A spirocyclic compound, a pharmaceutical composition comprising the same, and its applications

Abstract

This article discloses a spirocyclic compound, pharmaceutical compositions comprising the same, and their applications. The spirocyclic 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 class of spirocyclic compounds, pharmaceutical compositions containing them, and their applications. 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, the retrograde HOXD gene, especially HOXD13, is abnormally overexpressed, accompanied by high levels of menin and MLL1 proteins. HOXD13 is a downstream gene regulated by menin and MLL1.

[0005] Therefore, interfering with the interaction between menin and MLL proteins, especially through covalent binding, is a very promising strategy for treating tumors.

[0006] 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

[0007] 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.

[0008] To address the shortcomings of existing technologies, the present application aims to provide a spirocyclic compound, a pharmaceutical composition comprising the same, and the application thereof, wherein the spirocyclic compound and the pharmaceutical composition comprising the same are capable of interfering with the interaction between menin and MLL proteins.

[0009] To achieve this objective, the following technical solution is adopted in this application:

[0010] In a first aspect, this application provides a spirocyclic compound, the structural formula of which is shown in Formula I below:

[0011]

[0012] in,

[0013] R 1 Selected from -C(O)(NR) a R b )(Right now ); where R a R b Each is independently selected from H, optionally substituted C1-C6 alkyl groups, optionally substituted 3-8 membered cycloalkyl groups, and optionally substituted 4-8 membered heterocyclic groups, or R. a With R b It is connected to N to form an optionally substituted 4-8 membered heterocycle; wherein the heterocycle contains 1-3 heteroatoms selected from N, O, S, and P;

[0014] R 2 Selected from H, halogens, methyl and trifluoromethyl;

[0015] R 3 Selected from H and halogens;

[0016] R 4 Selected from H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, halogen, -NH2, -NO2, -COOH, -CN, -OH, optionally substituted C1-C6 alkylsulfonyl, optionally substituted C1-C6 alkylsulfoxide, optionally substituted C1-C6 alkylthio, -NHCOCR 4' =CH2 (i.e.) ), -NHCOCHR 4' R 4” (Right now -SO2C(R) 4' ) = CH2 (i.e.) ), -NHSO2CR 4' =CH2 (i.e.) ) and -NHSO2CHR 4' R 4” (Right now ); where R4' Selected from H, methyl, and fluorine; R 4” Selected from chlorine and bromine atoms;

[0017] Y and Z are independently selected from N and CH, respectively, and at least one of Y and Z is N;

[0018] W is selected from N and C;

[0019] V is selected from N and CR V , where R V It can be H, halogen, -CN, -OH, -NH2, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino or optionally substituted (C1-C4 alkyl)2amino;

[0020] U 1 U 2 U 3 U 4 U 5 U 6 U 7 U 8 Each is independently selected from: -C(R')(R”)-(i.e. ), -C(R')(R”)-C(R”')(R””)-(i.e. -C(=O)- (i.e.) ), -C(R')(R”)-C(=O)-(i.e. ), -C(R')(R”)-O- (i.e. ), -C(R')(R”)-NR”'- (i.e. ) and -N=C(NH2)- (i.e. ), and U 1 U 2 U 3 U 4 At most one of them is -C(=O)-, -C(R')(R”)-C(=O)-, -C(R')(R”)-O-, or -C(R')(R”)-NR”'-, U 5 U 6 At most one of them is -C(=O)-, -C(R')(R”)-C(=O)-, -C(R')(R”)-O-, -C(R')(R”)-NR”'-, or -N=C(NH2)-, U 7 U 8 At most one of them is -C(=O)-, -C(R')(R”)-C(=O)-, -C(R')(R”)-O-, -C(R')(R”)-NR”'-, or -N=C(NH2)-;

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

[0022] Each "R" is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, and cyano;

[0023] Each R”' is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy and cyano;

[0024] Each R”' is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy and cyano;

[0025] A is an optionally substituted 6-16 membered aromatic ring or an optionally substituted 5-16 membered heteroaromatic ring; wherein the heteroaromatic ring contains 1-3 heteroatoms selected from N, O, S, and P;

[0026] L 1 -CR does not exist L1 'R L1 "-(Right now ), -CO- (i.e.) -SO2- (i.e.) ), -SO-(i.e. -C(N=N)- (i.e.) ), oxygen or -NH-; where R L1 '、R L1 "Each is independently selected from: H, any substituted C1-C4 alkyl group and halogen, or R." L1 'With R L1 "To form optionally substituted 3-8 membered saturated or unsaturated cycloalkanes or optionally substituted 4-8 membered saturated or unsaturated heterocycles with the attached carbon atom; wherein,

[0027] The heterocycle contains 1-3 heteroatoms selected from N, O, S, and P;

[0028] L 2 Selected from: -SO2-, -SO-, -CO-, -CF2- and -C(N=N)-;

[0029] L 3 Selected from: oxygen atom, sulfur atom, -SO2-, -SO-, -CO-, -CR L3 'R L3 "-(Right now ) and -NR L3 "'-(Right now ); where R L3 '、R L3"Each is independently selected from: H, any substituted C1-C4 alkyl group and halogen, or R." L3 'and R L3 "Forms optionally substituted 3-8 membered saturated or unsaturated cycloalkanes or optionally substituted 4-8 membered saturated or unsaturated heterocycles with the attached carbon atom; wherein the heterocycle contains 1-3 heteroatoms selected from N, O, S, and P; R" L3 "Selected from: H, any substituted C1-C4 alkyl, any substituted 3-8 member saturated or unsaturated cycloalkanes and any substituted 4-8 member saturated or unsaturated heterocycles; wherein the heterocycle contains 1-3 heteroatoms selected from N, O, S, and P;

[0030] X is selected from: carbon atom, -S- and -SO-;

[0031] R 5 Selected from: -CH2R 5 '、 Among them, R 5 ' is a fluorine or chlorine atom; R 5 " is an H, methyl, or fluorine atom; R 5”' Selected from: H, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, optionally substituted (C1-C4 alkyl)2amino, optionally substituted C1-C4 alkylthio, optionally substituted 3-8 member saturated or unsaturated cycloalkyl, optionally substituted 4-8 member saturated or unsaturated heterocyclic groups and substituted or unsubstituted C2-C4 acyl groups; wherein the heterocyclic group comprises 1-3 heteroatoms selected from N, O, S, P;

[0032] Indicates the connection position of the group.

[0033] Preferably, the R 2 Selected from H and halogens; more preferably, the R 2 It is fluorine.

[0034] Preferably, the R 3 It is an H or fluorine atom; more preferably, the R 3 For H.

[0035] Preferably, the R 4 It can be H, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C4 alkoxy, an optionally substituted C1-C4 alkylamino, -NH2, or -CN.

[0036] Preferably, Y and Z are N respectively.

[0037] Preferably, W is C.

[0038] Preferably, V is N.

[0039] Preferably, the U 1 U 2 U 3 U 4 U 5 U 6 U 7 U 8 Each of the following is independently selected from: -C(R')(R”)-, -C(R')(R”)-C(R”')(R””), -C(=O)-, and -C(R')(R”)-C(=O)-, and U 1 U 2 U 3 U 4 At most one of them is -C(=O)- or -C(R')(R”)-C(=O)-, U 5 U 6 At most one of them is -C(=O)- or -C(R')(R”)-C(=O)-, U 7 U 8 At most one of them is -C(=O)- or -C(R')(R”)-C(=O)-; wherein each R' is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy and cyano; each R” is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy and cyano; each R”' is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy and cyano; each R”' is independently selected from: H, halogen, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy and cyano.

[0040] Preferably, A is an optionally substituted 6-10 membered aromatic ring or an optionally substituted 5-12 membered heteroaromatic ring; wherein the heteroaromatic ring contains 1-3 heteroatoms selected from N, O, S, and P; more preferably, A is an optionally substituted benzene ring, an optionally substituted pyridine ring, an optionally substituted pyridazine ring, an optionally substituted pyrimidine ring, an optionally substituted triazine ring, an optionally substituted thiophene ring, an optionally substituted thiazole ring, an optionally substituted imidazole ring, an optionally substituted pyrrole ring, an optionally substituted pyrazole ring, an optionally substituted oxazole ring, an optionally substituted isoxazole ring, or an optionally substituted triazole ring.

[0041] Preferably, the L 1 The L is absent or -CH2-; more preferably, the L 1 It is -CH2-.

[0042] Preferably, the L 2Selected from: -SO2-, -SO- and -CO-; more preferably, the L 2 It is -SO2.

[0043] Preferably, the L 3 Selected from: oxygen atom, sulfur atom, -CR L3 'R L3 "- and -NR L3 "'-; where R L3 '、R L3 "Each is independently selected from: H, any substituted C1-C4 alkyl group and halogen, or R." L3 'and R L3 "Forms optionally substituted 3-8 membered saturated or unsaturated cycloalkanes or optionally substituted 4-8 membered saturated or unsaturated heterocycles with the attached carbon atom; wherein the heterocycle contains 1-3 heteroatoms selected from N, O, S, and P; R" L3 "The L is selected from: H, any substituted C1-C4 alkyl, any substituted 3-8 member saturated or unsaturated cycloalkanes, and any substituted 4-8 member saturated or unsaturated heterocycles; wherein the heterocycle contains 1-3 heteroatoms selected from N, O, S, and P; more preferably, the L..." 3 It consists of oxygen or sulfur atoms.

[0044] Preferably, X is selected from carbon atoms and -SO-; more preferably, X is a carbon atom.

[0045] Preferably, the R 5 Selected from: -CH2R 5 '、 Among them, R 5 ' is a fluorine or chlorine atom; R 5 " is an H, methyl, or fluorine atom; R 5 "Selected from: H, optional substituted C1-C4 alkyl groups."

[0046] Preferably, in Formula I The spirocyclic ring shown is selected from any one of the following groups:

[0047]

[0048] Preferably, in Formula I The spirocyclic ring shown is selected from any one of the following groups:

[0049]

[0050] Preferably, in Formula I The cyclic moiety represented is selected from any one of the following groups:

[0051]

[0052] Among them, R e R f Each of the following 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, and diethylcarbamoyl.

[0053] Preferably, the structural formula in Formula I is: The cyclic portion shown is selected from any one of the following groups:

[0054] Preferably, the R 5 Selected from: -CH2F, -CH2F, -CH2Cl

[0055] Preferably, the compound represented by Formula I is selected from any one of the following compounds:

[0056]

[0057]

[0058]

[0059]

[0060] Preferably, the spirocyclic compound further includes any one of the following: a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, polymorph, or deuterated form of the compound represented by Formula I.

[0061] In a second aspect, this application provides a pharmaceutical composition comprising a spirocyclic compound as described in the first aspect and a pharmaceutically acceptable carrier.

[0062] 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.

[0063] Thirdly, this application provides the use of a spirocyclic 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:

[0064] (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;

[0065] (b) To prepare inhibitors for in vitro non-therapeutic use related to the activity of MLL1, MLL2, MLL fusion protein, and / or menin protein;

[0066] (c) Preparation of proliferation inhibitors for non-therapeutic tumor cells in vitro.

[0067] 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.

[0068] "Other diseases" include, but are not limited to, autoimmune diseases, non-alcoholic hepatitis, etc.

[0069] Terminology Explanation

[0070] 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.

[0071] 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.).

[0072] 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”.

[0073] In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0074] "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, propionate, hexanoate, octanoate, decanoate, undecenoate, glycolate, gluconate, lactate, sebate, adipate, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylic acid, and naphthalenedisulfonate. These salts can be prepared using methods known in this field.

[0075] "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.

[0076] 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.

[0077] 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.

[0078] 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.

[0079] 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.

[0080] The terms “preventive,” “prevention,” and “avoidance” used in this article include reducing the likelihood of a patient developing or worsening a disease or condition.

[0081] The term "treatment" and other similar synonyms used in this article include the following meanings:

[0082] (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;

[0083] (ii) To suppress a disease or symptom, that is, to curb its development;

[0084] (iii) To alleviate a disease or symptom, that is, to cause the condition of the disease or symptom to subside; or

[0085] (iv) To alleviate the symptoms caused by the disease or condition.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] Group definition

[0090] 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.

[0091] 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-.

[0092] 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.

[0093] 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.

[0094] Indicates the connection position of the group.

[0095] Except as otherwise specified, when used in the specification and claims of this application, the following terms shall have the following meanings.

[0096] In this article, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0097] "Hydroxy group" refers to the -OH group.

[0098] "Hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group (-OH) as defined below.

[0099] "Carbonyl" refers to the -C(=O)- group.

[0100] "Nitro" refers to -NO2.

[0101] "Cyano" refers to -CN.

[0102] "Amino" refers to -NH2.

[0103] "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.

[0104] The "carboxyl group" refers to -COOH.

[0105] 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.

[0106] 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.

[0107] 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.

[0108] 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.

[0109] 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.

[0110] 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.

[0111] 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.

[0112] 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.

[0113] 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.

[0114] In this application, "optionally" means that the event or condition described below may or may not occur, and the description includes both the occurrence and non-occurrence of the event or condition. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl groups. For example, where no substituent is explicitly listed, the terms "substituted" or "replaced" 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... Each of the groups may optionally be further substituted by one or more substituents selected from the following: 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(C Substituents include -1-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.

[0115] "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".

[0116] "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.

[0117] 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.

[0118] "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.

[0119] 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.

[0120] In this application, (C1-C4 alkyl)2amino represents an amine substituted with two C1-C4 alkyl groups, for example, it could be... 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'SENCYCLOPEDIAO FPRACTICAL 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 under nitrogen protection. 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) δ 5.24-5.15 (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 under nitrogen protection. 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) δ 4.41-4.32 (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 mixture was stirred at 25 °C for 0.5 hours under nitrogen protection. 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) 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (268 g, 705 mmol), diisopropylethylamine (114 g, 882 mmol), and compound B-2 (119 g, 1.18 mol) were added to a dichloromethane (1300 mL) solution of compound B-1 (100 g, 588 mmol) at 0 °C. The reaction mixture was stirred at 25 °C for 3 hours. Water (200 mL) was added, and the mixture was extracted with dichloromethane (200 mL × 3). The combined organic phases were washed with saturated brine (200 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 = 10:0 to 0:1) to obtain compound B-3.

[0149] MS-ESI[M+H] + The calculated value is 254, and the measured value is 254.

[0150] (2) Boron tribromide (257 g, 1.03 mol) was added to a dichloromethane (2.0 L) solution of compound B-3 (130 g, 513 mmol) at -78 °C. The reaction mixture was stirred at 25 °C for 4 hours. The reaction was quenched with ice water, neutralized to pH 8 with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane (300 mL × 3), and the combined organic phases were washed with saturated brine (300 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 = 10:0 to 1:1) to obtain compound B-4.

