CN116507618A

CN116507618A - Spiro compound, pharmaceutical composition containing spiro compound and application of spiro compound

Info

Publication number: CN116507618A
Application number: CN202180068196.6A
Authority: CN
Inventors: 陈旭星; 李京; 陈艳红; 赵兆
Original assignee: Shanghai Youli Huisheng Pharmaceutical Co ltd
Current assignee: Shanghai Youli Huisheng Pharmaceutical Co ltd
Priority date: 2020-11-18
Filing date: 2021-11-09
Publication date: 2023-07-28
Also published as: CN114516873A; TWI815210B; TW202220991A; WO2022105636A1

Abstract

Disclosed herein are spiro compounds, pharmaceutical compositions comprising the same, and uses thereof. The spiro compounds interfere with interactions between the mentin proteins and MLL1 or MLL2 or MLL-fusion oncoproteins, and are expected to be drugs for treating tumors, diabetes and other diseases that depend on the activities of the MLL1, MLL2, MLL fusion proteins, and/or the menin proteins.

Description

Spiro compound, pharmaceutical composition containing spiro compound and application of spiro compound

Technical Field

The application belongs to the field of pharmaceutical chemistry, and particularly relates to a spiro compound, a pharmaceutical composition containing the spiro compound and application of the spiro compound.

Background

Mixed-lineage leukemia (MLL) protein is a histone methyltransferase that plays an important role in the regulation of gene transcription. Most acute leukemias, including acute myelogenous leukemia (acute myeloblastic leukemia, AML), acute lymphoblastic leukemia (acute lymphoblastic leukemia, ALL) and mixed leukemia, are found to be translocated by the MLL gene located at the q23 band position of chromosome 11, forming a MLL fusion (MLL-r) protein with one of about 80 proteins (e.g., AF4, AF9, ENL, AF10, ELL, AF6, AF1p, GAS7, etc.). The MLL-r protein retains approximately 1400 amino acid sequences at the N-terminus of the MLL protein, lacks the C-terminal methyltransferase active region, and is capable of abnormally regulating transcription of various oncogenes including HOX and MEIS1, promoting cell proliferation, and ultimately leading to the occurrence of cancer. Leukemia patients with chromosomal translocation of the MLL gene generally have a poor prognosis and a 5-year survival rate of less than 40% (Slay, haemato Logica,2009,94,984-993).

The Menin protein, encoded by the multiple endocrine gland tumor (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 metal Res,2005,37,369-374). The Menin protein binds to the N-terminus of MLL proteins, including MLL1, MLL2 and MLL-r proteins, which binding is necessary for the oncogenic activity of the MLL proteins (Yokoyama et al, cell,2005,123,207-218;Cierpicki and Grembecka,Future Med.Chem, 2014,6,447-462). Interference with the interaction between the men and MLL-r proteins can selectively inhibit proliferation of MLL-r leukemia cells in vitro and in vivo (Gremebecka et al, nat. Chem. Biol.,2012,8,277-284; borkin et al, cancer cell,2015,27,589-602).

In specific hematological neoplasms, there are certain specific gene abnormalities or mutations, such as nuclear pore protein 98 (NUP 98) gene fusion, nuclear phosphoprotein (NPM 1) gene mutation, DNA methyltransferase 3A (DNMT 3A) mutation, MLL gene amplification, etc., which are often accompanied by high levels of HOX gene expression. The backward HOXD gene, especially HOXD13, is abnormally over-expressed in ewing's sarcoma, accompanied by high levels of meinin and MLL1 proteins, while HOXD13 is a downstream gene regulated by menin and MLL 1.

Thus, interfering with the interaction between the gin and the MLL protein, especially by covalent binding, is a very promising strategy for treating tumors.

Thus, there is an urgent need in the art to develop effective drugs capable of interfering with the interaction of the men and MLL proteins.

Disclosure of Invention

The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the claims.

In view of the shortcomings of the prior art, the present application aims to provide a spiro compound, a pharmaceutical composition containing the spiro compound and application thereof, wherein the spiro compound and the pharmaceutical composition containing the spiro compound can interfere interaction between the menin and the MLL protein.

To achieve the purpose, the application adopts the following technical scheme:

in a first aspect, the present application provides a spiro compound, where the structural formula of the spiro compound is shown in formula I below:

wherein,

R ¹ selected from-C (O) (NR) ^a R ^b ) (i.e) The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is ^a 、R ^b Each independently selected from H, optionally substituted C1-C6 alkyl, optionally substituted 3-8 membered cycloalkyl and optionally substituted 4-8 membered heterocyclyl, or R ^a And R is R ^b Is linked to N to form an optionally substituted 4-8 membered heterocyclic ring; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P;

R ² Selected from H, halogen, methyl and trifluoromethyl;

R ³ selected from H and halogen;

R ⁴ selected from H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, halogen, -NH ₂ 、-NO ₂ -COOH, -CN, -OH, optionally substituted C1-C6 alkyl sulphonyl, optionally substituted C1-C6 alkyl sulphoxide, optionally substituted C1-C6 alkylthio, -NHCOCR ^4' ＝CH ₂ (i.e)、-NHCOCHR ^4' R ^4” (i.e)、-SO ₂ C(R ^4' )＝CH ₂ (i.e)、-NHSO ₂ CR ^4' ＝CH ₂ (i.e) and-NHSO ₂ CHR ^4' R ^4” (i.e) The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is ^4' Selected from H, methyl and fluoro; r is R ^4” Selected from chlorine and bromine atoms;

y, Z are each independently selected from N and CH, and at least one of Y and Z is N;

w is selected from N and C;

v is selected from N and CR ^V Wherein R is ^V Is H, halogen, -CN, -OH, -NH ₂ Optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino or optionally substituted (C1-C4 alkyl) ₂ An amino group;

U ¹ 、U ² 、U ³ 、U ⁴ 、U ⁵ 、U ⁶ 、U ⁷ 、U ⁸ each 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 (NH ₂ ) - (i.e.)) And U is as follows ¹ 、U ² 、U ³ 、U ⁴ At most one of them is-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 ") -O-, -C (R ') (R") -NR ' "-, or-N=c (NH) ₂ )-，U ⁷ 、U ⁸ At most one of them is-C (=o) -, -C (R ') (R ") -O-, -C (R ') (R") -NR ' "-, or-N=c (NH) ₂ )-；

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;

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 to 3 heteroatoms selected from N, O, S, P;

L ¹ is absent, -CR ^L1' R ^L1” - (i.e.)) (CO) - (i.e.))、-SO ₂ - (i.e.)) (i.e.)) -C (n=n) - (i.e.)) Oxygen or-NH-; wherein R is ^L1' 、R ^L1” Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R ^L1' And R is R ^L1” Forming an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the attached carbon atom; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P;

L ² Selected from: -SO ₂ -、-SO-、-CO-、-CF ₂ -and-C (n=n) -;

L ³ selected from: oxygen atom, sulfur atom, -SO ₂ -、-SO-、-CO-、-CR ^L3' R ^L3” - (i.e.)) and-NR ^L3”' - (i.e.)) The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is ^L3' 、R ^L3” Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R ^L3' And R is ^L3” Forming an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the attached carbon atom; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P; r is R ^L3”' Selected from: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkyl and optionally substituted 4-8 membered saturated or unsaturated heterocycle; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P;

x is selected from: carbon atoms, -S-, and-SO-;

R ⁵ selected from: -CH ₂ R ^5' 、 Wherein R is ^5' Is a fluorine or chlorine atom; r is R ^5” Is H, methyl or fluorine atom; r is 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) ₂ Amino, optionally substituted C1-C4 alkylthio, optionally substituted 3-8 membered saturated or unsaturated cycloalkyl, optionally substituted 4-8 membered saturated or unsaturated heterocyclyl and substituted or unsubstituted C2-C4 acyl; wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O, S, P;

Represents the attachment position of the group.

Preferably, said R ² Selected from H and halogen; further preferably, the R ² Is fluorine.

Preferably, said R ³ Is H or fluorine atom; further preferably, the R ³ H.

Preferably, said R ⁴ Is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, -NH ₂ or-CN.

Preferably, each of the Y, Z is N.

Preferably, W is C.

Preferably, V is N.

Preferably, the U ¹ 、U ² 、U ³ 、U ⁴ 、U ⁵ 、U ⁶ 、U ⁷ 、U ⁸ Each 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.

Preferably, the 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 to 3 heteroatoms selected from N, O, S, P; further preferred, the 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 triazazine 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.

Preferably, the L ¹ Is absent or-CH ₂ -; further preferably, the L ¹ is-CH ₂ -。

Preferably, the L ² Selected from: -SO ₂ -, -SO-and-CO-; further preferably, the L ² is-SO ₂ 。

Preferably, the L ³ Selected from: oxygen atom, sulfur atom, -CR ^L3' R ^L3” -and-NR ^L3”' -; wherein R is ^L3' 、R ^L3” Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R ^L3' And R is ^L3” Forming an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the attached carbon atom; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P; r is R ^L3”' Selected from: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkyl and optionally substituted 4-8 membered saturated or unsaturated heterocycle; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P; further preferably, the L ³ Is an oxygen atom or a sulfur atom.

Preferably, said X is selected from: carbon atoms and-SO-; further preferably, X is a carbon atom.

Preferably, said R ⁵ Selected from: -CH ₂ R ^5' 、 Wherein R is ^5' Is a fluorine or chlorine atom; r is R ^5” Is H, methyl or fluorine atom; r is R ^5”' Selected from: H. optionally substituted C1-C4 alkyl.

Preferably, in the formula IThe spiro ring is selected from any one of the following groups:

preferably, in the formula IThe representative ring combining part is selected from any one of the following groups:

wherein R is ^e 、R ^f Each independently selected from: H. methyl, trifluoromethyl, difluoromethyl, methoxy, ethoxy, trifluoromethoxy, halogen, hydroxy, amino, cyano, methylaminoDimethylamino, ethylamino, methylamino, diethylamino, trifluoroethylamino, carboxy, methoxycarbonyl, ethoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, methylcarbamoyl and diethylcarbamoyl.

Preferably, the structural formula in the formula I isThe cyclic moiety shown is selected from any one of the following groups:

preferably, said R ⁵ Selected from: -CH ₂ F、-CH ₂ F、-CH ₂ Cl、

Preferably, the compound shown in the formula I is selected from any one of the following compounds:

preferably, the spiro compound further comprises any one of pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, cis-trans isomers, solvates or polymorphs or deuterated compounds of the compound shown in formula I.

In a second aspect, the present application provides a pharmaceutical composition comprising a spirocyclic compound according to the first aspect and a pharmaceutically acceptable carrier.

Preferably, the pharmaceutical composition further comprises other pharmaceutically acceptable therapeutic agents, in particular other antitumor agents. Such therapeutic agents include, but are not limited to: antitumor drugs acting on the chemical structure of DNA such as cisplatin, antitumor drugs affecting nucleic acid synthesis such as Methotrexate (MTX), 5-fluorouracil (5 FU) and the like, antitumor drugs affecting nucleic acid transcription such as doxorubicin, epirubicin, aclacinomycin, mithramycin and the like, antitumor drugs acting on tubulin synthesis such as paclitaxel, vinorelbine and the like, aromatase inhibitors such as aminoglutethimide, orchiron, letrozole, ryanodine and the like, cell signal pathway inhibitors such as epidermal growth factor receptor inhibitors Imatinib (Imatinib), gefitinib (Gefitinib), erlotinib (Erlotinib), lapatinib (Lapatinib) and the like.

In a third aspect, the present application provides a use of a spirocyclic compound as described in the first aspect or a pharmaceutical composition as described in the second aspect, said use being selected from any one of the following (a) - (c):

(a) Preparing a medicament for preventing or treating tumors, diabetes and other diseases related to the activity of MLL1, MLL2, MLL fusion proteins, and/or the menin protein;

(b) Preparing an inhibitor for in vitro non-therapeutic association with the activity of MLL1, MLL2, MLL fusion proteins, and/or grin proteins;

(c) Preparing proliferation inhibitor for in vitro non-therapeutic tumor cells.

In a preferred embodiment, the tumor associated with MLL1, MLL2, MLL fusion protein, and/or the activity of the grin protein is selected from the group consisting of: leukemia, ewing's sarcoma, breast cancer, prostate cancer, T-cell lymphoma, B-cell lymphoma, malignant rhabdomyoma, synovial sarcoma, colorectal cancer, endometrial tumor, gastric cancer, liver cancer, renal cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, brain glioma, cholangiocarcinoma, nasopharyngeal cancer, cervical cancer, head and neck cancer, esophageal cancer, thyroid cancer, and bladder cancer.

"other diseases" include, but are not limited to, autoimmune diseases, nonalcoholic hepatitis, and the like.

Description of the terms

Unless defined otherwise, 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 belongs.

As used herein, when used in reference to a specifically recited value, the term "about" means that the value can vary no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes 99 and 101 and all values therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the term "comprising" or "including" can be open, semi-closed, and closed. In other words, the term also includes "consisting essentially of …," or "consisting of ….

In the present application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

By "pharmaceutically acceptable acid addition salt" is meant a salt with an inorganic or organic acid that retains the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, and the like; organic acid salts include, but are not limited to, formate, acetate, 2-dichloroacetate, trifluoroacetate, propionate, hexanoate, octanoate, decanoate, undecylenate, glycolate, gluconate, lactate, sebacate, 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-aminosalicylate, naphthalenedisulfonate, and the like. These salts can be prepared by methods known in the art.

By "pharmaceutically acceptable base addition salt" is meant a salt formed with an inorganic or organic base that is capable of maintaining 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, aluminum, and the like. 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, secondary and tertiary amines, substituted amines including natural 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, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

In this application, "pharmaceutical composition" refers to a formulation of a compound of the present application with a medium commonly accepted in the art for delivery of biologically active compounds to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of organisms, facilitate the absorption of active ingredients and further exert biological activity.

