 202 μmol) in dichloromethane (8.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate salt of compound 3-8. The crude product is used directly in the next reaction step;

[0223] MS-ESI[M+H] + Calculated value 607, measured value 607.

[0224] (7) Compound 3-9 (16 mg, 99 μmol), diisopropylethylamine (13 mg, 99 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (38 mg, 99 μmol) were added to a solution of trifluoroacetate (60 mg, 99 μmol) of intermediate 3-8 in N,N-dimethylformamide (5.0 mL). The reaction solution was stirred at 25 °C for 1 hour. The reaction was quenched with saturated brine (50.0 mL), extracted with dichloromethane (50.0 mL × 1), and the combined organic phases were washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The trifluoroacetate of compound 3 was obtained by high performance liquid chromatography (ACQUITY C18, 4.6 mm × 30 mm 1.7 μm, A: water (0.01% trifluoroacetic acid); B: acetonitrile (0.01% trifluoroacetic acid), 5%-95%: 1.5 min; 95%: 1 min; 5%: 0.01 min) to prepare the crude product.

[0225] MS-ESI[M+H] + Calculated value 718, measured value 718.

[0226] 1H NMR (400MHz, MeOD) δ8.30-8.47(m,1H),7.24-7.52(m,4H),6.62-6.89(m,1H),6.50(d,J=15.2Hz,1H),4.55-4.67(m ,4H),4.11-4.49(m,9H),3.84-3.99(m,4H),3.67(brt,J=6.0Hz,2H),3.45-3.55(m,2H),3.07-3.18(m,2H),3.01(br t,J=5.2Hz,2H),2.85-2.94(m,1H),2.90(s,5H),2.32(br d,J=14.0Hz,2H),2.07(br s,2H).

[0227] Example 4

[0228] This embodiment provides a compound 4 represented by Formula I, the structural formula of which is shown below:

[0229]

[0230] The synthetic route for compound 4 is shown below:

[0231]

[0232] Triethylamine (10.0 mg, 99 μmol) was added to a solution of trifluoroacetate (60 mg, 99 μmol) of intermediate 3-8 from Example 3 in dichloromethane (5.0 mL). Compound 4-1 (9 mg, 99 μmol, 8.1 μL) was then added, and the reaction mixture was stirred at -78 °C for 20 minutes. The reaction was quenched with saturated brine (10.0 mL), extracted with dichloromethane (20.0 mL × 1), and the combined organic phases were washed with saturated brine (10.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (ACQUITY C18, 4.6 mm × 30 mm 1.7 μm, A: water (0.01% trifluoroacetic acid); B: acetonitrile (0.01% trifluoroacetic acid), 5%-95%: 1.5 min; 95%: 1 min; 5%: 0.01 min) to separate the trifluoroacetate of compound 4.

[0233] MS-ESI[M+H] + Calculated value: 661, Measured value: 661.

[0234] 1H NMR(400MHz,MeOD)δ8.42(s,1H),7.46(s,1H),7.29-7.38(m,3H),6.18-6.44(m,2H),5.72-5.83( m,1H),4.49-4.72(m,5H),4.14-4.41(m,8H),3.93(q,J=10.4Hz,2H),3.64-3.71(m,2H),3.51(br d,J=11.2Hz,2H),3.07-3.18(m,2H),3.00(br t,J=5.6Hz,2H),2.33(br d,J=14.4Hz,2H),2.03-2.14(m,2H).

[0235] Example 5

[0236] This embodiment provides a compound 5 represented by Formula I, the structural formula of which is shown below:

[0237]

[0238] The synthetic route for compound 5 is shown below:

[0239]

[0240] (1) Triethylamine (44.3 mg, 438 μmol), compound 5-1 (115 mg, 438 μmol), and sodium cyanoborohydride (82.6 mg, 1.31 mmol) were added to a methanol (5.0 mL) solution of the hydrochloride salt of compound 1-3 in Example 1 (200 mg, 438 μmol) and stirred at 25 °C for 2 hours. The reaction was quenched with saturated brine (50.0 mL), extracted with dichloromethane (50.0 mL × 1), the organic phase was washed with saturated brine (50.0 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1 to 0:1) to obtain compound 5-2;

[0241] MS-ESI[M+H] + Calculated value 589, measured value 589.

[0242] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 5-2 (72 mg, 122 μmol) in dichloromethane (4.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 5-3. The crude product is used directly in the next reaction step.

[0243] MS-ESI[M+H] + Calculated value 489, measured value 489.

[0244] (3) Triethylamine (35 mg, 348 μmol) was added to a solution of trifluoroacetate (70 mg, 116 μmol) in N,N-dimethylformamide (8.0 mL), followed by intermediate A (30 mg, 116 μmol). The reaction mixture was stirred at 25 °C for 5 minutes. The reaction was quenched with saturated brine (20.0 mL), extracted with dichloromethane (20.0 mL × 1), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 5-4 was obtained by silica gel column chromatography (dichloromethane / methanol = 100:1 to 10:1).

[0245] MS-ESI[M+H] + Calculated value 708, measured value 708.

[0246] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 5-4 (53 mg, 74.9 μmol) in dichloromethane (4.0 mL), and stir the reaction mixture at 25 °C for 20 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain the trifluoroacetate of compound 5-5. The crude product is used directly in the next reaction step.

[0247] MS-ESI[M+H] + Calculated value 608, measured value 608.

[0248] (5) Add triethylamine (7.0 mg, 69.3 μmol), trifluoroacetate of compound 5-5 (9.0 mg, 69.3 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (26.3 mg, 69.3 μmol) to a dichloromethane (5.0 mL) solution of compound 5-6 (50 mg, 69.3 μmol). Stir the reaction solution at 25 °C for 0.5 hours. Quench the reaction with saturated brine (20.0 mL), extract with dichloromethane (20.0 mL × 1), wash the organic phase with saturated brine (20.0 mL × 2), dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Phenomenexluna C18, 100mm×40mm 3μm, A: water (0.225% formic acid); B: acetonitrile, 0%-35%: 10 min) to separate the formate salt of compound 5.

[0249] MS-ESI[M+H] + Calculated value 719, measured value 719.

[0250] 1H NMR(400MHz,MeOD)δ8.40-8.33(m,1H),8.31-8.22(m,1H),7.70-7.58(m,2H),6.87-6.70(m,1H),6.3 3-6.20(m,1H),4.61-4.51(m,4H),4.44-4.37(m,1H),4.34-4.23(m,2H),3.95-3.79(m,5H),3.74(br s,5H),3.38-3.33(m,2H),3.01-2.94(m,2H),2.85-2.79(m,1H),2.76-2.67(m,2 H),2.62-2.54(m,1H),2.47-2.37(m,6H),2.16-2.02(m,2H),1.99-1.90(m,2H).

[0251] Example 6

[0252] This embodiment provides a compound 6 represented by Formula I, the structural formula of which is shown below:

[0253]

[0254] The synthetic route for compound 6 is shown below:

[0255]

[0256] (1) Triethylamine (160 mg, 1.58 mmol) and compound 6-1 (247 mg, 947 μmol) were added to a methanol (15.0 mL) solution of trifluoroacetate (360 mg, 789 μmol) of compound 3-3 in Example 3, and stirred at 25 °C for 15 minutes. Sodium cyanoborohydride (198 mg, 3.16 mmol) was added, and stirred at 25 °C for 1 hour. The reaction was quenched with saturated brine (50.0 mL), extracted with dichloromethane (50.0 mL × 1), washed with saturated brine (25.0 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 20:1 to 0:1) to obtain compound 6-2.

[0257] MS-ESI[M+H] + Calculated value 588, measured value 588.

[0258] 1H NMR(400MHz, CDCl3)δ8.42(s,1H),7.36(s,1H),7.13-7.18(m,1H),7.06-7.12(m,2 H),4.56(s,2H),3.78-3.93(m,3H),3.72-3.77(m,1H),3.59-3.69(m,6H),2.76(br s,4H),2.55-2.72(m,2H),2.06-2.17(m,2H),1.90-2.01(m,2H),1.45-1.50(m,9H).

[0259] (2) Add 2.0 mL of trifluoroacetic acid to a solution of compound 6-2 (220 mg, 220 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 15 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 6-3. The crude product is used directly in the next reaction step.

[0260] MS-ESI[M+H] + Calculated value 488, measured value 488.

[0261] (3) Triethylamine was added to a solution of trifluoroacetate (220 mg, 366 μmol) of compound 6-3 in dichloromethane (5.0 mL) to adjust the pH to 8. Intermediate A (140 mg, 549 μmol) was added, and the reaction mixture was stirred at 25 °C for 0.5 hours. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 3), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 6-4 was obtained by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1).

[0262] MS-ESI[M+H] + Calculated value 707, measured value 707.

[0263] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 6-4 (125 mg, 177 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 6-5. The crude product is used directly in the next reaction step.

[0264] MS-ESI[M+H] + Calculated value 607, measured value 607.

[0265] (5) Triethylamine was added to a solution of trifluoroacetate (60 mg, 83.2 μmol) of intermediate 6-5 in dichloromethane (3.0 mL) to adjust the pH to 7. Compound 6-6 (16 mg, 99 μmol) was then added, and the reaction mixture was stirred at -78 °C for 15 minutes. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (20.0 mL × 2), and the combined organic phases were washed with saturated brine (20.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Phenomenexluna C18, 100 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 0%-50%: 10 min) to obtain the formate salt of compound 6.

[0266] MS-ESI[M+H] + Calculated value: 661, Measured value: 661.

[0267] 1 H NMR(400MHz,MeOD)δ8.32-8.26(m,1H),7.63-7.58(m,1H),7.33-7.24(m,2H) ,7.22-7.14(m,1H),6.36-6.20(m,2H),5.80-5.73(m,1H),4.58-4.50(m,4H) ,4.37-4.29(m,1H),4.23-4.22(m,2H),4.11-4.00(m,2H),4.00-3.74(m,6H) ,3.70-3.60(m,2H),3.24-3.08(m,3H),3.05-2.91(m,3H),2.24-1.99(m,4H).

[0268] Example 7

[0269] This embodiment provides a compound 7 represented by Formula I, the structural formula of which is shown below:

[0270]

[0271] The synthetic route for compound 7 is shown below:

[0272]

[0273] In a solution of trifluoroacetate (60 mg, 83.3 μmol) of compound 6-5 in Example 6, in dichloromethane (3.0 mL), triethylamine (8.4 mg, 83.3 μmol), compound 7-1 (27.6 mg, 167 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (63.3 mg, 167 μmol) were added. The reaction mixture was stirred at 25 °C for 0.5 h. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 3), washed with saturated brine (20.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The formate salt of compound 7 was obtained by high performance liquid chromatography (1-Welch Ultimate, 70 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to prepare the crude product.

[0274] MS-ESI[M+H] + Calculated value 719, measured value 719.