[0151] MS-ESI[M+H] + The calculated value is 240, and the measured value is 240.

[0152] (3) Cesium carbonate (470 g, 1.44 mol) and compound B-5 (29.5 g, 186 mmol) were added to a solution of compound B-4 (115 g, 41.0 mmol) in N,N-dimethylformamide (1500 mL). The reaction mixture was stirred at 130 °C for 12 hours. After cooling to room temperature, water (500 mL) was added and the mixture was extracted with ethyl acetate (400 mL × 3). The combined organic phases were washed with saturated brine (400 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 = 20:1 to 1:1) to obtain compound B-6.

[0153] MS-ESI[M+H] + The calculated value is 318, and the measured value is 318.

[0154] (4) At 0°C, m-chloroperoxybenzoic acid (211 g, 1.04 mol, 85% purity) was added to a dichloromethane (300.0 mL) solution of compound B-6 (110 g, 347 mmol). The reaction solution was stirred at 25°C for 12 hours under nitrogen protection. The reaction was quenched with saturated sodium thiosulfate solution until the starch-potassium iodide test paper no longer turned blue. Then, it was extracted with dichloromethane (1000 mL). The organic phase was washed with saturated sodium bicarbonate aqueous solution (500 mL × 2) and saturated brine (500 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 = 10:1 to 0:1) to obtain compound B-7.

[0155] MS-ESI[M+H] + The calculated value is 334, and the measured value is 334.

[0156] (5) Triethylamine (22.8 g, 225 mmol) and compound B-7 (50.0 g, 150 mmol) were added to a solution of phosphorus oxychloride (46.0 g, 255 mmol) in chloroform (700 mL) at 0 °C. The reaction mixture was stirred at 65 °C for 12 hours. The reaction was quenched with ice water, neutralized to pH 8 with saturated sodium bicarbonate solution, extracted with dichloromethane (200 mL × 2), washed with saturated brine (200 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 = 10:1 to 0:1) to obtain compound B.

[0157] 3. Preparation of intermediate C

[0158]

[0159] The synthetic route for intermediate C is shown below:

[0160]

[0161] (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.

[0162] MS-ESI[M+H] + The calculated value is 261, and the measured value is 261.

[0163] 1 H 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).

[0164] (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.

[0165] 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).

[0166] 4. Preparation of intermediate D

[0167]

[0168] The synthetic route for intermediate D is shown below:

[0169]

[0170] (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 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 D-3.

[0171] MS-ESI[M+H] + The calculated value is 322, and the measured value is 322.

[0172] 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).

[0173] (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.

[0174] 1 H NMR (400MHz, MeOD) δ 10.05 (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).

[0175] 5. Preparation of intermediate E

[0176]

[0177] The synthetic route for intermediate E is shown below:

[0178]

[0179] 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), and extraction 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 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.

[0180] 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).

[0181] 6. Preparation of intermediate F

[0182]

[0183] The synthetic route for intermediate F is shown below:

[0184]

[0185] (1) 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (101 g, 265 mmol), diisopropylethylamine (68.4 g, 529 mmol), and compound F-2 (18.4 g, 212 mmol) were added to a dichloromethane (300 mL) solution of compound F-1 (30.0 g, 176 mmol) at 0 °C. The reaction solution was stirred at 25 °C for 1 hour. Dichloromethane (400 mL) was added, and the organic phase was washed with water (250 mL × 1) and saturated brine (220 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 3:1) to obtain compound F-3.

[0186] MS-ESI[M+H] + The calculated value is 240, and the measured value is 240.

[0187] (2) Boron tribromide (88.0 g, 351 mmol) was added to a dichloromethane (500.0 mL) solution of compound F-3 (48.0 g, 176 mmol) at -78 °C. The reaction mixture was stirred at 25 °C for 4 hours. The reaction was quenched with ice water, neutralized to pH 8 with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane (500 mL), and the combined organic phases were washed with saturated sodium bicarbonate aqueous solution (500 mL × 2) and saturated brine (500 mL × 1). 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 = 1:0 to 1:1) to obtain compound F-4.

[0188] MS-ESI[M+H] + The calculated value is 226, and the measured value is 226.

[0189] 1 H NMR(400MHz, CDCl3)δ8.97(br s, 1H), 6.94-7.03 (m, 1H), 6.90-6.93 (m, 1H), 6.87-6.90 (m, 1H), 4.29-4.40 (m, 1H), 3.41 (q, J=7.2Hz, 2H), 1.22-1.26 (m, 9H).

[0190] (3) Cesium carbonate (121 g, 371 mmol) and compound F-5 (29.5 g, 186 mmol) were added to a solution of compound F-4 (27.9 g, 124 mmol) in N,N-dimethylformamide (350 mL). The reaction mixture was stirred at 130 °C for 12 hours. After cooling to room temperature, ethyl acetate (600 mL) was added. The organic phase was washed with water (500 mL × 1) and saturated brine (300 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 2:1) to obtain compound F-6.

[0191] MS-ESI[M+H] + The calculated value is 304, and the measured value is 304.

[0192] 1H NMR (400MHz, CDCl3) δ8.93 (s, 1H), 8.43 (s, 2H), 7.12-7.04 (m, 2H), 6.99 (dd, J=4.4 , 8.8Hz, 1H), 3.81 (m, 1H), 3.51-3.35 (m, 1H), 3.31-3.16 (m, 1H), 1.28-1.02 (m, 9H).

[0193] (4) At 0°C, m-chloroperoxybenzoic acid (53.5 g, 264 mmol, 85% purity) was added to a dichloromethane (300.0 mL) solution of compound F-6 (25.0 g, 82.4 mmol). The reaction solution was stirred at 25°C for 12 hours under nitrogen protection. The reaction was quenched by adding saturated sodium sulfite solution (100 mL), followed by adding saturated sodium bicarbonate aqueous solution (500 mL). The mixture was extracted with dichloromethane (300 mL). The organic phase was washed with saturated sodium bicarbonate aqueous solution (500 mL × 2) and saturated brine (500 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 25:1) to obtain compound F-7.

[0194] MS-ESI[M+H] + The calculated value is 320, and the measured value is 320.

[0195] 1 H NMR (400MHz, CDCl3) δ8.68 (d, J=1.6Hz, 1H), 8.09 (s, 1H), 7.92 (d, J=2.4Hz, 1H), 7.03-7.13 (m, 3H), 3.76 (m, 1H), 3.34-3.45 (m, 1H), 3.23-3.34 (m, 1H), 1.17-1.25 (m, 3H), 1.14 (m, 6H).

[0196] (5) Triethylamine (8.7 g, 85.5 mmol) and compound F-7 (18.2 g, 57 mmol) were added to a solution of phosphorus oxychloride (14.9 g, 97.2 mmol) in chloroform (100.0 mL) at 0 °C. The reaction mixture was stirred at 25 °C for 12 hours. The reaction was quenched with ice water, neutralized to pH 8 with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane (500 mL), and the organic phase was washed with saturated sodium bicarbonate aqueous solution (500 mL × 2) and saturated brine (500 mL × 1). 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 = 1:0 to 20:1) to obtain compound F.

[0197] MS-ESI[M+H] +The calculated value is 338, and the measured value is 338.

[0198] 1 H NMR (400MHz, CDCl3) δ8.72 (s, 1H), 8.23-8.25 (m, 1H), 7.12-7.15 (m, 1H), 7.06-7.10 (m, 1H), 6.97-7. 01 (m, 1H), 3.83 (m, 1H), 3.39-3.48 (m, 1H), 3.25-3.30 (m, 1H), 1.22-1.25 (m, 3H), 1.12-1.17 (m, 6H).

[0199] Example 1

[0200] This embodiment provides a compound 1 represented by Formula I, the structural formula of which is shown below:

[0201]

[0202] The synthetic route for compound 1 is shown below:

[0203]

[0204] (1) Potassium carbonate (3.54 g, 25.6 mmol) was added to a solution of intermediate B (3.0 g, 8.53 mmol) and compound 1-1 (2.32 g, 10.2 mmol) in N,N-dimethylformamide (30.0 mL). The reaction solution was stirred at 70 °C under nitrogen protection for 3 hours. Water (50.0 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50.0 mL × 2). The organic phase was washed with saturated brine (200.0 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 = 1:0 to 0:1) to obtain compound 1-2.

[0205] 1 H NMR (400MHz, CDCl3) δ8.37 (s, 1H), 7.77 (s, 1H), 6.95-7.01 (m, 2H), 6.73-6.78 (m, 1H), 4.00 (br s, 2H), 3.87-3.93 (m, 2H), 3.78 (dt, J=13.6, 6.8Hz, 1H), 3.48 (dt,J=13.6, 6.8Hz, 1H), 3.31-3.38 (m, 4H), 1.66-1. 71 (m, 4H), 1.53 (d, J = 6.8 Hz, 3H), 1.47 (d, J = 6.8 Hz, 3H), 1.44 (s, 9H), 1.12 (d, J = 6.8 Hz, 3H), 1.09 (d, J = 6.8 Hz, 3H).

[0206] (2) Trifluoroacetic acid (7.7 g, 67.5 mmol) was added to a solution of compounds 1-2 (4.4 g, 8.12 mmol) in dichloromethane (20.0 mL), and the reaction solution was stirred at 25 °C for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain trifluoroacetate of compounds 1-3.

[0207] MS-ESI[M+H] + The calculated value is 442, and the measured value is 442.

[0208] (3) Triethylamine (206 mg, 2.04 mmol), compounds 1-4 (267 mg, 1.02 mmol), and sodium cyanoborohydride (427 mg, 6.79 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (300 mg, 679 μmol) of compounds 1-3. The mixture was stirred at 25 °C for 4 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate (20.0 mL) was added. The mixture was washed with water (20.0 mL × 1) and saturated brine (20.0 mL × 1). The organic phase was 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 24:1) to obtain compounds 1-5.

[0209] 1 H NMR (400MHz, CDCl3) δ8.36 (s, 1H), 7.75 (s, 1H), 7.03-7.16 (m, 3H), 6.96-7.00 (m, 2H), 6.75-6.80 (m, 1H), 4.55 (s, 2H), 3.97 (br s, 2H), 3.86-3.90 (m, 2H), 3.78 (dt, J=13.2, 6.4Hz, 1H), 3.63 (br s, 2H), 3.48 (dt, J=13.2, 6.8Hz, 3H), 2.81 (br s, 2H), 2.29-2.55 (m, 3H), 1.82 (br s, 3H), 1.54 (d, J=6.8Hz, 3H), 1.46-1.49 (m, 12H), 1.13 (d, J=6.8Hz, 3H), 1.08 (d, J=6.8Hz, 3H), 0.80-0.90 (m, 2H).

[0210] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compounds 1-5 (268 mg, 390 μmol) in dichloromethane (3.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain trifluoroacetates of compounds 1-6. The crude product is used directly in the next reaction step.

[0211] (5) Triethylamine (398 mg, 3.93 mmol) and intermediate A (200 mg, 782 μmol) were added to a solution of trifluoroacetate (273 mg, 390 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 16 hours under nitrogen protection. Dichloromethane (20.0 mL) was added, and the organic phase was washed with water (20.0 mL × 1) and saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into compounds 1-7 by silica gel column chromatography (dichloromethane / methanol = 1:0 to 32:1).

[0212] MS-ESI[M+H] + The calculated value is 806, and the measured value is 806.

[0213] 1 H NMR (400MHz, MeOD) δ8.23 (s, 1H), 7.74 (s, 1H), 7.10-7.20 (m, 5H), 6.99 (dd, J=8.8, 4.0Hz, 1H), 4.59 (s, 4H), 4.50 (s, 2H), 4.20-4.25 (m, 1H), 4.13 (br s, 3H), 3.93 (br d, J=8.8Hz, 2H), 3.82 (dt, J=13.2, 6.8Hz, 1H), 3.62 (br t, J=6.4Hz, 2H), 3.55 (br s, 2H), 2.92 (br t, J=6.0Hz, 2H), 2.49 (br s, 2H), 1.83(br s, 4H), 1.54 (d, J=6.8Hz, 3H), 1.43-1.47 (m, 12H), 1.28-1.31 (m, 2H), 1.18 (d, J=6.8Hz, 3H), 1.08 (d, J=6.8Hz, 3H).

[0214] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compounds 1-7 (213 mg, 264 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 60 minutes. Concentrate the reaction mixture under reduced pressure to obtain trifluoroacetates of compounds 1-8. The crude product is used directly in the next reaction step.

[0215] MS-ESI[M+H] + The calculated value is 706, and the measured value is 706.

[0216] (7) Triethylamine (66.6 mg, 659 μmol) was added to a solution of trifluoroacetate (108 mg, 132 μmol) of compounds 1-8 in dichloromethane (3.0 mL). Then, compounds 1-9 (17.9 mg, 198 μmol, 16.1 μL) were added, and the reaction mixture was stirred at -78 °C for 60 minutes under nitrogen protection. Dichloromethane (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 preparative high-performance liquid chromatography (Phenomenexluna C18, 100 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min) to obtain the formate salt of compound 1.

[0217] MS-ESI[M+H] + The calculated value is 760, and the measured value is 760.

[0218] 1 H NMR (400MHz, MeOD) δ8.25 (s, 1H), 7.76 (s, 1H), 7.12-7.24 (m, 5H), 6.98 (dd, J=8.8, 4.4Hz, 1H), 6.19-6.39(m, 2H), 5.76(dd, J=9.6, 2.8Hz, 1H), 4.50-4.57(m, 4H), 4.30-4.37(m, 1H), 4.24(br d, J=6.8Hz, 2H), 3.96-4.13(m, 2H), 3.94(br d, J=9.2Hz, 2H), 3.83 (dt, J=13.2, 6.8Hz, 1H), 3.73 (s, 2H), 3.59-3.68 (m, 3H), 2.95 (br t, J=5.6Hz, 2H), 2.67 (br d, J=4.4Hz, 4H), 1.83-1.94 (m, 4H), 1.54 (d, J=6.8Hz, 3H), 1.45 (d, J=6.8Hz, 3H), 1.18 (d, J=6.8Hz, 3H), 1.09 (d, J=6.8Hz, 3H).

[0219] Example 2

[0220] This embodiment provides a compound 2 represented by Formula I, the structural formula of which is shown below:

[0221]

[0222] The synthetic route for compound 2 is shown below:

[0223]

[0224] Triethylamine (133 mg, 1.31 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (100 mg, 263 μmol) were added to a dichloromethane (3.0 mL) solution of compound 2-1 hydrochloride (43.6 mg, 263 μmol). The mixture was stirred at 25 °C for 0.5 h. Trifluoroacetate of compounds 1-8 from Example 1 (108 mg, 132 μmol) was then added to the reaction mixture, and the mixture was stirred at 25 °C for 0.5 h. Dichloromethane (20.0 mL) was added, and the mixture was washed with water (20.0 mL × 1) and saturated brine (20.0 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 2 by preparative high performance liquid chromatography (Phenomenexluna C18, 100mm×40mm 3μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min).