The term "pharmaceutically acceptable" as used herein refers to a material (e.g., carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application, and is relatively non-toxic, i.e., the material can be administered to an individual without causing an adverse biological reaction or interacting in an adverse manner with any of the components contained in the composition.

Herein, "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonizing agent, solvent, or emulsifying agent that is approved by the relevant government regulatory agency as acceptable for human or livestock use.

The term "tumor" as used herein includes, but is not limited to, glioma, sarcoma, melanoma, joint chondrioma, cholangiocarcinoma, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, lung squamous carcinoma, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, and the like.

The terms "prevent", "preventing" and "preventing" as used herein include reducing the likelihood of a patient from developing or worsening a disease or condition.

The term "treatment" and other similar synonyms as used herein include the following meanings:

(i) Preventing the occurrence of a disease or disorder in a mammal, particularly when such mammal is susceptible to the disease or disorder, but has not been diagnosed as having the disease or disorder;

(ii) Inhibiting the disease or disorder, i.e., inhibiting its progression;

(iii) Alleviating a disease or condition, i.e., causing the state of the disease or condition to subside; or alternatively

(iv) Alleviating symptoms caused by the disease or condition.

The term "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein refers to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is required to provide clinically significant relief from a disorder. Effective amounts suitable in any individual case can be determined using techniques such as a dose escalation test.

The terms "administering," "administering," and the like as used herein refer to a method capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, duodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Application techniques useful in the compounds and methods described herein are well known to those skilled in the art, for example, at Goodman and Gilman, the Pharmacological Basis of Therapeutics, current ed.; pergamon; and Remington's, pharmaceutical Sciences (current edition), mack Publishing co., easton, pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

The terms "pharmaceutical combination", "co-administration", "administration of other treatments", "administration of other therapeutic agents" and the like as used herein refer to a pharmaceutical treatment obtained by mixing or combining more than one active ingredient, which includes both fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient in the form of a single entity or single dosage form. The term "ambulatory combination" refers to the simultaneous administration, co-administration, or sequential administration of at least one compound described herein and at least one synergistic formulation as separate entities to a patient at variable intervals. These also apply to cocktail therapies, for example, administration of three or more active ingredients.

Definition of groups

The definition of standard chemical terms can be found in references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY TH ED." vols. A (2000) and B (2001), plenum Press, new York). Unless otherwise indicated, conventional methods within the skill of the art, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy, and pharmacological methods, are employed. Unless specifically defined otherwise, the terms used herein in the description of analytical chemistry, organic synthetic chemistry, and pharmaceutical chemistry are known in the art. Standard techniques may be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, the reaction and purification can be carried out using the manufacturer's instructions for the kit, or in a manner well known in the art or as described herein. The techniques and methods described above may generally be practiced according to conventional methods well known in the art, based on a number of general and more specific descriptions in the literature cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by one skilled in the art to provide stable moieties and compounds.

When substituents are described by conventional formulas written from left to right, the substituents also include chemically equivalent substituents obtained when writing formulas from right to left. For example, -CH ₂ O-is equivalent to-OCH ₂ -。

The section headings used herein are for purposes of organizing articles only and should not be construed as limiting the subject matter. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and treatises, are hereby incorporated by reference in their entirety.

Certain chemical groups defined herein are preceded by a simplified symbol to indicate the total number of carbon atoms present in the group. For example, C1-C6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the reduced notation does not include carbon that may be present in a substituent of the group.

Represents the attachment position of the group.

In addition to the foregoing, when used in the specification and claims of this application, the following terms have the meanings indicated below, unless specifically indicated otherwise.

The term "halogen" herein refers to fluorine, chlorine, bromine or iodine.

"hydroxy" refers to an-OH group.

"hydroxyalkyl" refers to an alkyl group as defined below substituted with a hydroxyl (-OH).

"carbonyl" refers to a-C (=o) -group.

"nitro" means-NO ₂ 。

"cyano" refers to-CN.

"amino" means-NH ₂ 。

"substituted amino" refers to an amino group substituted with one or two alkyl, alkylcarbonyl, aralkyl, heteroaralkyl groups as defined below, e.g., mono-alkylamino, di-alkylamino, alkylamido, aralkylamino, heteroaralkylamino.

"carboxy" refers to-COOH.

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

The term "alkenyl" as part of a group or other group herein means a straight or branched hydrocarbon chain group consisting of only carbon and hydrogen atoms, containing at least one double bond, having, for example, from 2 to 14 (preferably from 2 to 10, more preferably from 2 to 6) carbon atoms, and being linked to the remainder of the molecule by a single bond, such as, but not limited to, ethenyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1, 4-dienyl, and the like.

The term "alkynyl" as part of a group or other group herein means a straight or branched hydrocarbon chain group consisting of only 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, and being attached to the remainder of the molecule by a single bond, such as, but not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

The term "cycloalkyl" as part of a group or other group herein means a stable, non-aromatic, mono-or polycyclic hydrocarbon group consisting of only carbon and hydrogen atoms, which may include fused ring systems, bridged ring systems, or spiro ring systems, having from 3 to 15 carbon atoms, preferably from 3 to 10 carbon atoms, more preferably from 3 to 8 carbon atoms, and which is saturated or unsaturated and may be attached to the remainder of the molecule by a single bond via any suitable carbon atom. Unless otherwise specifically indicated in the present specification, carbon atoms in the cycloalkyl group may optionally be oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexanyl, cyclooctyl, 1H-indenyl, 2, 3-indanyl, 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, 5,6,7,8,9, 10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo [2.2.1] heptyl, 7-dimethyl-bicyclo [2.2.1] heptyl, bicyclo [2.2.1] heptenyl, bicyclo [ 2.2.2.2 ] octyl, bicyclo [3.1.1] heptyl, bicyclo [3.2.1] octyl, bicyclo [2.2.2] octenyl, bicyclo [ 2.1.1 ] octadienyl, adamantylene, and the like.

The term "heterocyclyl" as part of a group or other group herein means 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 specifically indicated otherwise in the present specification, a heterocyclyl group may be a monocyclic, bicyclic, tricyclic or more cyclic ring system, which may include fused, bridged or spiro ring systems; the nitrogen, carbon or sulfur atoms in the heterocyclyl may optionally be oxidized; the nitrogen atom may optionally be quaternized; and the heterocyclyl may be partially or fully saturated. The heterocyclic group may be attached to the remainder of the molecule via a carbon atom or a heteroatom and by a single bond. In heterocyclyl groups containing fused rings, one or more of the rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the remainder of the molecule is a non-aromatic ring atom. For the purposes of this application, a heterocyclyl group is preferably a stable 4-to 11-membered non-aromatic monocyclic, bicyclic, bridged or spiro ring group comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 4-to 8-membered non-aromatic monocyclic, bicyclic, bridged or spiro ring group comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclyl groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2, 7-diaza-spiro [3.5] nonan-7-yl, 2-oxa-6-aza-spiro [3.3] heptan-6-yl, 2, 5-diaza-bicyclo [2.2.1] heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxacyclopentyl, tetrahydroisoquinolyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolyl, octahydroisoindolyl, pyrazolinyl, pyrazolidinyl, phthalimidyl, and the like.

The term "aryl" as part of a group or other group herein means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably having 6 to 10 carbon atoms. For the purposes of this application, aryl groups may be monocyclic, bicyclic, tricyclic or more ring systems, and may also be fused to cycloalkyl or heterocyclyl groups as defined above, provided that the aryl groups are attached to the remainder of the molecule via a single bond through an atom on an aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthryl, phenanthryl, fluorenyl, 2, 3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-benzoxazin-3 (4H) -one-7-yl, and the like.

The term "arylalkyl" herein refers to an alkyl group as defined above substituted with an aryl group as defined above.

The term "heteroaryl" as part of a group or other group herein means a 5-to 16-membered conjugated ring system group having 1 to 15 carbon atoms (preferably 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur within the ring. Unless otherwise specifically indicated in the present specification, heteroaryl groups may be monocyclic, bicyclic, tricyclic or more ring systems, and may also be fused to cycloalkyl or heterocyclyl groups as defined above, provided that heteroaryl groups are attached to the remainder of the molecule via an atom on an aromatic ring by a single bond. The nitrogen, carbon, or sulfur atoms in the heteroaryl group may optionally be oxidized; the nitrogen atom may optionally be quaternized. For the purposes of the present application, heteroaryl groups are preferably stable 5-to 12-membered aromatic groups comprising 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably stable 5-to 10-membered aromatic groups comprising 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur or 5-to 6-membered aromatic groups comprising 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furanyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoindolyl, indazolyl, isoindazolyl, purinyl, quinolinyl, isoquinolinyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, phenanthrolinyl, acridinyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxazolyl, cinnolinyl, quinazolinyl, -indolizinyl, phenanthroline, isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6, 7-tetrahydrobenzo [ b ] thienyl, naphthazinyl, [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-b ] pyridazine, imidazo [1,2-a ] pyrazine, and the like.

The term "heteroarylalkyl" herein refers to an alkyl group as defined above substituted with a heteroaryl group as defined above.

In this application, "optionally" means that the subsequently described event or condition may or may not occur, and that the description includes both cases where the event or condition occurs and where it does not. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted aryl groups and unsubstituted aryl groups. For example, where substituents are not explicitly listed, the term "substituted" or "substituted with … …" as used herein means that one or more hydrogen atoms on a given atom or group are independently substituted with one or more, e.g., 1, 2, 3, or 4 substituents independently selected from: deuterium (D), halogen, -OH, mercapto, cyano, -CD ₃ 、-C ₁ -C ₆ Alkyl (preferably-C _1-3 Alkyl group, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, cycloalkyl (preferably 3-8 membered cycloalkyl), aryl, heterocyclyl (preferably 3-8 membered heterocyclyl), heteroaryl, aryl-C ₁ -C ₆ Alkyl-, heteroaryl-C ₁ -C ₆ Alkyl-, C ₁ -C ₆ Haloalkyl-, -OC ₁ -C ₆ Alkyl (preferably-OC) ₁ -C ₃ Alkyl) -OC ₂ -C ₆ Alkenyl, -OC ₁ -C ₆ Alkylphenyl radicals C ₁ -C ₆ alkyl-OH (preferably-C) ₁ -C ₄ alkyl-OH), -C ₁ -C ₆ alkyl-SH, -C ₁ -C ₆ alkyl-O-C ₁ -C ₆ Alkyl, -OC ₁ -C ₆ Halogenated compoundsAlkyl, -NH ₂ 、-C ₁ -C ₆ alkyl-NH ₂ (preferably-C) ₁ -C ₃ alkyl-NH ₂ )、-N(C ₁ -C ₆ Alkyl group ₂ (preferably-N (C) ₁ -C ₃ Alkyl group ₂ )、-NH(C ₁ -C ₆ Alkyl) (preferably-NH (C) ₁ -C ₃ Alkyl)), -N (C) ₁ -C ₆ Alkyl) (C) ₁ -C ₆ Alkylphenyl), -NH (C) ₁ -C ₆ Alkylphenyl), nitro, -C (O) -OH, -C (O) OC ₁ -C ₆ Alkyl (preferably-C (O) OC) ₁ -C ₃ Alkyl), -CONRiRiri (where Ri and Rii are H, D and C) _1-6 Alkyl, preferably C _1-3 Alkyl), -NHC (O) (C) ₁ -C ₆ Alkyl), -NHC (O) (phenyl), -N (C) ₁ -C ₆ Alkyl) C (O) (C ₁ -C ₆ Alkyl), -N (C) ₁ -C ₆ Alkyl) C (O) (phenyl), -C (O) C ₁ -C ₆ Alkyl, -C (O) heteroaryl (preferably-C (O) -5-7 membered heteroaryl), -C (O) C ₁ -C ₆ Alkylphenyl, -C (O) C ₁ -C ₆ Haloalkyl, -OC (O) C ₁ -C ₆ Alkyl (preferably-OC (O) C) ₁ -C ₃ Alkyl), -S (O) ₂ -C ₁ -C ₆ Alkyl, -S (O) -C ₁ -C ₆ Alkyl, -S (O) ₂ -phenyl, -S (O) ₂ -C ₁ -C ₆ Haloalkyl, -S (O) ₂ NH ₂ 、-S(O) ₂ NH(C ₁ -C ₆ Alkyl), -S (O) ₂ NH (phenyl), -NHS (O) ₂ (C ₁ -C ₆ Alkyl), -NHS (O) ₂ (phenyl) and-NHS (O) ₂ (C ₁ -C ₆ Haloalkyl), wherein said alkyl, cycloalkyl, phenyl, aryl, heteroarylEach of the cyclic and heteroaryl groups is optionally further substituted with one or more substituents selected from the group consisting of: halogen, -OH, -NH ₂ Cycloalkyl, 3-8 membered heterocyclyl, C ₁ -C ₄ Alkyl, C ₁ -C ₄ Haloalkyl-, -OC ₁ -C ₄ Alkyl, -C ₁ -C ₄ alkyl-OH, -C ₁ -C ₄ alkyl-O-C ₁ -C ₄ Alkyl, -OC ₁ -C ₄ Haloalkyl, cyano, nitro, -C (O) -OH, -C (O) OC ₁ -C ₆ Alkyl, -CON (C) ₁ -C ₆ Alkyl group ₂ 、-CONH(C ₁ -C ₆ Alkyl), -CONH ₂ 、-NHC(O)(C ₁ -C ₆ Alkyl group), NH (C) ₁ -C ₆ Alkyl) C (O) (C ₁ -C ₆ Alkyl), -SO ₂ (C ₁ -C ₆ Alkyl), -SO ₂ (phenyl) -SO ₂ (C ₁ -C ₆ Haloalkyl) -SO ₂ NH ₂ 、-SO ₂ NH(C ₁ -C ₆ Alkyl), -SO ₂ NH (phenyl) -NHSO ₂ (C ₁ -C ₆ Alkyl), -NHSO ₂ (phenyl) and-NHSO ₂ (C ₁ -C ₆ Haloalkyl). When an atom or group is substituted with multiple substituents, the substituents may be the same or different. The terms "moiety", "structural moiety", "chemical moiety", "group", "chemical group" as used herein refer to a particular fragment or functional group in a molecule. Chemical moieties are generally considered to be chemical entities that are embedded or attached to a molecule.