[0275] 1 H NMR(400MHz,MeOD)δ8.32-8.25(m,1H),7.63-7.56(m,1H),7.36-7.25(m,2H),7.2 3-7.16(m,1H),6.82-6.71(m,1H),6.42-6.31(m,1H),4.62-4.50(m,4H),4.40-4. 32(m,1H),4.28-4.21(m,2H),4.18-4.08(m,2H),3.99-3.78(m,6H),3.73-3.59(m ,4H),3.28-3.05(m,4H),3.00-2.91(m,2H),2.73-2.60(m,6H),2.26-2.04(m,4H).

[0276] Example 8

[0277] This embodiment provides a compound 8 represented by Formula I, the structural formula of which is shown below:

[0278]

[0279] The synthetic route for compound 8 is shown below:

[0280]

[0281] (1) Potassium carbonate (219 mg, 1.58 mmol) was added to an N,N-dimethylformamide (8.0 mL) solution of intermediate B (200 mg, 792 μmol) and compound 8-1 (179 mg, 792 μmol), and the reaction solution was stirred at 80 °C for 2 hours. Saturated ammonium chloride aqueous solution (20.0 mL) was added, and the mixture was extracted with dichloromethane (50.0 mL). The organic phase was then washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1 to 1:1) to obtain compound 8-2.

[0282] MS-ESI[M+H] + Calculated value 443, measured value 443.

[0283] (2) Trifluoroacetic acid (1.0 mL) was added to a solution of compound 8-2 (218 mg, 493 μmol) in dichloromethane (3.0 mL), and the reaction solution was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 8-3.

[0284] MS-ESI[M+H] + Calculated value 343, measured value 343.

[0285] (3) Triethylamine (149 mg, 1.47 mmol) was added to a methanol (10.0 mL) solution of trifluoroacetate (224 mg, 491 μmol) of compound 8-3. Then, compound 8-4 (129 mg, 494 μmol) and sodium cyanoborohydride (308 mg, 4.90 mmol) were added, and the reaction mixture was stirred at 25 °C for 2 hours. Ethyl acetate (20.0 mL) was added, and the organic phase was washed with water (20.0 mL) and saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 8-5.

[0286] MS-ESI[M+H] + Calculated value 588, measured value 588.

[0287] 1H NMR(400MHz,MeOD)δ8.35(s,1H),7.51(s,1H),7.17-7.24(m,3H),4.61(s,2H),4.56(br s,2H),4.00(br s,2H),3.85-3.91(m,6H),3.63(br d,J=5.2Hz,2H),3.59(br s,2H),2.86(t,J=5.6Hz,2H),1.95-1.99(m,4H),1.49(s,9H).

[0288] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 8-5 (94.3 mg, 160 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate it under reduced pressure to obtain the trifluoroacetate of compound 8-6. The crude product is used directly in the next reaction step.

[0289] MS-ESI[M+H] + Calculated value 488, measured value 488.

[0290] (5) Triethylamine (81.1 mg, 801 μmol) was added to a solution of trifluoroacetate (96.5 mg, 160 μmol) in dichloromethane (5.0 mL), followed by intermediate A (61.7 mg, 241 μmol). The reaction mixture was stirred at 25 °C for 2 hours. The reaction was quenched with water (15.0 mL), and extracted with dichloromethane (20.0 mL). The organic phase was washed with water (20.0 mL) and saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain compound 8-7.

[0291] MS-ESI[M+H] + Calculated value 707, measured value 707.

[0292] 1 H NMR(400MHz,MeOD)δ8.33(s,1H),7.51(s,1H),7.15(dt,J=16.0,8.0Hz,3H),4.49(s,2H),4.19-4.33(m,1H),4.09-4.16(m,4H),3.90-3 .94(m,1H),3.85-3.89(m,5H),3.70(s,2H),3.62(t,J=6.0Hz,2H),3.24(s,4H),2.94(t,J=5.6Hz,2H),1.89-1.93(m,4H),1.43(s,9H).

[0293] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 8-7 (56.0 mg, 79.2 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate under reduced pressure to obtain the trifluoroacetate of compound 8-8. The crude product is used directly in the next reaction step.

[0294] MS-ESI[M+H] + Calculated value 607, measured value 607.

[0295] (7) Compound 8-9 (10.7 mg, 118 μmol) and triethylamine (40.0 mg, 395 μmol) were added to a solution of trifluoroacetate (57.1 mg, 79.2 μmol) in dichloromethane (3.0 mL). The reaction mixture was stirred at -78 °C for 1 hour. Dichloromethane (20.0 mL) was added, and the organic phase was washed sequentially with water (20.0 mL) and saturated brine (20.0 mL × 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18, 100 mm × 30 mm × 10 μm, A: water (0.225% formic acid); B: acetonitrile, 15%–45%: 10 min) to obtain the formate salt of compound 8.

[0296] MS-ESI[M+H] + Calculated value: 661, Measured value: 661.

[0297] 1 H NMR(400MHz,MeOD)δ8.35(s,1H),7.50(s,1H),7.24-7.30(m,2H),7.18-7.22(m,1H),6.21-6.36(m,2H),5.74-5.80(m,1H),4.49-4.5 6(m,4H),4.31-4.37(m,1H),4.24(d,J=7.2Hz,2H),4.09(s,2H),3.85-3.93(m,6H),3.70(s,4H),3.65(td,J=6.0,2.4Hz,2H),2.98(br t,J=6.0Hz,2H),1.94-2.02(m,4H).

[0298] Example 9

[0299] This embodiment provides a compound 9 represented by Formula I, the structural formula of which is shown below:

[0300]

[0301] The synthetic route for compound 9 is shown below:

[0302]

[0303] (1) Potassium carbonate (219 mg, 1.58 mmol) was added to a solution of intermediate B (200 mg, 792 μmol) and compound 9-1 (188 mg, 950 μmol) in N,N-dimethylformamide (10.0 mL). The reaction mixture was stirred at 25 °C for 2 hours. Saturated brine (50.0 mL) was added, and the mixture was extracted with dichloromethane (50.0 mL). The organic phase was then washed with saturated brine (50.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1 to 1:1) to obtain compound 9-2.

[0304] MS-ESI[M+H] + Calculated value 415, measured value 415.

[0305] (2) Trifluoroacetic acid (1.0 mL) was added to a solution of compound 9-2 (274 mg, 661 μmol) in dichloromethane (4.0 mL), and the reaction solution was stirred at 25 °C for 0.5 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 9-3.

[0306] MS-ESI[M+H] + Calculated value 315, measured value 315.

[0307] (3) Triethylamine (90.1 mg, 891 μmol) was added to a methanol (10.0 mL) solution of trifluoroacetate (280 mg, 891 μmol) of compound 9-3. Then, compound 9-4 (349 mg, 1.34 mmol) and sodium cyanoborohydride (55.9 mg, 891 μmol) were added, and the reaction mixture was stirred at 25 °C for 1 hour. Ethyl acetate (40.0 mL) was added, and the organic phase was washed successively with water (20.0 mL) and saturated brine (20.0 mL × 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 100:1 to 10:1) to obtain compound 9-5.

[0308] MS-ESI[M+H] + Calculated value 560, measured value 560.

[0309] 1H NMR(400MHz,MeOD)δ8.43-8.47(m,1H),7.08-7.13(m,4H),4.56-4.60(m,2H),4.42- 4.54(m,4H),3.60-3.69(m,6H),3.46-3.57(m,4H),2.82-2.87(m,2H),1.51(s,9H).

[0310] (4) Add 2.5 mL of trifluoroacetic acid to a solution of compound 9-5 (100 mg, 179 μmol) in dichloromethane (10.0 mL), and stir the reaction mixture at 25 °C for 0.5 hours. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 9-6. The crude product is used directly in the next reaction step.

[0311] MS-ESI[M+H] + Calculated value 460, measured value 460.

[0312] (5) Triethylamine (70.6 mg, 697 μmol) was added to a solution of trifluoroacetate (100 mg, 174 μmol) in dichloromethane (5.0 mL), followed by intermediate A (66.9 mg, 262 μmol). The reaction mixture was stirred at 25 °C for 5 minutes. The reaction was quenched with water (30.0 mL), and extracted with dichloromethane (40.0 mL). The organic phase was washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 100:1 to 10:1) to obtain compound 9-7.

[0313] MS-ESI[M+H] + Calculated value 679, measured value 679.

[0314] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 9-7 (77.0 mg, 113 μmol) in dichloromethane (4.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 9-8. The crude product is used directly in the next reaction step.

[0315] MS-ESI[M+H] + Calculated value 579, measured value 579.

[0316] (7) Compound 9-9 (9.8 mg, 108 μmol) and triethylamine (32.9 mg, 325 μmol) were added to a solution of trifluoroacetate (75.0 mg, 108 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at -78 °C for 5 minutes. Dichloromethane (50.0 mL) was added, and the organic phase was washed with saturated brine (50.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18, 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to obtain the formate salt of compound 9.

[0317] MS-ESI[M+H] + Calculated value: 633, Measured value: 633.

[0318] 1 H NMR(400MHz,MeOD)δ8.24-8.31(m,1H),7.32-7.42(m,1H),7.15-7.26(m,3H),6.19-6.38(m,2H),5.71-5.81(m,1H) ,4.49-4.64(m,8H),4.29-4.36(m,1H),4.20-4.27(m,2H),3.83-3.97(m,8H),3.61-3.69(m,2H),2.92-3.01(m,2H).

[0319] Example 10

[0320] This embodiment provides a compound 10 represented by Formula I, the structural formula of which is shown below:

[0321]

[0322] The synthetic route for compound 10 is shown below:

[0323]

[0324] (1) Triethylamine (221.7 mg, 2.19 mmol) was added to a methanol (15.0 mL) solution of trifluoroacetate (500 mg, 1.10 mmol) of compounds 1-3 in Example 1. Subsequently, compound 10-1 (343 mg, 1.31 mmol) and sodium cyanoborohydride (275 mg, 4.38 mmol) were added, and the reaction mixture was stirred at 25 °C for 1 hour. Water (20.0 mL) was added, and the mixture was extracted with dichloromethane (20.0 mL × 2). The combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 10-2.

[0325] MS-ESI[M+H] + Calculated value 588, measured value 588.

[0326] (2) Add 2.0 mL of trifluoroacetic acid to a solution of compound 10⁻² (330 mg, 561 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 10⁻³. The crude product is used directly in the next reaction step.

[0327] MS-ESI[M+H] + Calculated value 488, measured value 488.

[0328] (3) Triethylamine (111 mg, 1.1 mmol, 153 μL) was added to a solution of trifluoroacetate (330 mg, 548 μmol) of compound 10-2 in dichloromethane (5.0 mL), followed by intermediate A (210 mg, 823 μmol). The reaction mixture was stirred at 25 °C for 10 minutes. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 2), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 9:1) to obtain compound 10-4.

[0329] MS-ESI[M+H] + Calculated value 707, measured value 707.

[0330] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 10⁻⁴ (120 mg, 170 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 10 minutes. Filter the reaction mixture and concentrate it under reduced pressure to obtain trifluoroacetate of compound 10⁻⁵. The crude product is used directly in the next reaction step.

[0331] MS-ESI[M+H] + Calculated value 607, measured value 607.

