CN114516873A

CN114516873A - Spiro compound, pharmaceutical composition containing same and application thereof

Info

Publication number: CN114516873A
Application number: CN202011299036.6A
Authority: CN
Inventors: 陈旭星; 李京; 陈艳红; 赵兆
Original assignee: Suzhou Youli Biomedical Technology Co ltd
Current assignee: Suzhou Youli Biomedical Technology Co ltd
Priority date: 2020-11-18
Filing date: 2020-11-18
Publication date: 2022-05-20
Also published as: TWI815210B; CN116507618A; TW202220991A; WO2022105636A1

Abstract

The invention provides a spiro compound, a pharmaceutical composition containing the same and application thereof. The spiro compound interferes the interaction between the menin protein and MLL1 or MLL2 or MLL-fusion oncoprotein, and is expected to be a drug for treating tumors, diabetes and other diseases depending on MLL1, MLL2, MLL fusion protein and/or the activity of the menin protein.

Description

Spiro compound, pharmaceutical composition containing same and application thereof

Technical Field

The invention 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

The mixed-line leukemia (MLL) protein is a histone methyltransferase, and plays an important role in the process of gene transcription regulation. Most acute leukemias including Acute Myelogenous Leukemia (AML), Acute Lymphocytic Leukemia (ALL) and mixed lineage leukemia are found to have a translocation of the MLL gene located at the position of chromosome 11, q23, forming MLL fusion (MLL-r) proteins with one of approximately 80 proteins (e.g., AF4, AF9, ENL, AF10, ELL, AF6, AF1p, GAS7, etc.). The MLL-r protein approximately retains 1400 amino acid sequences at the N-terminal of the MLL protein, lacks the C-terminal methyltransferase active region, and can abnormally regulate the transcription of various oncogenes including HOX and MEIS1, promote cell proliferation, and finally cause cancer. Leukemia patients with chromosomal translocations of the MLL gene usually have a poor prognosis with a 5-year survival rate of less than 40% (Slany, Haematologica, 2009, 94, 984-.

The Menin protein, encoded by the Multiple Endocrine Neoplasms (MEN) gene, is a widely expressed nuclear protein that interacts with DNA replication and repair proteins, chromatin modification proteins, and various transcription factors (Agarwal et al, Horm Metab Res, 2005, 37, 369-. The Menin protein binds to the N-terminus of MLL proteins, including MLL1, MLL2, and MLL-r proteins, as is necessary for the oncogenic activity of MLL proteins (Yokoyama et al, Cell, 2005, 123, 207-. Interference with the interaction between menin and MLL-r proteins enables selective inhibition of MLL-r leukemia cell proliferation in vitro and in vivo (Grembecka et al, nat. chem. biol., 2012, 8, 277-284; Borkin et al, Cancer cell, 2015, 27, 589-602.)

In particular hematological neoplasms, there are certain genetic abnormalities or mutations, such as nuclear porin 98(NUP98) gene fusion, nuclear phosphoprotein (NPM1) gene mutation, DNA methyltransferase 3A (DNMT3A) mutation, MLL gene amplification, etc., which are often accompanied by high levels of HOX gene expression. The retro HOXD genes in ewing's sarcoma, particularly HOXD13, were abnormally over-expressed with high levels of meinin and MLL1 protein, whereas HOXD13 is a downstream gene regulated by menin and MLL 1.

Thus, interfering with the interaction between menin and MLL proteins, especially by covalent binding, is a very promising strategy for treating tumors. There is still a great need in the art to develop effective drugs that can interfere with the interaction of menin and MLL proteins.

Disclosure of Invention

In view of the defects of the prior art, the present invention aims to provide a spiro compound, a pharmaceutical composition containing the same and applications thereof, wherein the spiro compound and the pharmaceutical composition containing the same can interfere with the interaction between menin and MLL proteins.

In order to achieve the purpose, the invention adopts the following technical scheme:

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

wherein the content of the first and second substances,

R¹selected from the group consisting of-C (O) (NR)^aR^b) (i.e. the

) (ii) a Wherein R is^a、R^bEach independently selected from H, optionally substituted C1-C6 alkyl, optionally substituted 3-8 membered cycloalkyl and optionally substituted 4-8 membered heterocyclyl, or R^aAnd R^bLinked to N to form an optionally substituted 4-8 membered heterocyclic ring; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P;

R²selected from H, halogen, methyl and trifluoromethyl;

R³selected from H and halogen;

R⁴selected from H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, halogen, -NH₂、-NO₂-COOH, -CN, -OH, optionally substituted C1-C6 alkyl sulfone group, optionally substituted C1-C6 alkyl sulfoxide group, optionally substituted C1-C6 alkylthio group, -NHCORR⁴'＝CH₂(i.e. the

)、-NHCOCHR^4'R^4”(i.e. the

)、-SO₂C(R^4')＝CH₂(i.e. the

)、-NHSO₂ CR^4'＝CH₂(i.e. the

) and-NHSO₂CHR^4'R^4”(i.e. the

) (ii) a Wherein R is^4'Selected from H, methyl and fluoro; r is^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^VWherein R is^VIs H, halogen, -CN, -OH, -NH₂Optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino or optionally substituted (C1-C4 alkyl)₂An amino group;

U¹、U²、U³、U⁴、U⁵、U⁶、U⁷、U⁸each independently selected from: -C (R ') (R ') - (i.e. R ')

) -C (R ') (R') -C (R ') - (i.e., a compound of formula (I)' -)

) -C (═ O) - (i.e.)

) -C (R') (R ") -C (═ O) - (i.e., carbon monoxide-carbon monoxide

) -C (R ') (R') -O- (i.e., a compound of formula (I))

) -C (R ') (R') -NR '- (i.e., R') -, in a single reaction vessel

) and-N ═ C (NH)₂) - (i.e. using

) And U is¹、U²、U³、U⁴At most one of which is-C (═ O) -, -C (R ') (R ") -O-, or-C (R ') (R") -NR ' "-, U⁵、U⁶At most one of which is-C (═ O) -, -C (R ') (R ") -O-, -C (R ') (R") -NR ' "-, or-N ═ C (NH)₂)-，U⁷、U⁸Of which at most one 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;

a is an optionally substituted 6-16 membered aromatic ring or an optionally substituted 5-16 membered heteroaromatic ring; wherein said heteroaromatic ring contains 1-3 heteroatoms selected from N, O, S, P;

L¹is absent, -CR^L1'R^L1”- (i.e. using

) -CO- (i.e. a

)、-SO₂- (i.e. using

) -SO- (i.e. a

) 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^L1”Form an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the carbon atom to which it is attached; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P;

L²selected from: -SO₂-、-SO-、-CO-、-CF₂-and-C (N ═ N) -;

L³selected from: oxygen atom, sulfur atom, -SO₂-、-SO-、-CO-、-CR^L3'R^L3”- (i.e. of

) and-NR^L3”'- (i.e. using

) (ii) a Wherein R is^L3'、R^L3”Each independently selected from: H. optionally substituted C1-C4 alkyl and halogen, or R^L3' and R^L3”Form an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the carbon atom to which it is attached; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P; r^L3”'Selected from: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkane, and optionally substituted 4-8 membered saturated or unsaturated heterocycle; wherein the heterocyclic ring contains 1-3 groups selected from N,O, S, P;

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

R⁵selected from: -CH₂R^5'、

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

indicates the position of attachment of the group.

Preferably, said R is²Selected from H and halogen; further preferably, said R²Is fluorine.

Preferably, said R is³Is H or fluorine atom; further preferably, said R³Is H.

Preferably, said R is⁴Is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, -NH₂or-CN.

Preferably, the Y, Z are each N.

Preferably, W is C.

Preferably, said V is N.

Preferably, said U is¹、U²、U³、U⁴、U⁵、U⁶、U⁷、U⁸Each independently selected from: -C (R ') (R ") -, -C (R') (R") -C (R '") (R" "), -C (═ O) -, and-C (R') (R") -C (═ O) -, and U¹、U²、U³、U⁴At most one of them is-C (═ O) -, or-C (R') (R ") -C (═ O) -, U⁵、U⁶At most one of which is-C (═ O) -, or-C (R') (R ") -C (═ O) -, U⁷、U⁸At most one of which 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 said heteroaromatic ring contains 1-3 heteroatoms selected from N, O, S, P; further preferably, a is an optionally substituted benzene ring, an optionally substituted pyridine ring, an optionally substituted pyridazine ring, an optionally substituted pyrimidine ring, an optionally substituted 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 triazazole ring.

Preferably, said L¹Is absent or-CH₂-; further preferably, said L¹is-CH₂-。

Preferably, said L²Selected from: -SO₂-, -SO-and-CO-; further preferably, said L²is-SO₂。

Preferably, said L³Selected 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^L3”Form, with the carbon atom to which it is attached, an optionally substituted 3-to 8-membered saturated or unsaturated cycloalkane, an optionally substituted 4-to 8-membered saturated or unsaturated cycloalkaneA saturated heterocycle; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P; r^L3”'Selected from the group consisting of: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkane, and optionally substituted 4-8 membered saturated or unsaturated heterocycle; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P; further preferably, said L³Is an oxygen atom or a sulfur atom.