[0225] MS-ESI[M+H] + The calculated value is 817, and the measured value is 817.

[0226] 1 H NMR (400MHz, MeOD) δ8.28 (s, 1H), 7.80 (s, 1H), 7.21-7.32 (m, 3H), 7.13-7.21 (m, 2H), 6.97 (dd, J=8.8, 4.0Hz, 1H), 6.71-6.82 (m, 1H), 6.41 (br d, J=15.2Hz, 1H), 4.52-4.65 (m, 4H), 4.36 (br d, J=6.0Hz, 1H), 4.25 (br d, J=6.4Hz, 2H), 4.07 (br s, 4H), 3.97 (br d, J=9.2Hz, 2H), 3.83 (dt, J=13.2, 6.4Hz, 1H), 3.74 (br d, J=6.4Hz, 2H), 3.60-3.69 (m, 3H), 2.89-3.09 (m, 6H), 2.73 (s, 6H), 2.01 (br s, 4H), 1.55 (br d, J=6.4Hz, 3H), 1.45 (br d, J=6.8Hz, 3H), 1.18 (br d, J=6.4Hz, 3H), 1.10 (br d, J=6.4Hz, 3H).

[0227] Example 3

[0228] This embodiment provides a compound 3 represented by Formula I, the structural formula of which is shown below:

[0229]

[0230] The synthetic route for compound 3 is shown below:

[0231]

[0232] (1) Potassium carbonate (236 mg, 1.71 mmol) was added to an N,N-dimethylformamide (8.0 mL) solution of intermediate B (300 mg, 853 μmol) and compound 3-1 (213 mg, 941 μmol). The reaction solution was stirred at 70 °C under nitrogen protection for 2 hours. Ethyl acetate (40.0 mL) was added to the reaction solution, and the mixture was washed with water (40.0 mL × 1) and saturated brine (40.0 mL × 5). The organic phase was 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 = 100:1 to 1:1) to obtain compound 3-2.

[0233] MS-ESI[M+H] + The calculated value is 542, and the measured value is 542.

[0234] 1 H NMR (400MHz, MeOD) δ8.30 (d, J=4.8Hz, 1H), 7.78-7.87 (m, 1H), 7.09-7.17 (m, 2H), 6.79-6.94 (m, 1H) , 4.58(s, 2H), 3.81-3.91(m, 2H), 3.74-3.80(m, 1H), 3.53-3.66(m, 2H), 3.39-3.50(m, 2H), 3.28(br d, J=5.6Hz, 1H), 1.87-1.98 (m, 4H), 1.52-1.56 (m, 3H), 1.44-1.47 (m, 12H), 1.13-1.20 (m, 6H).

[0235] (2) Trifluoroacetic acid (2.0 mL) was added to a solution of compound 3-2 (435 mg, 803 μmol) in dichloromethane (4.0 mL), and the reaction solution was stirred at 25 °C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 3-3.

[0236] MS-ESI[M+H] + The calculated value is 442, and the measured value is 442.

[0237] (3) Triethylamine (285 mg, 2.82 mmol), compound 3-4 (369 mg, 1.41 mmol), and sodium cyanoborohydride (592 mg, 9.42 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (450 mg, 941 μmol) of compound 3-3. The mixture was stirred at 25 °C for 12 hours. Ethyl acetate (40.0 mL) was added to the reaction solution. The organic phase was washed with saturated ammonium chloride aqueous solution (40.0 mL × 2) and saturated brine (40.0 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 = 100:1 to 20:1) to obtain compound 3-5.

[0238] MS-ESI[M+H] + The calculated value is 687, and the measured value is 687.

[0239] 1 H NMR (400MHz, MeOD) δ8.29 (s, 1H), 7.82 (d, J=1.6Hz, 1H), 7.10-7.21 (m, 5H), 6.87 (dt, J=9.2, 4.8Hz, 1H), 4.61 (s, 6H), 4.54 (br s, 2H), 3.84-3.89 (m, 1H), 3.76 (br s, 2H), 3.61-3.65 (m, 3H), 2.83 (br t, J=5.6Hz, 2H), 2.71 (br s, 2H), 1.85-1.99 (m, 4H), 1.54 (d, J=6.8Hz, 3H), 1.49 (s, 9H), 1.40-1.45 (m, 3H), 1.18 (d, J=6.4Hz, 3H), 1.13 (dd, J=6.4, 4.4Hz, 3H).

[0240] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 3-5 (282 mg, 411 μmol) in dichloromethane (3.0 mL), and stir the reaction solution at 25 °C for 60 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.

[0241] MS-ESI[M+H] + The calculated value is 587, and the measured value is 587.

[0242] (5) Triethylamine (400 mg, 3.95 mmol) and intermediate A (157 mg, 614 μmol) were added to a solution of trifluoroacetate (286 mg, 408 μmol) in dichloromethane (3.0 mL). The reaction mixture was stirred at 25 °C for 16 hours under nitrogen protection. Dichloromethane (20.0 mL) was added, and 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 32:1) to obtain compounds 3-7.

[0243] MS-ESI[M+H] + The calculated value is 806, and the measured value is 806.

[0244] 1 H NMR (400MHz, MeOD) δ8.27 (s, 1H), 7.80 (d, J=1.6Hz, 1H), 7.09-7.19 (m, 5H), 6.84-6.91 (m, 1H), 4.61 (s, 2H), 4.49 (s, 2H), 4.22 (br d, J=6.8Hz, 1H), 4.13 (br s, 4H), 3.82-3.87 (m, 1H), 3.76 (br s, 2H), 3.62 (br d, J=5.2Hz, 5H), 2.90-2.94 (m, 2H), 2.70 (brs, 2H), 2.55 (br s, 2H), 1.93(br s, 2H), 1.80-1.87 (m, 2H), 1.54 (d, J=6.8Hz, 3H), 1.43 (s, 12H), 1.18 (d, J=6.4Hz, 3H), 1.13 (t, J=6.4Hz, 3H).

[0245] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 3-7 (250 mg, 310 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 3-8. The crude product is used directly in the next reaction step.

[0246] MS-ESI[M+H] + The calculated value is 706, and the measured value is 706.

[0247] (7) Triethylamine (78.4 mg, 775 μmol) was added to a solution of trifluoroacetate (127 mg, 155 μmol) of compounds 3-8 in dichloromethane (3.0 mL). Then, a solution of compounds 3-9 (29.5 mg, 326 μmol, 26.6 μL) in dichloromethane (1.0 mL) was added. The reaction mixture was stirred at -78 °C for 60 minutes under nitrogen protection. Dichloromethane (20.0 mL) was added, and the organic phase was washed 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 preparative high-performance liquid chromatography (Phenomenexluna C18, 100 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 0%–40%: 10 min) to obtain the formate salt of compound 3.

[0248] MS-ESI[M+H] + The calculated value is 760, and the measured value is 760.

[0249] 1 H NMR (400MHz, MeOD) δ8.29 (s, 1H), 7.83 (s, 1H), 7.18-7.24 (m, 2H), 7.08-7.17 (m, 3H), 6.84-6.91(m, 1H), 6.21-6.36(m, 2H), 5.76(dd, J=9.6, 2.4Hz, 1H), 4.62(br s, 1H), 4.49-4.57 (m, 4H), 4.30-4.38 (m, 1H), 4.25 (br d, J=7.2Hz, 2H), 3.77-3.91 (m, 4H), 3.55-3.75 (m, 6H), 2.94 (br t, J=5.6Hz, 3H), 2.76 (br s, 2H), 1.84-2.02 (m, 4H), 1.54 (d, J=6.8Hz, 3H), 1.38-1.47 (m, 3H), 1.18 (d, J=6.4Hz, 3H), 1.13 (dd, J=6.4, 2.8Hz, 3H).

[0250] Example 4

[0251] This embodiment provides a compound 4 represented by Formula I, the structural formula of which is shown below:

[0252]

[0253] The synthetic route for compound 4 is shown below:

[0254]

[0255] Triethylamine (78.8 mg, 779 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (118 mg, 310 μmol) were added to a dichloromethane (3.0 mL) solution of compound 4-1 hydrochloride (51.3 mg, 310 μmol). The mixture was stirred at 25 °C for 0.5 h. Trifluoroacetate of compound 3-8 from Example 3 (127 mg, 155 μmol) was added to the reaction mixture, and the mixture was stirred at 25 °C for 0.5 h. Dichloromethane (20.0 mL) was added. The organic phase was washed with water (20.0 mL × 1) and saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 4 by preparative high performance liquid chromatography (Phenomenexluna C18, 100mm×40mm 3μm, A: water (0.225% formic acid); B: acetonitrile, 0%-30%: 10 min).

[0256] MS-ESI[M+H] + The calculated value is 817, and the measured value is 817.

[0257] 1 H NMR (400MHz, MeOD) δ8.31 (s, 1H), 7.86 (s, 1H), 7.29 (br s, 2H), 7.12-7.22 (m, 3H), 6.88 (dt, J=9.6, 4.0Hz, 1H), 6.77 (br s, 1H), 6.34 (br d, J=13.6Hz, 1H), 4.54 (br s, 4H), 4.36 (br s, 1H), 4.26 (br d, J=5.6Hz, 2H), 4.11 (br s, 2H), 3.76-3.91 (m, 3H), 3.55-3.73 (m, 7H), 3.13-3.28 (m, 2H), 3.07 (br s, 2H), 2.97 (br s, 2H), 2.60 (br s, 6H), 2.00 (brs, 4H), 1.54 (d, J = 6.8Hz, 3H), 1.42 (d, J = 6.8Hz, 3H), 1.19 (d, J = 6.4Hz, 3H), 1.14 (br d, J=6.4Hz, 3H).

[0258] Example 5

[0259] This embodiment provides a compound 5 represented by Formula I, the structural formula of which is shown below:

[0260]

[0261] The synthetic route for compound 5 is shown below:

[0262]

[0263] (1) Potassium carbonate (118 mg, 853 μmol) was added to a solution of intermediate B (100 mg, 284 μmol) and compound 5-1 (81.3 mg, 341 μmol) in N,N-dimethylformamide (5.0 mL). The reaction solution was stirred at 70 °C under nitrogen protection for 3 hours. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (25.0 mL × 2). The organic phase was washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (petroleum ether / ethyl acetate = 1:1) to obtain compound 5-2.

[0264] 1 H NMR (400MHz, CDCl3) δ8.44-8.49(m, 1H), 7.89-7.94(m, 1H), 6.90-7.01(m, 2H), 6.54-6.61(m, 1H), 3.78-3.88(m, 1H), 3.59-3.77(m, 8H) , 3.47-3.57(m, 1H), 1.66-1.76(m, 4H), 1.53-1.57(m, 3H), 1.45-1.50(m, 3H), 1.41-1.45(m, 9H), 1.18-1.22(m, 3H), 1.13-1.17(m, 3H).

[0265] MS-ESI[M+H] + The calculated value is 542, and the measured value is 542.

[0266] (2) Trifluoroacetic acid (1.54 g, 13.5 mmol) was added to a solution of compound 5-2 (127 mg, 234 μmol) in dichloromethane (3.0 mL), and the reaction solution was stirred at 25 °C for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 5-3.

[0267] MS-ESI[M+H] + The calculated value is 442, and the measured value is 442.

[0268] (3) Triethylamine (71.0 mg, 702 μmol), compound 5-4 (61.2 mg, 234 μmol), and sodium cyanoborohydride (147 mg, 2.34 mmol) were added to a methanol (5.0 mL) solution of trifluoroacetate (130 mg, 234 μmol) of compound 5-3. The mixture was stirred at 25 °C for 2 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate (20.0 mL) was added. The organic phase was washed with water (20.0 mL × 1) and 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 5-5.

[0269] 1 H NMR (400MHz, CDCl3) δ8.35 (s, 1H), 7.86 (s, 1H), 7.08-7.15 (m, 5H), 6.82 (dd, J=9.6, 4.0Hz, 1H), 4.61 (br s, 4H), 4.53 (br s, 2H), 3.86 (dt, J=13.2, 6.8Hz, 1H), 3.71-3.76 (m, 3H), 3.67 (s, 2H), 3.61-3.64 (m, 2H), 3.17 (s, 2H), 2.82 (t, J=5.6Hz, 2H), 1 .75 (t, J=5.6Hz, 4H), 1.54 (d, J=6.8Hz, 3H), 1.49 (s, 9H), 1.45 (d, J=6.8Hz, 3H), 1.19 (d, J=6.8Hz, 3H), 1.18 (d, J=6.8Hz, 3H).

[0270] MS-ESI[M+H] + The calculated value is 687, and the measured value is 687.

[0271] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 5-5 (74.4 mg, 108 μmol) in dichloromethane (3.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 5-6.

[0272] MS-ESI[M+H] + The calculated value is 587, and the measured value is 587.

[0273] (5) Triethylamine (54.8 mg, 542 μmol) and intermediate A (55.4 mg, 217 μmol) were added to a solution of trifluoroacetate (75.9 mg, 108 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 2 hours under nitrogen protection. Dichloromethane (20.0 mL) was added, and the organic phase was washed with water (20.0 mL × 1) 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 5-7.

[0274] MS-ESI[M+H] + The calculated value is 806, and the measured value is 806.

[0275] 1 H NMR (400MHz, MeOD) δ8.35 (s, 1H), 7.87 (s, 1H), 7.10-7.15 (m, 5H), 6.82 (dd, J=9.6, 4.4Hz, 1H), 4.48 (s, 2H), 4.18-4.22 (m, 1H), 4.12 (br s, 4H), 3.84-3.88 (m, 1H), 3.71-3.76 (m, 4H), 3.67 (br s, 2H), 3.64 (d, J=6.4Hz, 2H), 3.60-3.61 (m, 1H), 3.13-3.19 (m, 4H), 2.93 (t, J=6.0Hz, 2H), 1.75 (br t, J=5.6Hz, 4H), 1.54 (d, J=6.8Hz, 3H), 1.45 (d, J=6.8Hz, 3H), 1.43 (s, 9H), 1.20 (d, J=6.4Hz, 3H), 1.19 (d, J=6.4Hz, 3H).

[0276] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 5-7 (56.0 mg, 69.5 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 5-8. The crude product is used directly in the next reaction step.

[0277] MS-ESI[M+H] + The calculated value is 706, and the measured value is 706.

[0278] (7) Triethylamine (35.1 mg, 347 μmol) was added to a solution of trifluoroacetate (56.9 mg, 69.4 μmol) of compounds 5-8 in dichloromethane (5.0 mL). Then, compounds 5-9 (9.4 mg, 104 μmol, 8.5 μL) were added, and the reaction mixture was stirred at -78 °C for 30 minutes under nitrogen protection. Dichloromethane (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 into formate salts of compound 5 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) and preparative thin-layer chromatography (dichloromethane / methanol = 10:1).