[0332] (5) Triethylamine (8.42 mg, 83.3 μmol, 11.6 μL) was added to a solution of trifluoroacetate (60.0 mg, 83.3 μmol) in dichloromethane (3.0 mL). Then, compound 10-6 (11.3 mg, 125 μmol, 10.2 μL) was added, and the reaction mixture was stirred at -78 °C for 5 minutes. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 2), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high-performance liquid chromatography (Welch Ultimate, 75 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 10%–40%: 10 min) to obtain the formate salt of compound 10.

[0333] MS-ESI[M+H] + Calculated value 661, measured value 661.

[0334] 1 H NMR (400MHz, MeOD) δ8.25-8.33(m,1H),7.36-7.45(m,1H),7.22-7.30(m,2H),7.16-7.21(m,1H),6.21-6.37(m,2H),5.73-5.80(m,1H),4.48-4. 61(m,3H),4.31-4.40(m,2H),4.04-4.30(m,6H),3.79-3.93(m,4H),3.6 1-3.73(m,2H),2.93-3.02(m,2H),2.62-2.92(m,4H),1.96-2.10(m,4H).

[0335] Example 11

[0336] This embodiment provides a compound 11 represented by Formula I, the structural formula of which is shown below:

[0337]

[0338] The synthetic route for compound 11 is shown below:

[0339]

[0340] (1) Triethylamine (8.42 mg, 83.3 μmol, 11.6 μL) was added to a solution of trifluoroacetate (60.0 mg, 83.3 μmol) of compound 10-5 in dichloromethane (3.0 mL), followed by 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (63.3 mg, 166 μmol) and hydrochloride of compound 11-1 (27.6 mg, 166 μmol). The reaction mixture was stirred at 25 °C for 0.5 hours. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 2), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The formate salt of compound 11 was obtained by high performance liquid chromatography (Welch Xtimate, 75 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 5%-35%: 10 min) to prepare the crude product.

[0341] MS-ESI[M+H]+, calculated value 718, measured value 718.

[0342] 1 H NMR(400MHz,MeOD)δ8.26-8.31(m,1H),7.38-7.42(m,1H),7.24-7.31(m,2H),7.1 7-7.22(m,1H),6.70-6.85(m,1H),6.27-6.41(m,1H),4.57-4.62(m,2H),4.32-4. 42(m,2H),4.06-4.30(m,6H),3.83-3.93(m,4H),3.64-3.70(m,2H),3.57-3.62(m ,2H),2.94-3.02(m,3H),2.77-2.93(m,4H),2.59-2.65(m,6H),1.96-2.12(m,4H).

[0343] Example 12

[0344] This embodiment provides a compound 12 represented by Formula I, the structural formula of which is shown below:

[0345]

[0346] The synthetic route for compound 12 is shown below:

[0347]

[0348] (1) Triethylamine (33.3 mg, 329 μmol, 45.74 μL) was added to a methanol (5.0 mL) solution of trifluoroacetate (150 mg, 329 μmol) of compounds 1-3 in Example 1. Compound E (132 mg, 493 μmol) was added, and the reaction mixture was stirred at 25 °C for 15 minutes. Sodium cyanoborohydride (82.6 mg, 1.31 mmol) was then added, and the mixture was stirred at 25 °C for 12 hours. The reaction was quenched with water (30.0 mL), extracted with ethyl acetate (30.0 mL × 3), the organic phase was washed with saturated brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound 12-1.

[0349] MS-ESI[M+H] + Calculated value 594, measured value 594.

[0350] (2) Add 2.0 mL of trifluoroacetic acid to a solution of compound 12-1 (190 mg, 320 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 10 minutes. Filter the reaction mixture and concentrate it under reduced pressure to obtain the trifluoroacetate of compound 12-2. The crude product is used directly in the next reaction step.

[0351] MS-ESI[M+H] + Calculated value 494, measured value 494.

[0352] (3) Triethylamine (31.6 mg, 313 μmol, 43.5 μL) was added to a solution of trifluoroacetate (190 mg, 313 μmol) of compound 12-2 in dichloromethane (5.0 mL), followed by intermediate A (80.0 mg, 313 μmol). The reaction mixture was stirred at 25 °C for 30 minutes. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (30.0 mL × 2), and the combined organic phases were washed with saturated brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 9:1) to obtain compound 12-3.

[0353] MS-ESI[M+H] + Calculated value 713, measured value 713.

[0354] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 12-3 (76.0 mg, 107 μmol) in dichloromethane (4.0 mL), and stir the reaction mixture at 25 °C for 10 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain the trifluoroacetate salt of compound 12-4. The crude product was used directly in the next reaction; MS-ESI [M+2H] 2+ Calculated value 307, measured value 307.

[0355] (5) Triethylamine (5.29 mg, 52.3 μmol, 7.28 μL) was added to a solution of the hydrochloride salt of compound 12-5 (17.3 mg, 104 μmol) in dichloromethane (3.0 mL), followed by the addition of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (39.8 mg, 105 μmol) and the trifluoroacetate salt of compound 12-4 (38.0 mg, 52.3 μmol). The reaction mixture was stirred at 25 °C for 0.5 hours. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (30.0 mL × 2), and the combined organic phases were washed with saturated brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 0%-30%: 10 min) to separate the formate salt of compound 12.

[0356] MS-ESI[M+H] + Calculated value 724, measured value 724.

[0357] 1 H NMR(400MHz,MeOD)δ8.18(s,1H),7.51(s,1H),6.74(s,1H),6.61-6.70(m,1H),6.23-6.32(m,1H),4.38-4.52(m,2H),4.28-4.34(m,2H),4.17-4. 27(m,1H),4.06-4.15(m,2H),3.93-4.05(m,2H),3.66-3.91(m,6H),3.5 4-3.65(m,4H),2.74-3.04(m,6H),2.55-2.65(m,6H),1.89-2.13(m,4H).

[0358] Example 13

[0359] This embodiment provides a compound 13 represented by Formula I, the structural formula of which is shown below:

[0360]

[0361] The synthetic route for compound 13 is shown below:

[0362]

[0363] (1) Triethylamine (5.29 mg, 52.3 μmol) was added to a solution of trifluoroacetate (38.0 mg, 52.3 μmol) of compound 12-5 in dichloromethane (3.0 mL). Then, compound 13-1 (7.10 mg, 78.4 μmol) was added, and the reaction mixture was stirred at -78 °C for 5 minutes. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (30.0 mL × 2), and the combined organic phases were washed with saturated brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high-performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 15%–35%: 10 min) to obtain the formate salt of compound 13.

[0364] MS-ESI[M+H] + Calculated value 667, measured value 667.

[0365] 1 H NMR(400MHz,MeOD)δ8.18(s,1H),7.51(s,1H),6.68(s,1H),6.09-6.25(m,2H),5.61-5.69(m,1H),4.35-4.47(m,2H),4.30 (s,2H),4.15-4.25(m,1H),4.06-4.14(m,2H),3.67-3.93(m,8H),3.50-3.60(m,2H),2.63-2.92(m,6H),1.85-2.10(m,4H).

[0366] Example 14

[0367] This embodiment provides a compound 14 represented by Formula I, the structural formula of which is shown below:

[0368]

[0369] The synthetic route for compound 14 is shown below:

[0370]

[0371] (1) Triethylamine (130 mg, 1.29 mmol, 179 μL) was added to a methanol (5.0 mL) solution of trifluoroacetate (176 mg, 386 μmol) of compounds 1-3 in Example 1. Then, compound C (101 mg, 386 μmol) and sodium cyanoborohydride (162 mg, 2.57 mmol) were added, and the mixture was stirred at 25 °C for 12 hours. The reaction solution was concentrated under reduced pressure, dissolved in ethyl acetate (150 mL), washed with saturated brine (120.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 16:1) to obtain compound 14-1.

[0372] MS-ESI[M+H] + Calculated value 589, measured value 589.

[0373] (2) Trifluoroacetic acid (441 mg, 3.87 mmol, 287 μL) was added to a solution of compound 14-1 (70.0 mg, 119 μmol) in dichloromethane (3.0 mL), and the reaction mixture was stirred at 25 °C for 30 minutes. The reaction mixture was filtered and concentrated under reduced pressure to obtain the trifluoroacetate of compound 14-2. The crude product was used directly in the next step of the reaction.

[0374] MS-ESI[M+H] + Calculated value 489, measured value 489.

[0375] (3) Triethylamine (353 mg, 3.48 mmol, 485 μL) was added to a solution of trifluoroacetate (70.0 mg, 116 μmol) in dichloromethane (3.0 mL), followed by intermediate A (44.6 mg, 174 μmol). The reaction mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (50.0 mL), extracted with dichloromethane (40.0 mL), and the organic phase was washed with saturated brine (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain compound 14-3.

[0376] MS-ESI[M+H] + Calculated value 708, measured value 708.

[0377] 1H NMR (400MHz, CDCl3) δ8.22-8.27(m,1H),7.52-7.62(m,2H),7.36(d,J=8.0Hz,1H),4.51(s,2H),4.22-4.30(m,1H),4.14(br s,4H),3.86(q,J=10.8Hz,4H),3.76(s,2H),3.69(br t,J=6.0Hz,2H),3.01(br t,J=6.0Hz,2H),2.80-2.88(m,1H),2.73(br d,J=9.6Hz,2H),2.59(br d,J=9.6Hz,1H),2.00-2.13(m,2H),1.88-1.98(m,2H),1.42(s,9H),1.27-1.34(m,2H)

[0378] (4) Trifluoroacetic acid (353 mg, 3.10 mmol, 229 μL) was added to a solution of compound 14-3 (39.0 mg, 55.1 μmol) in dichloromethane (2.0 mL), and the reaction mixture was stirred at 25 °C for 30 minutes. The reaction mixture was filtered and concentrated under reduced pressure to obtain the trifluoroacetate of compound 14-4. The crude product was used directly in the next step of the reaction.

[0379] MS-ESI[M+H] + Calculated value 608, measured value 608.

[0380] (5) Triethylamine (16.4 mg, 162 μmol, 22.6 μL) was added to a 2.0 mL solution of compound 14-6 hydrochloride (17.9 mg, 108 μmol) in dichloromethane. Then, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (41.1 mg, 108 μmol) was added, and the reaction mixture was stirred at 25 °C for 0.5 h. Then, a 2.0 mL solution of compound 14-4 trifluoroacetate (39.0 mg, 54.0 μmol) and triethylamine (8.20 mg, 81.1 μmol, 11.3 μL) in dichloromethane was added, and the reaction mixture was stirred at 25 °C for 0.5 h. The reaction was quenched with water (50.0 mL), extracted with dichloromethane (60.0 mL), and the organic phase was washed with saturated brine (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high-performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 15%–45%: 10 min) to obtain the formate salt of compound 14.

[0381] MS-ESI[M+H]+, calculated value 719, measured value 719.

[0382] 1 H NMR(400MHz,MeOD)δ8.29(s,1H),7.63(br s,2H),7.38(br s,1H),6.77(br s,1H),6.33(br d,J=14.0Hz,1H),4.58-4.68(m,2H),4.24-4.41(m,3H),4.00-4.08(m,2H),3.82-3.96(m,6H),3.75(br s,2H),3.55(m,2H),2.88-3.19(m,7H),2.59(br s,6H),1.79-2.41(m,5H).