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

Preferably, said R is⁵Selected from: -CH₂R^5'、

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

Preferably, in said formula I

The spiro ring is selected from any one of the following groups:

preferably, in said formula I

The spiro ring is selected from any one of the following groups:

preferably, in said formula I

The cyclic moiety represented by is selected from any one of the following groups:

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

Preferably, the structural formula in the formula I is

The cyclic part is selected from any one of the following groups:

preferably, said R is⁵Selected from: -CH₂F、-CH₂F、-CH₂Cl、

Preferably, the compound represented by 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 deuterations of the compound shown in formula I.

In a second aspect, the present invention provides a pharmaceutical composition comprising a spiro 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 anti-tumor drugs. Such therapeutic agents include, but are not limited to: an antitumor drug acting on a DNA chemical structure, such as cisplatin, an antitumor drug affecting nucleic acid synthesis, such as Methotrexate (MTX), 5-fluorouracil (5FU) and the like, an antitumor drug affecting nucleic acid transcription, such as doxorubicin, epirubicin, aclarubicin, mithramycin and the like, an antitumor drug affecting tubulin synthesis, such as paclitaxel, vinorelbine and the like, an aromatase inhibitor, such as aminoglutethimide, landetron, letrozole, ryanodine and the like, a cell signaling pathway inhibitor, such as epidermal growth factor receptor inhibitor Imatinib (Imatinib), Gefitinib (Gefitinib), Erlotinib (Erlotinib), Lapatinib (Lapatinib) and the like.

In a third aspect, the present invention provides a use of the spirocyclic compound according to the first aspect or the pharmaceutical composition according to the second aspect, wherein the use is selected from any one of the following (a) to (c):

(a) preparing a medicament for preventing or treating tumors, diabetes and other diseases associated with MLL1, MLL2, MLL fusion proteins, and/or menin protein activity;

(b) preparing an inhibitor associated with activity of MLL1, MLL2, MLL fusion protein, and/or menin protein for non-therapeutic use in vitro;

(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 menin protein activity 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, endometrioma, gastric cancer, liver cancer, kidney cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, brain glioma, bile duct cancer, 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 invention belongs.

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

As used herein, the term "comprising" or "includes" 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 salts" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"pharmaceutically acceptable acid addition salts" refers to salts with inorganic or organic acids which retain 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, formates, acetates, 2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, caprates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbates, salicylates, 4-aminosalicylates, napadisylates, and the like. These salts can be prepared by methods known in the art.

"pharmaceutically acceptable base addition salts" refers to salts with inorganic or organic bases which maintain the biological effectiveness 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 naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, 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 the present application, a "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally 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 facilitate administration to an organism, facilitate absorption of active ingredients and exert biological activity.

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

As used herein, "pharmaceutically acceptable excipient" includes, but is 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 emulsifier that is approved by the relevant governmental regulatory agency for human or livestock use.

The "tumor" of the present invention includes, but is not limited to, glioma, sarcoma, melanoma, chondroma of joint, cholangioma, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, squamous carcinoma of lung, adenocarcinoma of lung, 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, renal cancer, oral cancer, etc.

The terms "preventing," "prevention," and "prevention" as used herein include reducing the likelihood of occurrence or worsening of a disease or disorder in a patient.

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

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

(ii) inhibiting the disease or disorder, i.e., arresting its development;

(iii) alleviating the disease or condition, i.e., causing regression of the state of the disease or condition; or

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

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein, refer 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, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The terms "administering," "administration," "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, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Administration techniques useful for The compounds and methods described herein are well known to those skilled in The art, for example, in Goodman and Gilman, The pharmaceutical Basis of Therapeutics, current ed.; pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing co. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

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

Radical definitions

Definitions for the terms of the standardization sector can be found in the literature references including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols.A (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods within the skill of the art are employed, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy, and pharmacological methods. Unless a specific definition is set forth, the terms used herein in the pertinent description of analytical chemistry, organic synthetic chemistry, and pharmaceutical chemistry are known in the art. Standard techniques can 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 instructions of the kit from the manufacturer, or according to the methods known in the art or the instructions of the present invention. The techniques and methods described above can generally be practiced according to conventional methods well known in the art, as described in various general and more specific documents referred to and discussed in this specification. In the present specification, groups and substituents thereof may be selected by one skilled in the art to provide stable moieties and compounds.

When described by a conventional formula written from left to rightWhen a substituent is used, the substituent also includes chemically equivalent substituents obtained when the formula is written from right to left. For example, -CH₂O-is equivalent to-OCH₂-。

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. 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 shorthand notation 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 shorthand notation excludes carbons that may be present in a substituent of the group.

Indicates the position of attachment of the group.

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

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

"hydroxy" means an-OH group.

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

"carbonyl" refers to a-C (═ O) -or-CO-group.

"nitro" means-NO₂。

"cyano" means-CN.

"amino" means-NH₂。

"substituted amino" refers to amino substituted with one or two alkyl, alkylcarbonyl, aralkyl, heteroaralkyl groups as defined below, for example, monoalkylamino, dialkylamino, alkylamido, aralkylamino, heteroaralkylamino.

"carboxyl" means-COOH.

The term "alkyl" as used herein as a group or part of another group (e.g., in halo-substituted alkyl and the like) refers to a fully saturated straight or branched hydrocarbon chain radical consisting only of 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 attached to the remainder of the molecule by a single bond, including, for example, 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 the present invention, the term "alkyl" refers to alkyl groups containing from 1 to 6 carbon atoms.

The term "alkenyl" as used herein as a group or part of another group means a straight or branched hydrocarbon chain group consisting solely of 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 attached to the rest of the molecule by a single bond, such as, but not limited to, vinyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1, 4-dienyl, and the like.

The term "alkynyl" as a group or part of another group herein means a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, having, for example, from 2 to 14 (preferably from 2 to 10, more preferably from 2 to 6) carbon atoms, and attached to the rest of the molecule by single bonds, such as, but not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

The term "cyclic hydrocarbon group" as used herein as a group or part of another group means a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting of only carbon and hydrogen atoms, which may include fused, bridged 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 rest of the molecule by a single bond via any suitable carbon atom. Unless otherwise specifically indicated in the specification, carbon atoms in the cyclic hydrocarbon group may be optionally oxidized. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, 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] octyl, bicyclo [3.1.1] heptyl, bicyclo [3.2.1] octyl, bicyclo [2.2 ] octenyl, cyclooctenyl, Bicyclo [3.2.1] octenyl, adamantyl, octahydro-4, 7-methylene-1H-indenyl, octahydro-2, 5-methylene-pentalenyl and the like.

The term "heterocyclyl" as used herein as a group or part of another group 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 otherwise specifically indicated in the specification, a heterocyclyl group may be a monocyclic, bicyclic, tricyclic, or higher ring system, which may include fused, bridged, or spiro ring systems; wherein the nitrogen, carbon or sulfur atom in the heterocyclic group may be optionally oxidized; the nitrogen atoms may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be attached to the rest of the molecule via a carbon atom or a heteroatom and by a single bond. In heterocyclic 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 rest of the molecule is a non-aromatic ring atom. For the purposes of the present invention, heterocyclyl is preferably a stable 4-to 11-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 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 group containing 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, dioxolanyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolyl, octahydroisoindolyl, pyrazolinyl, pyrazolidinyl, phthalimido, and the like.

The term "aryl" or "aromatic ring" as used herein as a group or part of another group means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms. For the purposes of the present invention, an aryl group may be a monocyclic, bicyclic, tricyclic or higher polycyclic ring system and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the aryl group is attached to the remainder of the molecule by a single bond via an atom on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, 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" or "heteroaromatic ring" as used herein as a group or part of another group 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 in the ring. Unless otherwise specifically indicated in the specification, a heteroaryl group may be a monocyclic, bicyclic, tricyclic or higher ring system, and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the heteroaryl group is attached to the rest of the molecule by a single bond via an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group may be optionally oxidized; the nitrogen atoms may optionally be quaternized. For the purposes of the present invention, heteroaryl is preferably a stable 5-to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 5-to 10-membered aromatic group containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur or a 5-to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoindolyl, indazolyl, isoindolyl, purinyl, quinolyl, isoquinolyl, diazonaphthyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, phenanthrolinyl, acridinyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxazolyl, cinnolinyl, quinazolinyl, -indolizinyl, o-diazaphenanthryl, isoxazolyl, phenazinyl, phenothiazinyl, 4,5,6, 7-tetrahydrobenzo [ b ] thienyl, naphthopyridyl, [1,2,4] triazolo [4,3-b ] pyridazine, [1,2,4] triazolo [4,3-a ] pyrazine, [1,2,4] triazolo [4,3-c ] pyrimidine, [1,2,4] triazolo [4,3-a ] pyridine, imidazo [1,2-b ] pyridazine, imidazo [1,2-a ] pyrazine and the like.