[0279] MS-ESI[M+H] + The calculated value is 760, and the measured value is 760.

[0280] 1 H NMR (400MHz, MeOD) δ8.35 (s, 1H), 7.87 (s, 1H), 7.09-7.16 (m, 5H), 6.82 (dd, J=9.6, 4.4Hz, 1H), 6.22-6.34 (m, 2H), 5.76 (dd, J=9.6, 2.8Hz, 1H), 4.62 (br s, 2H), 4.48-4.53 (m, 4H), 4.29-4.35 (m, 1H), 4.21 -4.25 (m, 2H), 3.86 (dt, J=13.2, 6.8Hz, 1H), 3.70-3.77 (m, 4H), 3.68 (s, 2H), 3.63-3.66 (m, 2H), 3.18 (s, 3H), 2.94 (t, J=6.0 Hz, 2H), 1.75 (brt, J=5.6Hz, 4H), 1.54 (d, J=6.8Hz, 3H), 1.45 (d, J=6.8Hz, 3H), 1.20 (d, J=6.8Hz, 3H), 1.19 (d, J=6.8Hz, 3H).

[0281] Example 6

[0282] This embodiment provides a compound 6 represented by Formula I, the structural formula of which is shown below:

[0283]

[0284] The synthetic route for compound 6 is shown below:

[0285]

[0286] (1) Potassium carbonate (236 mg, 1.71 mmol) was added to a solution of intermediate B (200 mg, 569 μmol) and compound 6-1 (121 mg, 569 μmol) in N,N-dimethylformamide (5.0 mL). The reaction solution was stirred at 80 °C under nitrogen protection for 2 hours. Saturated brine (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50.0 mL × 1). The organic phase was washed with saturated brine (25.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (petroleum ether / ethyl acetate = 50:1 to 1:1) to obtain compound 6-2.

[0287] MS-ESI[M+H] + The calculated value is 528, and the measured value is 528.

[0288] (2) Trifluoroacetic acid (1.54 g, 13.5 mmol) was added to a solution of compound 6-2 (229 mg, 434 μmol) in dichloromethane (4.0 mL), and the reaction solution was stirred at 25 °C for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 6-3.

[0289] MS-ESI[M+H] + The calculated value is 428, and the measured value is 428.

[0290] (3) Triethylamine (85.9 mg, 849 μmol), compound 6-4 (111 mg, 425 μmol), and sodium cyanoborohydride (133 mg, 2.12 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (230 mg, 425 μmol) of compound 6-3. The mixture was stirred at 25 °C for 2 hours. The reaction solution was concentrated under reduced pressure, and dichloromethane (50.0 mL) was added. The organic phase was washed with saturated brine (25.0 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 = 20:1 to 0:1) to obtain compound 6-5.

[0291] MS-ESI[M+H] + The calculated value is 673, and the measured value is 673.

[0292] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 6-5 (220 mg, 326 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 6-6.

[0293] MS-ESI[M+H] + The calculated value is 573, and the measured value is 573.

[0294] (5) Triethylamine (32.4 mg, 320 μmol) and intermediate A (81.9 mg, 320 μmol) were added to a solution of trifluoroacetate (220 mg, 320 μmol) in dichloromethane (10.0 mL). The reaction mixture was stirred at 25 °C for 5 minutes under nitrogen protection. Dichloromethane (20.0 mL) was added, and the organic phase was washed with saturated ammonium chloride aqueous solution (20.0 mL × 1) 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 = 50:1 to 10:1) to obtain compound 6-7.

[0295] MS-ESI[M+H] + The calculated value is 792, and the measured value is 792.

[0296] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 6-7 (180 mg, 227 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 trifluoroacetate of compound 6-8. The crude product is used directly in the next reaction step.

[0297] MS-ESI[M+H] + The calculated value is 692, and the measured value is 692.

[0298] (7) Triethylamine (67.8 mg, 670 μmol) was added to a solution of trifluoroacetate (180 mg, 223 μmol) of compounds 6-8 in dichloromethane (5.0 mL). Then, compounds 6-9 (20.2 mg, 223 μmol, 18.2 μL) were added, and the reaction mixture was stirred at -78°C for 5 minutes under nitrogen protection. Dichloromethane (50.0 mL) was added, and the organic phase was washed with saturated brine (50.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative chiral high-performance liquid chromatography (DAICELCHIRALPAK AS, 250 mm × 30 mm 10 μm, A: water (0.1% ammonia); B: ethanol, 35%–35%: 48 min) to obtain compound 6.

[0299] MS-ESI[M+H] + The calculated value is 746, and the measured value is 746.

[0300] 1H NMR (400MHz, MeOD) δ8.31 (s, 1H), 7.80 (s, 1H), 7.17-7.09 (m, 5H), 6.93-6.87 (m, 1H), 6. 35-6.21(m, 2H), 5.80-5.72(m, 1H), 4.53-4.49(m, 4H), 4.35-4.30(m, 1H), 4.26-4.21(m, 2H), 3.90-3.71(m, 4H), 3.66-3.60(m, 6H), 3.27-3.22(m, 4H), 2.96-2.91(m, 2H), 2.18-2 .10 (m, 2H), 1.59-1.52 (m, 3H), 1.50-1.42 (m, 3H), 1.21-1.18 (m, 3H), 1.15-1.12 (m, 3H).

[0301] Example 7

[0302] This embodiment provides a compound 7 represented by Formula I, the structural formula of which is shown below:

[0303]

[0304] The synthetic route for compound 7 is shown below:

[0305]

[0306] (1) Potassium carbonate (235 mg, 1.71 mmol) was added to an N,N-dimethylformamide (8.0 mL) solution of intermediate B (300 mg, 852 μmol) and compound 7-1 (217 mg, 1.02 mmol). The reaction solution was stirred at 80 °C under nitrogen protection for 2 hours. The reaction solution was quenched with water (5.0 mL), extracted with ethyl acetate (10.0 mL × 3), washed with saturated brine (10.0 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain compound 7-2.

[0307] MS-ESI[M+H] + The calculated value is 528, and the measured value is 528.

[0308] (2) Trifluoroacetic acid (4.0 mL) was added to a solution of compound 7-2 (400 mg, 758 μmol) in dichloromethane (12.0 mL), and the reaction solution was stirred at 25 °C for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 7-3.

[0309] MS-ESI[M+H] + The calculated value is 428, and the measured value is 428.

[0310] (3) Triethylamine (74.7 mg, 738 μmol), compound 7-4 (96.5 mg, 369 μmol), and sodium cyanoborohydride (92.8 mg, 1.48 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (400 mg, 738 μmol) of compound 7-3. The mixture was stirred at 25 °C for 12 hours. The reaction solution was quenched with water (5.0 mL), extracted with ethyl acetate (10.0 mL × 3), washed with saturated brine (10.0 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain compound 7-5.

[0311] MS-ESI[M+H] + The calculated value is 673, and the measured value is 673.

[0312] (4) Add 3.0 mL of trifluoroacetic acid to a solution of compound 7-5 (220 mg, 326 μmol) in dichloromethane (9.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 7-6.

[0313] MS-ESI[M+H] + The calculated value is 573, and the measured value is 573.

[0314] (5) Triethylamine (32.4 mg, 320 μmol) and intermediate A (163 mg, 640 μmol) were added to a solution of trifluoroacetate (220 mg, 320 μmol) in dichloromethane (6.0 mL). The reaction mixture was stirred at 25 °C for 60 minutes under nitrogen protection. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL × 2), 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 = 50:1 to 10:1) to obtain compound 7-7.

[0315] MS-ESI[M+H] + The calculated value is 792, and the measured value is 792.

[0316] (6) Add 1.0 mL of trifluoroacetic acid to a solution of compound 7-7 (62.0 mg, 78.2 μmol) in dichloromethane (3.0 mL), and stir the reaction mixture at 25 °C for 60 minutes. Concentrate the reaction mixture under reduced pressure to obtain the trifluoroacetate of compound 7-8. The crude product is used directly in the next reaction step.

[0317] MS-ESI[M+H]+ The calculated value is 692, and the measured value is 692.

[0318] (7) Triethylamine (7.5 mg, 74.4 μmol) was added to a solution of trifluoroacetate (60 mg, 74.4 μmol) of compounds 7-8 in dichloromethane (5.0 mL). Then, compounds 7-9 (10.1 mg, 111 μmol, 9.1 μL) were added. The reaction mixture was stirred at -78 °C for 60 minutes under nitrogen protection. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL × 2), 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 preparative 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 7.

[0319] MS-ESI[M+H] + The calculated value is 746, and the measured value is 746.

[0320] 1 H NMR (400MHz, MeOD) δ8.25-8.28(m, 1H), 7.77-7.81(m, 1H), 7.23-7.27(m, 2H), 7.14-7.20(m, 3H), 6.94-6.98(m , 1H), 6.23-6.35(m, 2H), 5.74-5.78(m, 1H), 4.60-4.64(m, 1H), 4.51-4.56(m, 4H), 4.29-4.38(m, 2H), 4.24-4. 27(m, 2H), 4.14-4.19(m, 2H), 3.92-3.98(m, 2H), 3.79-3.85(m, 1H), 3.61-3.69(m, 3H), 3.12-3.19(m, 2H), 2.9 5-3.03 (m, 4H), 2.22-2.28 (m, 2H), 1.52-1.55 (m, 3H), 1.42-1.45 (m, 3H), 1.16-1.19 (m, 3H), 1.07-1.11 (m, 3H).

[0321] Example 8

[0322] This embodiment provides a compound 8 represented by Formula I, the structural formula of which is shown below:

[0323]

[0324] The synthetic route for compound 8 is shown below:

[0325]

[0326] (1) Trifluoroacetic acid (1.45 g, 14.4 mmol, 2.0 mL) was added to a solution of compound 8-1 (240 mg, 467 μmol) in dichloromethane (6.0 mL), and the reaction solution was stirred at 25 °C for 0.5 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain the trifluoroacetate of compound 8-2.

[0327] MS-ESI[M+H] + The calculated value is 414, and the measured value is 414.

[0328] (2) Triethylamine (46.0 mg, 455 mmol) was added to a methanol (10.0 mL) solution of trifluoroacetate (240 mg, 455 μmol) of compound 8-2. Compound 8-3 (238 mg, 910 μmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 15 minutes. Then sodium cyanoborohydride (143 mg, 2.27 mmol) was added, and the mixture was stirred at 25 °C for 3 hours. The reaction solution was concentrated under reduced pressure, poured into water (40.0 mL), and extracted with dichloromethane (40 mL × 3). The organic phases were combined, washed with saturated brine (40 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 = 0:1 to 10:1) to obtain compound 8-4.

[0329] MS-ESI[M+H] + The calculated value is 659, and the measured value is 659.

[0330] (3) Add 2.0 mL of trifluoroacetic acid to a solution of compound 8-4 (210 mg, 319 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 the trifluoroacetate of compound 8-5. The crude product is used directly in the next reaction step.

[0331] MS-ESI[M+H] + The calculated value is 559, and the measured value is 559.

[0332] (4) Triethylamine (63.2 mg, 624 μmol, 86.9 μL) and intermediate A (136 mg, 531 μmol) were added to a solution of trifluoroacetate (210 mg, 312 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 10 minutes under nitrogen protection. The reaction mixture was poured into water (20.0 mL) and extracted with dichloromethane (20.0 mL × 2). The organic phases were combined, 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 = 0:1 to 10:1) to obtain compound 8-6.

[0333] MS-ESI[M+H] + The calculated value is 779, and the measured value is 779.

[0334] (5) Add 2.0 mL of trifluoroacetic acid to a solution of compound 8-6 (120 mg, 154 μmol) in dichloromethane (6.0 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 of compound 8-7. The crude product is used directly in the next reaction step.

[0335] MS-ESI[M+H] + The calculated value is 678, and the measured value is 678.

[0336] (6) Triethylamine (7.67 mg, 75.8 μmol, 10.6 μL) was added to a solution of trifluoroacetate (60.0 mg, 75.8 μmol) in dichloromethane (3.0 mL). Then, compound 8-8 (10.3 mg, 114 μmol, 9.27 μL) was added, and the reaction mixture was stirred at -78 °C for 10 minutes under nitrogen protection. The reaction mixture was poured into water (20.0 mL), extracted with dichloromethane (20.0 mL × 2), and the organic phases were combined, 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 preparative 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 8.

[0337] MS-ESI[M+H] + The calculated value is 733, and the measured value is 733.

[0338] 1H NMR: (400MHz, MeOD) δ8.23-8.27(m, 1H), 7.73-7.80(m, 1H), 7.11-7.21(m, 5H), 6.94-6.99( m, 1H), 6.22-6.36 (m, 2H), 5.71-5.80 (m, 1H), 4.50-4.55 (m, 4H), 4.37-4.46 (m, 2H), 4.28-4. 34(m, 4H), 4.22-4.27(m, 2H), 3.82-3.82(m, 1H), 3.71-3.78(m, 2H), 3.59-3.68(m, 6H), 2.9 0-2.98 (m, 2H), 1.52-1.57 (m, 3H), 1.42-1.47 (m, 3H), 1.16-1.20 (m, 3H), 1.09-1.13 (m, 3H).

[0339] Example 9

[0340] This embodiment provides a compound 9 represented by Formula I, the structural formula of which is shown below:

[0341]

[0342] The synthetic route for compound 9 is shown below:

[0343]

[0344] To a solution of trifluoroacetate (60.0 mg, 75.8 μmol) of compounds 8-7 in Example 8 in dichloromethane (3.0 mL), triethylamine (7.7 mg, 75.8 μmol, 10.6 μL), compound 9-1 (25.1 mg, 151 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (57.6 mg, 151 μmol) were added, and the mixture was stirred at 25 °C for 0.5 h. The reaction solution was poured into water (20.0 mL) and extracted with dichloromethane (20.0 mL × 2). The organic phases were combined, washed with saturated brine (20.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 9 by preparative high performance liquid chromatography (Welch Ultimate, 75 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 5%-35%: 10 min).

[0345] MS-ESI[M+H] + The calculated value is 790, and the measured value is 790.

[0346] 1H NMR: (400MHz, MeOD) δ8.25-8.29(m, 1H), 7.77-7.82(m, 1H), 7.12-7.27(m, 5H), 6.92-6.97(m, 1H), 6. 72-6.82(m, 1H), 6.30-6.40(m, 1H), 4.90-4.93(m, 2H), 4.50-4.62(m, 4H), 4.40-4.49(m, 2H), 4.32-4. 35(m, 2H), 4.22-4.29(m, 2H), 3.94-4.08(m, 6H), 3.79-3.87(m, 1H), 3.58-3.67(m, 4H), 2.93-3.00(m , 2H), 2.60-2.68(m, 6H), 1.52-1.57(m, 3H), 1.42-1.47(m, 3H), 1.16-1.21(m, 3H), 1.09-1.14(m, 3H).