[0383] Example 15

[0384] This embodiment provides a compound 15 represented by Formula I, the structural formula of which is shown below:

[0385]

[0386] The synthetic route for compound 15 is shown below:

[0387]

[0388] (1) Triethylamine (1.96 mg, 19.4 μmol, 2.70 μL) and compound 15-1 (3.51 mg, 38.8 μmol, 3.16 μL) were added to a dichloromethane (2.0 mL) solution of trifluoroacetate (17.0 mg, 23.6 μmol) of compound 14-4 from Example 14. The reaction mixture was stirred at -78 °C for 3 hours. The reaction was quenched with water (25.0 mL), extracted with dichloromethane (30.0 mL), washed with saturated brine (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 15%-35%: 10 min) to obtain the formate salt of compound 15.

[0389] MS-ESI[M+H] + Calculated value 662, measured value 662.

[0390] 1H NMR(400MHz,MeOD)δ8.30(s,1H),7.60-7.65(m,2H),7.33-7.38(m,1H),6.26-6.32(m,1H),5.77(dd,J=9.6,2.5Hz,1H),4.61-4.66( m,2H),4.49-4.56(m,2H),4.38(ddd,J=13.7,8.0,5.7Hz,1H),4.22-4.32(m,2H),4.13-4.19(m,2H),3.88(q,J=10.6Hz,6H),3.80(br t,J=6.1Hz,1H),3.74-3.77(m,2H),3.13-3.26(m,3H),3.07(br t,J=5.7Hz,3H),2.18(br d,J=5.6Hz,2H),2.07-2.13(m,2H).

[0391] Example 16

[0392] This embodiment provides a compound 16 represented by Formula I, the structural formula of which is shown below:

[0393]

[0394] The synthetic route for compound 16 is shown below:

[0395]

[0396] (1) Triethylamine (68.7 mg, 679 μmol, 94.5 μL) was added to a methanol (5.0 mL) solution of trifluoroacetate (310 mg, 679 μmol) of compounds 1-3 in Example 1. Then, compound 16-1 (272 mg, 1.02 mmol) and sodium cyanoborohydride (171 mg, 2.72 mmol) were added, and the mixture was stirred at 25 °C for 12 hours. The reaction solution was poured into water (40.0 mL), extracted with ethyl acetate (40 mL × 3), the organic phases were combined, washed with saturated brine (40.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 9:1) to obtain compound 16-2.

[0397] MS-ESI[M+H] + Calculated value 594, measured value 594.

[0398] (2) Trifluoroacetic acid (3.08 g, 27.0 mmol, 2 mL) was added to a solution of compound 16-2 (350 mg, 589 μmol) in dichloromethane (8.0 mL). The reaction mixture was stirred at 25 °C for 10 minutes. The reaction mixture was filtered and concentrated under reduced pressure to obtain trifluoroacetate of compound 16-3. The crude product was used directly in the next reaction step.

[0399] MS-ESI[M+H] + Calculated value 494, measured value 494.

[0400] (3) Triethylamine (58.3 mg, 576 μmol, 80.2 μL) was added to a solution of trifluoroacetate (350 mg, 576 μmol) in dichloromethane (5.0 mL), followed by intermediate A (221 mg, 864 μmol). The reaction mixture was stirred at 25 °C for 0.5 hours. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (30.0 mL × 3), and the organic phases were combined, washed with saturated brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 9:1) to obtain compound 16-4.

[0401] MS-ESI[M+H] + Calculated value 713, measured value 713.

[0402] (4) Trifluoroacetic acid (3.08 g, 27.0 mmol, 2 mL) was added to a solution of compound 16-4 (210 mg, 295 μmol) in dichloromethane (6.0 mL). The reaction mixture was stirred at 25 °C for 30 minutes. The reaction mixture was filtered and concentrated under reduced pressure to obtain the trifluoroacetate of compound 16-5. The crude product was used directly in the next reaction step.

[0403] MS-ESI[M+2H] 2+ Calculated value 307, measured value 307.

[0404] (5) Triethylamine (14.6 mg, 144 μmol, 20.11 μL) was added to a solution of 47.9 mg, 289 μmol of the hydrochloride salt of compound 16-6 in dichloromethane (5.0 mL), followed by the addition of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (110 mg, 289 μmol) and trifluoroacetate of compound 16-5 (105 mg, 144 μmol). The reaction mixture was stirred at 25 °C for 0.5 h. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 2), washed with saturated brine (20.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 0%-30%: 10 min) to separate the formate salt of compound 16.

[0405] MS-ESI[M+H] + Calculated value 724, measured value 724.

[0406] 1 H NMR(400MHz,MeOD)δ8.28(s,1H),7.61(s,1H),6.71-6.84(m,2H),6.25-6.38(m,1H),4.53-4.56(m,2H),4.27-4.35(m,1 H),4.18-4.24(m,2H),3.98-4.04(m,2H),3.59-3.91(m,12H),2.73-3.07(m,6H),2.65-2.67(m,6H),1.93-2.21(m,4H).

[0407] Example 17

[0408] This embodiment provides a compound 17 represented by Formula I, the structural formula of which is shown below:

[0409]

[0410] The synthetic route for compound 17 is shown below:

[0411]

[0412] (1) Triethylamine (14.6 mg, 144 μmol, 20.1 μL) was added to a dichloromethane (5.0 mL) solution of trifluoroacetate (105 mg, 144 μmol) of compound 16-5 from Example 16, followed by compound 17-1 (19.6 mg, 217 μmol, 17.7 μL). The reaction mixture was stirred at -78 °C for 5 minutes. The reaction was quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL × 2), washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to obtain the formate salt of compound 17.

[0413] MS-ESI[M+H] + Calculated value 667, measured value 667.

[0414] 1 H NMR(400MHz,MeOD)δ8.27(s,1H),7.59(s,1H),6.84(s,1H),6.18-6.33(m,2H),5.71-5.79(m,1H),4.43-4.61(m,2H),4.25-4.34(m,1H),4.18-4. 24(m,2H),4.03-4.12(m,2H),3.74-3.97(m,7H),3.52-3.72(m,3H),2.9 6-3.16(m,3H),2.84-2.91(m,1H),2.71-2.79(m,2H),1.96-2.22(m,4H).

[0415] Example 18

[0416] This embodiment provides a compound 18 represented by Formula I, the structural formula of which is shown below:

[0417]

[0418] The synthetic route for compound 18 is shown below:

[0419]

[0420] (1) Triethylamine (76.1 mg, 752 μmol), compound D (65.7 mg, 250 μmol), and sodium cyanoborohydride (63.0 mg, 1.0 mmol) were added to a methanol (5.0 mL) solution of trifluoroacetate (95.0 mg, 250 μmol) of compound 3-3 in Example 3. The mixture was stirred at 25 °C for 16 hours. Ammonium chloride (10.0 mL) was added, and the mixture was extracted with dichloromethane (10.0 mL × 2). The organic phase was washed with saturated brine (10.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 20:1 to 1:0) to obtain compound 18-1.

[0421] MS-ESI[M+H] + Calculated value 589, measured value 589.

[0422] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 18-1 (89 mg, 151 μmol) in dichloromethane (4.0 mL), and stir the reaction solution at 25 °C for 20 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 18-2. The crude product is used directly in the next reaction step.

[0423] (3) Triethylamine (15.1 mg, 149 μmol) was added to a solution of trifluoroacetate (73.0 mg, 149 μmol) of compound 18-2 in N,N-dimethylformamide (2.0 mL). Intermediate A (38.2 mg, 149 μmol) was added, and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was poured into water (30.0 mL), extracted with dichloromethane (15.0 mL × 2), and the combined organic phases were washed with saturated brine (10.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 50:1 to 10:1) to obtain compound 18-3.

[0424] MS-ESI[M+H] + Calculated value 708, measured value 708.

[0425] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 18-3 (98 mg, 138 μmol) in dichloromethane (4.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate under reduced pressure to obtain the trifluoroacetate of compound 18-4. The crude product is used directly in the next reaction step.

[0426] MS-ESI[M+H] + Calculated value: 608, Actual value: 608.

[0427] (5) To a solution of trifluoroacetate (84 mg, 116 μmol) of intermediate 18-4 in dichloromethane (2.0 mL), N,N-diisopropylethylamine (45.1 mg, 349 μmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (88.5 mg, 232 μmol), and compound 19-6 (15.0 mg, 99 μmol) were added. The reaction mixture was stirred at 25 °C for 30 minutes. Water (30.0 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50.0 mL). The combined organic phases were washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The trifluoroacetate of compound 18 was obtained by high performance liquid chromatography (Kromasil C18, 150 mm × 30 mm 5 μm, A: water (0.2% trifluoroacetic acid); B: acetonitrile, 5%-65%: 20 min; 100%: 5 min) to prepare the crude product.

[0428] MS-ESI[M+H] + Calculated value 719, measured value 719.

[0429] 1 H NMR(400MHz,MeOD)δ8.29-8.24(m,1H),8.20-8.16(m,1H),7.63-7.54(m,1H),7.33-7.27 (m,1H),6.76-6.60(m,1H),6.33-6.20(m,1H),5.31-5.19(m,1H),4.54-4.50(m,1H),4.3 5-4.30(m,1H),4.23-4.17(m,2H),3.82-3.74(m,2H),3.60-3.50(m,5H),2.94-2.89(m,2 H),2.55-2.46(m,8H),2.12-2.07(m,1H),1.96-1.83(m,5H),1.55-1.46(m,1H),1.23(br s,7H).

[0430] Example 19

[0431] This embodiment provides a compound 19 represented by Formula I, the structural formula of which is shown below:

[0432]

[0433] The synthetic route for compound 19 is shown below:

[0434]

[0435] (1) To a tetrahydrofuran (100 mL) solution of compound B-4 (17.5 g, 78.1 mmol), N,N'-carbonyldiimidazole (37.9 g, 234 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (35.7 g, 234 mmol) were added, and the reaction mixture was stirred at 70 °C for 4 hours. The reaction was quenched with water (50.0 mL), extracted with ethyl acetate (50.0 mL × 1), and the aqueous phase was concentrated under reduced pressure to obtain compound 19-1.

[0436] MS-ESI[M+H] + Calculated value 251, measured value 251.

[0437] (2) Compound 19-1 (9.12 g, 36.45 mmol) was dissolved in phosphorus oxychloride (34.0 mL), and the reaction solution was stirred at 115 °C for 8 hours. The reaction solution was directly concentrated under reduced pressure to remove phosphorus oxychloride, and the pH was adjusted to greater than 7 by adding ice and saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (30.0 mL × 2), and the combined organic phases were washed with saturated brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound 19-2.