The term "heteroarylalkyl" as used 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 circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl groups. For example, as used herein, the term "substituted" or "substituted with … …" without explicitly listing substituents 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, -CD₃、-C₁-C₆Alkyl (preferably-C)_1-3Alkyl group), C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, cycloalkyl (preferably 3-to 8-membered cycloalkyl), aryl, heterocyclyl (preferably 3-to 8-membered heterocyclyl), heteroaryl, aryl C₁-C₆Alkyl, heteroaryl C₁-C₆Alkyl radical, C₁-C₆Haloalkyl-, OC₁-C₆Alkyl (preferably-OC)₁-C₃Alkyl), -OC₂-C₆Alkenyl radical, OC₁-C₆Alkylphenyl, -C₁-C₆Alkyl OH (preferably-C)₁-C₄Alkyl OH), -C₁-C₆Alkyl SH, -C₁-C₆Alkyl radical O-C₁-C₆Alkyl, OC₁-C₆Haloalkyl, NH₂、C₁-C₆Alkyl NH₂(preferably C)₁-C₃alkyl-NH₂)、N(C₁-C₆Alkyl radical)₂(preferably N (C)₁-C₃Alkyl radical)₂)、NH(C₁-C₆Alkyl) (preferably NH (C)₁-C₃Alkyl)), N (C)₁-C₆Alkyl) (C)₁-C₆Alkylphenyl), NH (C)₁-C₆Alkylphenyl), nitro, C (O) -OH, C (O) OC₁-C₆Alkyl (preferably-C (O) OC₁-C₃Alkyl), -CONRiri (where Ri and Rii are H, D and C)_1-6Alkyl, preferably C_1-3Alkyl), -NHC (O) (C)₁-C₆Alkyl), NHC (O) (phenyl), N (C)₁-C₆Alkyl radical C (O) (C)₁-C₆Alkyl group), N (C)₁-C₆Alkyl group C (O) (phenyl), C (O) C₁-C₆Alkyl, C (O) heteroaryl (preferably C (O) -5-7 membered heteroaryl), C (O) C₁-C₆Alkylphenyl, C (O) C₁-C₆Haloalkyl, OC (O) C₁-C₆Alkyl (preferably OC (O) C)₁-C₃Alkyl), -S (O)₂-C₁-C₆Alkyl, -S (O) -C₁-C₆Alkyl, -S (O)₂-phenyl, -S (O)₂-C₁-C₆Haloalkyl, -S (O)₂NH₂、S(O)₂NH(C₁-C₆Alkyl), S (O)₂NH (phenyl), -NHS (O)₂(C₁-C₆Alkyl), -NHS (O)₂(phenyl) and NHS (O)₂(C₁-C₆Haloalkyl), wherein each of said alkyl, cycloalkyl, phenyl, aryl, heterocyclyl and heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of: halogen, -OH, -NH₂Cycloalkyl, 3-8 membered heterocyclyl, C₁-C₄Alkyl radical, C₁-C₄Haloalkyl-, -OC₁-C₄Alkyl, -C₁-C₄Alkyl OH, -C₁-C₄Alkyl radical O-C₁-C₄Alkyl, OC₁-C₄Haloalkyl, cyano, nitro, -C (O) -OH, -C (O) OC₁-C₆Alkyl, CON (C)₁-C₆Alkyl radical)₂、CONH(C₁-C₆Alkyl), CONH₂、NHC(O)(C₁-C₆Alkyl), NH (C)₁-C₆Alkyl radical C (O) (C)₁-C₆Alkyl), -SO₂(C₁-C₆Alkyl), -SO₂(phenyl), -SO₂(C₁-C₆Haloalkyl), -SO₂NH₂、SO₂NH(C₁-C₆Alkyl), SO₂NH (phenyl), -NHSO₂(C₁-C₆Alkyl), -NHSO₂(phenyl) and-NHSO₂(C₁-C₆Haloalkyl). When an atom or group is substituted with a plurality of 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 specific fragment or functional group in a molecule. Chemical moieties are generally considered to be chemical entities that are embedded in or attached to a molecule.

"absent" means directly linked by a chemical bond to both sides of a group as defined above. For example, "B is absent in A-B-C" means "A-C".

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

When the compounds of the present invention contain olefinic double bonds, unless otherwise indicated, the compounds of the present invention are intended to include both E-and Z-geometric isomers.

"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 invention are also intended to be included within the scope of the invention.

The compounds of the present invention or pharmaceutically acceptable salts thereof may contain one or more chiral carbon atoms and may therefore give rise to enantiomers, diastereomers, and other stereoisomeric forms. Each chiral carbon atom may be defined as (R) -or (S) -, based on stereochemistry. The present invention is intended to include all possible isomers, as well as racemates and optically pure forms thereof. The compounds of the invention may be prepared by selecting as starting materials or intermediates racemates, diastereomers or enantiomers. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, e.g., crystallization and chiral chromatography.

In the present invention, (C1-C4 alkyl)₂Amino, represents 2C 1-C4 alkyl-substituted amines and may be, for example

And the like.

In the present invention, "each R '", "each R" "refer to" each R' "," each R "" each.

Conventional techniques for the preparation/separation of 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, as described, for example, in Gerald Gubitz and Martin G.Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol.243, 2004; m.stalup, Chiral Separations, annu.rev.anal.chem.3:341-63, 2010; fumiss et al (eds.), VOGEL' S ENCYCOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5. 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 processes described below, the functional groups of the intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl groups (e.g.tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino and guanidino include t-butyloxycarbonyl, benzyloxycarbonyl and the like. Suitable thiol protecting groups include-C (O) -R "(where 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 organic Synthesis, (1999), 4th Ed., Wiley. The protecting group may also be a polymeric resin.

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

(1) the invention provides a compound shown as a formula I or a pharmaceutically acceptable salt thereof;

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

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitation of the present invention.

The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are percentages and parts by weight.

The test materials and reagents used in the following examples are commercially available without specific reference.

In each of the embodiments of the present invention,¹h NMR was recorded by a BRUKER AVANCE NEO model 400MHz NMR spectrometer with chemical shifts expressed in δ (ppm); liquid chromatography-mass spectrometry (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 of the intermediate A is shown as follows:

(1) to a solution of compound A-1(5.0g, 28.9mmol) in dichloromethane (25.0mL) was added triethylamine (5.84g, 57.7mmol) and compound A-2(7.27g, 63.5mmol), and the reaction was stirred at 0 deg.C under nitrogen for 1 hour. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate (40.0mL), 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 concentrated under reduced pressure to give Compound A-3.

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

(2) To a solution of compound A-3(7.2g, 28.6mmol) in N, N-dimethylformamide (70.0mL) was added compound A-4(6.54g, 57.3mmol), and the reaction mixture was stirred at 85 ℃ for 12 hours under nitrogen. The reaction was quenched by the addition of water (50.0mL), 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.

¹H 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(5.0g, 28.9mmol) was dissolved in acetic acid (20.0mL) and water (2.0mL), N-chlorosuccinimide (865.9mg, 6.48mmol) was added, and the reaction was stirred at 25 ℃ for 0.5 h under nitrogen. The reaction was quenched by the addition of water (10.0mL), 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 concentrated under reduced pressure to give intermediate A.

¹H NMR(400MHz，CDCl₃)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 of intermediate B is as follows:

(1) to a solution of compound B-1(100g, 588mmol) in dichloromethane (1300mL) at 0 deg.C were added 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (268g, 705mmol), diisopropylethylamine (114g, 882mmol) and compound B-2(119g, 1.18mol), and the reaction was stirred at 25 deg.C for 3 hours. Water (200mL) was added, extraction was performed with dichloromethane (200mL × 3), the combined organic phases were washed with saturated brine (200mL × 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 ═ 10:0 to 0:1) to give compound B-3.

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

(2) Boron tribromide (257g, 1.03mol) was added to a solution of compound B-3(130g, 513mmol) in dichloromethane (2.0L) at-78 deg.C, and the reaction solution was stirred at 25 deg.C for 4 hours. The reaction was quenched by addition of ice water, neutralized with saturated aqueous sodium bicarbonate to pH 8, 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 was 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 value 240, found value 240.

(3) To a solution of compound B-4(115g, 41.0mmol) in N, N-dimethylformamide (1500mL) were added cesium carbonate (470g, 1.44mol) and compound B-5(29.5g, 186mmol), and the reaction solution was stirred at 130 ℃ for 12 hours. After cooling to room temperature, water (500mL) was added and extracted with ethyl acetate (400mL × 3), the combined organic phases were washed with saturated brine (400mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 20:1 to 1:1) to give compound B-6.

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

(4) To a solution of Compound B-6(110g, 347mmol) in dichloromethane (300.0mL) at 0 deg.C was added m-chloroperoxybenzoic acid (211g, 1.04mol, 85% purity) and the reaction was stirred at 25 deg.C under nitrogen for 12 hours. The reaction was quenched by addition of saturated sodium thiosulfate solution until the starch potassium iodide paper did not turn blue any more, followed by extraction with dichloromethane (1000mL), and the organic phase was washed with saturated aqueous sodium bicarbonate (500mL × 2) and saturated brine (500mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure by the organic phase. 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, found value 334.

(5) Triethylamine (22.8g, 225mmol) and compound B-7(50.0g, 150mmol) were added to a solution of phosphorus oxychloride (46.0g, 255mmol) in chloroform (700mL) at 0 deg.C, and the reaction mixture was stirred at 65 deg.C for 12 hours. The reaction was quenched by addition of ice water, neutralized with saturated aqueous sodium bicarbonate to pH 8, 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 of intermediate C is shown below:

(1) compound C-1(5.0g, 18.6mmol), compound C-2(3.74g, 27.9mmol), cesium carbonate (12.1g, 37.2mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (1.36g, 1.86mmol) were dissolved in dioxane (40.0mL) and water (40.0 mL). The reaction solution was stirred at 110 ℃ for 1 hour under nitrogen protection. Aqueous solution (100mL) was added, extraction was performed with dichloromethane (100mL × 2), the combined organic phases were washed with saturated brine (100mL × 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, found 261.