[0347] Example 10

[0348] This embodiment provides a compound 10 represented by Formula I, the structural formula of which is shown below:

[0349]

[0350] The synthetic route for compound 10 is shown below:

[0351]

[0352] (1) Triethylamine (273 mg, 2.70 mmol) was added to a methanol (10.0 mL) solution of trifluoroacetate (500 mg, 0.95 mmol) of compounds 1-3 from Example 1. Compound 10-1 (282 mg, 1.08 mmol) and sodium cyanoborohydride (169 mg, 2.70 mmol) were added to the reaction solution, and the mixture was stirred at 25 °C for 16 hours. The mixture was then poured into a saturated ammonium chloride aqueous solution (40.0 mL) and extracted with ethyl acetate (50.0 mL × 1). The organic phases were combined, washed with saturated brine (25.0 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 = 0:1 to 10:1) to obtain compound 10-2.

[0353] MS-ESI[M+H] + The calculated value is 687, and the measured value is 687.

[0354] (1) Add 2.0 mL of trifluoroacetic acid to a solution of compound 10⁻² (120 mg, 0.175 mmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 15 minutes. Filter the reaction mixture and concentrate it under reduced pressure to obtain trifluoroacetate of compound 10⁻³. The crude product is used directly in the next reaction step.

[0355] MS-ESI[M+H] + The calculated value is 587, and the measured value is 587.

[0356] (2) Triethylamine was added to a solution of trifluoroacetate (120 mg, 0.171 mmol) of compound 10⁻³ in dichloromethane (5.0 mL) to adjust the pH to 8. Then, intermediate A (87.6 mg, 0.342 mmol) was added. The reaction mixture was stirred at 25 °C for 0.5 hours under nitrogen protection. The reaction mixture was poured into water (15.0 mL) and extracted with dichloromethane (10.0 mL × 3). The organic phases were combined, 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 = 0:1 to 10:1) to obtain compound 10⁻⁴.

[0357] MS-ESI[M+H] + The calculated value is 806, and the measured value is 806.

[0358] (3) Add 2.0 mL of trifluoroacetic acid to a solution of compound 10⁻⁴ (130 mg, 0.16 mmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 30 minutes. Filter the reaction mixture and concentrate it under reduced pressure to obtain trifluoroacetate of compound 10⁻⁵. The crude product is used directly in the next reaction step.

[0359] MS-ESI[M+H] + The calculated value is 706, and the measured value is 706.

[0360] (4) Triethylamine (4.94 mg, 48.8 μmol) was added to a solution of trifluoroacetate (40.0 mg, 48.8 μmol) of compound 10-5 in dichloromethane (3.0 mL). Then, compound 10-6 (5.30 mg, 58.5 μmol, 4.77 μL) was added, and the reaction mixture was stirred at -78 °C for 15 minutes under nitrogen protection. The reaction mixture was poured into water (20.0 mL) and extracted with dichloromethane (20.0 mL × 2). The organic phases were combined, washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative 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 10.

[0361] MS-ESI[M+H] + The calculated value is 760, and the measured value is 760.

[0362] 1 H NMR: (400MHz, MeOD) δ8.24 (s, 1H), 7.75 (s, 1H), 7.21-7.10 (m, 5H), 6.99 (dd, J=4.2, 8.8Hz, 1H), 6.29-6.27 (d, J=9 .2Hz, 2H), 5.78-5.74(m, 1H), 4.79-4.61(m, 2H), 4.54-4.51(m, 4H), 4.49-4.29(m, 1H), 4.26-4.24(m, 2H), 3.93(br d, J=9.2Hz, 1H), 3.82-3.75(m, 2H), 3.71–3.65(m, 1H), 3.63-3.53(m, 5H), 2.95(br t, J=6.0Hz, 2H), 2.58 (s, 3H), 1.91-1.79 (m, 4H), 1.54 (d, J=6.8Hz, 3H), 1.45 (d, J=6.8Hz, 3H), 1.18 (d, J=6.4Hz, 3H), 1.09 (d, J=6.4Hz, 3H).

[0363] Example 11

[0364] This embodiment provides a compound 11 represented by Formula I, the structural formula of which is shown below:

[0365]

[0366] The synthetic route for compound 11 is shown below:

[0367]

[0368] To a solution of trifluoroacetate (40.0 mg, 48.8 μmol) of compound 10-4 from Example 10 in dichloromethane (3.0 mL), triethylamine (4.94 mg, 48.8 μmol, 6.79 μL), compound 11-1 (16.2 mg, 97.6 μmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (37.1 mg, 97.6 μmol) were added, and the mixture was stirred at 25 °C for 0.5 h. The reaction solution was poured into water (20.0 mL) and extracted with dichloromethane (20.0 mL × 3). The organic phases were combined, washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 11 by preparative high performance liquid chromatography (Phenomenex luna C18, 100mm×40mm 3μm, A: water (0.225% formic acid); B: acetonitrile, 0%-40%: 10 min).

[0369] MS-ESI[M+H] + The calculated value is 817, and the measured value is 817.

[0370] 1 H NMR: (400MHz, MeOD) δ8.28-8.22(m, 1H), 7.82-7.75(m, 1H), 7.29-7.20(m, 2H), 7.19-7.12(m, 2H), 7.01-6.95 (m, 1H), 6.83-6.72 (m, 1H), 6.36-6.25 (m, 1H), 4.51 (s, 4H), 4.41-4.32 (m, 1H), 4.29-4.22 (m, 2H), 4.14-3.92 (m, 4H), 3.89-3.80 (m, 3H), 3.70-3.59 (m, 3H), 3.55-3.44 (m, 2H), 3.01-2.93 (m, 2H), 2.88-2.65 (m, 4H), 2.60 -2.46(m, 6H), 2.00-1.85(m, 4H), 1.57-1.52(m, 3H), 1.48-1.43(m, 3H), 1.21-1.16(m, 3H), 1.12-1.07(m, 3H).

[0371] Example 12

[0372] This embodiment provides a compound 12 represented by Formula I, the structural formula of which is shown below:

[0373]

[0374] The synthetic route for compound 12 is shown below:

[0375]

[0376] (1) Triethylamine (45.8 mg, 452 μmol), compound 12-1 (121 mg, 452 μmol), and sodium cyanoborohydride (142 mg, 2.26 mmol) were added to a methanol (5.0 mL) solution of trifluoroacetate (200 mg, 452 μmol) of compounds 1-3 in Example 1. The mixture was stirred at 25 °C for 12 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate (80.0 mL) was added. The organic phase was washed with water (40.0 mL) and saturated brine (50.0 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 = 1:0 to 0:1) to obtain compound 12-2.

[0377] MS-ESI[M+H] + The calculated value is 693, and the measured value is 693.

[0378] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 12-2 (90.0 mg, 129 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 12-3. The crude product is used directly in the next reaction step.

[0379] MS-ESI[M+H] + The calculated value is 593, and the measured value is 593.

[0380] (3) Triethylamine (12.9 mg, 127 μmol) and intermediate A (32.5 mg, 127 μmol) were added to a solution of trifluoroacetate (90 mg, 127 μmol) in dichloromethane (10.0 mL). The reaction mixture was stirred at 25 °C for 5 minutes under nitrogen protection. Dichloromethane (20.0 mL) was added, followed by washing with saturated brine (70.0 mL), drying with anhydrous sodium sulfate, filtration, and concentration of the organic phase under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane / methanol = 100:1 to 10:1) to obtain compound 12-4.

[0381] MS-ESI[M+H] + The calculated value is 812, and the measured value is 812.

[0382] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 12-4 (70.0 mg, 75.6 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 12-5. The crude product is used directly in the next reaction step.

[0383] MS-ESI[M+H] + The calculated value is 713, and the measured value is 713.

[0384] (5) Triethylamine (8.53 mg, 84.3 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (27.6 mg, 72.6 μmol) were added to a dichloromethane (10.0 mL) solution of compound 12-5 hydrochloride (18.1 mg, 109 μmol). The mixture was stirred at 25 °C for 30 minutes. Trifluoroacetate of compound 12-6 (60.0 mg, 72.6 μmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 30 minutes. Dichloromethane (50.0 mL) was added. The organic phase was washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into formate salts of compound 12 by preparative high performance liquid chromatography (Phenomenex luna C18, 100mm×30mm 3μm, A: water (0.225% formic acid); B: acetonitrile, 0%-30%: 8 min).

[0385] MS-ESI[M+H] + The calculated value is 824, and the measured value is 824.

[0386] 1 H NMR (400MHz, MeOD) δ8.26-8.21(m, 1H), 7.77-7.72(m, 1H), 7.21-7.11(m, 2H), 7.03-6.96(m, 1H), 6.83-6. 72(m, 2H), 6.31-6.21(m, 1H), 4.54-4.47(m, 3H), 4.37-4.24(m, 2H), 4.23-4.19(m, 2H), 3.95-3.90(m, 2H) , 3.87-3.76(m, 2H), 3.75-3.72(m, 2H), 3.70-3.56(m, 4H), 3.44-3.39(m, 2H), 2.80-2.72(m, 3H), 2.56-2. 43 (m, 9H), 1.87-1.81 (m, 4H), 1.56-1.52 (m, 3H), 1.47-1.43 (m, 3H), 1.20-1.16 (m, 3H), 1.11-1.07 (m, 3H).

[0387] Example 13

[0388] This embodiment provides a compound 13 represented by Formula I, the structural formula of which is shown below:

[0389]

[0390] The synthetic route for compound 13 is shown below:

[0391]

[0392] (1) To a methanol (3.0 mL) solution of trifluoroacetate (200 mg, 453 μmol) of compounds 1-3 in Example 1, triethylamine (91.7 mg, 906 μmol) and compound E (145 mg, 544 μmol) were added. The mixture was stirred at 25 °C for 15 minutes, and sodium cyanoborohydride (142 mg, 2.26 mmol) was added. The mixture was stirred at 25 °C for 12 hours. The reaction solution was concentrated under reduced pressure, and dichloromethane (10.0 mL) was added. The organic phase was washed with water (3.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by preparative thin-layer chromatography (dichloromethane / methanol = 10:1) to obtain compound 13-1.

[0393] MS-ESI[M+H] + The calculated value is 693, and the measured value is 693.

[0394] (2) Add 1.0 mL of trifluoroacetic acid to a solution of compound 13-1 (140 mg, 202 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 13-2. The crude product is used directly in the next reaction step.

[0395] MS-ESI[M+H] + The calculated value is 593, and the measured value is 593.

[0396] (3) Triethylamine (40.1 mg, 396 μmol) and intermediate A (101 mg, 396 μmol) were added to a solution of trifluoroacetate (140 mg, 198 μmol) in dichloromethane (3.0 mL). The reaction mixture was stirred at 25 °C for 10 minutes under nitrogen protection. Dichloromethane (20.0 mL) and water (2.0 mL) were added, and the mixture was dried over anhydrous sodium sulfate. The mixture was 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 8:1) to obtain compound 12-3.

[0397] MS-ESI[M+H] + The calculated value is 812, and the measured value is 812.

[0398] (4) Add 10.9 μL of trifluoroacetic acid to a solution of compound 13-3 (120 mg, 148 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 13-4. The crude product is used directly in the next reaction step.

[0399] MS-ESI[M+H] + The calculated value is 712, and the measured value is 712.

[0400] (5) Triethylamine (14.7 mg, 145 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (41.4 mg, 109 μmol) were added to a dichloromethane (2.0 mL) solution of the hydrochloride salt of compound 13-5 (36.1 mg, 218 μmol). The mixture was stirred at 25 °C for 10 minutes. Then, trifluoroacetate of compound 13-4 (60.0 mg, 72.7 μmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 60 minutes. The organic phase was concentrated under reduced pressure. The crude product was prepared by high performance liquid chromatography (Phenomenex luna C18, 100 mm × 30 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 8 min) to separate the formate salt of compound 13.

[0401] MS-ESI[M+H] + The calculated value is 823, and the measured value is 823.

[0402] 1 H NMR (400MHz, MeOD) δ8.24 (s, 1H), 7.76 (s, 1H), 7.10-7.21 (m, 2H), 6.99 (dd, J=9.2, 4.4Hz, 1H), 6.73-6.84 (m, 2H), 6.35 (br d, J=15.2Hz, 1H), 4.48-4.61(m, 2H), 4.42(s, 2H), 4.30-4.37(m, 1H), 4.24(br d, J=6.8Hz, 2H), 4.04(br s, 2H), 3.77-3.96 (m, 5H), 3.55-3.73 (m, 5H), 2.91 (br s, 3H), 2.64 (s, 9H), 1.89 (br s, 4H), 1.54 (d, J=6.8Hz, 3H), 1.45 (d, J=6.8Hz, 3H), 1.18 (d, J=6.8Hz, 3H), 1.09 (d, J=6.8Hz, 3H).

[0403] Example 14

[0404] This embodiment provides a compound 14 represented by Formula I, the structural formula of which is shown below:

[0405]

[0406] The synthetic route for compound 14 is shown below:

[0407]

[0408] Triethylamine (14.7 mg, 145 μmol) was added to a solution of trifluoroacetate (60 mg, 72.7 μmol) of compound 13-4 in dichloromethane (3.0 mL). Then, compound 14-2 (7.23 mg, 79.9 μmol, 6.52 μL) was added, and the reaction mixture was stirred at -78°C for 10 minutes under nitrogen protection. The crude product was separated into formate salts of compound 14 by preparative high-performance liquid chromatography (Phenomenex Luna C18, 100 mm × 30 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 10%–40%: 8 min).

[0409] MS-ESI[M+H] + The calculated value is 766, and the measured value is 766.

[0410] 1 H NMR (400MHz, MeOD) δ8.25 (s, 1H), 7.77 (br s, 1H), 7.10-7.23 (m, 2H), 6.98 (dd, J=8.8, 4.0Hz, 1H), 6.82 (br s, 1H), 6.17-6.37 (m, 2H), 5.77 (dd, J=9.2, 2.3Hz, 1H), 4.41-4.60 (m, 4H), 4.18-4.38 (m, 3H), 3.87-4.13(m, 6H), 3.77-3.87(m, 1H), 3.55-3.76(m, 3H), 2.58-2.98(m, 6H), 1.91(br s, 4H), 1.54(br d, J=6.8Hz, 3H), 1.45 (br d, J=6.8Hz, 3H), 1.18 (br d, J=6.8Hz, 3H), 1.09 (br d, J=6.8Hz, 3H).