By "absent" is meant that both sides of the groups defined above are directly linked by chemical bonds. For example, the absence of B in "A-B-C" means "A-C". .

"stereoisomers" refer to compounds that consist of the same atoms, are bonded by the same bonds, but have different three-dimensional structures. The present application will cover various stereoisomers and mixtures thereof.

When an olefinic double bond is contained in a compound of the present application, the compound of the present application is intended to include both E-and Z-geometric isomers unless otherwise specified.

"tautomer" refers to an isomer formed by the transfer of a proton from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the present application are also intended to be included within the scope of the present application.

The compounds of the present application, or pharmaceutically acceptable salts thereof, may contain one or more chiral carbon atoms and thus may produce enantiomers, diastereomers, and other stereoisomeric forms. Each chiral carbon atom may be defined as (R) -or (S) -, based on stereochemistry. The present application is intended to include all possible isomers, as well as racemates and optically pure forms thereof. The compounds of the present application may be prepared by selecting racemates, diastereomers 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.

In the present application, (C1-C4 alkyl) ₂ Amino, representing 2C 1-C4 alkyl-substituted amines, may be, for exampleEtc.

In the present application, "each R '", "each R" "" refer to "each R '", "each R" "", each R ' "appearing in the thienopyrimidine compound represented by formula I.

Conventional techniques for preparing/separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high performance liquid chromatography, see, for example, gerald gabiz and Martin g.schmid (eds.), chiral Separations, methods and Protocols, methods in Molecular Biology, vol.243,2004; m.stalcup, chiral Separations, annu.rev.animal.chem.3:341-63, 2010; fumigs et al (EDs.), VOGEL' S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY.sup.TH ED, longman Scientific and Technical Ltd., essex,1991,809-816; heller, acc.chem.Res.1990,23,128.

It will also be appreciated by those skilled in the art that in the methods described below, the intermediate compound functional groups may need to be protected by appropriate protecting groups. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl groups (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino and guanidino groups include t-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable mercapto-protecting groups include-C (O) -R "(wherein R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl, and the like. Suitable carboxyl protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups may 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 described in detail in Greene, t.w. and p.g.m. wuts, protective Groups in Organi Synthesis, (1999), 4th Ed. The protecting group may also be a polymeric resin.

Compared with the prior art, the application has the following beneficial effects:

(1) The application provides a compound shown as a formula I or pharmaceutically acceptable salt thereof

(2) The application provides a compound shown as a formula I for preparing a pharmaceutical composition for preventing and treating diseases related to MLL1, MLL2, MLL fusion protein and/or menin protein activity.

Still other aspects will be apparent upon reading and understanding the detailed description.

Detailed Description

The technical scheme of the application is further described through the following specific embodiments. It should be apparent to those skilled in the art that the examples are merely provided to aid in understanding the present application and should not be construed as limiting the present application in any way.

The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. Percentages and parts are weight percentages and parts unless otherwise indicated.

The experimental materials and reagents used in the following examples were obtained from commercial sources unless otherwise specified.

In the context of the various embodiments of the present invention, ¹ h NMR was recorded by BRUKER AVANCE NEO 400.400 MHz NMR and chemical shifts were expressed as δ (ppm); LC-MS (LCMS) was recorded by Shimadzu LC-20AD, SIL-20A, CTO-20AC, SPD-M20A, CBM-20A, LCMS-2020 type mass spectrometer; preparative HPLC separation was performed using a Gilson-281 model liquid chromatograph.

Preparation of intermediates

1. Preparation of intermediate A

The synthetic route for intermediate a is shown below:

(1) To a solution of compound A-1 (5.0 g,28.9 mmol) in methylene chloride (25.0 mL) were added triethylamine (5.84 g,57.7 mmol) and compound A-2 (7.27 g,63.5 mmol), and the reaction was stirred under nitrogen at 0deg.C for 1 hour. The reaction was quenched with saturated aqueous sodium bicarbonate (40.0 mL), extracted with dichloromethane (40.0 mL. Times.3), and the combined organic phases were washed with saturated brine (30.0 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure to give Compound A-3.

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

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

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

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

¹ H NMR(400MHz，CDCl ₃ )δ4.56-4.47(m，1H)，4.42-4.30(m，4H)，1.48-1.44(m，9H)。

2. Preparation of intermediate B

The synthetic route for intermediate B is shown below:

(1) To a solution of compound B-1 (100 g,588 mmol) in methylene chloride (1300 mL) was added 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (268 g, 704 mmol), diisopropylethylamine (114 g,882 mmol) and compound B-2 (119 g,1.18 mol) at 0deg.C, and the reaction was stirred at 25deg.C for 3 hours. Water (200 mL) was added, extracted with dichloromethane (200 mL. Times.3), the combined organic phases were washed with saturated brine (200 mL. Times.3), dried over anhydrous sodium sulfate, filtered, the organic phase concentrated under reduced pressure, and the crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=10:0 to 0:1) to give compound B-3.

MS-ESI[M+H] ⁺ Calculated 254, measured 254.

(2) To a solution of compound B-3 (130 g,513 mmol) in methylene chloride (2.0L) at-78℃was added boron tribromide (257 g,1.03 mol), and the reaction was stirred at 25℃for 4 hours. The reaction was quenched with ice, neutralized to pH 8 with saturated aqueous sodium bicarbonate, extracted with dichloromethane (300 mL. Times.3), the combined organic phases were washed with saturated brine (300 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=10:0 to 1:1) to give compound B-4.

MS-ESI[M+H] ⁺ Calculated 240, measured 240.

(3) To a solution of compound B-4 (115 g,41.0 mmol) in N, N-dimethylformamide (1500 mL) was added cesium carbonate (470 g,1.44 mol) and compound B-5 (29.5 g,186 mmol), and the reaction was stirred at 130℃for 12 hours. Cooled to room temperature, water (500 mL) was added and 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, the organic phase concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=20:1 to 1:1) to give compound B-6.

MS-ESI[M+H] ⁺ Calculated 318, measured 318.

(4) To a solution of Compound B-6 (110 g, 277 mmol) in methylene chloride (300.0 mL) at 0deg.C was added m-chloroperoxybenzoic acid (211 g,1.04mol,85% purity), and the reaction was stirred under nitrogen at 25deg.C for 12 hours. The reaction was quenched by addition of saturated sodium thiosulfate solution until the starch potassium iodide paper became no longer blue, followed by extraction with methylene chloride (1000 mL), and the organic phase was washed with saturated aqueous sodium bicarbonate (500 mL. Times.2) and saturated brine (500 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=10:1 to 0:1) to give compound B-7.

MS-ESI[M+H] ⁺ Calculated value 334, measured value 334.

(5) To a solution of phosphorus oxychloride (46.0 g,255 mmol) in chloroform (700 mL) was added triethylamine (22.8 g,225 mmol) and compound B-7 (50.0 g,150 mmol) at 0deg.C, and the reaction was stirred at 65deg.C for 12 hours. The reaction mixture was quenched with ice, neutralized to pH 8 with saturated aqueous sodium bicarbonate, extracted with dichloromethane (200 mL. Times.2), and the organic phase was washed with saturated brine (200 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=10:1 to 0:1) to give compound B.

3. Preparation of intermediate C

The synthetic route for intermediate C is shown below:

(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 (diphenylphosphino) ferrocene ] palladium dichloride (1.36 g,1.86 mmol) were dissolved in dioxane (40.0 mL) and water (40.0 mL). The reaction solution was stirred for 1 hour at 110℃under nitrogen. Aqueous solution (100 mL) was added, extracted with dichloromethane (100 ml×2), the combined organic phases were washed with saturated brine (100 ml×1), dried over anhydrous sodium sulfate, filtered, the organic phase was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 0:1) to give compound C-3.

MS-ESI[M+H] ⁺ Calculated 261, measured 261.

¹ H NMR(400MHz，CDCl ₃ )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)。

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

¹ H NMR(400MHz，CDCl ₃ )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)。

4. Preparation of intermediate D

The synthetic route for intermediate D is shown below:

(1) To a solution of compound D-1 (390 mg,1.40 mmol) in methylene chloride (15 mL) was added 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), and the reaction was stirred at 25℃for 16 hours. Saturated aqueous sodium bicarbonate (20.0 mL) was added, 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, the organic phase 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 give compound D-3.

MS-ESI[M+H] ⁺ Calculated 322, measured 322.

¹ H NMR(400MHz，MeOD)δ8.64(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)。

(2) To a solution of compound D-3 (400 mg,1.24 mmol) in tetrahydrofuran (15.0 mL) at-78℃was added diisobutylaluminum hydride (1.5 mol/L,2.49mL,3.74 mmol), and the reaction was stirred at 0℃for 0.5 hours under nitrogen. The reaction was quenched by the addition of hydrochloric acid (1 mol/L), the pH was adjusted to 7, followed by the addition of water (20.0 mL), extraction with ethyl acetate (20.0 mL. Times.2), washing of the combined organic phases with saturated brine (20.0 mL. Times.1), drying over anhydrous sodium sulfate, filtration, and concentration of the organic phase under reduced pressure. The crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 0:1) to give intermediate D.

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

5. Preparation of intermediate E

The synthetic route for intermediate E is shown below:

to a solution of Compound E-1 (1.0 g,3.14 mmol) in tetrahydrofuran (15.0 mL) at-78deg.C under nitrogen was added N, N-dimethylformamide (344 mg,4.71 mmol), followed by dropwise addition of N-butyllithium (2.5 mol/L,1.89mL,4.73 mmol), and the reaction was stirred at-78deg.C under nitrogen for 2 hours. The reaction was quenched by the addition of saturated aqueous ammonium chloride (30.0 mL) at 0deg.C, followed by the addition of water (40.0 mL), extraction with ethyl acetate (40.0 mL. Times.2), washing of the combined organic phases with saturated brine (50.0 mL. Times.1), drying over anhydrous sodium sulfate, filtration, and concentration of the organic phase under reduced pressure. The crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 0:1) to give intermediate E.

¹ H NMR(400MHz，CDCl ₃ )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)。

6. Preparation of intermediate F

The synthetic route for intermediate F is shown below:

(1) To a solution of compound F-1 (30.0 g,176 mmol) in methylene chloride (300 mL) at 0deg.C was added 2- (7-azabenzotriazol) -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), and the reaction was stirred at 25deg.C for 1 hour. Dichloromethane (400 mL) was added, the organic phase was washed with water (250 ml×1) and saturated brine (220 ml×3), dried over anhydrous sodium sulfate, filtered, the organic phase concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 3:1) to give compound F-3.

MS-ESI[M+H] ⁺ Calculated 240, measured 240.

(2) To a solution of compound F-3 (48.0 g,176 mmol) in methylene chloride (500.0 mL) at-78℃was added boron tribromide (88.0 g,351 mmol), and the reaction was stirred at 25℃for 4 hours. The reaction was quenched with ice, neutralized to pH 8 with saturated aqueous sodium bicarbonate, extracted with dichloromethane (500 mL), and the combined organic phases were washed with saturated aqueous sodium bicarbonate (500 mL. Times.2) and saturated brine (500 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 1:1) to give compound F-4.

MS-ESI[M+H] ⁺ Calculated 226, measured 226.

¹ H NMR(400MHz，CDCl ₃ )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)。

(3) To a solution of compound F-4 (27.9 g,124 mmol) in N, N-dimethylformamide (350 mL) was added cesium carbonate (121 g, 375 mmol) and compound F-5 (29.5 g,186 mmol), and the reaction was stirred at 130℃for 12 hours. Cooled 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, the organic phase was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 2:1) to give compound F-6.

MS-ESI[M+H] ⁺ Calculated 304, measured 304.

¹ H NMR(400MHz，CDCl ₃ )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)。

(4) To a solution of compound F-6 (25.0 g,82.4 mmol) in methylene chloride (300.0 mL) at 0deg.C was added m-chloroperoxybenzoic acid (53.5 g,264mmol,85% purity), and the reaction was stirred under nitrogen at 25deg.C for 12 hours. The reaction was quenched by the addition of saturated sodium sulfite solution (100 mL), followed by saturated aqueous sodium bicarbonate solution (500 mL), extracted with dichloromethane (300 mL), and the organic phase was washed with saturated aqueous sodium bicarbonate solution (500 mL. Times.2) and saturated brine (500 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 25:1) to give compound F-7.

MS-ESI[M+H] ⁺ Calculated 320, measured 320.

¹ H NMR(400MHz，CDCl ₃ )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)。

(5) To a solution of phosphorus oxychloride (14.9 g,97.2 mmol) in chloroform (100.0 mL) at 0deg.C were added triethylamine (8.7 g,85.5 mmol) and compound F-7 (18.2 g,57 mmol), and the reaction was stirred at 25deg.C for 12 hours. The reaction mixture was quenched with ice, neutralized to pH 8 with saturated aqueous sodium bicarbonate, extracted with dichloromethane (500 mL), and the organic phase was washed with saturated aqueous sodium bicarbonate (500 mL. Times.2) and saturated brine (500 mL. Times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 20:1) to give compound F.

MS-ESI[M+H] ⁺ Calculated 338, measured 338.