[0438] (3) Potassium carbonate (2.89 g, 20.9 mmol) was added to a solution of intermediate 19-2 (2.15 g, 4.51 mmol) and compound 19-3 (2.01 mg, 7.66 mmol) in N,N-dimethylformamide (20.0 mL). The reaction mixture was stirred at 50 °C for 3 hours. Saturated brine (50.0 mL) was added, and the mixture was extracted with ethyl acetate (50.0 mL). The organic phase was then washed with saturated brine (50.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 19-4.

[0439] MS-ESI[M+H] + Calculated value 477, measured value 477.

[0440] (4) Methylamine (21.5 mL, 2 mol / L, tetrahydrofuran solution) was added to a tetrahydrofuran (20.0 mL) solution of compound 19-4 (2.0 g, 6.97 mmol), and the reaction mixture was stirred at 50 °C for 12 hours. The mixture was filtered, the organic phase was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound 19-5.

[0441] 1H NMR(400MHz,DMSO-d6)δ7.01-7.25(s,1H),6.51-6.66(m,1H),3.80-3.88(m,2H),3.3 6(m,4H),3.28-3.33(m,4H),2.69-2.84(m,3H),1.65-1.78(m,4H),1.34-1.43(m,9H).

[0442] (5) Trifluoroacetic acid (3.0 mL) was added to a solution of compound 19-5 (1.26 g, 2.67 mmol) in dichloromethane (15.0 mL), and the reaction solution was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 19-6.

[0443] MS-ESI[M+H] + Calculated value 372, measured value 372.

[0444] (6) Triethylamine (3.45 mg, 3.41 mmol) was added to a methanol (20.0 mL) solution of trifluoroacetate (950 mg, 2.56 mmol) of compound 19-6. Then, compound 19-7 (534 mg, 2.05 mmol) and sodium cyanoborohydride (642 mg, 10.2 mmol) were added, and the reaction mixture was stirred at 25 °C for 12 hours. Ethyl acetate (40.0 mL) was added, and the organic phase was washed successively with water (20.0 mL) and saturated brine (20.0 mL × 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 100:1 to 10:1) to obtain compound 19-8.

[0445] 1 H NMR(400MHz, DMSO-d6)δ7.13(s,4H),6.61(d,J=4.4Hz,1H),4.50(s,2H),4.08-4.11(m,2H),3.85(q,J=10.8Hz,3H),3.5 5(s,3H),3.17(d,J=5.2Hz,4H),3.07-3.12(m,2H),2.78(s,2H),2.76(d,J=4.4Hz,3H),1.43(s,9H),1.14-1.25(m,4H).

[0446] (7) Add 3.0 mL of trifluoroacetic acid to a solution of compound 19-8 (700 mg, 1.14 mmol) in dichloromethane (15.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 19-9. The crude product is used directly in the next reaction step.

[0447] MS-ESI[M+H] + Calculated value 517, measured value 517.

[0448] (8) Triethylamine (1.74 g, 17.2 mmol) was added to a solution of trifluoroacetate (480 mg, 929 μmol) of compound 19-9 in N,N-dimethylformamide (10.0 mL), followed by intermediate A (475 mg, 1.86 mmol). The reaction mixture was stirred at 25 °C for 30 minutes. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (40.0 mL), washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 19-10.

[0449] MS-ESI[M+H] + Calculated value 736, measured value 736.

[0450] (9) Add 1.0 mL of trifluoroacetic acid to a solution of compound 19-10 (100 mg, 135 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate it under reduced pressure to obtain the trifluoroacetate of compound 19-11. The crude product is used directly in the next reaction step.

[0451] MS-ESI[M+H] + Calculated value 636, measured value 636.

[0452] (10) Triethylamine (363 mg, 3.59 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (59.8 mg, 157 μmol), and compound 19-12 (50.0 mg, 78.6 μmol) were added to a dichloromethane (5.0 mL) solution of trifluoroacetate (19.5 mg, 117 μmol) of intermediate 19-11. The reaction solution was stirred at 25 °C for 0.5 h. Water (30.0 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (50.0 mL). The combined organic phases were washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 0%-30%: 10 min) to separate the formate salt of compound 19.

[0453] MS-ESI[M+H] + Calculated value: 747, measured value: 747.

[0454] 1H NMR(400MHz,MeOD)δ7.25-7.31(m,2H),7.20(d,J=8.0Hz,1H),7.10(s,1H),6 .73-6.82(m,1H),6.35(d,J=15.2Hz,1H),4.51-4.60(m,4H),4.32-4.39(m,1H ),4.26(d,J=6.8Hz,2H),4.09(s,4H),3.94(s,2H),3.64-3.74(m,4H),3.61(d ,J=6.8Hz,2H),2.98(t,J=5.6Hz,3H),2.85(s,6H),2.63(s,6H),2.02(s,4H).

[0455] Example 20

[0456] This embodiment provides a compound 20 represented by Formula I, the structural formula of which is shown below:

[0457]

[0458] The synthetic route for compound 20 is shown below:

[0459]

[0460] Triethylamine (23.8 mg, 235 μmol), propylphosphonic anhydride (200 mg, 314 μmol, 50% ethyl acetate solution), 4A molecular sieve (50.0 mg), and compound 20-1 (21.2 mg, 235 μmol) were added to a solution of trifluoroacetate (50.0 mg, 78.6 μmol) in ethyl acetate (5.0 mL) from intermediate 19-11 in Example 19. The reaction mixture was stirred at 25 °C for 0.5 h. Water (50.0 mL) was added, and the mixture was extracted with ethyl acetate (50.0 mL). The combined organic phases were washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 15%–35%: 10 min) to obtain the formate salt of compound 20. .

[0461] MS-ESI[M+H] + Calculated value 708, measured value 708.

[0462] 1H NMR(400MHz,MeOD)δ7.18-7.28(m,3H),7.09-7.12(s,1H),5.47-5.64(m,1 H),5.19-5.26(m,1H),4.58-4.70(m,3H),4.54-4.56(m,2H),4.33-4.39(m ,1H),4.24-4.31(m,2H),3.97-4.15(s,4H),3.79-3.86(s,2H),3.64-3.73 (m,4H),2.95-3.00(m,2H),2.89-2.91(s,3H),2.74-2.87(s,3H),1.99(br s,4H)).

[0463] Example 21

[0464] This embodiment provides a compound 21 represented by Formula I, the structural formula of which is shown below:

[0465]

[0466] The synthetic route for compound 21 is shown below:

[0467]

[0468] Triethylamine (363 mg, 3.59 mmol) was added to a dichloromethane (5.0 mL) solution of trifluoroacetate (50.0 mg, 78.7 μmol) of intermediate 19-11 from Example 19, followed by compound 21-1 (14.2 mg, 157 μmol). The reaction mixture was stirred at -78°C for 30 minutes. The reaction mixture was concentrated under reduced pressure to obtain a crude product. The crude product was then separated into formate salts by preparative high-performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min).

[0469] MS-ESI[M+H] + The calculated value is 690, and the measured value is 690.

[0470] 1H NMR(400MHz,MeOD)δ7.37-7.42(m,2H),7.25-7.32(m,2H),6.19-6.36(m,2H),5.72 -5.82(m,1H),4.61-4.58(m,3H),4.46-4.57(m,2H),4.35-4.44(m,2H),4.29-4.35 (m,3H),4.21-4.29(m,3H),3.81(m,J=10.8Hz,2H),3.63-3.71(m,2H),3.37-3.54( m,2H),3.07-3.24(m,2H),2.99-3.04(m,2H),2.94-2.99(m,3H),2.07-2.34(m,4H).

[0471] Example 22

[0472] This embodiment provides a compound 22 represented by Formula I, the structural formula of which is shown below:

[0473]

[0474] The synthetic route for compound 22 is shown below:

[0475]

[0476] (1) Triethylamine (24.9 mg, 246 μmol), compound 22-1 (59.8 mg, 223 μmol), and sodium cyanoborohydride (70.2 mg, 1.12 mmol) were added to a methanol (2.0 mL) solution of trifluoroacetate (102.0 mg, 223 μmol) of compound 3-3 in Example 3. The mixture was stirred at 25 °C for 12 hours. Water (3.0 mL) was added, and the mixture was extracted with dichloromethane (10.0 mL). The extract was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain compound 22-2.

[0477] MS-ESI[M+H] + Calculated value 594, measured value 594.

[0478] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 22-2 (90 mg, 152 μmol) in dichloromethane (3.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 22-3. The crude product is used directly in the next reaction step.

[0479] MS-ESI[M+H] + Calculated value 494, measured value 494.

[0480] (3) Triethylamine (45.0 mg, 444 μmol) was added to a solution of trifluoroacetate (90 mg, 148 μmol) in dichloromethane (3.0 mL). Intermediate A (56.8 mg, 222 μmol) was added, and the reaction mixture was stirred at 25 °C for 10 minutes. The reaction was quenched with water (2.0 mL), and the mixture was extracted with dichloromethane (10.0 mL × 2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 100:1 to 10:1) to obtain compound 22-4.

[0481] MS-ESI[M+H] + Calculated value 713, measured value 713.

[0482] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 22-4 (90 mg, 126 μmol) in dichloromethane (3.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Filter the reaction solution and concentrate under reduced pressure to obtain the trifluoroacetate of compound 22-5. The crude product is used directly in the next reaction step.

[0483] MS-ESI[M+H] + Calculated value: 613, Measured value: 613.

[0484] (5) Add triethylamine (8.35 mg, 82.6 μmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (27.21 mg, 71.55 μmol), and compound 24-6 (27.4 mg, 165 μmol) to a dichloromethane (2.0 mL) solution of trifluoroacetate (40.0 mg, 55 μmol) of intermediate 22-5. Stir the reaction solution at 25 °C for 3 hours. Add water (30.0 mL) to the reaction solution, extract with dichloromethane (50.0 mL), combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Phenomenex Luna C18 100mm×30mm 3μm, A: water (0.225% formic acid); B: acetonitrile, 0%-30%: 8 min) to separate the formate salt of compound 22.

[0485] MS-ESI[M+H] + The calculated value is 724, and the measured value is 724.

[0486] 1H NMR(400MHz,MeOD)δ8.26-8.32(m,1H),7.37-7.44(m,1H),6.70-6.85(m,2H),6.33-6.44(m ,1H),4.50-4.63(m,4H),4.05-4.39(m,6H),3.84-3.93(m,4H),3.62-3.74(m,4H),3.32(br s,2H),2.74-2.84(m,3H),2.60-2.72(m,8H),1.95-2.05(m,4H).

[0487] Example 23

[0488] This embodiment provides a compound 23 represented by Formula I, the structural formula of which is shown below:

[0489]

[0490] The synthetic route for compound 23 is shown below:

[0491]

[0492] Triethylamine (15.9 mg, 157 μmol) was added to a solution of trifluoroacetate (57.0 mg, 78.4 μmol) in dichloromethane (3.0 mL), followed by compound 23-1 (7.95 mg, 87.8 μmol). The reaction mixture was stirred at -78 °C for 10 minutes. The reaction mixture was concentrated under reduced pressure, and the crude product was separated by high performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 10%–40%: 10 min) to obtain the formate salt of compound 23.

[0493] MS-ESI[M+H] + The calculated value is 667, and the measured value is 667.