[0411] Example 15

[0412] This embodiment provides a compound 15 represented by Formula I, the structural formula of which is shown below:

[0413]

[0414] The synthetic route for compound 15 is shown below:

[0415]

[0416] (1) To a methanol (10.0 mL) solution of trifluoroacetate (200 mg, 359 μmol) of compound 3-3 from Example 3, triethylamine (72.8 mg, 720 μmol), compound C (188 mg, 719 μmol), and sodium cyanoborohydride (90.4 mg, 1.44 mmol) were added, and the mixture was stirred at 25 °C for 12 hours. Water (10.0 mL) and dichloromethane (20.0 mL) were added to the reaction solution. The organic phase was washed with saturated brine (10.0 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 (dichloromethane / methanol = 1:0 to 0:1) to obtain compound 15-1.

[0417] MS-ESI[M+H] + The calculated value is 688, and the measured value is 688.

[0418] (2) Add 3.0 mL of trifluoroacetic acid to a solution of compound 15-1 (200 mg, 290 μmol) in dichloromethane (9.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 15-2. The crude product is used directly in the next reaction step.

[0419] MS-ESI[M+H] + The calculated value is 589, and the measured value is 589.

[0420] (3) Triethylamine (28.8 mg, 285 μmol) and intermediate A (145 mg, 570 μmol) were added to a solution of trifluoroacetate (200 mg, 285 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 1 hour under nitrogen protection. Water (10.0 mL) and dichloromethane (20.0 mL) were added. The organic phase was washed with saturated brine (10.0 mL), 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 15-3.

[0421] MS-ESI[M+H] + The calculated value is 807, and the measured value is 807.

[0422] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 15-3 (120 mg, 148 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 15-4. The crude product is used directly in the next reaction step.

[0423] MS-ESI[M+H] + The calculated value is 708, and the measured value is 708.

[0424] (5) Triethylamine (14.7 mg, 146 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (55.5 mg, 146 μmol) were added to a dichloromethane (4.0 mL) solution of compound 15-5 hydrochloride (28.3 mg, 219 μmol). The mixture was stirred at 25 °C for 1 hour. Trifluoroacetate of compound 15-4 (60 mg, 73.0 μmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 1 hour. Dichloromethane (20.0 mL) was added. The organic phase was washed with water (10.0 mL) and saturated brine (10.0 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, 10%-40%: 10 min), and the formate salt of compound 15 was separated.

[0425] MS-ESI[M+H] + The calculated value is 818, and the measured value is 818.

[0426] 1 H NMR (400MHz, MeOD) δ8.29-8.36 (s, 1H), 7.84-7.89 (s, 1H), 7.58-7.68 (m, 1H), 7.29-7.40 (m, 1H), 7.12-7.19 (m , 2H), 6.86-6.91(m, 1H), 6.70-6.71(m, 1H), 6.29-6.45(m, 1H), 4.51-4.67(m, 4H), 4.38-4.46(m, 1H), 4.21-4. 36(m,4H),3.79-3.94(m,3H),3.71-3.79(m,3H),3.60-3.70(m,4H),3.17-3.31(m,2H),3.01-3.17(m,4H),2.5 8-2.76 (m, 6H), 1.98-2.09 (m, 4H), 1.51-1.56 (m, 3H), 1.40-1.45 (m, 3H), 1.17-1.22 (m, 3H), 1.11-1.16 (m, 3H).

[0427] Example 16

[0428] This embodiment provides a compound 16 represented by Formula I, the structural formula of which is shown below:

[0429]

[0430] The synthetic route for compound 16 is shown below:

[0431]

[0432] (1) Triethylamine (72.8 mg, 719 μmol), compound C (188 mg, 719 μmol), and sodium cyanoborohydride (90.4 mg, 1.44 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (200 mg, 359 μmol) of compound 3-3 in Example 3, and the mixture was stirred at 25 °C for 12 hours. Water (100.0 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (100.0 mL × 2). The organic phase was washed with saturated brine (100.0 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 (dichloromethane / methanol = 1:0 to 0:1) to obtain compound 16-1.

[0433] MS-ESI[M+H] + The calculated value is 688, and the measured value is 688.

[0434] (2) Trifluoroacetic acid (2.0 mL) was added to a solution of compound 16-1 (170 mg, 247 μmol) in dichloromethane (6.0 mL), and the reaction solution was stirred at 25 °C for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 16-2.

[0435] MS-ESI[M+H] + The calculated value is 588, and the measured value is 588.

[0436] (3) Triethylamine (24.5 mg, 242 μmol) and intermediate A (123 mg, 484 μmol) were added to a solution of trifluoroacetate (170 mg, 242 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 60 minutes under nitrogen protection. Water (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), 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 16-3.

[0437] MS-ESI[M+H]+ The calculated value is 807, and the measured value is 807.

[0438] (4) Add 3.0 mL of trifluoroacetic acid to a solution of compound 16-3 (137 mg, 169 μmol) in dichloromethane (9.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 16-4. The crude product is used directly in the next reaction step.

[0439] MS-ESI[M+H] + The calculated value is 707, and the measured value is 707.

[0440] (5) Triethylamine (9.25 mg, 91.3 μmol) was added to a solution of trifluoroacetate (75.0 mg, 91.3 μmol) in dichloromethane (5.0 mL). Then, compound 16-5 (12.4 mg, 137 μmol, 11.1 μL) was added, and the reaction mixture was stirred at -78 °C for 60 minutes under nitrogen protection. Water (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), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated 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) to obtain the formate of compound 16.

[0441] MS-ESI[M+H] + The calculated value is 761, and the measured value is 761.

[0442] 1 H NMR (400MHz, MeOD) δ8.27-8.34(m, 1H), 7.80-7.88(m, 1H), 7.59-7.66(m, 1H), 7.31-7.37(m, 1H), 7.11-7. 18(m, 2H), 6.84-6.91(m, 1H), 6.13-6.38(m, 2H), 5.70-5.81(m, 1H), 4.53-4.60(m, 4H), 4.36-4.43(m, 1H) , 4.25-4.33(m, 2H), 4.01-4.14(m, 2H), 3.71-3.90(m, 6H), 3.60-3.65(m, 2H), 3.03-3.16(m, 4H), 2.87-2. 99 (m, 2H), 1.93-2.05 (m, 4H), 1.52-1.56 (m, 3H), 1.41-1.45 (m, 3H), 1.17-1.21 (m, 3H), 1.12-1.16 (m, 3H).

[0443] Example 17

[0444] This embodiment provides a compound 17 represented by Formula I, the structural formula of which is shown below:

[0445]

[0446] The synthetic route for compound 17 is shown below:

[0447]

[0448] (1) Triethylamine (45.8 mg, 452 μmol), compound C (237 mg, 905 μmol), and sodium cyanoborohydride (113.8 mg, 1.81 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (200 mg, 452 μmol) of compounds 1-3 in Example 1, and the mixture was stirred at 25 °C for 1 hour. Water (100.0 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (100.0 mL × 2). The organic phase was washed with saturated brine (100.0 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 (dichloromethane / methanol = 1:0 to 0:1) to obtain compound 17-1.

[0449] MS-ESI[M+H] + The calculated value is 688, and the measured value is 688.

[0450] (2) Trifluoroacetic acid (3.0 mL) was added to a solution of compound 17-1 (160 mg, 232 μmol) in dichloromethane (9.0 mL), and the reaction solution was stirred at 25 °C for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 17-2.

[0451] MS-ESI[M+H] + The calculated value is 588, and the measured value is 588.

[0452] (3) Triethylamine (23.0 mg, 228 μmol) and intermediate A (116 mg, 456 μmol) were added to a solution of trifluoroacetate (160 mg, 228 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 60 minutes under nitrogen protection. Water (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), 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 17-3.

[0453] MS-ESI[M+H] +The calculated value is 807, and the measured value is 807.

[0454] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 17-3 (80 mg, 99.1 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 17-5. The crude product is used directly in the next reaction step.

[0455] MS-ESI[M+H] + The calculated value is 707, and the measured value is 707.

[0456] (5) To a solution of trifluoroacetate (35.0 mg, 49.5 μmol) of compound 17-4 in dichloromethane (5.0 mL), triethylamine (10.0 mg, 99.1 μmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (37.7 mg, 99.1 μmol) and compound 17-5 (19.2 mg, 148 μmol) were added, and the mixture was stirred at 25 °C for 1 hour. Water (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), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 17 by preparative high performance liquid chromatography (Xtimate C18, 100 mm × 40 mm 3 μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min).

[0457] MS-ESI[M+H] + The calculated value is 818, and the measured value is 818.

[0458] 1H NMR (400MHz, MeOD) δ8.23-8.30 (m, 1H), 7.74-7.80 (m, 1H), 7.60-7.67 (m, 1H), 7.34-7.41 (m, 1H), 7.11-7.21 (m, 2H) , 6.94-7.03(m, 1H), 6.72-6.83(m, 1H), 6.33-6.44(m, 1H), 4.52-4.66(m, 4H), 4.37-4.45(m, 1H), 4.24-4.35(m, 2H) , 3.88-4.16(m, 6H), 3.80-3.87(m, 1H), 3.72-3.79(m, 2H), 3.55-3.69(m, 3H), 3.04-3.11(m, 2H), 2.72-2.92(m, 4H) , 2.52-2.72(m, 6H), 1.86-2.02(m, 4H), 1.50-1.59(m, 3H), 1.41-1.49(m, 3H), 1.15-1.24(m, 3H), 1.05-1.14(m, 3H).

[0459] Example 18

[0460] This embodiment provides a compound 18 represented by Formula I, the structural formula of which is shown below:

[0461]

[0462] The synthetic route for compound 18 is shown below:

[0463]

[0464] Triethylamine (5.02 mg, 49.5 μmol) was added to a solution of trifluoroacetate (35.0 mg, 49.5 μmol) of compound 17-4 in dichloromethane (3.0 mL). Then, compound 18-1 (6.73 mg, 74.3 μmol, 6.1 μL) was added, and the reaction mixture was stirred for 60 minutes under nitrogen protection at -78 °C. Water (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), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated 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) to obtain the formate salt of compound 18.

[0465] MS-ESI[M+H] + The calculated value is 761, and the measured value is 761.

[0466] 1 H NMR (400MHz, MeOD) δ8.22-8.28(m, 1H), 7.73-7.80(m, 1H), 7.59-7.67(m, 1H), 7.32-7.40(m, 1H), 7.13-7.21(m, 2H) , 6.95-7.02(m, 1H), 6.15-6.39(m, 2H), 5.70-5.81(m, 1H), 4.51-4.61(m, 4H), 4.36-4.43(m, 1H), 4.24-4.33(m, 2H) , 3.99-4.19(m, 2H), 3.90-3.99(m, 4H), 3.81-3.87(m, 1H), 3.73-3.78(m, 2H), 3.59-3.67(m, 1H), 3.02-3.11(m, 2H) , 2.61-2.92(m, 4H), 1.89-2.00(m, 4H), 1.52-1.57(m, 3H), 1.43-1.48(m, 3H), 1.15-1.21(m, 3H), 1.08-1.14(m, 3H).

[0467] Example 19

[0468] This embodiment provides a compound 19 represented by Formula I, the structural formula of which is shown below:

[0469]

[0470] The synthetic route for compound 19 is shown below:

[0471]

[0472] (1) Potassium carbonate (392 mg, 2.84 mmol) was added to a solution of intermediate B (500 mg, 1.42 mmol) and compound 19-1 (385 mg, 1.71 mmol) in N,N-dimethylformamide (10.0 mL). The reaction solution was stirred at 80 °C for 2 hours under nitrogen protection. The reaction solution was quenched with water (10.0 mL) and extracted with dichloromethane (10.0 mL × 2). The organic phases were combined, washed with saturated brine (10.0 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 (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 19-2.

[0473] MS-ESI[M+H] + The calculated value is 543, and the measured value is 543.

[0474] 1H NMR (400MHz, CDCl3) δ8.38-8.47(m, 1H), 7.77-7.86(m, 1H), 6.93-7.05(m, 2H), 6.62-6.79(m, 1H), 3.58-3.86(m, 4H) , 3.37-3.57(m, 4H), 3.21-3.35(m, 2H), 1.84-1.95(m, 4H), 1.52-1.57(m, 3H), 1.44-1.48(m, 12H), 1.10-1.18(m, 6H).

[0475] (2) Trifluoroacetic acid (9.24 g, 81.0 mmol, 6.0 mL) was added to a solution of compound 19-2 (673 mg, 1.24 mmol) in dichloromethane (18.0 mL). Under nitrogen protection, the reaction solution was stirred at 25 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 19-3.

[0476] MS-ESI[M+H] + The calculated value is 442, and the measured value is 442.

[0477] (3) The trifluoroacetate of compound 19-3 (100 mg, 180 μmol), compound 12-1 (57.7 mg, 216 μmol), and triethylamine (36.4 mg, 359 μmol, 50.1 μL) were suspended in methanol (5.0 mL), and sodium cyanoborohydride (45.2 mg, 719 μmol) was added. The mixture was stirred at 25 °C for 12 hours under nitrogen protection. The reaction was quenched with water (10.0 mL), and the mixture was extracted with dichloromethane (10.0 mL × 2). The organic phases were combined, washed with saturated brine (10.0 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 (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 19-4.

[0478] MS-ESI[M+H] + The calculated value is 693, and the measured value is 693.

[0479] (4) Trifluoroacetic acid (3.08 g, 27.0 mmol, 2.0 mL) was added to a solution of compound 19-4 (120 mg, 173 μmol) in dichloromethane (6.0 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to obtain the trifluoroacetate of compound 19-5. The crude product was used directly in the next reaction step;

[0480] MS-ESI[M+H] + The calculated value is 593, and the measured value is 593.

[0481] (5) To the trifluoroacetate of compound 19-5 (120 mg, 169 μmol), dichloromethane (5.0 mL) solution and triethylamine (34.3 mg, 339 μmol, 47.2 μL) were added, followed by intermediate A (86.8 mg, 339 μmol). Under nitrogen protection, 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), the organic phases were combined, washed with saturated brine (10.0 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 (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 19-6;

[0482] MS-ESI[M+H] + The calculated value is 812, and the measured value is 812.

[0483] (6) Trifluoroacetic acid (1.54 g, 13.5 mmol, 1.0 mL) was added to a solution of compound 19-6 (45.0 mg, 55.4 μmol) in dichloromethane (3.0 mL). Under nitrogen protection, 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 19-7. The crude product was used directly in the next reaction step.

[0484] MS-ESI[M+H] + The calculated value is 712, and the measured value is 712.

[0485] (7) The trifluoroacetate of compound 19-7 (45.0 mg, 54.4 μmol), compound 19-8 (27.0 mg, 163 μmol), triethylamine (5.51 mg, 54.4 μmol, 7.58 μL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (41.4 mg, 108 μmol) were dissolved in dichloromethane (3.0 mL) and stirred at 25 °C for 1 hour under nitrogen protection. The reaction solution was diluted with water (10.0 mL), extracted with dichloromethane (10.0 mL × 2), the organic phases were combined, washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 19 by preparative high performance liquid chromatography (Xtimate C18, 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 10%-40%: 10 min).