¹ H NMR(400MHz，CDCl ₃ )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)。

Example 1

The present embodiment provides a compound 1 shown in formula I, wherein the structural formula of the compound 1 is as follows:

the synthetic route for compound 1 is shown below:

(1) 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) was added potassium carbonate (3.54 g,25.6 mmol), and the reaction was stirred under nitrogen at 70℃for 3 hours. The reaction solution was added with water (50.0 mL), 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, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 0:1) to give compound 1-2.

¹ H NMR(400MHz，CDCl ₃ )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＝6.8Hz，3H)，1.47(d,J＝6.8Hz，3H)，1.44(s，9H)，1.12(d,J＝6.8Hz，3H)，1.09(d,J＝6.8Hz，3H)。

(2) To a solution of compound 1-2 (4.4 g,8.12 mmol) in dichloromethane (20.0 mL) was added trifluoroacetic acid (7.7 g,67.5 mmol), and the reaction was stirred at 25℃for 30 min. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 1-3.

MS-ESI[M+H] ⁺ Calculated 442, measured 442.

(3) To a solution of trifluoroacetate salt of compound 1-3 (300 mg, 679. Mu. Mol) in methanol (10.0 mL) was added triethylamine (206 mg,2.04 mmol), compound 1-4 (267 mg,1.02 mmol) and sodium cyanoborohydride (427 mg,6.79 mmol), and the mixture was stirred at 25℃for 4 hours. The reaction solution was concentrated under reduced pressure, ethyl acetate (20.0 mL) was added, washed with water (20.0 ml×1) and saturated brine (20.0 ml×1), and 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 give compounds 1 to 5.

¹ H NMR(400MHz，CDCl ₃ )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)。

(4) To a solution of compounds 1-5 (268 mg, 390. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain trifluoroacetate salts of the compounds 1 to 6. The crude product was used directly in the next reaction.

(5) To a solution of trifluoroacetate salt (279 mg, 390. Mu. Mol) of compounds 1-6 in methylene chloride (5.0 mL) were added triethylamine (390 mg,3.93 mmol) and intermediate A (200 mg, 782. Mu. Mol), and the reaction solution was stirred under nitrogen at 25℃for 16 hours. 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 the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 32:1) to give compounds 1-7.

MS-ESI[M+H] ⁺ Calculated 806, measured 806.

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)。

(6) To a solution of compounds 1-7 (213 mg, 264. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 1-8. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 706, measured 706.

(7) To a solution of trifluoroacetate salt (108 mg, 132. Mu. Mol) of compounds 1-8 in methylene chloride (3.0 mL) was added triethylamine (66.6 mg, 659. Mu. Mol). Compounds 1-9 (17.9 mg, 198. Mu. Mol, 16.1. Mu.L) were then added and the reaction stirred at-78℃for 60 minutes under nitrogen. 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 (Phenomenex luna C18, 100 mm. Times.40 mm 3. Mu.m, A: water (0.225% formic acid; B: acetonitrile, 10% -40% for 10 min) to give the formate of compound 1.

MS-ESI[M+H] ⁺ Calculated 760, measured 760.

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)。

Example 2

The present embodiment provides a compound 2 shown in formula I, wherein the structural formula of the compound 2 is as follows:

the synthetic route for compound 2 is shown below:

to a solution of the hydrochloride of compound 2-1 (43.6 mg, 263. Mu. Mol) in methylene chloride (3.0 mL) were added triethylamine (133 mg,1.31 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (100 mg, 263. Mu. Mol), and the mixture was stirred at 25℃for 0.5 hours, and the trifluoroacetate salt (108 mg, 132. Mu. Mol) of compound 1-8 in example 1 was added and the mixture was stirred at 25℃for 0.5 hours. Dichloromethane (20.0 mL) was added, washed with water (20.0 ml×1) and saturated brine (20.0 ml×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative high performance liquid chromatography (Phenomenex luna C18, 100 mm. Times.40 mm 3. Mu.m, A: water (0.225% formic acid; B: acetonitrile, 10% -40% for 10 min) to give the formate of compound 2.

MS-ESI[M+H] ⁺ Calculated 817, measured 817.

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)。

Example 3

The present embodiment provides a compound 3 shown in formula I, wherein the structural formula of the compound 3 is as follows:

the synthetic route for compound 3 is shown below:

(1) To a solution of intermediate B (300 mg, 853. Mu. Mol) and Compound 3-1 (213 mg, 941. Mu. Mol) in N, N-dimethylformamide (8.0 mL) was added potassium carbonate (236 mg,1.71 mmol), and the reaction was stirred under nitrogen at 70℃for 2 hours. Ethyl acetate (40.0 mL) was added to the reaction solution, which was washed with water (40.0 ml×1) and saturated brine (40.0 ml×5), and 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 give compound 3-2.

MS-ESI[M+H] ⁺ Calculated 542, measured 542.

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)。

(2) To a solution of compound 3-2 (435 mg, 803. Mu. Mol) in methylene chloride (4.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 2 hours. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 3-3.

MS-ESI[M+H] ⁺ Calculated 442, measured 442.

(3) To a solution of trifluoroacetate salt of compound 3-3 (450 mg, 941. Mu. Mol) in methanol (10.0 mL) was added triethylamine (284 mg,2.82 mmol), compound 3-4 (369 mg,1.41 mmol) and sodium cyanoborohydride (592 mg,9.42 mmol), and the mixture was stirred at 25℃for 12 hours. Ethyl acetate (40.0 mL) was added to the reaction solution, the organic phase was washed with saturated aqueous ammonium chloride (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, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=100:1 to 20:1) to give compound 3-5.

MS-ESI[M+H] ⁺ Calculated 687, measured 687.

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)。

(4) To a solution of compound 3-5 (282 mg, 411. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction was stirred at 25℃for 60 minutes. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 3-6. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 587, measured 587.

(5) To a solution of trifluoroacetate salt (284 mg, 408. Mu. Mol) of compound 3-6 in methylene chloride (3.0 mL) were added triethylamine (400 mg,3.95 mmol) and intermediate A (157 mg, 614. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 16 hours. 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 isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 32:1) to give compounds 3-7.

MS-ESI[M+H] ⁺ Calculated 806, measured 806.

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(br s，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)。

(6) To a solution of compound 3-7 (250 mg, 310. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 3-8. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated value 706, trueA measurement 706.

(7) To a solution of the trifluoroacetate salt of compound 3-8 (127 mg, 155. Mu. Mol) in methylene chloride (3.0 mL) was added triethylamine (78.4 mg, 775. Mu. Mol). A solution of Compound 3-9 (29.5 mg, 326. Mu. Mol, 26.6. Mu.L) in methylene chloride (1.0 mL) was then added, and the reaction was stirred at-78℃for 60 minutes under nitrogen. 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 (Phenomenex luna C18, 100 mm. Times.40 mm 3. Mu.m, A: water (0.225% formic acid; B: acetonitrile, 0% -40% for 10 min) to give the formate of compound 3.

MS-ESI[M+H] ⁺ Calculated 760, measured 760.

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)。

Example 4

The present embodiment provides a compound 4 shown in formula I, wherein the structural formula of the compound 4 is as follows:

the synthetic route for compound 4 is shown below:

to a solution of the hydrochloride (51.3 mg, 310. Mu. Mol) of the compound 4-1 in methylene chloride (3.0 mL) were added triethylamine (78.8 mg,779 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (118 mg, 310. Mu. Mol), and the mixture was stirred at 25℃for 0.5 hours, and the trifluoroacetate (127 mg, 155. Mu. Mol) of the compound 3-8 in example 3 was added, and the mixture was stirred at 25℃for 0.5 hours. 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 preparative high performance liquid chromatography (Phenomenex luna C18, 100 mm. Times.40 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 0% -30% for 10 min) to give formate of compound 4.

MS-ESI[M+H] ⁺ Calculated 817, measured 817.

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(br s，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)。

Example 5

The present embodiment provides a compound 5 shown in formula I, wherein the structural formula of the compound 5 is as follows:

the synthetic route for compound 5 is shown below:

(1) To a solution of intermediate B (100 mg, 284. Mu. Mol) and compound 5-1 (81.3 mg, 341. Mu. Mol) in N, N-dimethylformamide (5.0 mL) was added potassium carbonate (118 mg, 853. Mu. Mol), and the reaction mixture was stirred under nitrogen at 70℃for 3 hours. The reaction mixture was added with water (100 mL), extracted with ethyl acetate (25.0 ml×2), and 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, and the crude product was separated by preparative thin layer chromatography (petroleum ether/ethyl acetate=1:1) to give compound 5-2.

¹ H NMR(400MHz，CDCl ₃ )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)。

MS-ESI[M+H] ⁺ Calculated 542, measured 542.

(2) To a solution of compound 5-2 (127 mg, 234. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.54 g,13.5 mmol), and the reaction was stirred at 25℃for 60 minutes. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 5-3.

MS-ESI[M+H] ⁺ Calculated 442, measured 442.

(3) To a solution of the trifluoroacetate salt of compound 5-3 (130 mg, 234. Mu. Mol) in methanol (5.0 mL) was added triethylamine (71.0 mg, 702. Mu. Mol), compound 5-4 (61.2 mg, 234. Mu. Mol) and sodium cyanoborohydride (147 mg,2.34 mmol), and the mixture was stirred at 25℃for 2 hours. The reaction solution was concentrated under reduced pressure, 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, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 5-5.

¹ H NMR(400MHz，CDCl ₃ )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)。

MS-ESI[M+H] ⁺ Calculated 687, measured 687.

(4) To a solution of compound 5-5 (74.4 mg, 108. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 5-6.

MS-ESI[M+H] ⁺ Calculated 587, measured 587.

(5) To a solution of trifluoroacetate salt (75.9 mg, 108. Mu. Mol) of compound 5-6 in methylene chloride (5.0 mL) were added triethylamine (54.8 mg, 542. Mu. Mol) and intermediate A (55.4 mg, 217. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 2 hours. 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 isolated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compounds 5-7.

MS-ESI[M+H] ⁺ Calculated 806, measured 806.

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)。

(6) To a solution of compound 5-7 (56.0 mg, 69.5. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction solution was stirred at 25℃for 30 minutes. Concentrating the reaction solution under reduced pressure to obtain the trifluoroacetate salt of the compound 5-8. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated value 706, measured 706.

(7) To a solution of the trifluoroacetate salt of compound 5-8 (56.9 mg, 69.4. Mu. Mol) in methylene chloride (5.0 mL) was added triethylamine (35.1 mg, 347. Mu. Mol). Then, compound 5-9 (9.4 mg, 104. Mu. Mol, 8.5. Mu.L) was added thereto, and the reaction mixture was stirred at-78℃for 30 minutes under nitrogen. 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 (Xtime C18, 100 mm. Times.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) to give the formate salt of compound 5.

MS-ESI[M+H] ⁺ Calculated 760, measured 760.

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＝6.0Hz，2H)，1.75(br t，J＝5.6Hz，4H)，1.54(d，J＝6.8Hz，3H)，1.45(d，J＝6.8Hz，3H)，1.20(d，J＝6.8Hz，3H)，1.19(d，J＝6.8Hz，3H)。

Example 6

The present embodiment provides a compound 6 shown in formula I, wherein the structural formula of the compound 6 is as follows:

The synthetic route for compound 6 is shown below:

(1) To a solution of intermediate B (200 mg, 569. Mu. Mol) and Compound 6-1 (121 mg, 569. Mu. Mol) in N, N-dimethylformamide (5.0 mL) was added potassium carbonate (236 mg,1.71 mmol), and the reaction solution was stirred under nitrogen at 80℃for 2 hours. The reaction solution was added to saturated brine (50 mL), 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, and the crude product was separated by preparative thin layer chromatography (petroleum ether/ethyl acetate=50:1 to 1:1) to give compound 6-2.

MS-ESI[M+H] ⁺ Calculated 528, measured 528.

(2) To a solution of Compound 6-2 (229 mg, 434. Mu. Mol) in methylene chloride (4.0 mL) was added trifluoroacetic acid (1.54 g,13.5 mmol), and the reaction was stirred at 25℃for 30 minutes. The reaction solution is concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 6-3.

MS-ESI[M+H] ⁺ Calculated 428, measured 428.

(3) To a solution of the trifluoroacetate salt of compound 6-3 (230 mg, 425. Mu. Mol) in methanol (10.0 mL) were added triethylamine (85.9 mg, 849. Mu. Mol), compound 6-4 (111 mg, 425. Mu. Mol) and sodium cyanoborohydride (133 mg,2.12 mmol), and the mixture was stirred at 25℃for 2 hours. The reaction solution was concentrated under reduced pressure, methylene chloride (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, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=20:1 to 0:1) to give compound 6-5.

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

(4) To a solution of compound 6-5 (220 mg, 326. Mu. Mol) in methylene chloride (4.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 6-6.

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

(5) To a solution of trifluoroacetate salt (220 mg, 320. Mu. Mol) of compound 6-6 in methylene chloride (10.0 mL) were added triethylamine (32.4 mg, 320. Mu. Mol) and intermediate A (81.9 mg, 320. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 5 minutes. Dichloromethane (20.0 mL) was added, and the organic phase was washed with saturated aqueous ammonium chloride (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 isolated by silica gel column chromatography (dichloromethane/methanol=50:1 to 10:1) to give compound 6-7.

MS-ESI[M+H] ⁺ Calculated 792, found 792.

(6) To a solution of Compound 6-7 (180 mg, 227. Mu. Mol) in methylene chloride (4.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution is concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 6-8. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 692, measured 692.

(7) To a solution of trifluoroacetate salt of Compound 6-8 (180 mg, 223. Mu. Mol) in methylene chloride (5.0 mL) was added triethylamine (67.8 mg, 670. Mu. Mol). Then, compound 6-9 (20.2 mg, 223. Mu. Mol, 18.2. Mu.L) was added, and the reaction mixture was stirred at-78℃for 5 minutes under nitrogen. 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 (DAICEL CHIRALPAK AS,250 mm. Times.30 mm 10 μm, A: water (0.1% ammonia; B: ethanol, 35% -35%:48 min) to give compound 6.