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

(2) Compound C-3(1.6g, 6.15mmol), sodium periodate (3.94g, 18.4mmol) and potassium osmate (452mg, 1.23mmol) were dissolved in tetrahydrofuran (15.0mL) and water (24.0 mL). The reaction solution was stirred for 1 hour at 25 ℃ under nitrogen. The reaction was quenched by the addition of water (100mL), 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.

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

4. Preparation of intermediate D

The synthetic route of intermediate D is as follows:

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

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

¹H 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(400mg, 1.24mmol) in tetrahydrofuran (15.0mL) at-78 deg.C was added diisobutylaluminum hydride (1.5mol/L, 2.49mL, 3.74mmol), and the reaction was stirred at 0 deg.C under nitrogen for 0.5 h. The reaction was quenched by the addition of hydrochloric acid (1mol/L), the pH was adjusted to 7, water (20.0mL) was then added, extraction was performed with ethyl acetate (20.0 mL. times.2), the combined organic phases were washed with saturated brine (20.0 mL. times.1), 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 (petroleum ether/ethyl acetate 1:0 to 0:1) to afford intermediate D.

¹H 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 of intermediate E is shown below:

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

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

6. Preparation of intermediate F

The synthetic route of intermediate F is shown below:

(1) to a solution of compound F-1(30.0g, 176mmol) in dichloromethane (300mL) at 0 deg.C were added 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (101g, 265mmol), diisopropylethylamine (68.4g, 529mmol) and compound F-2(18.4g, 212mmol), and the reaction was stirred at 25 deg.C for 1 hour. Dichloromethane (400mL) was added, the organic phase was washed with water (250mL × 1) and saturated brine (220mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:0 to 3:1) to give compound F-3.

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

(2) Boron tribromide (88.0g, 351mmol) was added to a solution of compound F-3(48.0g, 176mmol) in dichloromethane (500.0mL) at-78 deg.C, and the reaction was stirred at 25 deg.C for 4 hours. The reaction was quenched by addition of ice water, neutralized to pH 8 with saturated aqueous sodium bicarbonate solution, extracted with dichloromethane (500mL), and the combined organic phases were 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 concentrated under reduced pressure with organic phase. 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, found 226.

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

(3) To a solution of compound F-4(27.9g, 124mmol) in N, N-dimethylformamide (350mL) were added cesium carbonate (121g, 371mmol) and compound F-5(29.5g, 186mmol), and the reaction solution was stirred at 130 ℃ for 12 hours. After cooling to room temperature, ethyl acetate (600mL) was added, the organic phase was washed with water (500mL × 1) and saturated brine (300mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:0 to 2:1) to give compound F-6.

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

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

(4) To a solution of compound F-6(25.0g, 82.4mmol) in dichloromethane (300.0mL) at 0 deg.C was added m-chloroperoxybenzoic acid (53.5g, 264mmol, 85% purity) and the reaction was stirred at 25 deg.C under nitrogen for 12 hours. The reaction was quenched by the addition of saturated sodium sulfite solution (100mL), followed by the addition of saturated aqueous sodium bicarbonate solution (500mL), extraction with dichloromethane (300mL), washing of the organic phase with saturated aqueous sodium bicarbonate solution (500 mL. times.2) and saturated brine (500 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 (dichloromethane/methanol ═ 1:0 to 25:1) to give compound F-7.

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

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

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

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

¹H NMR(400MHz，CDCl₃)δ8.72(s，1H)，8.23-8.25(m，1H)，7.12-7.15(m，1H)，7.06-7.10(m，1H)，6.97-7.01(m，1H)，3.83(m，1H)，3.39-3.48(m，1H)，3.25-3.30(m，1H)，1.22-1.25(m，3H)，1.12-1.17(m，6H)。

Example 1

This example provides a compound 1 of formula I, the structural formula of compound 1 being shown below:

the synthetic route of compound 1 is shown below:

(1) to a solution of intermediate B (3.0g, 8.53mmol) and compound 1-1(2.32g, 10.2mmol) in N, N-dimethylformamide (30.0mL) was added potassium carbonate (3.54g, 25.6mmol), and the reaction was stirred at 70 ℃ under nitrogen for 3 hours. The reaction mixture was added with water (50.0mL), extracted with ethyl acetate (50.0mL × 2), the organic phase was washed with saturated brine (200.0mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:0 to 0:1) to give compound 1-2.

¹H NMR(400MHz，CDCl₃)δ8.37(s，1H)，7.77(s，1H)，6.95-7.01(m，2H)，6.73-6.78(m，1H)，4.00(br s，2H)，3.87-3.93(m，2H)，3.78(dt,J＝13.6，6.8Hz，1H)，3.48(dt,J＝13.6，6.8Hz，1H)，3.31-3.38(m，4H)，1.66-1.71(m，4H)，1.53(d,J＝6.8Hz，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(4.4g, 8.12mmol) in dichloromethane (20.0mL) was added trifluoroacetic acid (7.7g, 67.5mmol), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 1-3.

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

(3) To a solution of trifluoroacetate salt of compound 1-3 (300mg, 679. mu. mol) in methanol (10.0mL) were added triethylamine (206mg, 2.04mmol), compound 1-4(267mg, 1.02mmol) and sodium cyanoborohydride (427mg, 6.79mmol), and the mixture was stirred at 25 ℃ for 4 hours. The reaction mixture was concentrated under reduced pressure, ethyl acetate (20.0mL) was added, the mixture was washed with water (20.0mL × 1) and saturated brine (20.0mL × 1), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 24:1) to give compound 1-5.

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

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

(5) To a solution of the trifluoroacetate salt of compounds 1-6 (273mg, 390. mu. mol) in dichloromethane (5.0mL) was added triethylamine (398mg, 3.93mmol) and intermediate A (200mg, 782. mu. mol), and the reaction was stirred under nitrogen at 25 ℃ for 16 h. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0 mL. times.1) and saturated brine (20.0 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 ═ 1:0 to 32:1) to give compound 1-7.

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

¹H 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 compound 1-7(213mg, 264. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain trifluoroacetate salts of compounds 1 to 8. The crude product was used directly in the next reaction.

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

(7) To a solution of the trifluoroacetate salt of compounds 1-8 (108mg, 132. mu. mol) in dichloromethane (3.0mL) was added triethylamine (66.6mg, 659. mu. mol). Compound 1-9(17.9mg, 198. mu. mol, 16.1. mu.L) was then added and the reaction stirred under nitrogen at-78 ℃ for 60 minutes. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0mL) and 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 (Phenomenexlunna C18, 100 mm. times.40 mm, 3 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of Compound 1.

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

¹H 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

This example provides compound 2 of formula I, wherein the structural formula of compound 2 is as follows:

the synthetic route of compound 2 is shown below:

triethylamine (133mg, 1.31mmol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (100mg, 263. mu. mol) were added to a solution of the hydrochloride of the compound 2-1 (43.6mg, 263. mu. mol) in dichloromethane (3.0mL), and the mixture was stirred at 25 ℃ for 0.5 hour, and the trifluoroacetate (108mg, 132. mu. mol) of the compounds 1 to 8 in example 1 was added to the reaction mixture, and the reaction mixture was stirred at 25 ℃ for 0.5 hour. Dichloromethane (20.0mL) was added, and the mixture was washed with water (20.0 mL. times.1) and saturated brine (20.0 mL. times.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 (Phenomenexlunna C18, 100 mm. times.40 mm, 3 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of Compound 2.

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

¹H 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

This example provides a compound 3 of formula I, the structural formula of compound 3 being shown below:

the synthetic route of compound 3 is shown below:

(1) to a solution of intermediate B (300mg, 853. mu. mol) and compound 3-1(213mg, 941. mu. mol) in N, N-dimethylformamide (8.0mL) was added potassium carbonate (236mg, 1.71mmol), and the reaction was stirred at 70 ℃ under nitrogen for 2 hours. Ethyl acetate (40.0mL) was added to the reaction mixture, which was washed with water (40.0mL × 1) and saturated brine (40.0mL × 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 value 542, found value 542.

¹H 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(435mg, 803. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 3-3.

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

(3) To a solution of the trifluoroacetate salt of compound 3-3 (450mg, 941. mu. mol) in methanol (10.0mL) were added triethylamine (285mg, 2.82mmol), compound 3-4(369mg, 1.41mmol) and sodium cyanoborohydride (592mg, 9.42mmol), and the mixture was stirred at 25 ℃ for 12 hours. Ethyl acetate (40.0mL) was added to the reaction solution, and the organic phase was washed with a saturated aqueous ammonium chloride solution (40.0mL × 2) and a saturated brine (40.0mL × 1), dried over anhydrous sodium sulfate, filtered, 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]⁺687 was calculated and 687 was found.

¹H 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(282mg, 411. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 3-6. The crude product was used directly in the next reaction.

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

(5) To a solution of the trifluoroacetate salt of compound 3-6 (286mg, 408. mu. mol) in dichloromethane (3.0mL) was added triethylamine (400mg, 3.95mmol) and intermediate A (157mg, 614. mu. mol), and the reaction was stirred under nitrogen at 25 ℃ for 16 hours. Dichloromethane (20.0mL) was added, and the organic phase was washed with saturated brine (20.0 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 ═ 1:0 to 32:1) to give compound 3-7.