[0494] 1 H NMR (400MHz, MeOD) δ8.25-8.28(m,1H),7.37-7.44(m,1H),6.76-6.81(m,1H),6.20-6.36(m,2H),5.72-5.79(m,1H),4.58-4.69(m,8H),4.46-4. 55(m,2H),4.26-4.35(m,1H),4.17-4.25(m,2H),3.80-3.94(m,4H),3.6 1-3.72(m,2H),2.73-2.82(m,2H),2.49-2.72(m,2H),1.88-2.01(m,4H)

[0495] Example 24

[0496] This embodiment provides a compound 24 represented by Formula I, the structural formula of which is shown below:

[0497]

[0498] The synthetic route for compound 24 is shown below:

[0499]

[0500] (1) To a methanol (5.0 mL) solution of trifluoroacetate (100.0 mg, 219 μmol) of compound 3-3 from Example 3, triethylamine (44.3 mg, 438 μmol), compound E (87.8 mg, 328 μmol), and sodium cyanoborohydride (55.1 mg, 876 μmol) were added, and the mixture was stirred at 25 °C for 12 hours. Water (40.0 mL) was added, and the mixture was extracted with ethyl acetate (40.0 mL × 2). The organic phase was washed with saturated brine (40.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound 24-1.

[0501] MS-ESI[M+H] + Calculated value 594, measured value 594.

[0502] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 24-1 (120 mg, 202 μmol) in dichloromethane (4.0 mL), and stir the reaction solution at 25 °C for 10 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 24-2. The crude product is used directly in the next reaction step.

[0503] MS-ESI[M+H] + Calculated value 494, measured value 494.

[0504] (3) Triethylamine (20.0 mg, 198 μmol) was added to a solution of trifluoroacetate (120.0 mg, 198 μmol) in dichloromethane (5.0 mL). Intermediate A (101 mg, 395 μmol) was added, and the reaction mixture was stirred at 25 °C for 30 minutes. The reaction was quenched with water (20.0 mL), extracted with ethyl acetate (20.0 mL × 3), and the combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 24-3 was obtained by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1).

[0505] MS-ESI[M+H] + Calculated value 713, measured value 713.

[0506] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 24-3 (80.0 mg, 112 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Filter the reaction solution and concentrate under reduced pressure to obtain the trifluoroacetate of compound 24-4. The crude product is used directly in the next reaction step.

[0507] (5) Add triethylamine (10.4 mg, 103 μmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (78.5 mg, 206 μmol), and compound 24-5 (75.0 mg, 103 μmol) to a dichloromethane (5.0 mL) solution of trifluoroacetate (34.2 mg, 206 μmol) of intermediate 24-4. Stir the reaction solution at 25 °C for 30 minutes. Add water (30.0 mL) to the reaction solution, extract with ethyl acetate (20.0 mL × 3), combine the organic phases, wash with saturated brine (20.0 mL × 2), dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to separate the formate salt of compound 24.

[0508] MS-ESI[M+H] + The calculated value is 724, and the measured value is 724.

[0509] 1 H NMR(400MHz,MeOD)δ8.27(s,1H),7.40(s,1H),6.69-6.89(m,2H),6.29-6.41(m,1H),5.49(s,1H),4.48-4.62(m,2H),4.40-4. 47(m,2H),4.29-4.38(m,2H),4.06-4.27(m,6H),3.81-3.94(m,4H),3.56-3.76(m,4H),2.93(s,2H),2.63(s,9H),2.01(s,4H)

[0510] Example 25

[0511] This embodiment provides a compound 25 represented by Formula I, the structural formula of which is shown below:

[0512]

[0513] The synthetic route for compound 25 is shown below:

[0514]

[0515] (1) Triethylamine (66.5 mg, 657 μmol), compound C (344 mg, 1.31 mmol), and sodium cyanoborohydride (165 mg, 2.63 mmol) were added to a methanol (5.0 mL) solution of trifluoroacetate (300.0 mg, 657 μmol) of compound 3-3 in Example 3. The mixture was stirred at 25 °C for 12 hours. Water (40.0 mL) was added, and the mixture was extracted with ethyl acetate (40.0 mL × 2). The organic phase was washed with saturated brine (40.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound 25-1.

[0516] MS-ESI[M+H] + Calculated value 589, measured value 589.

[0517] (2) Add 2.5 mL of trifluoroacetic acid to a solution of compound 25-1 (383 mg, 651 μmol) in dichloromethane (4.0 mL), and stir the reaction solution at 25 °C for 30 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 25-2. The crude product is used directly in the next reaction step.

[0518] MS-ESI[M+H] + Calculated value 489, measured value 489.

[0519] (3) Triethylamine (33.6 mg, 332 μmol) was added to a solution of trifluoroacetate (200 mg, 332 μmol) in dichloromethane (5.0 mL). Intermediate A (127 mg, 498 μmol) was added, and the reaction mixture was stirred at 25 °C for 1 hour. Water (30.0 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (50.0 mL). The combined organic phases were washed with saturated brine (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 25-3 was obtained by silica gel column chromatography (dichloromethane / methanol = 1:0 to 45:1).

[0520] MS-ESI[M+H] + Calculated value 708, measured value 708.

[0521] (4) Add 0.5 mL of trifluoroacetic acid to a solution of compound 25-3 (125 mg, 177 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate it under reduced pressure to obtain the trifluoroacetate of compound 25-4. The crude product is used directly in the next reaction step.

[0522] MS-ESI[M+H] + Calculated value: 608, Actual value: 608.

[0523] (5) Add triethylamine (25.3 mg, 250 μmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (63.2 mg, 166 μmol), and compound 25-5 (27.6 mg, 167 μmol) to a dichloromethane (2.0 mL) solution of trifluoroacetate (60.0 mg, 83.1 μmol) of intermediate 25-4. Stir the reaction solution at 25 °C for 30 minutes. Add water (30.0 mL) to the reaction solution, extract with dichloromethane (50.0 mL), combine the organic phases, wash with saturated brine (30 mL × 3), dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to separate the formate salt of compound 25.

[0524] MS-ESI[M+H] + Calculated value 719, measured value 719.

[0525] 1 H NMR (400MHz, MeOD) δ8.28 (s, 1H), 7.64 (br d, J = 7.6Hz, 1H), 7.36-7.45 (m, 2H), 6.70-6.86 (m, 1H), 6.40 (br d,J=15.6Hz,1H),4.55-4.66(m,4H),4.38-4.45(m,1H),4.24-4.36(m,3H),4.05-4.24(m,3H),3.96(s,2H),3.88(q,J=10.8Hz,2H),3.77(br t,J=5.6Hz,2H),3.71(br d,J=6.4Hz,2H),3.09(br t,J=5.6Hz,2H),2.87(br s,4H),2.71(s,6H),1.99-2.10(m,4H).

[0526] Example 26

[0527] This embodiment provides a compound 26 represented by Formula I, the structural formula of which is shown below:

[0528]

[0529] The synthetic route for compound 26 is shown below:

[0530]

[0531] (1) Triethylamine (25.2 mg, 249 μmol) was added to a solution of trifluoroacetate (60.0 mg, 83.1 μmol) of compound 25-4 in dichloromethane (2.0 mL), followed by compound 26-1 (15.1 mg, 166 μmol). The reaction mixture was stirred at -78 °C for 1 hour. Water (30.0 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50.0 mL). The combined organic phases were washed with saturated brine (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 2%-25%: 10 min) to obtain the formate salt of compound 26.

[0532] MS-ESI[M+H] + Calculated value: 662, Actual value: 662.

[0533] 1 H NMR(400MHz,MeOD)δ8.29(s,1H),7.65(br d,J=8.0Hz,1H),7.37-7.41(m,2H),6.22-6.38(m,2H),5.77(dd,J=9.6,2. 4Hz,1H),4.50-4.59(m,2H),4.38-4.43(m,1H),4.25-4.33(m,3H),4.23(br d,J=11.6Hz,3H),4.02(s,2H),3.88(q,J=10.8Hz,3H),3.76(br d,J=8.8Hz,2H),3.10(br t,J=5.6Hz,2H),2.98-3.04(m,1H),2.94(br s,4H), 2.07(br s,4H)

[0534] Example 27

[0535] This embodiment provides a compound 27 represented by Formula I, the structural formula of which is shown below:

[0536]

[0537] The synthetic route for compound 27 is shown below:

[0538]

[0539] (1) Triethylamine (177 mg, 1.75 mmol), compound 27-1 (112 mg, 525 μmol), and sodium cyanoborohydride (55.0 mg, 876 μmol) were added to a solution of trifluoroacetate (200.0 mg, 438 μmol) of compound 3-3 in dichloromethane (10.0 mL). The mixture was stirred at 25 °C for 12 hours. Water (10.0 mL) was added, and the mixture was extracted with ethyl acetate (10.0 mL × 2). The organic phase was washed with saturated brine (10.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 0:1) to obtain compound 27-2.

[0540] MS-ESI[M+H] + Calculated value 540, measured value 540.

[0541] (2) Add 3.0 mL of trifluoroacetic acid to a solution of compound 27-2 (200 mg, 370 μmol) in dichloromethane (9.0 mL), and stir the reaction solution at 25 °C for 1 hour. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 27-3. The crude product is used directly in the next reaction step.

[0542] MS-ESI[M+H] + Calculated value 440, measured value 440.

[0543] (3) Triethylamine (73.1 mg, 722 μmol) was added to a solution of trifluoroacetate (200.0 mg, 361 μmol) in dichloromethane (6.0 mL). Intermediate A (184 mg, 722 μmol) was added, and the reaction mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL × 2), and the combined organic phases were washed with saturated brine (10.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 0:1) to obtain compound 27-4.

[0544] MS-ESI[M+H] + Calculated value: 659, Actual value: 659.

[0545] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 27-4 (143 mg, 217 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate under reduced pressure to obtain the trifluoroacetate of compound 27-5. The crude product is used directly in the next reaction step.

[0546] MS-ESI[M+H] + Calculated value 559, measured value 559.

[0547] (5) Add triethylamine (21.0 mg, 208 μmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (79.1 mg, 208 μmol), and compound 27-6 (51.7 mg, 312 μmol) to a dichloromethane (5.0 mL) solution of trifluoroacetate (70.0 mg, 104 μmol) of compound 27-5. Stir the reaction mixture at 25 °C for 1 hour. Add water (10.0 mL), extract with dichloromethane (10.0 mL × 2), combine the organic phases, wash with saturated brine (10.0 mL × 2), dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100mm×30mm10μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to separate the formate salt of compound 27.

[0548] MS-ESI[M+H] + The calculated value is 670, and the measured value is 670.

[0549] 1 H NMR(400MHz,MeOD)δ8.29-8.31(m,1H),7.38-7.41(m,1H),6.73-6.83(m,1H),6.35-6.47(m,1H),4.58-4.67(m,1H),4.19-4.38(m,6H) ,3.66-3.96(m,7H),2.97-3.16(m,4H),2.76-2.96(m,5H),2.70-2.76(m,6H),2.07-2.20(m,4H),1.88-2.02(m,3H),1.27-1.36(m,2H).