MS-ESI[M+H] ⁺ Calculated 746, measured 746.

¹ H 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)。

Example 7

This example provides a compound 7 of formula I, the structural formula of the compound 7 is shown below:

the synthetic route for compound 7 is shown below:

(1) To a solution of intermediate B (300 mg, 852. Mu. Mol) and Compound 7-1 (217 mg,1.02 mmol) in N, N-dimethylformamide (8.0 mL) was added potassium carbonate (235 mg,1.71 mmol), and the reaction was stirred under nitrogen at 80℃for 2 hours. The reaction mixture was quenched with water (5.0 mL), extracted with ethyl acetate (10.0 ml×3), the organic phase was washed with saturated brine (10.0 ml×3), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 7-2.

MS-ESI[M+H] ⁺ Calculated 528, measured 528.

(2) To a solution of compound 7-2 (400 mg, 758. Mu. Mol) in methylene chloride (12.0 mL) was added trifluoroacetic acid (4.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 7-3.

MS-ESI[M+H] ⁺ Calculated 428, measured 428.

(3) To a solution of compound 7-3 in trifluoroacetic acid salt (400 mg, 738. Mu. Mol) in methanol (10.0 mL) were added triethylamine (74.7 mg, 738. Mu. Mol), compound 7-4 (96.5 mg, 369. Mu. Mol) and sodium cyanoborohydride (92.8 mg,1.48 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction mixture was quenched with water (5.0 mL), extracted with ethyl acetate (10.0 ml×3), the organic phase was 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 give compound 7-5.

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

(4) To a solution of compound 7-5 (220 mg, 326. Mu. Mol) in methylene chloride (9.0 mL) was added trifluoroacetic acid (3.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 7-6.

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

(5) To a solution of trifluoroacetate salt (220 mg, 320. Mu. Mol) of compound 7-6 in methylene chloride (6.0 mL) were added triethylamine (32.4 mg, 320. Mu. Mol) and intermediate A (163 mg, 640. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 60 minutes. The reaction mixture was quenched with water (10.0 mL), extracted with methylene chloride (10.0 mL. Times.2), and the organic phase was washed with saturated brine (10.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=50:1 to 10:1) to give compound 7-7.

MS-ESI[M+H] ⁺ Calculated 792, found 792.

(6) To a solution of compound 7-7 (62.0 mg, 78.2. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction solution was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 7-8. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 692, measured 692.

(7) To a solution of trifluoroacetate salt of Compound 7-8 (60 mg, 74.4. Mu. Mol) in dichloromethane (5.0 mL) was added triethylamine (7.5 mg, 74.4. Mu. Mol). Then, compound 7-9 (10.1 mg, 111. Mu. Mol, 9.1. Mu.L) was added thereto, and the reaction mixture was stirred at-78℃for 60 minutes under nitrogen. The reaction mixture was quenched with water (10.0 mL), extracted with methylene chloride (10.0 mL. Times.2), and the organic phase was washed with saturated brine (10.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative high performance liquid chromatography (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 15% -45%:10 min) to give the formate of compound 7.

MS-ESI[M+H] ⁺ Calculated 746, measured 746.

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)，2.95-3.03(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)。

Example 8

The present embodiment provides a compound 8 shown in formula I, wherein the structural formula of the compound 8 is as follows:

the synthetic route for compound 8 is shown below:

(1) To a solution of Compound 8-1 (240 mg, 467. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (1.45 g,14.4mmol,2.0 mL), and the reaction was stirred at 25℃for 0.5 hours. The reaction solution is filtered and concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 8-2.

MS-ESI[M+H] ⁺ Calculated 414, measured 414.

(2) To a solution of compound 8-2 in trifluoroacetic acid salt (240 mg, 455. Mu. Mol) in methanol (10.0 mL) was added triethylamine (46.0 mg,45 mmol). To the reaction mixture was added compound 8-3 (238 mg, 910. Mu. Mol), and the mixture was stirred at 25℃for 15 minutes. Sodium cyanoborohydride (143 mg,2.27 mmol) was then added and stirred at 25℃for 3 hours. The reaction mixture was concentrated under reduced pressure, poured into water (40.0 mL), and extracted with methylene chloride (40 mL. Times.3). The organic phases were combined, washed with saturated brine (40 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=0:1 to 10:1) to give compound 8-4.

MS-ESI[M+H] ⁺ Calculated 659, measured 659.

(3) To a solution of Compound 8-4 (210 mg, 319. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution is filtered and concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 8-5. The crude product was used directly in the next reaction.

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

(4) To a solution of trifluoroacetate salt (210 mg, 312. Mu. Mol) of compound 8-5 in methylene chloride (5.0 mL) were added triethylamine (63.2 mg, 264. Mu. Mol, 86.9. Mu.L) and intermediate A (136 mg, 531. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 10 minutes. The reaction mixture was poured into water (20.0 mL) and extracted with methylene chloride (20.0 mL. Times.2). The organic phases were combined, washed with saturated brine (20.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=0:1 to 10:1) to give compound 8-6.

MS-ESI[M+H] ⁺ Calculated 779, measured 779.

(5) To a solution of compound 8-6 (120 mg, 154. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 10 minutes. The reaction solution is filtered and concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 8-7. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 678, measured 678.

(6) To a solution of trifluoroacetate salt of Compound 8-7 (60.0 mg, 75.8. Mu. Mol) in methylene chloride (3.0 mL) was added triethylamine (7.67 mg, 75.8. Mu. Mol, 10.6. Mu. L). Then, compound 8-8 (10.3 mg, 114. Mu. Mol, 9.27. Mu.L) was added thereto, and the reaction mixture was stirred at-78℃for 10 minutes under nitrogen. The reaction solution was poured into water (20.0 mL), extracted with methylene chloride (20.0 mL. Times.2), and the organic phases were combined, washed with saturated brine (20.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by preparative high performance liquid chromatography (Welch Xtime, 75 mm. Times.40 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40% for 10 min) to give the formate of compound 8.

MS-ESI[M+H] ⁺ Calculated 733, measured 733.

¹ H 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)，2.90-2.98(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)。

Example 9

This example provides a compound 9 of formula I, the structural formula of the compound 9 is shown below:

the synthetic route for compound 9 is shown below:

to a solution of the trifluoroacetate salt (60.0 mg, 75.8. Mu. Mol) of compound 8-7 of example 8 in methylene chloride (3.0 mL) were added triethylamine (7.7 mg, 75.8. Mu. Mol, 10.6. Mu. L), compound 9-1 (25.1 mg, 151. Mu. Mol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (57.6 mg, 151. Mu. Mol), and the mixture was stirred at 25℃for 0.5 hours. The reaction mixture was poured into water (20.0 mL) and extracted with methylene chloride (20.0 mL. Times.2). The organic phases were combined, washed with saturated brine (20.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Welch Xtime, 75 mm. Times.40 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 5% -35% for 10 min) to give the formate salt of Compound 9.

MS-ESI[M+H] ⁺ Calculated 790, measured 790.

¹ H 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)。

Example 10

The present embodiment provides a compound 10 shown in formula I, wherein the structural formula of the compound 10 is as follows:

the synthetic route for compound 10 is shown below:

(1) To a solution of the trifluoroacetate salt of compounds 1-3 of example 1 (500 mg,0.95 mmol) in methanol (10.0 mL) was added triethylamine (279 mg,2.70 mmol). To the reaction solution was added compound 10-1 (282 mg,1.08 mmol), sodium cyanoborohydride (169 mg,2.70 mmol), and the mixture was stirred at 25℃for 16 hours. Poured into saturated aqueous ammonium chloride (40.0 mL) and extracted with ethyl acetate (50.0 mL. Times.1). The organic phases were combined, washed with saturated brine (25.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=0:1 to 10:1) to give compound 10-2.

MS-ESI[M+H] ⁺ Calculated 687, measured 687.

(1) To a solution of compound 10-2 (120 mg,0.175 mmol) in dichloromethane (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction was stirred at 25℃for 15 min. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of Compound 10-3. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 587, measured 587.

(2) To a solution of trifluoroacetate salt (120 mg,0.171 mmol) of compound 10-3 in dichloromethane (5.0. 5.0 mL) was added triethylamine to adjust the pH to 8, then intermediate A (87.6 mg, 0.348 mmol) was added, and the reaction solution was stirred under nitrogen at 25℃for 0.5 hours. The reaction mixture was poured into water (15.0 mL) and extracted with methylene chloride (10.0 mL. Times.3). The organic phases were combined, washed with saturated brine (10.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=0:1 to 10:1) to give compound 10-4.

MS-ESI[M+H] ⁺ Calculated 806, measured 806.

(3) To a solution of compound 10-4 (130 mg,0.16 mmol) in dichloromethane (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction was stirred at 25℃for 30 min. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of Compound 10-5. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 706, measured 706.

(4) To a solution of the trifluoroacetate salt of compound 10-5 (40.0 mg, 48.8. Mu. Mol) in methylene chloride (3.0 mL) was added triethylamine (4.94 mg, 48.8. Mu. Mol). Then, compound 10-6 (5.30 mg, 58.5. Mu. Mol, 4.77. Mu.L) was added thereto, and the reaction solution was stirred at-78℃for 15 minutes under nitrogen. The reaction mixture was poured into water (20.0 mL) and extracted with methylene chloride (20.0 mL. Times.2). The organic phases were combined, washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by preparative high performance liquid chromatography (Phenomenex luna C18, 100 mm. Times.40 mm 3. Mu.m, A: water (0.225% formic acid; B: acetonitrile, 0% -50% for 10 min) to give the formate of compound 10.

MS-ESI[M+H] ⁺ Calculated 760, measured 760.

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)。

Example 11

The present embodiment provides a compound 11 shown in formula I, wherein the structural formula of the compound 11 is as follows:

The synthetic route for compound 11 is shown below:

to a solution of the trifluoroacetate salt (40.0 mg, 48.8. Mu. Mol) of compound 10-4 of example 10 in methylene chloride (3.0 mL) were added triethylamine (4.94 mg, 48.8. Mu. Mol, 6.79. Mu. L), compound 11-1 (16.2 mg, 97.6. Mu. Mol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (37.1 mg, 97.6. Mu. Mol), and the mixture was stirred at 25℃for 0.5 hours. The reaction mixture was poured into water (20.0 mL) and extracted with methylene chloride (20.0 mL. Times.3). The organic phases were combined, washed with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated by preparative high performance liquid chromatography (Phenomenex luna C, 100 mm. Times.40 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 0% -40% for 10 min) to give the formate of compound 11.

MS-ESI[M+H] ⁺ Calculated 817, measured 817.

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)。

Example 12

The present embodiment provides a compound 12 shown in formula I, wherein the structural formula of the compound 12 is as follows:

the synthetic route for compound 12 is shown below:

(1) To a solution of the trifluoroacetate salt (200 mg, 452. Mu. Mol) of compound 1-3 in example 1 in methanol (5.0 mL) was added triethylamine (45.8 mg, 452. Mu. Mol), compound 12-1 (121 mg, 452. Mu. Mol) and sodium cyanoborohydride (142 mg,2.26 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction solution was concentrated under reduced pressure, 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, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate=1:0 to 0:1) to give compound 12-2.

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

(2) To a solution of compound 12-2 (90.0 mg, 129. Mu. Mol) in methylene chloride (4.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 12-3. The crude product was used directly in the next reaction.

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

(3) To a solution of trifluoroacetate salt (90 mg, 127. Mu. Mol) of compound 12-3 in methylene chloride (10.0 mL) were added triethylamine (12.9 mg, 127. Mu. Mol) and intermediate A (32.5 mg, 127. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 5 minutes. Dichloromethane (20.0 mL) was added thereto, and the mixture was washed with saturated brine (70.0. 70.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=100:1 to 10:1) to give compound 12-4.

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

(4) To a solution of compound 12-4 (70.0 mg, 75.6. Mu. Mol) in methylene chloride (4.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 12-5. The crude product was used directly in the next reaction.

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

(5) To a solution of the hydrochloride (18.1 mg, 109. Mu. Mol) of compound 12-5 in methylene chloride (10.0 mL) were added triethylamine (8.53 mg, 84.3. Mu. Mol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (27.6 mg, 72.6. Mu. Mol), and the mixture was stirred at 25℃for 30 minutes, and the trifluoroacetate (60.0 mg, 72.6. Mu. Mol) of compound 12-6 was added to the reaction mixture, and the reaction mixture was stirred at 25℃for 30 minutes. Dichloromethane (50.0 mL) was added, and 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 by preparative high performance liquid chromatography (Phenomenex luna C, 100 mm. Times.30 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 0% -30%:8 min) to give formate of compound 12.

MS-ESI[M+H] ⁺ Calculated value 824, measured value 824.

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)。

Example 13

This example provides a compound 13 of formula I, the structural formula of the compound 13 is shown below:

the synthetic route for compound 13 is shown below:

(1) To a solution of the trifluoroacetate salt (200 mg, 453. Mu. Mol) of the compound 1-3 in example 1 in methanol (3.0 mL) was added triethylamine (91.7 mg, 906. Mu. Mol), the compound E (145 mg, 544. Mu. Mol), stirred at 25℃for 15 minutes, sodium cyanoborohydride (142 mg,2.26 mmol) was added, and stirred at 25℃for 12 hours. The reaction solution was concentrated under reduced pressure, 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, and the crude product was separated by preparative thin layer chromatography (dichloromethane/methanol=10:1) to give compound 13-1.