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

¹H 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(250mg, 310. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 3-8. The crude product was used directly in the next reaction.

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

(7) To a solution of the trifluoroacetate salt of compound 3-8 (127mg, 155. mu. mol) in dichloromethane (3.0mL) was added triethylamine (78.4mg, 775. mu. mol). Then, a solution of compound 3-9(29.5mg, 326. mu. mol, 26.6. mu.L) in dichloromethane (1.0mL) was added and the reaction was stirred under nitrogen at-78 ℃ for 60 minutes. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0mL) and 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 (Phenomenexlunna C18, 100 mm. times.40 mm 3 μm, A: water (0.225% formic acid), B: acetonitrile, 0% -40% for 10 minutes) to give the formate salt of Compound 3.

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

¹H 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

This example provides a compound 4 of formula I, wherein the structural formula of compound 4 is as follows:

the synthetic route of compound 4 is shown below:

triethylamine (78.8mg, 779mmol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (118mg, 310. mu. mol) were added to a solution of the hydrochloride of the compound 4-1 (51.3mg, 310. mu. mol) in methylene chloride (3.0mL), and the mixture was stirred at 25 ℃ for 0.5 hour, and the trifluoroacetate (127mg, 155. mu. mol) of the compound 3-8 of example 3 was added to the reaction mixture and the reaction mixture was stirred at 25 ℃ for 0.5 hour. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0 mL. times.1) and saturated brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was subjected to preparative high performance liquid chromatography (Phenomenexlunna C18, 100 mm. times.40 mm, 3 μm, A: water (0.225% formic acid), B: acetonitrile, 0% -30% for 10 minutes) to isolate the formate salt of Compound 4.

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

¹H 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

This example provides a compound 5 of formula I, the structural formula of compound 5 being shown below:

the synthetic route of compound 5 is shown below:

(1) to a solution of intermediate B (100mg, 284. mu. mol) and compound 5-1(81.3mg, 341. mu. mol) in N, N-dimethylformamide (5.0mL) was added potassium carbonate (118mg, 853. mu. mol), and the reaction solution was stirred at 70 ℃ under nitrogen for 3 hours. Water (100mL) was added to the reaction mixture, and extraction was performed with ethyl acetate (25.0mL × 2), and the organic phase was washed with saturated brine (50.0mL), dried over anhydrous sodium sulfate, filtered, and 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.

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

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

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

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

(3) To a solution of the trifluoroacetate salt of compound 5-3 (130mg, 234. mu. mol) in methanol (5.0mL) were added triethylamine (71.0mg, 702. mu. mol), compound 5-4(61.2mg, 234. mu. mol) and sodium cyanoborohydride (147mg, 2.34mmol), and the mixture was stirred at 25 ℃ for 2 hours. The reaction mixture was concentrated under reduced pressure, ethyl acetate (20.0mL) was added, the organic phase was washed with water (20.0mL × 1) and saturated brine (20.0mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 5-5.

¹H NMR(400MHz，CDCl₃)δ8.35(s，1H)，7.86(s，1H)，7.08-7.15(m，5H)，6.82(dd，J＝9.6，4.0Hz，1H)，4.61(br s，4H)，4.53(br s，2H)，3.86(dt，J＝13.2，6.8Hz，1H)，3.71-3.76(m，3H)，3.67(s，2H)，3.61-3.64(m，2H)，3.17(s，2H)，2.82(t，J＝5.6Hz，2H)，1.75(t，J＝5.6Hz，4H)，1.54(d，J＝6.8Hz，3H)，1.49(s，9H)，1.45(d，J＝6.8Hz，3H)，1.19(d，J＝6.8Hz，3H)，1.18(d，J＝6.8Hz，3H)。

MS-ESI[M+H]⁺687 was calculated and 687 was found.

(4) To a solution of compound 5-5(74.4mg, 108. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 60 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 5-6.

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

(5) To a solution of the trifluoroacetate salt of compound 5-6 (75.9mg, 108. mu. mol) in dichloromethane (5.0mL) were added triethylamine (54.8mg, 542. mu. mol) and intermediate A (55.4mg, 217. mu. mol), and the reaction solution was stirred under nitrogen at 25 ℃ for 2 hours. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0 mL. times.1) and saturated brine (20.0 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 ═ 10:1) to afford compounds 5-7.

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

¹H 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) Trifluoroacetic acid (1.0mL) was added to a solution of compound 5-7(56.0mg, 69.5. mu. mol) in dichloromethane (3.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 5-8. The crude product was used directly in the next reaction.

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

(7) To a solution of the trifluoroacetate salt of compound 5-8 (56.9mg, 69.4. mu. mol) in dichloromethane (5.0mL) was added triethylamine (35.1mg, 347. mu. mol). Compound 5-9(9.4mg, 104. mu. mol, 8.5. mu.L) was then added and the reaction stirred at-78 ℃ for 30 minutes under nitrogen. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0mL) and saturated brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 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 compound 5.

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

¹H 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

This example provides a compound 6 of formula I, the structural formula of compound 6 being shown below:

the synthetic route of compound 6 is shown below:

(1) to a solution of intermediate B (200mg, 569. mu. mol) and compound 6-1(121mg, 569. mu. mol) in N, N-dimethylformamide (5.0mL) was added potassium carbonate (236mg, 1.71mmol), and the reaction mixture was stirred at 80 ℃ under nitrogen for 2 hours. The reaction mixture was added with saturated brine (50mL), extracted with ethyl acetate (50.0mL × 1), the organic phase was washed with saturated brine (25.0mL × 2), dried over anhydrous sodium sulfate, filtered, 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, found 528.

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

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

(3) To a solution of the trifluoroacetate salt of compound 6-3 (230mg, 425. mu. mol) in methanol (10.0mL) were added triethylamine (85.9mg, 849. mu. mol), compound 6-4(111mg, 425. mu. mol) and sodium cyanoborohydride (133mg, 2.12mmol), and the mixture was stirred at 25 ℃ for 2 hours. The reaction mixture was concentrated under reduced pressure, methylene chloride (50.0mL) was added, the organic phase was washed with saturated brine (25.0mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 20:1 to 0:1) to give compound 6-5.

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

(4) To a solution of Compound 6-5(220mg, 326. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction 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 value 573, found value 573.

(5) To a solution of the trifluoroacetate salt of compound 6-6 (220mg, 320. mu. mol) in dichloromethane (10.0mL) were added triethylamine (32.4mg, 320. mu. mol) and intermediate A (81.9mg, 320. mu. mol), and the reaction solution was stirred under nitrogen at 25 ℃ for 5 minutes. Dichloromethane (20.0mL) was added, and the organic phase was washed with a saturated aqueous ammonium chloride solution (20.0 mL. times.1) and a saturated brine (20.0 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 afford compound 6-7.

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

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

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

(7) To a solution of the trifluoroacetate salt of compound 6-8 (180mg, 223. mu. mol) in dichloromethane (5.0mL) was added triethylamine (67.8mg, 670. mu. mol). Subsequently, compound 6-9(20.2mg, 223. mu. mol, 18.2. mu.L) was added and the reaction was stirred at-78 ℃ for 5 minutes under a nitrogen blanket. Dichloromethane (50.0mL) was added, and the organic phase was washed with saturated brine (50.0 mL. times.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% for 48 min) to give compound 6.

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

¹H NMR(400MHz，MeOD)δ8.31(s，1H)，7.80(s，1H)，7.17-7.09(m，5H)，6.93-6.87(m，1H)，6.35-6.21(m，2H)，5.80-5.72(m，1H)，4.53-4.49(m，4H)，4.35-4.30(m，1H)，4.26-4.21(m，2H)，3.90-3.71(m，4H)，3.66-3.60(m，6H)，3.27-3.22(m，4H)，2.96-2.91(m，2H)，2.18-2.10(m，2H)，1.59-1.52(m，3H)，1.50-1.42(m，3H)，1.21-1.18(m，3H)，1.15-1.12(m，3H)。

Example 7

This example provides a compound 7 of formula I, wherein the structural formula of compound 7 is as follows:

the synthetic route of compound 7 is shown below:

(1) to a solution of intermediate B (300mg, 852. mu. mol) and compound 7-1(217mg, 1.02mmol) in N, N-dimethylformamide (8.0mL) was added potassium carbonate (235mg, 1.71mmol), and the reaction solution was stirred at 80 ℃ under nitrogen for 2 hours. The reaction mixture was quenched by addition of water (5.0mL), extracted with ethyl acetate (10.0mL × 3), the organic phase was washed with saturated brine (10.0mL × 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 7-2.

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

(2) To a solution of Compound 7-2(400mg, 758. mu. mol) in dichloromethane (12.0mL) was added trifluoroacetic acid (4.0mL), and the reaction 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, found 428.

(3) To a solution of the trifluoroacetate salt of compound 7-3 (400mg, 738. mu. mol) in methanol (10.0mL) were added triethylamine (74.7mg, 738. mu. mol), compound 7-4(96.5mg, 369. mu. mol) and sodium cyanoborohydride (92.8mg, 1.48mmol), and the mixture was stirred at 25 ℃ for 12 hours. The reaction mixture was quenched by addition of water (5.0mL), extracted with ethyl acetate (10.0mL × 3), the organic phase was washed with saturated brine (10.0mL × 3), 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 ═ 10:1) to give compound 7-5.