[0550] Example 28

[0551] This embodiment provides a compound 28 represented by Formula I, the structural formula of which is shown below:

[0552]

[0553] The synthetic route for compound 28 is shown below:

[0554]

[0555] (1) Triethylamine (21.0 mg, 208 μmol) and compound 28-1 (14.1 mg, 156 μmol) were added to a dichloromethane (5.0 mL) solution of trifluoroacetate (70.0 mg, 104 μmol) of compound 27-5 from Example 27. The reaction solution was stirred at -78 °C for 1 hour. Water (10.0 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (10 mL × 2). The combined organic phases were washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 5%-35%: 10 min) to obtain the formate salt of compound 28.

[0556] MS-ESI[M+H] + The calculated value is 613, and the measured value is 613.

[0557] 1 H NMR(400MHz,MeOD)δ8.28-8.30(m,1H),7.39-7.40(m,1H),6.25-6.36(m,2H),5.76-5.81(m,1H),4.58-4.62(m,1H),4.47-4.51(m,1H) ,4.19-4.37(m,6H),3.76-3.94(m,5H),2.84-2.98(m,6H),2.66-2.70(m,2H),2.05-2.10(m,4H),1.86-1.93(m,3H),1.22-1.31(m,2H)

[0558] Example 29

[0559] This embodiment provides a compound 29 represented by Formula I, the structural formula of which is shown below:

[0560]

[0561] The synthetic route for compound 29 is shown below:

[0562]

[0563] (1) Potassium carbonate (219 mg, 1.58 mmol) was added to a solution of intermediate B (200 mg, 792 μmol) and compound 29-1 (185 mg, 871 μmol) in N,N-dimethylformamide (5.0 mL). The reaction mixture was stirred at 80 °C for 12 hours. Water (20.0 mL) was added, and the mixture was extracted with ethyl acetate (20.0 mL × 2). The combined organic phases were washed with saturated brine (20.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 1:0 to 0:1) to obtain compound 29-2.

[0564] MS-ESI[M+H] + Calculated value 429, measured value 429.

[0565] (2) Trifluoroacetic acid (2.0 mL) was added to a solution of compound 29-2 (258 mg, 602 μmol) in dichloromethane (6.0 mL), and the reaction solution was stirred at 25 °C for 0.5 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 29-3.

[0566] MS-ESI[M+H] + Calculated value 329, measured value 329.

[0567] (3) Triethylamine (86.9 mg, 859 μmol) was added to a methanol (5.0 mL) solution of trifluoroacetate (190 mg, 430 μmol) of compound 29-3. Then, compound 30-4 (148 mg, 568 μmol) and sodium cyanoborohydride (108 mg, 1.72 mmol) were added, and the reaction mixture was stirred at 25 °C for 1 hour. Water (20.0 mL) was added, and the mixture was extracted with dichloromethane (20.0 mL × 2). The combined organic phases were washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 29-5.

[0568] MS-ESI[M+H] + Calculated value 574, measured value 574.

[0569] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 29-5 (210 mg, 366 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 0.5 hours. Filter the reaction solution and concentrate under reduced pressure to obtain trifluoroacetate of compound 29-6. The crude product is used directly in the next reaction step.

[0570] MS-ESI[M+H] + Calculated value 474, measured value 474.

[0571] (5) Triethylamine (36.2 mg, 357 μmol) was added to a solution of trifluoroacetate (210 mg, 357 μmol) in dichloromethane (5.0 mL), followed by intermediate A (183 mg, 715 μmol). The reaction mixture was stirred at 25 °C for 30 minutes. The reaction was quenched with water (20.0 mL), and extracted with dichloromethane (20.0 mL × 2). The organic phase was washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 29-7.

[0572] MS-ESI[M+H] + Calculated value 693, measured value 693.

[0573] (6) Add 2.0 mL of trifluoroacetic acid to a solution of compound 29-7 (195 mg, 274 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 29-8. The crude product is used directly in the next reaction step.

[0574] MS-ESI[M+H] + Calculated value 593, measured value 593.

[0575] (7) Compound 29-9 (18.3 mg, 202 μmol) and triethylamine (13.6 mg, 134 μmol) were added to a solution of trifluoroacetate (95.0 mg, 134 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at -78 °C for 5 minutes. Dichloromethane (50.0 mL) was added, and the organic phase was washed with saturated brine (50.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18, 100 mm × 30 mm 10 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to obtain the formate salt of compound 29.

[0576] MS-ESI[M+H] + Calculated value: 647, Measured value: 647.

[0577] 1H NMR(400MHz,MeOD)δ8.27(s,1H),7.36(s,1H),7.19-7.27(m,2H),7.12-7 .17(m,1H),6.19-6.34(m,2H),5.71-5.78(m,1H),4.49-4.55(m,4H),4.28 -4.47(m,5H),4.20-4.25(m,2H),3.83-3.91(m,2H),3.72-3.78(m,2H),3. 62-3.68(m,2H),2.93-3.03(m,4H),2.76-2.85(m,2H),2.24-2.32(m,2H).

[0578] Example 30

[0579] This embodiment provides a compound 30 represented by Formula I, the structural formula of which is shown below:

[0580]

[0581] The synthetic route for compound 30 is shown below:

[0582]

[0583] (1) Potassium carbonate (328 mg, 2.37 mmol) was added to a solution of intermediate B (317 mg, 1.42 mmol) and compound 30-1 (220 mg, 1.18 mmol) in N,N-dimethylformamide (5.0 mL). The reaction mixture was stirred at 80 °C for 2 hours. Water (40.0 mL) was added, and the mixture was extracted with ethyl acetate (40.0 mL × 2). The combined organic phases were washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 30-2.

[0584] MS-ESI[M+H] + Calculated value 403, measured value 403.

[0585] (2) Trifluoroacetic acid (5.0 mL) was added to a solution of compound 30-2 (470 mg, 1.17 μmol) in dichloromethane (15.0 mL), and the reaction solution was stirred at 25 °C for 0.5 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 30-3.

[0586] MS-ESI[M+H] + Calculated value 303, measured value 303.

[0587] (3) Triethylamine (48.6 mg, 480 μmol) was added to a methanol (5.0 mL) solution of trifluoroacetate (100 mg, 240 μmol) of compound 30-3. Then, compound 30-4 (79.4 mg, 288 μmol) and sodium cyanoborohydride (60.3 mg, 960 μmol) were added, and the reaction mixture was stirred at 25 °C for 12 hours. Water (40.0 mL) was added, and the mixture was extracted with ethyl acetate (40.0 mL × 2). The combined organic phases were washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 30-5.

[0588] MS-ESI[M+H] + Calculated value 554, measured value 554.

[0589] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 30-5 (78.0 mg, 140 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 0.5 hours. Filter the reaction mixture and concentrate it under reduced pressure to obtain trifluoroacetate of compound 30-6. The crude product is used directly in the next reaction step.

[0590] MS-ESI[M+H] + Calculated value 454, measured value 454.

[0591] (5) Triethylamine (13.9 mg, 137 μmol) was added to a solution of trifluoroacetate (78.0 mg, 137 μmol) in dichloromethane (3.0 mL), followed by intermediate A (70.2 mg, 274 μmol). The reaction mixture was stirred at 25 °C for 30 minutes. The reaction was quenched with water (40.0 mL), and extracted with ethyl acetate (40.0 mL × 2). The organic phase was washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 30-7.

[0592] MS-ESI[M+H] + Calculated value 673, measured value 673.

[0593] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 30-7 (57.0 mg, 84.7 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate under reduced pressure to obtain trifluoroacetate of compound 30-8. The crude product is used directly in the next reaction step.

[0594] MS-ESI[M+H]+ Calculated value 573, measured value 573.

[0595] (7) Add triethylamine (8.55 mg, 84.4 μmol), compound 30-9 (41.9 mg, 253 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (64.2 mg, 168 μmol) to a dichloromethane (3.0 mL) solution of trifluoroacetate of compound 30-8 (58.0 mg, 84.4 μmol), and the reaction mixture to a solution of triethylamine (8.55 mg, 84.4 μmol), compound 30-9 (41.9 mg, 253 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (64.2 mg, 168 μmol). Stir the reaction mixture at 25 °C for 1 hour. Quench the reaction with water (40.0 mL), extract with ethyl acetate (40.0 mL × 2), wash the organic phase with saturated brine (50.0 mL × 1), dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18, 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 15%-45%: 10 min) to separate the formate salt of compound 30.

[0596] MS-ESI[M+H] + Calculated value: 684, Measured value: 684.

[0597] 1 H NMR(400MHz,MeOD)δ8.34-8.38(m,1H),7.50-7.57(m,1H),6.70-6.82(m,2H),6.32-6.43(m,1H),4.50-4.62(m,4H),4.28-4.3 6(m,1H),4.19-4.26(m,2H),3.94-4.03(m,4H),3.85-3.93(m,2H),3.74-3.79(m,2H),3.61-3.73(m,4H),2.64-2.79(m,12H).

[0598] Example 31

[0599] This embodiment provides a compound 31 represented by Formula I, the structural formula of which is shown below:

[0600]

[0601] The synthetic route for compound 31 is shown below:

[0602]

[0603] (1) Triethylamine (48.6 mg, 480 μmol) was added to a methanol (5.0 mL) solution of trifluoroacetate (100 mg, 240 μmol) of compound 30-3 from Example 30. Then, compound E (79.4 mg, 288 μmol) and sodium cyanoborohydride (60.3 mg, 960 μmol) were added, and the reaction mixture was stirred at 25 °C for 12 hours. Water (40.0 mL) was added, and the mixture was extracted with ethyl acetate (40.0 mL × 2). The combined organic phases were washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 31-1.

[0604] MS-ESI[M+H] + Calculated value 554, measured value 554.

[0605] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 31-1 (72.0 mg, 130 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 0.5 hours. Filter the reaction mixture and concentrate it under reduced pressure to obtain trifluoroacetate of compound 31-2. The crude product was used directly in the next reaction step.

[0606] MS-ESI[M+H] + Calculated value 454, measured value 454.

[0607] (3) Triethylamine (12.4 mg, 123 μmol) was added to a solution of trifluoroacetate (70.0 mg, 123 μmol) in dichloromethane (5.0 mL), followed by intermediate A (63.0 mg, 246 μmol). The reaction mixture was stirred at 25 °C for 60 minutes. The reaction was quenched with water (40.0 mL), and extracted with ethyl acetate (40.0 mL × 2). The organic phase was washed with saturated brine (50.0 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 31-3.

[0608] MS-ESI[M+H] + Calculated value 673, measured value 673.

[0609] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 31-3 (46.0 mg, 68.3 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 1 hour. Filter the reaction mixture and concentrate it under reduced pressure to obtain the trifluoroacetate of compound 31-4. The crude product is used directly in the next reaction step.

[0610] MS-ESI[M+H]+ Calculated value 573, measured value 573.