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

(2) To a solution of compound 13-1 (140 mg, 202. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 13-2. The crude product was used directly in the next reaction.

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

(3) To a solution of trifluoroacetate salt (140 mg, 198. Mu. Mol) of compound 13-2 in methylene chloride (3.0 mL) were added triethylamine (40.1 mg, 396. Mu. Mol) and intermediate A (101 mg, 396. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 10 minutes. Dichloromethane (20.0 mL) was added, washed with water (2.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=100:1 to 8:1) to give compound 12-3.

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

(4) To a solution of compound 13-3 (120 mg, 148. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (10.9. Mu.L), and the reaction mixture was stirred at 25℃for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 13-4. The crude product was used directly in the next reaction.

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

(5) To a solution of the hydrochloride (36.1 mg, 218. Mu. Mol) of the compound 13-5 in methylene chloride (2.0 mL) were added triethylamine (14.7 mg, 145. Mu. Mol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (41.4 mg, 109. Mu. Mol), and the mixture was stirred at 25℃for 10 minutes, and to the reaction solution was added the trifluoroacetate (60.0 mg, 72.7. Mu. Mol) of the compound 13-4, and the reaction solution was stirred at 25℃for 60 minutes. The organic phase was concentrated under reduced pressure. High performance liquid chromatography (Phenomenex luna C, 100 mm. Times.30 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40% for 8 min) of the crude product was carried out to obtain formate of compound 13.

MS-ESI[M+H] ⁺ Calculated 823, measured 823.

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)。

Example 14

The present embodiment provides a compound 14 shown in formula I, wherein the structural formula of the compound 14 is as follows:

the synthetic route for compound 14 is shown below:

to a solution of the trifluoroacetate salt of compound 13-4 of example 13 (60 mg, 72.7. Mu. Mol) in methylene chloride (3.0 mL) was added triethylamine (14.7 mg, 145. Mu. Mol). Then, compound 14-2 (7.23 mg, 79.9. Mu. Mol, 6.52. Mu.L) was added thereto, and the reaction mixture was stirred at-78℃under nitrogen for 10 minutes. The crude product was separated by preparative high performance liquid chromatography (Phenomenex Luna C, 100 mm. Times.30 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:8 min) to give the formate of compound 14.

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

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)。

Example 15

This example provides a compound 15 of formula I, the structural formula of the compound 15 is shown below:

the synthetic route for compound 15 is shown below:

(1) To a solution of the trifluoroacetate salt of compound 3-3 (200 mg, 359. Mu. Mol) in methanol (10.0 mL) of example 3 were added triethylamine (72.8 mg, 720. Mu. Mol), compound C (188 mg, 719. Mu. Mol) and sodium cyanoborohydride (90.4 mg,1.44 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction solution was added with water (10.0 mL) and dichloromethane (20.0 mL), the organic phase was washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, the organic phase was concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 0:1) to give compound 15-1.

MS-ESI[M+H] ⁺ Calculated 688, measured 688.

(2) To a solution of compound 15-1 (200 mg, 290. Mu. Mol) in methylene chloride (9.0 mL) was added trifluoroacetic acid (3.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 15-2. The crude product was used directly in the next reaction.

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

(3) To a solution of the trifluoroacetate salt of compound 15-2 (200 mg, 285. Mu. Mol) in methylene chloride (5.0 mL) were added triethylamine (28.8 mg, 285. Mu. Mol) and intermediate A (145 mg, 570. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 1 hour. Water (10.0 mL) and methylene chloride (20.0 mL) were added, and 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 isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 0:1) to give compound 15-3.

MS-ESI[M+H] ⁺ Calculated value 807, measured value 807.

(4) To a solution of compound 15-3 (120 mg, 148. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 15-4. The crude product was used directly in the next reaction.

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

(5) To a solution of the hydrochloride (28.3 mg, 219. Mu. Mol) of the compound 15-5 in methylene chloride (4.0 mL) were added triethylamine (14.7 mg, 146. Mu. Mol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (55.5 mg, 146. Mu. Mol), and the mixture was stirred at 25℃for 1 hour, and to the reaction solution was added the trifluoroacetate (60 mg, 73.0. Mu. Mol) of the compound 15-4, and the reaction solution was stirred at 25℃for 1 hour. Dichloromethane (20.0 mL) was added, and 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. High performance liquid chromatography (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) of crude product, separation afforded formate of compound 15.

MS-ESI[M+H] ⁺ Calculated 818, measured 818.

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)，2.58-2.76(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)。

Example 16

The present embodiment provides a compound 16 shown in formula I, wherein the structural formula of the compound 16 is as follows:

The synthetic route for compound 16 is shown below:

(1) To a solution of the trifluoroacetate salt of compound 3-3 (200 mg, 359. Mu. Mol) in methanol (10.0 mL) of example 3 were added triethylamine (72.8 mg, 719. Mu. Mol), compound C (188 mg, 719. Mu. Mol) and sodium cyanoborohydride (90.4 mg,1.44 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction mixture was added with water (100.0 mL), 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, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 0:1) to give compound 16-1.

MS-ESI[M+H] ⁺ Calculated 688, measured 688.

(2) To a solution of compound 16-1 (170 mg, 247. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 16-2.

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

(3) To a solution of trifluoroacetate salt (170 mg, 242. Mu. Mol) of compound 16-2 in methylene chloride (5.0 mL) were added triethylamine (24.5 mg, 242. Mu. Mol) and intermediate A (123 mg, 484. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 60 minutes. Water (10.0 mL) was added, the mixture was extracted with dichloromethane (10.0 mL. Times.2), 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 isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 0:1) to give compound 16-3.

MS-ESI[M+H] ⁺ Calculated value 807, measured value 807.

(4) To a solution of compound 16-3 (137 mg, 169. Mu. Mol) in methylene chloride (9.0 mL) was added trifluoroacetic acid (3.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 16-4. The crude product was used directly in the next reaction.

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

(5) To a solution of the trifluoroacetate salt of Compound 16-4 (75.0 mg, 91.3. Mu. Mol) in methylene chloride (5.0 mL) was added triethylamine (9.25 mg, 91.3. Mu. Mol). Then, compound 16-5 (12.4 mg, 137. Mu. Mol, 11.1. Mu.L) was added thereto, and the reaction mixture was stirred at-78℃for 60 minutes under nitrogen. Water (10.0 mL) was added, the mixture was extracted with dichloromethane (10.0 mL. Times.2), 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 preparative high performance liquid chromatography (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) to give the formate salt of compound 16.

MS-ESI[M+H] ⁺ Calculated 761, measured 761.

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)。

Example 17

This example provides a compound 17 of formula I, the structural formula of compound 17 is shown below:

the synthetic route for compound 17 is shown below:

(1) To a solution of the trifluoroacetate salt (200 mg, 452. Mu. Mol) of the compound 1-3 in example 1 in methanol (10.0 mL) were added triethylamine (45.8 mg, 452. Mu. Mol), compound C (237 mg, 905. Mu. Mol) and sodium cyanoborohydride (113.8 mg,1.81 mmol), and the mixture was stirred at 25℃for 1 hour. The reaction mixture was added with water (100.0 mL), 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, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 0:1) to give compound 17-1.

MS-ESI[M+H] ⁺ Calculated 688, measured 688.

(2) To a solution of compound 17-1 (160 mg, 232. Mu. Mol) in methylene chloride (9.0 mL) was added trifluoroacetic acid (3.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 17-2.

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

(3) To a solution of the trifluoroacetate salt of compound 17-2 (160 mg, 228. Mu. Mol) in methylene chloride (5.0 mL) were added triethylamine (23.0 mg, 228. Mu. Mol) and intermediate A (116 mg, 456. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 60 minutes. Water (10.0 mL) was added, the mixture was extracted with dichloromethane (10.0 mL. Times.2), 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 isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 0:1) to give compound 17-3.

MS-ESI[M+H] ⁺ Calculated value 807, measured value 807.

(4) To a solution of compound 17-3 (80 mg, 99.1. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 17-5. The crude product was used directly in the next reaction.

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

(5) To a solution of the trifluoroacetate salt (35.0 mg, 49.5. Mu. Mol) of compound 17-4 in methylene chloride (5.0 mL) was added triethylamine (10.0 mg, 99.1. Mu. Mol), 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (37.7 mg, 99.1. Mu. Mol) and compound 17-5 (19.2 mg, 148. Mu. Mol), and the mixture was stirred at 25℃for 1 hour. Water (10.0 mL) was added, the mixture was extracted with dichloromethane (10.0 mL. Times.2), 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 preparative high performance liquid chromatography (Xtime C18, 100 mm. Times.40 mm 3 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40% for 10 min) to give formate of compound 17.

MS-ESI[M+H] ⁺ Calculated 818, measured 818.

¹ H 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)。

Example 18

The present embodiment provides a compound 18 of formula I, wherein the structural formula of the compound 18 is as follows:

The synthetic route for compound 18 is shown below:

to a solution of the trifluoroacetate salt of compound 17-4 of example 17 (35.0 mg, 49.5. Mu. Mol) in methylene chloride (3.0 mL) was added triethylamine (5.02 mg, 49.5. Mu. Mol). Then, compound 18-1 (6.73 mg, 74.3. Mu. Mol, 6.1. Mu.L) was added thereto, and the reaction mixture was stirred under nitrogen at-78℃for 60 minutes. Water (10.0 mL) was added, the mixture was extracted with dichloromethane (10.0 mL. Times.2), 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 preparative high performance liquid chromatography (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) to give the formate salt of compound 18.

MS-ESI[M+H] ⁺ Calculated 761, measured 761.

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)。

Example 19

This example provides a compound 19 of formula I, the structural formula of the compound 19 is shown below:

the synthetic route for compound 19 is shown below:

(1) 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) was added potassium carbonate (390 mg,2.84 mmol), and the reaction was stirred at 80℃for 2 hours under nitrogen. The reaction was quenched with water (10.0 mL) and extracted with dichloromethane (10.0 mL. Times.2). The organic phases were combined, washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 19-2;

MS-ESI[M+H] ⁺ Calculated value 543, measured value 543.

¹ H NMR(400MHz，CDCl ₃ )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)。

(2) To a solution of compound 19-2 (673 mg,1.24 mmol) in methylene chloride (18.0 mL) was added trifluoroacetic acid (9.24 g,81.0mmol,6.0 mL) and the reaction was stirred at 25℃for 1 hour under nitrogen. Concentrating the reaction solution under reduced pressure to obtain trifluoroacetate of the compound 19-3;

MS-ESI[M+H] ⁺ calculated 442, measured 442.

(3) The trifluoroacetate salt of Compound 19-3 (100 mg, 180. Mu. Mol), compound 12-1 (57.7 mg, 216. Mu. Mol) and triethylamine (36.4 mg, 359. Mu. Mol, 50.1. Mu. L) were suspended in methanol (5.0 mL), and sodium cyanoborohydride (45.2 mg, 719. Mu. Mol) was added thereto, followed by stirring at 25℃for 12 hours under nitrogen. The reaction was quenched by addition of 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 concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 19-4;

MS-ESI[M+H] ⁺ calculated 693, measured 693.

(4) To a solution of compound 19-4 (120 mg, 173. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (3.08 g,27.0mmol,2.0 mL), and the reaction was stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of Compound 19-5. The crude product is directly used for the next reaction;

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

(5) To the trifluoroacetate salt (120 mg, 169. Mu. Mol) of compound 19-5 were added a solution of methylene chloride (5.0 mL) and triethylamine (34.3 mg, 339. Mu. Mol, 47.2. Mu.L), followed by intermediate A (86.8 mg, 339. Mu. Mol) and stirring the reaction solution at 25℃for 1 hour under nitrogen. The reaction was quenched with water (10.0 mL), extracted with dichloromethane (10.0 mL. Times.2), the organic phases were combined, washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 19-6;

MS-ESI[M+H] ⁺ calculated 812, measured 812.

(6) To a solution of compound 19-6 (45.0 mg, 55.4. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.54 g,13.5mmol,1.0 mL), and the reaction was stirred at 25℃for 1 hour under nitrogen. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 19-7. The crude product was used directly in the next reaction.

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

(7) The trifluoroacetate salt (45.0 mg, 54.4. Mu. Mol) of compound 19-7, the compound 19-8 (27.0 mg, 163. Mu. Mol), triethylamine (5.51 mg, 54.4. Mu. Mol, 7.58. Mu. L) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (41.4 mg, 108. Mu. Mol) were dissolved in methylene chloride (3.0 mL), and stirred under nitrogen at 25℃for 1 hour. The reaction solution was diluted with water (10.0 mL), extracted with dichloromethane (10.0 ml×2), and 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 by preparative high performance liquid chromatography (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) to give the formate salt of compound 19.

MS-ESI[M+H] ⁺ Calculated 823, measured 823.

¹ H 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)

Example 20

The present embodiment provides a compound 20 shown in formula I, wherein the structural formula of the compound 20 is as follows:

the synthetic route for compound 20 is shown below:

(1) The trifluoroacetate salt (120 mg, 216. Mu. Mol) of compound 20-1 of example 20, compound E (69.2 mg, 259. Mu. Mol) and triethylamine (43.7 mg, 431. Mu. Mol, 60.1. Mu.L) were suspended in methanol (5.0 mL), and sodium cyanoborohydride (54.2 mg, 863. Mu. Mol) was added thereto, followed by stirring at 25℃for 12 hours under nitrogen. 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, the organic phase concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 20-2;

MS-ESI[M+H] ⁺ calculated 693, measured 693.

(2) To a solution of compound 20-2 (120 mg, 173. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (3.08 g,27.0mmol,2.0 mL), and the reaction was stirred at 25℃for 1 hour. The reaction solution is concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 20-3. The crude product is directly used for the next reaction;

MS-ESI[M+H] ⁺ calculated 593, measured 593.