[0486] MS-ESI[M+H] + The calculated value is 823, and the measured value is 823.

[0487] 1H NMR (400MHz, MeOD) δ8.29-8.33(m, 1H), 7.82-7.87(m, 1H), 7.13-7.18(m, 2H), 6.86-6.91(m, 2H), 6.72-6. 81(m, 1H), 6.33-6.43(m, 1H), 4.52-4.62(m, 4H), 4.30-4.37(m, 1H), 4.16-4.26(m, 4H), 3.85-3.91(m, 1H) , 3.76-3.84(m, 2H), 3.60-3.74(m, 7H), 3.06-3.19(m, 2H), 2.94-3.02(m, 2H), 2.75-2.80(m, 2H), 2.69-2. 74(m, 6H), 1.92-2.04(m, 4H), 1.53-1.57(m, 3H), 1.42-1.46(m, 3H), 1.18-1.21(m, 3H), 1.13-1.16(m, 3H)

[0488] Example 20

[0489] This embodiment provides a compound 20 represented by Formula I, the structural formula of which is shown below:

[0490]

[0491] The synthetic route for compound 20 is shown below:

[0492]

[0493] (1) The trifluoroacetate (120 mg, 216 μmol), compound E (69.2 mg, 259 μmol), and triethylamine (43.7 mg, 431 μmol, 60.1 μL) of compound 20-1 from Example 20 were suspended in methanol (5.0 mL), and sodium cyanoborohydride (54.2 mg, 863 μmol) was added. The mixture was stirred at 25 °C for 12 hours under nitrogen protection. The reaction was quenched with water (100.0 mL), and the mixture was extracted with dichloromethane (100.0 mL × 2). The organic phases were combined, washed with saturated brine (100.0 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 (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 20-2.

[0494] MS-ESI[M+H] + The calculated value is 693, and the measured value is 693.

[0495] (2) Trifluoroacetic acid (3.08 g, 27.0 mmol, 2.0 mL) was added to a solution of compound 20-2 (120 mg, 173 μmol) in dichloromethane (6.0 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 20-3. The crude product was used directly in the next reaction step;

[0496] MS-ESI[M+H] + The calculated value is 593, and the measured value is 593.

[0497] (3) To the trifluoroacetate of compound 20-3 (120 mg, 169 μmol), dichloromethane (5.0 mL) solution and triethylamine (34.3 mg, 339 μmol, 47.2 μL) were added, followed by intermediate A (86.8 mg, 339 μmol). Under nitrogen protection, the reaction mixture was stirred at 25 °C for 1 hour. The reaction was quenched with water (100.0 mL), extracted with dichloromethane (100.0 mL × 2), the organic phases were combined, washed with saturated brine (100.0 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 (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 20-4;

[0498] MS-ESI[M+H] + The calculated value is 812, and the measured value is 812.

[0499] (4) Add 1.0 mL of trifluoroacetic acid to a solution of compound 20-4 (23.0 mg, 28.3 μmol) in dichloromethane (3.0 mL). Under nitrogen protection, stir the reaction solution at 25 °C for 1 hour. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 20-5. The crude product is used directly in the next reaction step.

[0500] MS-ESI[M+H] + The calculated value is 712, and the measured value is 712.

[0501] (5) The trifluoroacetate of compound 20-5 (23.0 mg, 27.8 μmol), compound 20-6 (10.7 mg, 65.1 μmol), triethylamine (2.82 mg, 27.8 μmol, 3.88 μL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (21.1 mg, 55.6 μmol) were dissolved in dichloromethane (3.0 mL). The mixture was stirred at 25 °C for 1 hour under nitrogen protection. The reaction solution was diluted with water (10.0 mL), extracted with dichloromethane (10.0 mL × 2), the organic phases were combined, washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into formate salts of compound 20 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).

[0502] MS-ESI[M+H] + The calculated value is 823, and the measured value is 823.

[0503] 1 H NMR (400MHz, MeOD) δ8.27-8.32 (m, 1H), 7.81-7.86 (m, 1H), 7.12-7.19 (m, 2H), 6.85-6.91 (m, 1H), 6.72-6. 84(m, 2H), 6.28-6.40(m, 1H), 4.52-4.60(m, 2H), 4.39-4.43(m, 2H), 4.21-4.35(m, 3H), 4.00-4.09(m, 2H) , 3.74-3.91(m, 3H), 3.58-3.72(m, 7H), 2.95-3.06(m, 2H), 2.89-2.94(m, 2H), 2.81-2.88(m, 2H), 2.56-2. 67 (m, 6H), 1.89-2.04 (m, 4H), 1.52-1.57 (m, 3H), 1.41-1.47 (m, 3H), 1.17-1.21 (m, 3H), 1.12-1.17 (m, 3H).

[0504] Example 21

[0505] This embodiment provides a compound 21 represented by Formula I, the structural formula of which is shown below:

[0506]

[0507] The synthetic route for compound 21 is shown below:

[0508]

[0509] (1) Potassium carbonate (246 mg, 1.78 mmol) was added to an N,N-dimethylformamide (8.0 mL) solution of intermediate F (300 mg, 888 μmol) and compound 21-1 (241 mg, 1.07 μmol). The reaction solution was stirred at 80 °C for 12 hours under nitrogen protection. The reaction solution was quenched with water (30.0 mL) and extracted with ethyl acetate (40.0 mL × 2). The organic phases were combined, washed with saturated brine (40.0 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 = 1:0 to 0:1) to obtain compound 21-2;

[0510] MS-ESI[M+H] + The calculated value is 528, and the measured value is 528.

[0511] (2) Add 2 mL of trifluoroacetic acid to a solution of compound 21-2 (350 mg, 663 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 10 minutes. Filter the reaction solution and concentrate under reduced pressure to obtain the trifluoroacetate salt of compound 21-3;

[0512] MS-ESI[M+H] + The calculated value is 428, and the measured value is 428.

[0513] (3) The trifluoroacetate of compound 21-3 (237 mg, 555 μmol), compound C (291 mg, 1.11 mmol), and triethylamine (56.2 mg, 555 μmol, 77.3 μL) were suspended in methanol (5.0 mL), and sodium cyanoborohydride (140 mg, 2.22 mmol) was added. The mixture was stirred at 25 °C for 12 hours. The reaction was quenched with water (30.0 mL), and the mixture was extracted with dichloromethane (30.0 mL × 2). The organic phases were combined, 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 (dichloromethane / methanol = 1:0 to 9:1) to obtain compound 21-4.

[0514] MS-ESI[M+H] + The calculated value is 675, and the measured value is 675.

[0515] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 21-4 (370 mg, 549 μmol) in dichloromethane (6.0 mL), and stir the reaction mixture at 25 °C for 0.5 hours. Filter the reaction mixture and concentrate under reduced pressure to obtain the trifluoroacetate of compound 21-5. The crude product is used directly in the next reaction step;

[0516] MS-ESI[M+H] + The calculated value is 574, and the measured value is 574.

[0517] (5) To the trifluoroacetate of compound 21-5 (370 mg, 538 μmol), dichloromethane (8.0 mL) solution and triethylamine (54.4 mg, 538 μmol, 74.9 μL) were added, followed by intermediate A (206 mg, 807 μol). The reaction mixture was stirred at 25 °C for 1 hour under nitrogen protection. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (20.0 mL × 2), the organic phases were combined, 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 (dichloromethane / methanol = 1:0 to 9:1) to obtain compound 21-6;

[0518] MS-ESI[M+H] + The calculated value is 793, and the measured value is 793.

[0519] (6) Trifluoroacetic acid (3.08 g, 27.0 mmol, 2.0 mL) was added to a solution of compound 21-6 (210 mg, 264 μmol) in dichloromethane (6.0 mL), and the reaction mixture was stirred at 25 °C for 10 minutes. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate of compound 21-7. The crude product was used directly in the next reaction step.

[0520] MS-ESI[M+H] + The calculated value is 694, and the measured value is 694.

[0521] (7) The trifluoroacetate of compound 21-7 (41.1 mg, 248 μmol), compound 21-8 (100 mg, 124 μmol), triethylamine (12.5 mg, 124 μmol, 17.3 μL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (94.3 mg, 248 μmol) were dissolved in dichloromethane (5.0 mL) and stirred at 25 °C for 0.5 hours. The reaction solution was diluted with water (30.0 mL), extracted with dichloromethane (30.0 mL × 2), 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 formate salt of compound 21 was obtained 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 prepare the crude product.

[0522] MS-ESI[M+H] + The calculated value is 805, and the measured value is 805.

[0523] 1 H NMR (400MHz, MeOD) δ8.30-8.34 (m, 1H), 7.81-7.92 (m, 1H), 7.58-7.67 (m, 1H), 7.30-7.39 (m, 1H), 7.11-7.24 (m, 2H), 6.83-6.91 (m, 1H), 6.73-6.83 (m, 1H), 6.37-6.46 (m, 1H), 4.5 4-4.67(m, 4H), 4.43-4.29(m, 5H), 3.89-3.98(m, 1H), 3.77-3.87(m, 2H), 3.71-3.76(m, 4 H), 3.52-3.70(m, 2H), 3.36-3.51(m, 3H), 3.13-3.30(m, 3H), 3.04-3.12(m, 2H), 2.87(br d, J=4.4Hz, 1H), 2.69-2.76 (m, 5H), 1.99-2.12 (m, 3H), 1.26-1.35 (m, 5H), 1.14-1.10 (m, 5H).

[0524] Example 22

[0525] This embodiment provides a compound 22 represented by Formula I, the structural formula of which is shown below:

[0526]

[0527] The synthetic route of compound 22 is shown below.

[0528]

[0529] Triethylamine (12.5 mg, 124 μmol) was added to a solution of trifluoroacetate (100 mg, 124 μmol) of compound 21-7 in dichloromethane (5.0 mL). Compound 22-2 (16.9 mg, 186 μmol, 15.2 μL) was then added, and the reaction mixture was stirred at -78°C for 5 minutes under nitrogen protection. Dichloromethane (60 mL) was added, and the organic phase was washed with water (30 mL) and saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated 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) to obtain the formate salt of compound 22.

[0530] MS-ESI[M+H] + Calculated value: 747, Measured value: 747

[0531] 1 H NMR (400MHz, MeOD) δ8.27-8.34(m, 1H), 7.80-7.88(m, 1H), 7.58-7.65(m, 1H), 7.30-7.37(m, 1H) ,7.14-7.22(m,2H),6.83-6.91(m,1H),6.22-6.37(m,2H),5.71-5.80(m,1H),4.49-4.57(m,2H), 4.31-4.48(m, 2H), 4.22-4.31(m, 2H), 4.09-4.19(m, 2H), 3.89-3.97(m, 1H), 3.56-3.87(m, 6H), 3.45-3.55(m, 1H), 3.34-3.44(m, 1H), 2.92-3.29(m, 6H), 1.92-2.07(m, 4H), 1.08-1.33(m, 10H).

[0532] Example 23

[0533] This embodiment provides a compound represented by Formula I, the structural formula of which is shown below:

[0534]

[0535] The synthetic route for compound 23 is shown below:

[0536]

[0537] (1) Triethylamine (160 mg, 1.58 mmol), compound D (207 mg, 791 μmol), and sodium cyanoborohydride (199 mg, 3.17 mmol) were added to a methanol (10.0 mL) solution of the hydrochloride salt of intermediate compound 3-3 in Example 3 (440 mg, 791 μmol), and the mixture was stirred at 25 °C for 12 hours. Water (100 mL) and dichloromethane (200 mL) were added to the reaction solution. The organic phase was washed with saturated brine (100 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 (dichloromethane / methanol = 1:0 to 10:1) to obtain compound 23-1.

[0538] MS-ESI[M+H] + The calculated value is 688, and the measured value is 688.

[0539] (2) Trifluoroacetic acid (4.0 mL) was added to a solution of compound 23-1 (500 mg, 726 μmol) in dichloromethane (12.0 mL), and the reaction mixture was stirred at 25 °C for 60 minutes. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate of compound 23-2. The crude product was used directly in the next reaction step.

[0540] MS-ESI[M+H] + The calculated value is 588, and the measured value is 588.

[0541] (3) Triethylamine (72.1 mg, 712 μmol) and intermediate A (364 mg, 1.43 mmol) were added to a solution of trifluoroacetate (500 mg, 712 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at 25 °C for 1 hour under nitrogen protection. Water (10.0 mL) and dichloromethane (20.0 mL) were added. The organic phase was washed with saturated brine (10.0 mL), 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 23-3.

[0542] MS-ESI[M+H] + The calculated value is 807, and the measured value is 807.

[0543] (4) Add 2.0 mL of trifluoroacetic acid to a solution of compound 23-3 (100 mg, 123 μmol) in dichloromethane (6.0 mL), and stir the reaction solution at 25 °C for 60 minutes. Concentrate the reaction solution under reduced pressure to obtain the trifluoroacetate of compound 23-4. The crude product is used directly in the next reaction step.

[0544] MS-ESI[M+H] + The calculated value is 707, and the measured value is 707.

[0545] (5) Triethylamine (12.3 mg, 121 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (46.3 mg, 121 μmol) were added to a dichloromethane (3.0 mL) solution of the hydrochloride salt of compound 23-5 (23.6 mg, 182 μmol). The trifluoroacetate salt of compound 23-4 (50 mg, 60.9 μmol) was added to the reaction solution. The reaction solution was stirred at 25 °C for 1 hour. Dichloromethane (200 mL) was added. The organic phase was washed with water (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated into the formate salt of compound 23 by preparative high performance liquid chromatography (Xtimate C18, 100mm×30mm 10μm, A: water (0.225% formic acid); B: acetonitrile, 3%-33%: 10 min).

[0546] MS-ESI[M+H] + The calculated value is 818, and the measured value is 818.

[0547] 1 H NMR (400MHz, MeOD) δ8.37-8.42(m, 2H), 7.79-7.86(m, 1H), 7.59-7.78(m, 1H), 7.12-7.18(m , 2H), 6.86-6.91(m, 1H), 6.72-6.83(m, 1H), 6.31-6.47(m, 1H), 4.53-4.67(m, 4H), 4.39-4.4 6(m, 1H), 4.26-4.37(m, 2H), 3.59-3.89(m, 12H), 2.88-3.05(m, 4H), 2.68-2.83(m, 8H), 1.8 7-2.04(m, 4H), 1.52-1.56(m, 3H), 1.40-1.45(m, 3H), 1.17-1.22(m, 3H), 1.11-1.16(m, 3H).