[0611] (5) Add triethylamine (6.78 mg, 66.9 μmol), compound 31-5 (33.2 mg, 200 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (50.9 mg, 133 μmol) to a dichloromethane (3.0 mL) solution of trifluoroacetate of compound 31-4 (46.0 mg, 66.9 μmol), and the reaction mixture to a solution of triethylamine (6.78 mg, 66.9 μmol), compound 31-5 (33.2 mg, 200 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (50.9 mg, 133 μmol). Stir the reaction mixture at 25 °C for 1 hour. Quench the reaction with water (40.0 mL), extract with ethyl acetate (40.0 mL × 2), wash the organic phase with saturated brine (50.0 mL × 1), dry with anhydrous sodium sulfate, filter, and concentrate the organic phase under reduced pressure. The crude product was prepared by high performance liquid chromatography (Xtimate C18, 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 15%-45%: 10 min) to separate the formate salt of compound 31.

[0612] MS-ESI[M+H] + Calculated value: 684, Measured value: 684.

[0613] 1 H NMR(400MHz,MeOD)δ8.32-8.39(m,1H),7.49-7.58(m,1H),6.63-6.85(m,2H),6.29-6.48(m,1H),4.50-4.65(m,2H),4.37-4.4 7(m,2H),4.21-4.37(m,3H),3.93-4.05(m,4H),3.85-3.93(m,2H),3.60-3.82(m,6H),2.86-2.96(m,2H),2.56-2.81(m,10H).

[0614] Test case

[0615] Determination of the antiproliferative effect of the compound on MV-4-11 cells:

[0616] 1. Experimental Principle: MV-4-11 is a human leukemia cell line that carries an MLL translocation and expresses the MLL fusion protein MLL-AF4. The compound involved in this application inhibits the proliferation of MV-4-11 cells by interfering with the menin / MLL protein / protein interaction.

[0617] 2. Experimental materials: Cell Counting Kit-8 was purchased from Shanghai Liji Biotechnology Co., Ltd. (catalog number D3100L4057); 96-well clear white cell culture plates were purchased from Corning Costar (catalog number 3610); fetal bovine serum was purchased from GIBCO (catalog number #10099-141); Ishkov modified medium (IMDM) was purchased from Invitrogen (catalog number 12440046); SpectraMax i3X benchtop microplate reader was purchased from Molecular Devices.

[0618] 3. Experimental Methods: Cells in the logarithmic growth phase were resuspended in complete culture medium (IMDM + 10% fetal bovine serum (FBS)) and seeded into 96-well plates (100 μL of cell suspension per well, i.e., 15,000 cells per well). Cells were incubated at 37°C, 100% relative humidity, and 5% CO2 for 24 hours.

[0619] The test compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mM stock solution, which was then serially diluted 8 times with DMSO at a 4-fold gradient. It was then diluted 20-fold with culture medium. 25 μL / well was added to each well of a 96-well plate inoculated with cells, resulting in final compound concentrations of: 100 μM, 25 μM, 6.25 μM, 1.56 μM, 0.39 μM, 0.098 μM, 0.024 μM, 0.006 μM, and 0.0015 μM (4-fold dilutions, 9 concentrations).

[0620] Cells with the test compound added were incubated at 37°C, 100% relative humidity, and 5% CO2 for 72 hours. Cell viability was detected using the CCK-8 assay: 10 μL of CCK-8 assay reagent was added to each well, and the cells were incubated for approximately 4 hours. Cells were read using a benchtop microplate reader at a wavelength of 450 nm (reference wavelength 650 nm).

[0621] 4. Data Processing:

[0622] The inhibition rate of the drug on tumor cell growth is calculated using the following formula:

[0623] Tumor cell growth inhibition rate % = [(ODc-ODs) / (ODc-ODb)] × 100%

[0624] Wherein, ODs: OD of the sample (cells + CCK-8 + test compound), ODc: OD of the negative control (cells + CCK-8 + DMSO), ODb: OD of the blank control (culture medium + CCK-8 + DMSO).

[0625] DMSO).

[0626] The IC50 of the compound was calculated using Graphpad software. 50 .

[0627] The specific testing methods are shown in Table 1:

[0628] Table 1

[0629] Test compounds <![CDATA[MV 4-11IC 50 (nM)]]> Trifluoroacetate of Example 1 1819 Trifluoroacetate of Example 2 6193 Trifluoroacetate of Example 3 1087 Trifluoroacetate of Example 4 >10000 Formate of Example 5 409.9 Formate in Example 6 336.4 Formate in Example 7 41.68 Formate in Example 8 2406 Formate of Example 9 6062 Formate of Example 10 187.7 Formate from Example 11 104.4

[0630] As shown in Table 1, the compound represented by Formula I in this application has a good inhibitory effect on the growth of human myeloid monocytic leukemia MV-4-11 cells and has the potential to be used to prepare drugs for the treatment and prevention of leukemia.

[0631] Experimental Example 2

[0632] Determination of the antiproliferative effect of the compound on MV-4-11 cells (CTG method):

[0633] 1. Experimental Principle: MV-4-11 is a human leukemia cell line that carries an MLL translocation and expresses the MLL fusion protein MLL-AF4. The compound involved in this application inhibits the proliferation of MV-4-11 cells by interfering with the menin / MLL protein / protein interaction.

[0634] 2. Experimental materials: CellTiter-Glo was purchased from Promega (catalog number #G7571); IMDM medium was purchased from Gibco (catalog number #12440061); fetal bovine serum was purchased from Excel (catalog number #FND500); dimethyl sulfoxide (DMSO) was purchased from Sigma (catalog number #D2650); 384-well cell culture plates were purchased from Corning (catalog number #3756); automated cell counter was purchased from Lifetechnologies (model Countess II); microplate reader was purchased from PerkinElmer (model EnVisionMultilabel Reader).

[0635] 3. Experimental Method: Cells in the logarithmic growth phase were resuspended in growth medium (IMDM + 10% FBS) and diluted to the target density (50,000 / mL). The cell suspension was seeded into 384-well plates at a rate of 50 μL per well and incubated overnight at 37°C in a 5% CO2 incubator.

[0636] The test compound was dissolved in DMSO to prepare a 10 mM stock solution. The stock solution was first diluted to 2 mmol / L with DMSO, then serially diluted 3-fold to obtain 10 concentrations. 5.5 μL of each concentration was diluted with 94.5 μL of growth medium. Then, 5 μL of each solution was added to each well of a 384-well plate used for cell seeding.

[0637] Cells containing the test compound were incubated at 37°C in a 5% CO2 incubator for 72 hours. 384-well plates were equilibrated at room temperature, and 15 μL of CellTiter-Glo reagent was added to each well. The mixture was vortexed for 2 minutes and incubated at room temperature for 60 minutes. The luminescence values ​​were read using an EnVision Multilabel Reader, and the IC50 of the compound was calculated using GraphPad Prism 5.0 software. 50 .

[0638] 4. Experimental data:

[0639] The specific test results are shown in Table 2:

[0640] Table 2

[0641]

[0642]

[0643] As shown in Table 2, the spirocyclic compound represented by Formula I in this application has a good inhibitory effect on the growth of human myeloid monocytic leukemia MV-4-11 cells and has the potential to be used to prepare drugs for the treatment and prevention of leukemia.

[0644] The applicant declares that this application illustrates the thiophene-pyrimidine compounds, pharmaceutical compositions containing them, and their applications through the above embodiments, but this application is not limited to the above embodiments, that is, it does not mean that this application must rely on the above embodiments to be implemented.

Claims

1. A thiophene-pyrimidine compound, characterized in that, The structural formula of the thiophene-pyrimidine compound is shown in Formula I below: in, R 1 It is a halogen-substituted C1-C6 alkyl group; R 2 For H; R 3 Selected from H, C1-C4 alkylamino groups; Both Y and Z are N; W is C; U 1 U 2 U 3 U 4 U 5 U 6 Selected independently U 7 U 8 All in, Indicates the connection position of the group. Each R' is independently selected from: H, C1-C4 alkyl groups; Each "R" is independently selected from: H, C1-C4 alkyl; Each R”' is independently selected from: H, C1-C4 alkyl; Each R is independently selected from: H, C1-C4 alkyl; A is selected from a benzene ring and contains 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, which are 5-6 membered heteroaryl groups; L 1 It is -CH2-; L 2 for in, Indicates the connection position of the group; X is a carbon atom; R 4 Selected from Wherein, R4' is selected from fluorine or chlorine, R4” is selected from H, methyl or fluorine, R 4 "The substituent is selected from H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkylamino, substituted or unsubstituted (C1-C4 alkyl)2amino, substituted or unsubstituted 4-8 membered heterocyclic groups, wherein each of the substituted substituents is independently selected from -C." 1-3 Alkyl, -N(C1-C3 alkyl) 2, 3-8 membered heterocyclic groups, wherein, Indicates the connection position of the group.

2. The thiophene-pyrimidine compound according to claim 1, characterized in that, R 1 It is trifluoroethyl.

3. The thiophene-pyrimidine compound according to claim 1, characterized in that, A represents a benzene ring, a pyridine ring, or a thiophene ring.

4. The thiophene-pyrimidine compound according to claim 1, characterized in that, The R 4 Selected from Wherein, R4' is selected from fluorine or chlorine, R4" is selected from H, methyl or fluorine, R4"' is selected from H, substituted or unsubstituted C1-C4 alkyl groups, and each of the substituted substituents is independently selected from -C 1-3 Alkyl, -N(C1-C3 alkyl) 2, 3-8 membered heterocyclic groups, Indicates the connection position of the group.

5. The thiophene-pyrimidine compound according to claim 1, characterized in that, The spirocyclic portion of the compound represented by Formula I has the following structural formula: in, The position of the group is indicated by selecting any one of the following groups:

6. The thiophene-pyrimidine compound according to claim 1, characterized in that, The structural formula in equation I is: The loop portion shown, wherein The position of the group is indicated by selecting any one of the following groups: Among them, R e R f All are H.

7. The thiophene-pyrimidine compound according to claim 1, characterized in that, The structural formula in equation I is: The annular portion shown, wherein, Indicates the connection position of the group, selected from 8. The thiophene-pyrimidine compound according to claim 1, characterized in that, The R 4 Selected from -CH2F, -CH2Cl, 9. The thiophene-pyrimidine compound according to claim 1, characterized in that, The compound represented by Formula I is selected from any one of the following compounds:

10. The thiophene-pyrimidine compound according to claim 1, characterized in that, The thiophene-pyrimidine compounds also include pharmaceutically acceptable salts of the compounds represented by Formula I.

11. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a thiophene-pyrimidine compound as described in any one of claims 1-10 and a pharmaceutically acceptable carrier.

12. Use of the thienopyrimidine compound according to any one of claims 1-10 or the pharmaceutical composition according to claim 11, characterized in that, The intended use is selected from any one of the following (a)-(c): (a) To prepare medicines for the prevention or treatment of tumors, diabetes and other diseases associated with the activity of MLL1, MLL2, MLL fusion protein and / or menin protein; (b) To prepare inhibitors for in vitro non-therapeutic use related to the activity of MLL1, MLL2, MLL fusion protein, and / or menin protein; (c) Preparation of proliferation inhibitors for non-therapeutic tumor cells in vitro.

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