(3) To the trifluoroacetate salt (120 mg, 169. Mu. Mol) of the compound 20-3 were added a solution of methylene chloride (5.0 mL) and triethylamine (34.3 mg, 339. Mu. Mol, 47.2. Mu.L), followed by the addition of the intermediate A (86.8 mg, 339. Mu. Mol), and the reaction mixture was stirred at 25℃for 1 hour under nitrogen. The reaction was quenched with water (100.0 mL), extracted with dichloromethane (100.0 mL. Times.2), the organic phases were combined, washed with saturated brine (100.0 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure. Separating the crude product by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to obtain compound 20-4;

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

(4) To a solution of compound 20-4 (23.0 mg, 28.3. Mu. Mol) in methylene chloride (3.0 mL) was added trifluoroacetic acid (1.0 mL), and the reaction mixture was stirred at 25℃for 1 hour under nitrogen. The reaction solution is concentrated under reduced pressure to obtain the trifluoroacetate salt of the compound 20-5. The crude product was used directly in the next reaction.

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

(5) The trifluoroacetate salt (23.0 mg, 27.8. Mu. Mol) of compound 20-5, the compound 20-6 (10.7 mg, 65.1. Mu. Mol), triethylamine (2.82 mg, 27.8. Mu. Mol, 3.88. Mu. L) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (21.1 mg, 55.6. Mu. Mol) were dissolved in methylene chloride (3.0 mL) and stirred at 25℃for 1 hour under nitrogen. The reaction solution was diluted with water (10.0 mL), extracted with dichloromethane (10.0 ml×2), and 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 by preparative high performance liquid chromatography (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) to give the formate salt of compound 20.

MS-ESI[M+H] ⁺ Calculated 823, measured 823.

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)。

Example 21

The present embodiment provides a compound 21 shown in formula I, wherein the structural formula of the compound 21 is as follows:

The synthetic route for compound 21 is shown below:

(1) To a solution of intermediate F (300 mg, 888. Mu. Mol) and compound 21-1 (241 mg, 1.07. Mu. Mol) in N, N-dimethylformamide (8.0 mL) was added potassium carbonate (246 mg,1.78 mmol), and the reaction was stirred at 80℃for 12 hours under nitrogen. The reaction mixture was quenched with water (30.0 mL) and extracted with ethyl acetate (40.0 mL. Times.2). The organic phases were combined, washed with saturated brine (40.0 ml×2), dried over anhydrous sodium sulfate, filtered, and 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 give compound 21-2;

MS-ESI[M+H] ⁺ calculated 528, measured 528.

(2) To a solution of compound 21-2 (350 mg, 663. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2 mL), and the reaction mixture was stirred at 25℃for 10 minutes. Filtering the reaction solution, and concentrating under reduced pressure to obtain trifluoroacetate salt of the compound 21-3;

MS-ESI[M+H] ⁺ calculated 428, measured 428.

(3) The trifluoroacetate salt of Compound 21-3 (237 mg, 555. Mu. Mol), compound C (2911 mg,1.11 mmol) and triethylamine (56.2 mg, 555. Mu. Mol, 77.3. Mu.L) were suspended in methanol (5.0 mL), and sodium cyanoborohydride (140 mg,2.22 mmol) was added thereto and stirred at 25℃for 12 hours. The reaction was quenched by addition of 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 the organic phase concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 9:1) to give compound 21-4;

MS-ESI[M+H] ⁺ Calculated 675, measured 675.

(4) To a solution of compound 21-4 (370 mg, 549. Mu. Mol) in dichloromethane (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction solution was stirred at 25℃for 0.5 hours. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of Compound 21-5. The crude product is directly used for the next reaction;

MS-ESI[M+H] ⁺ calculated 574, measured 574.

(5) To the trifluoroacetate salt (370 mg, 538. Mu. Mol) of compound 21-5 were added a solution of methylene chloride (8.0 mL) and triethylamine (54.4 mg, 538. Mu. Mol, 74.9. Mu.L), followed by intermediate A (206 mg, 803. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 1 hour. The reaction was quenched with water (30.0 mL), extracted with dichloromethane (20.0 mL. Times.2), the organic phases were combined, washed with saturated brine (20.0 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure. Separating the crude product by silica gel column chromatography (dichloromethane/methanol=1:0 to 9:1) to obtain compound 21-6;

MS-ESI[M+H] ⁺ calculated 793, measured 793.

(6) To a solution of compound 21-6 (210 mg, 264. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (3.08 g,27.0mmol,2.0 mL), and the reaction was stirred at 25℃for 10 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 21-7. The crude product was used directly in the next reaction.

MS-ESI[M+H] ⁺ Calculated 694, measured 694.

(7) The trifluoroacetate salt (41.1 mg, 248. Mu. Mol) of compound 21-7, compound 21-8 (100 mg, 124. Mu. Mol), triethylamine (12.5 mg, 124. Mu. Mol, 17.3. Mu. L) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (94.3 mg, 248. Mu. Mol) were dissolved in methylene chloride (5.0 mL), and stirred at 25℃for 0.5 hours. The reaction solution was diluted with water (30.0 mL), extracted with dichloromethane (30.0 ml×2), and the organic phases were combined, washed with saturated brine (30.0 ml×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparation of crude product high performance liquid chromatography (Xtimate C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min.) afforded the formate salt of Compound 21.

MS-ESI[M+H] ⁺ Calculated value 805, measured value 805.

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)，4.54-4.67(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，4H)，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)。

Example 22

The present embodiment provides a compound 22 shown in formula I, wherein the structural formula of the compound 22 is as follows:

the synthetic route for compound 22 is shown below

To a solution of the trifluoroacetate salt (100 mg, 124. Mu. Mol) of compound 21-7 of example 21 in dichloromethane (5.0 mL) was added triethylamine (12.5 mg, 124. Mu. Mol). Compound 22-2 (16.9 mg, 186. Mu. Mol, 15.2. Mu.L) was then added, and the reaction was stirred under nitrogen at-78℃for 5 minutes. 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 (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) to give the formate salt of compound 22.

MS-ESI[M+H] ⁺ Calculated 747, measured 747

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)。

Example 23

The present embodiment provides a compound of formula I, wherein the structural formula of the compound 23 is as follows:

the synthetic route for compound 23 is shown below:

(1) To a solution of the hydrochloride salt of intermediate compound 3-3 (440 mg, 791. Mu. Mol) in example 3 (10.0 mL) was added triethylamine (160 mg,1.58 mmol), compound D (207 mg, 791. Mu. Mol) and sodium cyanoborohydride (199mg, 3.17 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction solution was added with water (100 mL) and dichloromethane (200 mL), 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, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 23-1.

MS-ESI[M+H] ⁺ Calculated value 688, realMeasured 688.

(2) To a solution of compound 23-1 (500 mg, 726. Mu. Mol) in methylene chloride (12.0 mL) was added trifluoroacetic acid (4.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 23-2. The crude product was used directly in the next reaction.

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

(3) To a solution of trifluoroacetate salt of compound 23-2 (500 mg, 712. Mu. Mol) in methylene chloride (5.0 mL) were added triethylamine (72.1 mg, 712. Mu. Mol) and intermediate A (264 mg,1.43 mmol), and the reaction mixture was stirred under nitrogen at 25℃for 1 hour. Water (10.0 mL) and methylene chloride (20.0 mL) were added, and 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 isolated by silica gel column chromatography (dichloromethane/methanol=1:0 to 10:1) to give compound 23-3.

MS-ESI[M+H] ⁺ Calculated value 807, measured value 807.

(4) To a solution of compound 23-3 (100 mg, 123. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.0 mL), and the reaction mixture was stirred at 25℃for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 23-4. The crude product was used directly in the next reaction.

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

(5) To a solution of the hydrochloride (23.6 mg, 182. Mu. Mol) of the compound 23-5 in methylene chloride (3.0 mL) were added triethylamine (12.3 mg, 121. Mu. Mol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (46.3 mg, 121. Mu. Mol), and to the reaction solution was added the trifluoroacetate (50 mg, 60.9. Mu. Mol) of the compound 23-4, and the reaction solution was stirred at 25℃for 1 hour. 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 (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 3% -33% for 10 min) to give the formate of compound 23.

MS-ESI[M+H] ⁺ Calculated 818, measured 818.

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)，4.39-4.46(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)，1.87-2.04(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)。

Example 24

This example provides a compound 24 of formula I, the structural formula of the compound 24 is shown below:

The synthetic route for compound 24 is shown below:

to a solution of the trifluoroacetate salt of compound 23-4 of example 23 (50 mg, 60.9. Mu. Mol) in methylene chloride (2.0 mL) was added triethylamine (6.16 mg, 60.9. Mu. Mol). Then, compound 24-1 (8.27 mg, 91.3. Mu. Mol, 7.45. Mu.L) was added thereto, and the reaction mixture was stirred at-78℃under nitrogen for 60 minutes. 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 (Xtime C18, 100 mm. Times.30 mm 10 μm, A: water (0.225% formic acid; B: acetonitrile, 10% -40%:10 min) to give the formate salt of compound 24.

MS-ESI[M+H] ⁺ Calculated 761, measured 761.

¹ H 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，1H)，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)。

Example 25

This example provides a compound 25 of formula I, the structural formula of the compound 25 is shown below:

the synthetic route for compound 25 is shown below:

(1) To a solution of trifluoroacetate salt of compound 3-3 (400 mg, 719. Mu. Mol) in methanol (10.0 mL) were added triethylamine (72.8 mg, 719. Mu. Mol), compound 25-1 (192 mg, 719. Mu. Mol) and sodium cyanoborohydride (361 mg,5.70 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction mixture was poured into water (5.0 mL), stirred for 5 minutes, and extracted with ethyl acetate (10.0 mL. Times.3). The organic phases were combined, washed with saturated brine (10.0 ml×3), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 25-2.

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

(2) To a solution of compound 25-2 (282 mg, 406. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (3.00 g,26.2 mmol), and the reaction was stirred at 25℃for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 25-3. The crude product was used directly in the next reaction.

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

(3) To a solution of the trifluoroacetate salt of compound 25-3 (280 mg, 396. Mu. Mol) in methylene chloride (8.0 mL) were added triethylamine (40.0 mg, 396. Mu. Mol) and intermediate A (151 mg, 594. Mu. Mol), and the reaction mixture was stirred under nitrogen at 25℃for 1 hour. Water (10.0 mL) was added to the reaction mixture, and the mixture was extracted with methylene chloride (10.0 mL. Times.2). The organic phases were combined, washed with saturated brine (10.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 25-4.

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

(4) To a solution of compound 25-4 (157 mg, 193. Mu. Mol) in methylene chloride (6.0 mL) was added trifluoroacetic acid (2.60 g,23.0 mmol), and the reaction was stirred at 25℃for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 25-5. The crude product was used directly in the next reaction.

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

(5) To a solution of the trifluoroacetate salt (135 mg, 163. Mu. Mol) of compound 25-5 in methylene chloride (5.0 mL) was added triethylamine (16.5 mg, 163. Mu. Mol) and compound 25-6 (22.1 mg, 245. Mu. Mol). The reaction solution was stirred at-78℃for 1 hour under nitrogen protection. The reaction mixture was poured into water (20 mL) and extracted with methylene chloride (20.0 mL. Times.2). The organic phases were combined, washed with saturated brine (20.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 25.

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

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)。

Example 26

This example provides a compound 26 of formula I, the structural formula of the compound 26 is shown below:

the synthetic route for compound 26 is shown below:

(1) To a solution of trifluoroacetate salt of compound 3-3 (1.00 g,1.80 mmol) in methanol (20.0 mL) was added triethylamine (182 mg,1.80 mmol), compound E (577 mg,2.10 mmol) and sodium cyanoborohydride (399 mg,5.40 mmol), and the mixture was stirred at 25℃for 12 hours. The reaction mixture was poured into water (10.0 mL), stirred for 5 minutes, and extracted with ethyl acetate (20.0 mL. Times.3). The organic phases were combined, washed with saturated brine (20.0 ml×3), dried over anhydrous sodium sulfate, filtered, and the organic phase concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 26-1.

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

(2) To a solution of compound 26-1 (1.00 g,1.50 mmol) in dichloromethane (21.0 mL) was added trifluoroacetic acid (10.7 g,94.5 mmol), and the reaction was stirred at 25℃for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 26-2. The crude product was used directly in the next reaction.

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

(3) To a solution of trifluoroacetate salt (1.00 g,1.40 mmol) of compound 26-2 in dichloromethane (20.0 mL) were added triethylamine (143 mg,1.40 mmol) and intermediate A (552 mg,2.10 mmol), and the reaction was stirred under nitrogen at 25℃for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with methylene chloride (10.0 mL. Times.2). The organic phases were combined, washed with saturated brine (10.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 26-3.

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

(4) To a solution of compound 26-3 (1.10 g,1.30 mmol) in dichloromethane (21.0 mL) was added trifluoroacetic acid (10.7 g,94.5 mmol), and the reaction was stirred at 25℃for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 26-4. The crude product was used directly in the next reaction.

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

(5) To a solution of trifluoroacetate salt of compound 26-4 (500 mg,605. Mu. Mol) in dichloromethane (5.0 mL) was added triethylamine (61.2 mg, 605.3. Mu. Mol) and compound 26-5 (82.1 mg, 908. Mu. Mol). The reaction solution was stirred at-78℃for 1 hour under nitrogen protection. The reaction mixture was poured into water (10.0 mL) and extracted with methylene chloride (10.0 mL. Times.2). The organic phases were combined, washed with saturated brine (10.0 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol=10:1) to give compound 26.

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

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),1.89-1.97(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)。

Test example 1

Determination of the antiproliferative effect of the Compounds on MV-4-11 cells (CCK method):

1. the experimental principle is that MV-4-11 is a human leukemia cell line with MLL translocation and expresses MLL fusion protein MLL-AF4. The compounds of the present invention inhibit MV-4-11 proliferation by interfering with the binding of the menin/MLL proteins/protein interactions.