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

(4) To a solution of compound 7-5(220mg, 326. mu. mol) in dichloromethane (9.0mL) was added trifluoroacetic acid (3.0mL), and the reaction 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 value 573, found value 573.

(5) To a solution of the trifluoroacetate salt of compound 7-6 (220mg, 320. mu. mol) in dichloromethane (6.0mL) were added triethylamine (32.4mg, 320. mu. mol) and intermediate A (163mg, 640. mu. mol), and the reaction solution was stirred under nitrogen at 25 ℃ for 60 minutes. The reaction mixture was quenched by addition of water (10.0mL), extracted with dichloromethane (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(62.0mg, 78.2. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction 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, found 692.

(7) To a solution of the trifluoroacetate salt of compound 7-8 (60mg, 74.4. mu. mol) in dichloromethane (5.0mL) was added triethylamine (7.5mg, 74.4. mu. mol). Then, compound 7-9(10.1mg, 111. mu. mol, 9.1. mu.L) was added and the reaction was stirred at-78 ℃ for 60 minutes under nitrogen. The reaction mixture was quenched by addition of water (10.0mL), extracted with dichloromethane (10.0 mL. times.2), 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 preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 15% -45% for 10 min) to give compound 7.

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

¹H 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

This example provides a compound 8 of formula I, the structural formula of compound 8 being shown below:

the synthetic route of compound 8 is shown below:

(1) to a solution of compound 8-1(240mg, 467. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (1.45g, 14.4mmol, 2.0mL), and the reaction was stirred at 25 ℃ for 0.5 hour. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 8-2.

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

(2) To a solution of the trifluoroacetate salt of compound 8-2 (240mg, 455. mu. mol) in methanol (10.0mL) was added triethylamine (46.0mg, 455 mmol). To the reaction mixture was added compound 8-3(238mg, 910. mu. mol), and the mixture was stirred at 25 ℃ for 15 minutes. Sodium cyanoborohydride (143mg, 2.27mmol) was then added and stirred at 25 ℃ for 3 hours. The reaction mixture was concentrated under reduced pressure, poured into water (40.0mL), 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 value 659, found value 659.

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

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

(4) To a solution of the trifluoroacetate salt of compound 8-5 (210mg, 312. mu. mol) in dichloromethane (5.0mL) were added triethylamine (63.2mg, 624umol, 86.9uL) and intermediate A (136mg, 531. mu. mol), and the reaction mixture was stirred at 25 ℃ for 10 minutes under nitrogen. The reaction mixture was poured into water (20.0mL) and extracted with dichloromethane (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]⁺779 is calculated and 779 is found.

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

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

(6) To a solution of the trifluoroacetate salt of compound 8-7 (60.0mg, 75.8. mu. mol) in dichloromethane (3.0mL) was added triethylamine (7.67mg, 75.8. mu. mol, 10.6. mu.L). Then, compound 8-8(10.3mg, 114. mu. mol, 9.27. mu.L) was added and the reaction was stirred at-78 ℃ for 10 minutes under nitrogen. The reaction mixture was poured into water (20.0mL), extracted with dichloromethane (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 isolated by preparative high performance liquid chromatography (Welch Xtimate, 75 mm. times.40 mm 3 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of Compound 8.

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

¹H NMR:(400MHz，MeOD)δ8.23-8.27(m，1H)，7.73-7.80(m，1H)，7.11-7.21(m，5H)，6.94-6.99(m，1H)，6.22-6.36(m，2H)，5.71-5.80(m，1H)，4.50-4.55(m，4H)，4.37-4.46(m，2H)，4.28-4.34(m，4H)，4.22-4.27(m，2H)，3.82-3.82(m，1H)，3.71-3.78(m，2H)，3.59-3.68(m，6H)，2.90-2.98(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 compound 9 being shown below:

the synthetic route of compound 9 is shown below:

to a solution of the trifluoroacetate salt (60.0mg, 75.8. mu. mol) of compounds 8 to 7 in example 8 in dichloromethane (3.0mL) were added triethylamine (7.7mg, 75.8. mu. mol, 10.6. mu.L), compound 9-1(25.1mg, 151. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (57.6mg, 151. mu. mol), and the mixture was stirred at 25 ℃ for 0.5 hour. The reaction mixture was poured into water (20.0mL) and extracted with dichloromethane (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 Xtimate, 75 mm. times.40 mm 3 μm, A: water (0.225% formic acid), B: acetonitrile, 5% -35% for 10 minutes) to give the formate salt of compound 9.

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

¹H NMR:(400MHz，MeOD)δ8.25-8.29(m，1H)，7.77-7.82(m，1H)，7.12-7.27(m，5H)，6.92-6.97(m，1H)，6.72-6.82(m，1H)，6.30-6.40(m，1H)，4.90-4.93(m，2H)，4.50-4.62(m，4H)，4.40-4.49(m，2H)，4.32-4.35(m，2H)，4.22-4.29(m，2H)，3.94-4.08(m，6H)，3.79-3.87(m，1H)，3.58-3.67(m，4H)，2.93-3.00(m，2H)，2.60-2.68(m，6H)，1.52-1.57(m，3H)，1.42-1.47(m，3H)，1.16-1.21(m，3H)，1.09-1.14(m，3H)。

Example 10

This example provides a compound 10 of formula I, the structural formula of compound 10 being shown below:

the synthetic route of compound 10 is shown below:

(1) to a solution of the trifluoroacetate salt of compounds 1-3 in example 1(500mg, 0.95mmol) in methanol (10.0mL) was added triethylamine (273mg, 2.70 mmol). To the reaction mixture were added compound 10-1(282mg, 1.08mmol), sodium cyanoborohydride (169mg, 2.70mmol), and the mixture was stirred at 25 ℃ for 16 hours. Poured into saturated aqueous ammonium chloride (40.0mL) 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]⁺687 was calculated and 687 was found.

(1) To a solution of compound 10-2(120mg, 0.175mmol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 15 minutes. 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 value 587, found value 587.

(2) To a solution of the trifluoroacetate salt of compound 10-3 (120mg, 0.171mmol) in dichloromethane (5.0mL) was added triethylamine to adjust the pH to 8, followed by intermediate A (87.6mg, 0.342mmol), and the reaction was stirred under nitrogen at 25 ℃ for 0.5 h. The reaction mixture was poured into water (15.0mL) and extracted with dichloromethane (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, found 806.

(3) To a solution of compound 10-4(130mg, 0.16mmol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. 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, found 706.

(4) To a solution of the trifluoroacetate salt of compound 10-5 (40.0mg, 48.8. mu. mol) in dichloromethane (3.0mL) was added triethylamine (4.94mg, 48.8. mu. mol). Compound 10-6(5.30mg, 58.5umol, 4.77uL) was then added and the reaction stirred at-78 ℃ for 15 minutes under nitrogen. The reaction mixture was poured into water (20.0mL) and extracted with dichloromethane (20.0 mL. times.2). The organic phases were combined, washed with brine (20.0mL), 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 (Phenomenexlunna C18, 100 mm. times.40 mm, 3 μm, A: water (0.225% formic acid), B: acetonitrile, 0% -50% for 10 minutes) to give the formate salt of Compound 10.

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

¹H 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

This example provides a compound 11 of formula I, the structural formula of compound 11 being shown below:

the synthetic route of compound 11 is shown below:

to a solution of the trifluoroacetate salt (40.0mg, 48.8. mu. mol) of compound 10-4 in example 10 in dichloromethane (3.0mL) were added triethylamine (4.94mg, 48.8. mu. mol, 6.79. mu.L), compound 11-1(16.2mg, 97.6. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (37.1mg, 97.6. mu. mol), and the mixture was stirred at 25 ℃ for 0.5 hour. The reaction mixture was poured into water (20.0mL) and extracted with dichloromethane (20.0 mL. times.3). The organic phases were combined, washed with brine (20.0mL), 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 μm, A: water (0.225% formic acid), B: acetonitrile, 0% -40% for 10 min) to give the formate salt of Compound 11.

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

¹H 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

This example provides a compound 12 of formula I, wherein the structural formula of compound 12 is as follows:

the synthetic route of compound 12 is shown below:

(1) to a solution of the trifluoroacetate salt (200mg, 452. mu. mol) of compounds 1 to 3 in example 1 in methanol (5.0mL) were added triethylamine (45.8mg, 452. mu. mol), compound 12-1(121mg, 452. mu. mol) and sodium cyanoborohydride (142mg, 2.26mmol), and the mixture was stirred at 25 ℃ for 12 hours. The reaction mixture was concentrated under reduced pressure, ethyl acetate (80.0mL) was added, the organic phase was washed with water (40.0mL) and saturated brine (50.0mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:0 to 0:1) to give compound 12-2.

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

(2) To a solution of compound 12-2(90.0mg, 129. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction 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, found 593.

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

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

(4) To a solution of compound 12-4(70.0mg, 75.6. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction 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 and found 713.