[0548] Example 24

[0549] This embodiment provides a compound 24 represented by Formula I, the structural formula of which is shown below:

[0550]

[0551] The synthetic route for compound 24 is shown below:

[0552]

[0553] Triethylamine (6.16 mg, 60.9 μmol) was added to a solution of trifluoroacetate (50 mg, 60.9 μmol) of compound 23-4 in dichloromethane (2.0 mL). Compound 24-1 (8.27 mg, 91.3 μmol, 7.45 μL) was then added, and the reaction mixture was stirred at -78°C for 60 minutes under nitrogen protection. Dichloromethane (200 mL) was added, and the organic phase was washed with water (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated 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) to obtain the formate salt of compound 24.

[0554] MS-ESI[M+H] + The calculated value is 761, and the measured value is 761.

[0555] 1H NMR (400MHz, MeOD) δ8.37-8.40 (m, 1H), 8.28-8.30 (m, 1H), 7.81-7.84 (m, 1H), 7.66-7.70 (m, 1H), 7.12-7 .16(m, 2H), 6.86-6.91(m, 1H), 6.19-6.39(m, 2H), 5.73-5.82(m, 1H), 4.54-4.60(m, 4H), 4.39-4.44(m, 1 H), 4.25-4.34(m, 2H), 3.80-3.89(m, 4H), 3.58-3.71(m, 6H), 2.97-3.02(m, 2H), 2.84-2.92(m, 2H), 2.69 -2.76 (m, 2H), 1.88-1.99 (m, 4H), 1.53-1.55 (m, 3H), 1.40-1.44 (m, 3H), 1.18 (m, 3H), 1.12-1.15 (m, 3H).

[0556] Example 25

[0557] This embodiment provides a compound 25 represented by Formula I, the structural formula of which is shown below:

[0558]

[0559] The synthetic route for compound 25 is shown below:

[0560]

[0561] (1) Triethylamine (72.8 mg, 719 μmol), compound 25-1 (192 mg, 719 μmol), and sodium cyanoborohydride (361 mg, 5.70 mmol) were added to a methanol (10.0 mL) solution of trifluoroacetate (400 mg, 719 μmol) of compound 3-3. The mixture was stirred at 25 °C for 12 hours. The reaction solution was poured into water (5.0 mL), stirred for 5 minutes, and extracted with ethyl acetate (10.0 mL × 3). The organic phases were combined, washed with saturated brine (10.0 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 25-2.

[0562] MS-ESI[M+H] + Calculated value: 693, Measured value: 693.

[0563] (2) Trifluoroacetic acid (3.00 g, 26.2 mmol) was added to a solution of compound 25-2 (282 mg, 406 μmol) in dichloromethane (6.0 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate of compound 25-3. The crude product was used directly in the next step of the reaction.

[0564] MS-ESI[M+H] + Calculated value 593, measured value 593.

[0565] (3) Triethylamine (40.0 mg, 396 μmol) and intermediate A (151 mg, 594 μmol) were added to a solution of trifluoroacetate (280 mg, 396 μmol) in dichloromethane (8.0 mL). The reaction mixture was stirred at 25 °C for 1 hour under nitrogen protection. Water (10.0 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10.0 mL × 2). The organic phases were combined, 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 = 10:1) to obtain compound 25-4.

[0566] MS-ESI[M+H] + Calculated value 812, measured value 812.

[0567] (4) Trifluoroacetic acid (2.60 g, 23.0 mmol) was added to a solution of compound 25-4 (157 mg, 193 μmol) in dichloromethane (6.0 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate of compound 25-5. The crude product was used directly in the next reaction step.

[0568] MS-ESI[M+H] + Calculated value 712, measured value 712.

[0569] (5) Triethylamine (16.5 mg, 163 μmol) and compound 25-6 (22.1 mg, 245 μmol) were added to a solution of trifluoroacetate (135 mg, 163 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at -78 °C for 1 hour under nitrogen protection. The reaction mixture was poured into water (20 mL) and extracted with dichloromethane (20.0 mL × 2). The organic phases were combined, 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 = 10:1) to obtain compound 25.

[0570] MS-ESI[M+H] +Calculated value 766, measured value 766.

[0571] 1 H NMR(400MHz,MeOD)δ8.25-8.28(m,1H),7.78-7.80(m,1H),7.09-7.14(m,2H),6.82-6.90(m,1H),6.67-6.69( m,1H),6.20-6.32(m,2H),5.73-5.76(m,1H),4.57-4.63(m,2H),4.52-4.54(m,2H),4.46-4.49(m,2H),4.25- 4.31(m,1H),4.19-4.22(m,2H),3.74-3.76(m,2H),3.57-3.70(m,6H),2.69-2.75(m,4H),2.49-2.57(m,2H), 1.88-1.95(m,2H),1.78-1.85(m,2H),1.51-1.54(m,3H),1.40-1.45(m,3H),1.17(m,3H),1.10-1.15(m,3H).

[0572] Example 26

[0573] This embodiment provides a compound 26 represented by Formula I, the structural formula of which is shown below:

[0574]

[0575] The synthetic route for compound 26 is shown below:

[0576]

[0577] (1) To a methanol (20.0 mL) solution of trifluoroacetate (1.00 g, 1.80 mmol) of compound 3-3, triethylamine (182 mg, 1.80 mmol), compound E (577 mg, 2.10 mmol), and sodium cyanoborohydride (339 mg, 5.40 mmol) were added, and the mixture was stirred at 25 °C for 12 hours. The reaction solution was poured into water (10.0 mL), stirred for 5 minutes, and extracted with ethyl acetate (20.0 mL × 3). The organic phases were combined, 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 (dichloromethane / methanol = 10:1) to obtain compound 26-1.

[0578] MS-ESI[M+H] + Calculated value: 693, Measured value: 693.

[0579] (2) Trifluoroacetic acid (10.7 g, 94.5 mmol) was added to a solution of compound 26-1 (1.00 g, 1.50 mmol) in dichloromethane (21.0 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate of compound 26-2. The crude product was used directly in the next step of the reaction.

[0580] MS-ESI[M+H] + Calculated value 593, measured value 593.

[0581] (3) Triethylamine (143 mg, 1.40 mmol) and intermediate A (542 mg, 2.10 mmol) were added to a solution of trifluoroacetate (1.00 g, 1.40 mmol) in dichloromethane (20.0 mL). The reaction mixture was stirred at 25 °C for 1 hour under nitrogen protection. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10.0 mL × 2). The organic phases were combined, 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 = 10:1) to obtain compound 26-3.

[0582] MS-ESI[M+H] + Calculated value 812, measured value 812.

[0583] (4) Trifluoroacetic acid (10.7 g, 94.5 mmol) was added to a solution of compound 26-3 (1.10 g, 1.30 mmol) in dichloromethane (21.0 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate of compound 26-4. The crude product was used directly in the next reaction step.

[0584] MS-ESI[M+H] + Calculated value 712, measured value 712.

[0585] (5) Triethylamine (61.2 mg, 605.3 μmol) and compound 26-5 (82.1 mg, 908 μmol) were added to a solution of trifluoroacetate (500 mg, 605 μmol) in dichloromethane (5.0 mL). The reaction mixture was stirred at -78 °C for 1 hour under nitrogen protection. The reaction mixture was poured into water (10.0 mL) and extracted with dichloromethane (10.0 mL × 2). The organic phases were combined, 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 = 10:1) to obtain compound 26.

[0586] MS-ESI[M+H] +Calculated value 766, measured value 766.

[0587] 1 H NMR(400MHz,MeOD)δ8.26-8.29(m,1H),7.79-7.82(m,1H),7.10-7.15(m,2H),6.85-6.92(m,1H),6.68-6.71(m ,1H),6.20-6.34(m,2H),5.73-5.79(m,1H),4.47-4.56(m,2H),4.38-4.42(m,2H),4.28-4.35(m,1H),4.19-4. 26(m,2H),3.71-3.81(m,4H),3.53-3.70(m,6H),2.87-2.91(m,2H),2.71-2.77(m,2H),2.53-2.61(m,2H),1.8 9-1.97(m,2H),1.80-1.87(m,2H),1.53-1.55(m,3H),1.42-1.46(m,3H),1.17-1.20(m,3H),1.12-1.16(m,3H).

[0588] Experimental Example 1

[0589] Determination of the antiproliferative effect of the compound on MV-4-11 cells (CCK method):

[0590] 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 compounds involved in this invention inhibit the proliferation of MV-4-11 cells by interfering with the menin / MLL protein / protein interaction.

[0591] 2. Experimental materials: Cell Counting Kit-8 was purchased from Shanghai Liji Biotechnology Co., Ltd. (item number D3100L4057); 96-well clear white cell culture plates were purchased from Corning Costar (item number 3610); fetal bovine serum was purchased from GIBCO (item number #10099-141); Ishkov modified medium (IMDM) was purchased from Invitrogen (item number 12440046); and the benchtop microplate reader SpectraMax i3X was purchased from Molecular Devices.

[0592] 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.

[0593] The test compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution with a concentration of 10 mmol / L. This stock solution was then serially diluted 8 times with DMSO at a 4-fold gradient. Finally, it was 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 concentrations of the compound as follows: 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).

[0594] 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).

[0595] 4. Data Processing:

[0596] The inhibition rate of drug on tumor cell growth is calculated using the following formula:

[0597] Tumor cell growth inhibition rate % = [(ODc-ODs) / (ODc-ODb)] × 100%

[0598] Wherein, ODs: OD of the sample (cells + CCK-8 + test compound), ODc: OD of the negative control (cells + CCK-8 + DMSO), and ODb: OD of the blank control (culture medium + CCK-8 + DMSO).

[0599] The IC50 of the compound was calculated using Graphpad software. 50 .

[0600] The specific test results are shown in Table 1:

[0601] Table 1

[0602]

[0603]

[0604] As shown in Table 1, the spirocyclic compound represented by Formula I of this invention 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.

[0605] Experimental Example 2

[0606] Determination of the antiproliferative effect of the compound on MV-4-11 cells (CTG method):

[0607] 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 compounds involved in this invention inhibit the proliferation of MV-4-11 cells by interfering with the menin / MLL protein / protein interaction.

[0608] 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).

[0609] 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.

[0610] The test compound was dissolved in DMSO to prepare a 10 mM stock solution. The stock solution was first diluted to 2 mM 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.

[0611] Cells with 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 .

[0612] 4. Experimental data:

[0613] The specific test results are shown in Table 2:

[0614] Table 2

[0615] Test compounds <![CDATA[MV 4-11IC 50 (nM)]]> Formate in Example 2 237.2 Formate in Example 3 8.36 Formate of Example 5 575.1 Example 6 422.5 Formate in Example 7 81.22 Formate of Example 12 124.2 Formate of Example 13 317.6 Formate of Example 14 485.7 Formate from Example 15 47.09 Formate from Example 16 91.37 Formate from Example 17 90.38 Formate of Example 18 167.2 Formate from Example 19 8.83 Formate of Example 20 19.36 Formate from Example 21 95.84 Formate of Example 22 78.54 Formate of Example 23 12.06 Formate of Example 24 29.70 Example 25 36.40 Example 26 75.73

[0616] As shown in Table 2, the spirocyclic compound represented by Formula I of this invention 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.

[0617] The applicant declares that this application illustrates spirocyclic 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 spiro compound, characterized by, The structural formula of the spiro compound is shown in the following formula I: wherein, R 1 -C(O)(NR a R b ); wherein R a , R b are each independently selected from H, C1-C6alkyl, and 3-8 membered cycloalkyl; R 2 is fluorine; R 3 is H; R 4 is H; Y and Z are N respectively; W is C; V is N; U 1 , U 2 , U 3 , U 4 , U 5 , U 6 are each independently selected from: -C(R ' )(R ” )- and -C(R ' )(R ” )-C(R ”' )(R ”” )-; U 7 , U 8 are each -C(R ' )(R ” )-; Each R ' Each is independently selected from: H and C1-C4 alkyl groups; Each R ” Each is independently selected from: H and C1-C4 alkyl groups; Each R ”' Each is independently selected from: H and C1-C4 alkyl groups; Each R ”” Each is independently selected from: H and C1-C4 alkyl groups; A is a benzene ring or a 5-6 membered heteroaromatic ring; wherein the heteroaromatic ring contains 1-3 heteroatoms selected from N, O and S; L 1 is -CH2-; L 2 -SO2-; L 3 is an oxygen atom; X is a carbon atom; R 5 is selected from the group consisting of: -CH2R 5' , wherein R 5' is a fluorine or chlorine atom; R 5” is H, methyl or a fluorine atom; R 5”' is selected from the group consisting of: H, optionally substituted C1-C4alkyl, C1-C4alkylamino, (C1-C4alkyl)2amino and C2-C4acyl; wherein the optional substituent of C1-C4alkyl is selected from -N(C1-C3alkyl)2; indicates the position of attachment of the group.

2. The spiro compound according to claim 1, characterized by The A is a benzene ring, a pyridine ring or a thiophene ring.

3. The spiro compound according to claim 1, wherein said R 5 is selected from the group consisting of: -CH2R 5' , wherein R 5' is a fluorine or chlorine atom; R 5” is H, a methyl group or a fluorine atom; R 5”' is selected from the group consisting of: H and optionally substituted C1-C4alkyl; wherein the optional substituents of C1-C4alkyl are selected from the group consisting of: -N(C1-C3alkyl)2.

4. The spiro compound of claim 1, wherein The spirocycle shown in formula I is selected from any one of the following groups: The spirocycle shown in formula I is selected from any one of the following groups:

5. The spiro compound of claim 1, wherein in the formula I represents a fused ring moiety selected from any one of the following groups: wherein R e , R f are each independently selected from: H.

6. The spiro compound of claim 1, wherein The structural formula in the formula I is The cyclic moiety shown in the formula I is selected from the following groups:

7. The spiro compound of claim 1, wherein said R 5 selected from: -CH2F, -CH2Cl, 8. The spiro compound of claim 1, wherein The compound shown in the formula I is selected from any one of the following compounds:

9. The spiro compound of claim 1, wherein The spiro compound further includes a pharmaceutically acceptable salt of the compound shown in the formula I.

10. A pharmaceutical composition, characterized by, The pharmaceutical composition includes the spiro compound according to any one of claims 1-9 and a pharmaceutically acceptable carrier.

11. Use of the spiro compound according to any one of claims 1-9 or the pharmaceutical composition according to claim 10, characterized in that, The use is selected from any one of the following (a)-(c): (a) preparing a drug for preventing or treating tumors, diabetes and other diseases related to the activity of MLL1, MLL2, MLL fusion protein and / or menin protein; (b) preparing an inhibitor for in vitro non-therapeutic activity related to MLL1, MLL2, MLL fusion protein and / or menin protein; (c) preparing a proliferation inhibitor for in vitro non-therapeutic tumor cells.

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