2. Experimental materials Cell Counting Kit-8 were purchased from Shanghai Li Ji Biotechnology Co., ltd (cat# D3100L 4057); 96 well clear bottom white cell culture plates were purchased from Corning Costar (cat No. 3610); fetal bovine serum was purchased from GIBCO (cat# 10099-141); modified Ichkoff culture broth (IMDM) medium was purchased from Invitrogen (cat. No. 12440046); table microplate reader SpectraMax i3X was purchased from Molecular Devices.

3. Experimental method cells in the logarithmic growth phase were resuspended in complete medium (imdm+10% Fetal Bovine Serum (FBS)) and plated into 96-well plates (100 μl of cell suspension per well, i.e. 15000 cells per well). Cells were incubated at 37℃with 100% relative humidity, 5% CO ₂ Incubate in incubator for 24 hours.

The compound to be tested is dissolved in dimethyl sulfoxide (DMSO) to prepare stock solution with the concentration of 10mmol/L, and the stock solution is diluted step by step for 8 times by using DMSO according to a 4-fold gradient. And diluted 20 times with medium. The final concentrations of the compounds were 100. Mu.M, 25. Mu.M, 6.25. Mu.M, 1.56. Mu.M, 0.39. Mu.M, 0.098. Mu.M, 0.024. Mu.M, 0.006. Mu.M, 0.0015. Mu.M (4-fold dilution, 9 concentrations) in the 96-well plates inoculated with cells at 25. Mu.L/well.

Cells to which test compounds were added were placed at 37℃with 100% relative humidity, 5% CO ₂ Incubating for 72 hours in an incubator respectively; cell viability was measured using the CCK-8 method by adding 10. Mu.L of CCK-8 assay reagent per well and placing in an incubator for a continuous incubation period of about 4 hours. The wavelength was read using a bench-top microplate reader (reference wavelength 650 nM).

4. And (3) data processing:

the inhibition of tumor cell growth by the drug was calculated as follows:

tumor cell growth inhibition ratio = [ (ODc-ODs)/(ODc-ODb) ]. Times.100%

Wherein ODs are the OD of the sample (cell+CCK-8+test compound), ODc is the OD of the negative control (cell+CCK-8+DMSO), ODb is the OD of the blank control (medium+CCK-8+DMSO).

IC of the compound was calculated with Graphpad software ₅₀ 。

The specific test results are shown in table 1:

TABLE 1

Test compounds	MV 4-11 IC ₅₀ (nM)
Formate salt of example 1	30.46
Formate salt of example 2	374.9
Formate salt of example 3	6.03
Formate salt of example 4	12.31
Example 6	362.3
Formate salt of example 8	142.0
Formate salt of example 9	67.18
Formate salt of example 10	196.9
Formate salt of example 11	102.5

As shown in the test data of Table 1, the spiro compound shown in the formula I has a better inhibition effect on the growth of human myelomonocytic leukemia MV-4-11 cells, and has potential for preparing medicines for treating and preventing leukemia.

Test example 2

Determination of the antiproliferative effect of the Compounds on MV-4-11 cells (CTG method):

2. Experimental materials CellTiter-Glo was purchased from Promega (cat# G7571); IMDM medium was purchased from Gibco (cat# 12440061); fetal bovine serum was purchased from excel (cat# FND 500); dimethyl sulfoxide (DMSO) was purchased from Sigma (cat#d2650); 384 well cell culture plates were purchased from Corning (cat# 3756); automatic cytometer is available from Life technologies (model Countess II); the microplate reader was purchased from PerkinElmer (model EnVisionMultilabel Reader).

3. Experimental methods cells in the logarithmic growth phase were resuspended in growth medium (IMDM+10% FBS) and diluted to the target density (50000/mL). Inoculating 50 mu L of the cell suspension into a 384-well plate; at 37 ℃,5% CO ₂ Incubate overnight in incubator.

Test compounds were dissolved in DMSO to prepare a stock solution at a concentration of 10 mM. Stock was first diluted to 2mM in DMSO and then diluted in a 3-fold gradient for a total of 10 concentrations. 5.5. Mu.L of the above solution was taken at each concentration, and diluted with 94.5. Mu.L of growth medium, respectively. Then 5. Mu.L/well was added to 384 well plates inoculated with cells.

Placing the cells added with the compound to be tested at 37 ℃ and 5% CO ₂ Incubate in incubator for 72 hours. 384 well plates were equilibrated at room temperature, 15 μl CellTiter-Glo reagent was added to each well, mixed on a vortexing device for 2 min, incubated at room temperature for 60 min, enVisionMultilabel Reader read luminescence, and the IC of the compound was calculated using GraphPad Prism 5.0 software ₅₀ 。

4. Experimental data:

the specific test results are shown in table 2:

TABLE 2

Test compounds	MV 4-11 IC ₅₀ (nM)
Formate salt of example 2	237.2
Formate salt of example 3	8.36

Formate salt of example 5	575.1
Example 6	422.5
Formate salt of example 7	81.22
Formate salt of example 12	124.2
Formate salt of example 13	317.6
Formate salt of example 14	485.7
Formate salt of example 15	47.09
Formate salt of example 16	91.37
Formate salt of example 17	90.38
Formate salt of example 18	167.2
Formate salt of example 19	8.83
Formate salt of example 20	19.36
Formate salt of example 21	95.84
Formate salt of example 22	78.54
Formate salt of example 23	12.06
Formate salt of example 24	29.70
Example 25	36.40
Example 26	75.73

As shown in the test data of Table 2, the spiro compound shown in the formula I has a better inhibition effect on the growth of human myelomonocytic leukemia MV-4-11 cells, and has potential for preparing medicines for treating and preventing leukemia.

The applicant states that the present application describes spiro compounds, pharmaceutical compositions comprising them and their use by way of the above examples, but the present application is not limited to, i.e. does not mean that the present application must rely on the above examples to be practiced.

Claims

The spiro compound is characterized by having a structural formula as shown in the following formula I:

wherein,

R ¹ selected from-C (O) (NR) ^a R ^b ) The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is ^a 、R ^b Each independently selected from H, optionally substituted C1-C6 alkyl, optionally substituted 3-8 membered cycloalkyl and optionally substituted 4-8 membered heterocyclyl, or R ^a And R is R ^b Is linked to N to form an optionally substituted 4-8 membered heterocyclic ring; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P;

R ² selected from H, halogen, methyl and trifluoromethyl;

R ³ selected from H and halogen;

R ⁴ selected from H, optionally substituted C1-C6 alkyl, optionally substitutedC1-C4 alkoxy, optionally substituted C1-C4 alkylamino, halogen, -NH ₂ 、-NO ₂ -COOH, -CN, -OH, optionally substituted C1-C6 alkyl sulphonyl, optionally substituted C1-C6 alkyl sulphoxide, optionally substituted C1-C6 alkylthio, -NHCOCR ^4' ＝CH ₂ 、-NHCOCHR ^4' R ^4” 、-SO ₂ C(R ^4' )＝CH ₂ 、-NHSO ₂ CR ^4' ＝CH ₂ and-NHSO ₂ CHR ^4' R ^4” The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is ^4' Selected from H, methyl and fluoro; r is R ^4” Selected from chlorine and bromine atoms;

y, Z are each independently selected from N and CH, and at least one of Y and Z is N;

w is selected from N and C;

v is selected from N and CR ^V Wherein R is ^V Is H, halogen, -CN, -OH, -NH ₂ Optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino or optionally substituted (C1-C4 alkyl) ₂ An amino group;

U ¹ 、U ² 、U ³ 、U ⁴ 、U ⁵ 、U ⁶ 、U ⁷ 、U ⁸ each independently selected from: -C (R ') (R ") -, -C (R') (R") -C (R '") (R" "-, -C (=o) -, -C (R') (R") -O-, -C (R ') (R ") -NR'" -and-N=c (NH) ₂ ) -, and U ¹ 、U ² 、U ³ 、U ⁴ At most one of them is-C (=o) -, -C (R ') (R ") -O-, or-C (R ') (R") -NR ' "-, U (U) ⁵ 、U ⁶ At most one of them is-C (=o) -, -C (R ') (R ") -O-, -C (R ') (R") -NR ' "-or-N=c (NH) ₂ )-，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 (NH) ₂ )-；

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;

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 to 3 heteroatoms selected from N, O, S, P;

L ¹ is absent, -CR ^L1' R ^L1” -、-CO-、-SO ₂ -SO-, -C (n=n) -, oxygen or-NH-; wherein R is ^L1' 、R ^L1” Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R ^L1' And R is R ^L1” Forming an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the attached carbon atom; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P;

L ² Selected from: -SO ₂ -、-SO-、-CO-、-CF ₂ -and-C (n=n) -;

L ³ selected from: oxygen atom, sulfur atom, -SO ₂ -、-SO-、-CO-、-CR ^L3' R ^L3” -and-NR ^L3”' -; wherein R is ^L3' 、R ^L3” Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R ^L3' And R is ^L3” Forming an optionally substituted 3-to 8-membered saturated or unsaturated with the attached carbon atomAnd cycloalkanes, optionally substituted 4-8 membered saturated or unsaturated heterocycles; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P; r is R ^L3”' Selected from: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkyl and optionally substituted 4-8 membered saturated or unsaturated heterocycle, wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P;

x is selected from: carbon atoms, -S-, and-SO-;

R ⁵ selected from: -CH ₂ R ^5' 、 Wherein R is ^5' Is a fluorine or chlorine atom; r is R ^5” Is H, methyl or fluorine atom; r is 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) ₂ Amino, optionally substituted C1-C4 alkylthio, optionally substituted 3-8 membered saturated or unsaturated cycloalkyl, optionally substituted 4-8 membered saturated or unsaturated heterocyclyl and substituted or unsubstituted C2-C4 acyl; wherein the heterocyclyl contains 1-3 heteroatoms selected from N, O, S, P;

Represents the attachment position of the group.
The spirocyclic compound of claim 1, wherein R ² Selected from H and halogen.
The spirocyclic compound of claim 2, wherein R ² Is fluorine.
The spirocyclic compound of claim 1The object is characterized in that R is ³ Is H or fluorine atom; further preferably, the R ³ Is H;

preferably, said R ⁴ Is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, -NH ₂ or-CN;

preferably, each of the Y, Z is N;

preferably, W is C;

preferably, V is N;

preferably, the U ¹ 、U ² 、U ³ 、U ⁴ 、U ⁵ 、U ⁶ 、U ⁷ 、U ⁸ Each 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;

Preferably, the 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 to 3 heteroatoms selected from N, O, S, P; further preferred, the 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 triazazine 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;

preferably, the L ¹ Is absent or-CH ₂ -; further preferably, the L ¹ is-CH ₂ -；

Preferably, the L ² Selected from: -SO ₂ -, -SO-and-CO-; further preferably, the L ² is-SO ₂ -；

Preferably, the L ³ Selected from: oxygen atom, sulfur atom, -CR ^L3' R ^L3” -and-NR ^L3”' -; wherein R is ^L3' 、 R ^L3” Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R ^L3' And R is ^L3” Forming an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the attached carbon atom; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P; r is R ^L3”' Selected from: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkyl and optionally substituted 4-8 membered saturated or unsaturated heterocycle; wherein the heterocycle comprises 1-3 heteroatoms selected from N, O, S, P; further preferably, the L ³ Is an oxygen atom or a sulfur atom;

preferably, said X is selected from: carbon atoms and-SO-; further preferably, X is a carbon atom;

preferably, said R ⁵ Selected from: -CH ₂ R ^5' 、 Wherein R is ^5' Is a fluorine or chlorine atom; r is R ^5” Is H, methyl or fluorine atom; r is R ^5”' Selected from: h and optionally substituted C1-C4 alkyl.
The spirocyclic compound of any one of claims 1 to 4, wherein in formula IThe spiro ring is selected from any one of the following groups:

preferably, in the formula IThe spiro ring is selected from any one of the following groups:
the spirocyclic compound of any one of claims 1-5, wherein in formula IThe representative ring combining part is selected from any one of the following groups:

wherein R is ^e 、R ^f Each independently selected from: H. methyl, trifluoromethyl, difluoromethyl, methoxy, ethoxy, trifluoromethoxy, halogen, hydroxy, amino, cyano, methylamino, dimethylamino, ethylamino, diethylamino, trifluoroethylamino, carboxy, methoxycarbonyl, ethoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, methylcarbamoyl and diethylcarbamoyl.
The spirocyclic compound of any one of claims 1-6, wherein formula I isThe cyclic moiety shown is selected from any one of the following groups:
the spirocyclic compound of any one of claims 1-7, wherein R ⁵ Selected from: -CH ₂ F、-CH ₂ F、-CH ₂ Cl、
The spirocyclic compound according to any one of claims 1 to 8, wherein said compound of formula I is selected from any one of the following compounds:
the spirocyclic compound of any one of claims 1-9, further comprising any one of a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate, or polymorph or deuterate of a compound of formula I.
A pharmaceutical composition comprising a spirocyclic compound according to any one of claims 1-10 and a pharmaceutically acceptable carrier.
Use of a spirocyclic compound according to any one of claims 1-10 or a pharmaceutical composition according to claim 11, wherein said use is selected from any one of the following (a) - (c):

(a) Preparing a medicament for preventing or treating tumors, diabetes and other diseases related to the activity of MLL1, MLL2, MLL fusion proteins, and/or the menin protein;

(b) Preparing an inhibitor for in vitro non-therapeutic association with the activity of MLL1, MLL2, MLL fusion proteins, and/or grin proteins;

(c) Preparing proliferation inhibitor for in vitro non-therapeutic tumor cells.