(5) Triethylamine (8.53mg, 84.3. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (27.6mg, 72.6. mu. mol) were added to a solution of the hydrochloride of compound 12-5 (18.1mg, 109. mu. mol) in methylene chloride (10.0mL), and the mixture was stirred at 25 ℃ for 30 minutes, and the trifluoroacetate of compound 12-6 (60.0mg, 72.6. mu. mol) was added to the reaction solution and the reaction solution was stirred at 25 ℃ for 30 minutes. Dichloromethane (50.0mL) was added, and the organic phase was washed with saturated brine (50.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Phenomenex luna C18, 100 mm. times.30 mm 3 μm, A: water (0.225% formic acid), B: acetonitrile, 0% -30% for 8 minutes) to give compound 12.

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

¹H 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 compound 13 being shown below:

the synthetic route of compound 13 is shown below:

(1) to a solution of the trifluoroacetate salt (200mg, 453. mu. mol) of compounds 1 to 3 in example 1 in methanol (3.0mL) was added triethylamine (91.7mg, 906. mu. mol) and compound E (145mg, 544. mu. mol), and the mixture was stirred at 25 ℃ for 15 minutes, added sodium cyanoborohydride (142mg, 2.26mmol) and stirred at 25 ℃ for 12 hours. The reaction solution was concentrated under reduced pressure, dichloromethane (10.0mL) was added, the organic phase was washed with water (3.0mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was isolated by preparative thin layer chromatography (dichloromethane/methanol ═ 10:1) to give compound 13-1.

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

(2) To a solution of compound 13-1(140mg, 202. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction solution 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, found 593.

(3) To a solution of the trifluoroacetate salt of compound 13-2 (140mg, 198. mu. mol) in dichloromethane (3.0mL) were added triethylamine (40.1mg, 396. mu. mol) and intermediate A (101mg, 396. mu. mol), and the reaction mixture was stirred at 25 ℃ for 10 minutes under nitrogen. Dichloromethane (20.0mL) was added, washed with water (2.0mL), 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 afford compound 12-3.

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

(4) To a solution of compound 13-3(120mg, 148. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (10.9. mu.L), and the reaction 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 value 712, found value 712.

(5) Triethylamine (14.7mg, 145. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (41.4mg, 109. mu. mol) were added to a solution of the hydrochloride of compound 13-5 (36.1mg, 218. mu. mol) in dichloromethane (2.0mL), and the mixture was stirred at 25 ℃ for 10 minutes, and the trifluoroacetate of compound 13-4 (60.0mg, 72.7. mu. mol) was added to the reaction solution and the reaction solution was stirred at 25 ℃ for 60 minutes. The organic phase is concentrated under reduced pressure. Preparation of crude product by high performance liquid chromatography (Phenomenex luna C18, 100 mm. times.30 mm, 3 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% for 8 min) to obtain compound 13.

MS-ESI[M+H]⁺Calculated 823 and found 823.

¹H 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

This example provides a compound 14 of formula I, wherein the structural formula of compound 14 is shown below:

the synthetic route of compound 14 is shown below:

to a solution of the trifluoroacetate salt of compound 13-4 in example 13 (60mg, 72.7. mu. mol) in dichloromethane (3.0mL) was added triethylamine (14.7mg, 145. mu. mol). Compound 14-2(7.23mg, 79.9. mu. mol, 6.52. mu.L) was then added and the reaction stirred at-78 ℃ for 10 minutes under nitrogen. The crude product is separated by preparative high performance liquid chromatography (Phenomenex Luna C18, 100mm x 30mm 3 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% and 8 min) to obtain compound 14.

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

¹H 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 compound 15 being shown below:

the synthetic route of compound 15 is shown below:

(1) to a solution of the trifluoroacetate salt (200mg, 359. mu. mol) of compound 3-3 in example 3 in methanol (10.0mL) were added triethylamine (72.8mg, 720. mu. mol), compound C (188mg, 719. mu. mol) and sodium cyanoborohydride (90.4mg, 1.44mmol), and the mixture was stirred at 25 ℃ for 12 hours. Water (10.0mL) and dichloromethane (20.0mL) were added to the reaction solution, the organic phase was washed with saturated brine (10.0mL), 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 15-1.

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

(2) To a solution of compound 15-1(200mg, 290. mu. mol) in dichloromethane (9.0mL) was added trifluoroacetic acid (3.0mL), and the reaction 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 value 589, found value 589.

(3) To a solution of the trifluoroacetate salt of compound 15-2 (200mg, 285. mu. mol) in dichloromethane (5.0mL) were added triethylamine (28.8mg, 285. mu. mol) and intermediate A (145mg, 570. mu. mol), and the reaction solution was stirred under nitrogen at 25 ℃ for 1 hour. Water (10.0mL) and methylene chloride (20.0mL) were added, and the organic phase was washed with saturated brine (10.0mL), 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 807, found 807.

(4) To a solution of compound 15-3(120mg, 148. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction 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 value 708, found value 708.

(5) Triethylamine (14.7mg, 146. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (55.5mg, 146. mu. mol) were added to a solution of the hydrochloride of the compound 15-5 (28.3mg, 219. mu. mol) in methylene chloride (4.0mL), and the mixture was stirred at 25 ℃ for 1 hour, and the trifluoroacetate of the compound 15-4 (60mg, 73.0. mu. mol) was added to the reaction solution and the reaction solution was stirred at 25 ℃ for 1 hour. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (10.0mL) and saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Crude preparation by HPLC (Xtimate C18, 100 mm. times.30 mm 10 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to isolate compound 15.

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

¹H 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

This example provides a compound 16 of formula I, wherein the structural formula of compound 16 is shown below:

the synthetic route for compound 16 is shown below:

(1) to a solution of the trifluoroacetate salt (200mg, 359. mu. mol) of compound 3-3 in example 3 in methanol (10.0mL) were added triethylamine (72.8mg, 719. mu. mol), compound C (188mg, 719. mu. mol) and sodium cyanoborohydride (90.4mg, 1.44mmol), and the mixture was stirred at 25 ℃ for 12 hours. Water (100.0mL) was added to the reaction mixture, and extraction was performed with dichloromethane (100.0mL × 2), and the organic phase was washed with saturated brine (100.0mL), 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 0:1) to obtain compound 16-1.

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

(2) To a solution of compound 16-1(170mg, 247. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction 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, found 588.

(3) Triethylamine (24.5mg, 242. mu. mol) and intermediate A (123mg, 484. mu. mol) were added to a solution of the trifluoroacetate salt (170mg, 242. mu. mol) of Compound 16-2 in dichloromethane (5.0mL), and the reaction solution was stirred at 25 ℃ for 60 minutes under nitrogen. Water (10.0mL) was added, extraction was performed with dichloromethane (10.0 mL. times.2), and the organic phase was washed with saturated brine (10.0mL), 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 16-3.

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

(4) To a solution of compound 16-3(137mg, 169. mu. mol) in dichloromethane (9.0mL) was added trifluoroacetic acid (3.0mL), and the reaction 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 and measured value 707.

(5) To a solution of the trifluoroacetate salt of compound 16-4 (75.0mg, 91.3. mu. mol) in dichloromethane (5.0mL) was added triethylamine (9.25mg, 91.3. mu. mol). Compound 16-5(12.4mg, 137. mu. mol, 11.1. mu.L) was then added and the reaction stirred under nitrogen at-78 ℃ for 60 minutes. Water (10.0mL) was added, extraction was performed with dichloromethane (10.0 mL. times.2), and the organic phase was washed with saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to afford compound 16.

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

¹H 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 being shown below:

the synthetic route of compound 17 is shown below:

(1) to a solution of the trifluoroacetate salt of compounds 1 to 3 in example 1(200mg, 452. mu. mol) in methanol (10.0mL) were added triethylamine (45.8mg, 452. mu. mol), compound C (237mg, 905. mu. mol) and sodium cyanoborohydride (113.8mg, 1.81mmol), and the mixture was stirred at 25 ℃ for 1 hour. Water (100.0mL) was added to the reaction mixture, and extraction was performed with dichloromethane (100.0mL × 2), and the organic phase was washed with saturated brine (100.0mL), 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 0:1) to obtain compound 17-1.

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

(2) To a solution of compound 17-1(160mg, 232. mu. mol) in dichloromethane (9.0mL) was added trifluoroacetic acid (3.0mL), and the reaction 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, found 588.

(3) To a solution of the trifluoroacetate salt of compound 17-2 (160mg, 228. mu. mol) in dichloromethane (5.0mL) were added triethylamine (23.0mg, 228. mu. mol) and intermediate A (116mg, 456. mu. mol), and the reaction solution was stirred under nitrogen at 25 ℃ for 60 minutes. Water (10.0mL) was added, extraction was performed with dichloromethane (10.0 mL. times.2), and the organic phase was washed with saturated brine (10.0mL), 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 17-3.

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

(4) To a solution of compound 17-3(80mg, 99.1. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction 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 and measured value 707.

(5) Triethylamine (10.0mg, 99.1. mu. mol), 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (37.7mg, 99.1. mu. mol) and compound 17-5(19.2mg, 148. mu. mol) were added to a solution of the trifluoroacetate salt of compound 17-4 (35.0mg, 49.5. mu. mol) in dichloromethane (5.0mL), and stirred at 25 ℃ for 1 hour. Water (10.0mL) was added, the mixture was extracted with dichloromethane (10.0 mL. times.2), and the organic phase was washed with saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 40mm 3 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of compound 17.

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

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

Example 18

This example provides a compound 18 of formula I, wherein the structural formula of compound 18 is shown below:

the synthetic route of compound 18 is shown below:

to a solution of the trifluoroacetate salt (35.0mg, 49.5. mu. mol) of compound 17-4 in example 17 in dichloromethane (3.0mL) was added triethylamine (5.02mg, 49.5. mu. mol). Compound 18-1(6.73mg, 74.3. mu. mol, 6.1. mu.L) was then added and the reaction stirred at-78 ℃ for 60 minutes under nitrogen. Water (10.0mL) was added, extraction was performed with dichloromethane (10.0 mL. times.2), and the organic phase was washed with saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of compound 18.

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

¹H 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 compound 19 is shown below:

the synthetic route for compound 19 is shown below:

(1) to a solution of intermediate B (500mg, 1.42mmol) and compound 19-1(385mg, 1.71mmol) in N, N-dimethylformamide (10.0mL) was added potassium carbonate (392mg, 2.84mmol), and the reaction was stirred at 80 ℃ for 2 hours under nitrogen. The reaction solution was quenched with water (10.0mL) and extracted with dichloromethane (10.0 mL. times.2). The organic phases were combined, washed with saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was isolated 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.

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

(2) To a solution of compound 19-2(673mg, 1.24mmol) in dichloromethane (18.0mL) was added trifluoroacetic acid (9.24g, 81.0mmol, 6.0mL), 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 and measured 442.

(3) The trifluoroacetate salt of compound 19-3 (100mg, 180. mu. mol), compound 12-1(57.7mg, 216. mu. mol) and triethylamine (36.4mg, 359. mu. mol, 50.1. mu.L) were suspended in methanol (5.0mL), and sodium cyanoborohydride (45.2mg, 719. mu. mol) was added and stirred at 25 ℃ for 12 hours under nitrogen. Quenching the reaction with water (10.0mL), extracting with dichloromethane (10.0mL × 2), combining the organic phases, washing with saturated brine (10.0mL), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and isolating the crude product by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 19-4;

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

(4) To a solution of compound 19-4(120mg, 173. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (3.08g, 27.0mmol, 2.0mL), 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]⁺calcd 593, found 593.

(5) To the trifluoroacetate salt of compound 19-5 (120mg, 169. mu. mol) were added a solution of dichloromethane (5.0mL) and triethylamine (34.3mg, 339. mu. mol, 47.2. mu.L), followed by the addition of intermediate A (86.8mg, 339. mu. mol), and the reaction was stirred at 25 ℃ for 1 hour under nitrogen. The reaction was quenched by the addition of water (10.0mL), extracted with dichloromethane (10.0 mL. times.2), and the organic phases were combined, washed with saturated brine (10.0mL), 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 19-6;

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

(6) To a solution of compound 19-6(45.0mg, 55.4umol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.54g, 13.5mmol, 1.0mL), 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 value 712, found value 712.

(7) The trifluoroacetate salt of compound 19-7 (45.0mg, 54.4. mu. mol), compound 19-8(27.0mg, 163. mu. mol), triethylamine (5.51mg, 54.4. mu. mol, 7.58. mu.L) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (41.4mg, 108. mu. mol) were dissolved in dichloromethane (3.0mL) and stirred at 25 ℃ for 1 hour under nitrogen. The reaction mixture was diluted with water (10.0mL), extracted with dichloromethane (10.0 mL. times.2), and the organic phases were combined, washed with saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of compound 19.

MS-ESI[M+H]⁺Calculated 823 and found 823.

¹H NMR(400MHz，MeOD)δ8.29-8.33(m，1H)，7.82-7.87(m，1H)，7.13-7.18(m，2H)，6.86-6.91(m，2H)，6.72-6.81(m，1H)，6.33-6.43(m，1H)，4.52-4.62(m，4H)，4.30-4.37(m，1H)，4.16-4.26(m，4H)，3.85-3.91(m，1H)，3.76-3.84(m，2H)，3.60-3.74(m，7H)，3.06-3.19(m，2H)，2.94-3.02(m，2H)，2.75-2.80(m，2H)，2.69-2.74(m，6H)，1.92-2.04(m，4H)，1.53-1.57(m，3H)，1.42-1.46(m，3H)，1.18-1.21(m，3H)，1.13-1.16(m，3H)

Example 20

This example provides a compound 20 of formula I, wherein the formula of compound 20 is shown below:

the synthetic route for compound 20 is shown below:

(1) the trifluoroacetate salt (120mg, 216. mu. mol) of compound 20-1 in example 20, compound E (69.2mg, 259. mu. mol) and triethylamine (43.7mg, 431. mu. mol, 60.1. mu.L) were suspended in methanol (5.0mL), and sodium cyanoborohydride (54.2mg, 863. mu. mol) was added and stirred at 25 ℃ for 12 hours under nitrogen. Quenching the reaction with water (100.0mL), extracting with dichloromethane (100.0mL × 2), combining the organic phases, washing with saturated brine (100.0mL), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and isolating the crude product by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 20-2;

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

(2) To a solution of compound 20-2(120mg, 173. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (3.08g, 27.0mmol, 2.0mL), 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 20-3. The crude product is directly used for the next reaction;

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

(3) To the trifluoroacetate salt of compound 20-3 (120mg, 169. mu. mol) were added a solution of dichloromethane (5.0mL) and triethylamine (34.3mg, 339. mu. mol, 47.2. mu.L), followed by the addition of intermediate A (86.8mg, 339. mu. mol), and the reaction was stirred at 25 ℃ for 1 hour under nitrogen. The reaction was quenched by the addition of water (100.0mL), extracted with dichloromethane (100.0 mL. times.2), and the organic phases were combined, washed with saturated brine (100.0mL), 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 20-4;

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

(4) Trifluoroacetic acid (1.0mL) was added to a solution of compound 20-4(23.0mg, 28.3. mu. mol) in dichloromethane (3.0mL), 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 20-5. The crude product was used directly in the next reaction.

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

(5) Compound 20-5 trifluoroacetate (23.0mg, 27.8. mu. mol), compound 20-6(10.7mg, 65.1. mu. mol), triethylamine (2.82mg, 27.8. mu. mol, 3.88. mu.L) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (21.1mg, 55.6. mu. mol) were dissolved in dichloromethane (3.0mL) and stirred under nitrogen at 25 ℃ for 1 hour. The reaction mixture was diluted with water (10.0mL), extracted with dichloromethane (10.0 mL. times.2), and the organic phases were combined, washed with saturated brine (10.0mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of compound 20.

MS-ESI[M+H]⁺Calculated 823 and found 823.

¹H 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

This example provides a compound 21 of formula I, the structural formula of compound 21 being shown below:

the synthetic route for compound 21 is shown below:

(1) to a solution of intermediate F (300mg, 888. mu. mol) and compound 21-1(241mg, 1.07. mu. mol) in N, N-dimethylformamide (8.0mL) was added potassium carbonate (246mg, 1.78mmol), and the reaction was stirred at 80 ℃ for 12 hours under nitrogen. The reaction solution was quenched with water (30.0mL) and extracted with ethyl acetate (40.0 mL. times.2). Combining the organic phases, washing with saturated brine (40.0mL × 2), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and separating the crude product by silica gel column chromatography (petroleum ether/ethyl acetate ═ 1:0 to 0:1) to give compound 21-2;

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

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

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

(3) The trifluoroacetate salt of compound 21-3 (237mg, 555. mu. mol), compound C (291mg, 1.11mmol) and triethylamine (56.2mg, 555. mu. mol, 77.3. mu.L) were suspended in methanol (5.0mL), and sodium cyanoborohydride (140mg, 2.22mmol) was added and stirred at 25 ℃ for 12 hours. Quenching the reaction with water (30.0mL), extracting with dichloromethane (30.0mL × 2), combining the organic phases, washing with saturated brine (30.0mL × 2), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and separating the crude product by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 9:1) to give compound 21-4;

MS-ESI[M+H]⁺calculated 675 and found 675.

(4) Trifluoroacetic acid (2.0mL) was added to a solution of compound 21-4(370mg, 549. mu. mol) in dichloromethane (6.0mL), and the reaction was stirred at 25 ℃ for 0.5 hour. 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 value 574, found value 574.

(5) To the trifluoroacetate salt of compound 21-5 (370mg, 538. mu. mol) were added a solution of dichloromethane (8.0mL) and triethylamine (54.4mg, 538. mu. mol, 74.9. mu.L), followed by intermediate A (206mg, 807. mu. mol), and the reaction was stirred at 25 ℃ for 1 hour under nitrogen. The reaction was quenched by the addition of water (30.0mL), extracted with dichloromethane (20.0 mL. times.2), and the organic phases were combined, washed with saturated brine (20.0 mL. times.3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 9:1) to give compound 21-6;

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

(6) To a solution of compound 21-6(210mg, 264. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (3.08g, 27.0mmol, 2.0mL), 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, found 694.

(7) The trifluoroacetate salt of compound 21-7 (41.1mg, 248. mu. mol), compound 21-8(100mg, 124. mu. mol), triethylamine (12.5mg, 124. mu. mol, 17.3. mu.L) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (94.3mg, 248. mu. mol) were dissolved in dichloromethane (5.0mL), and stirred at 25 ℃ for 0.5 hour. The reaction mixture was diluted with water (30.0mL), extracted with dichloromethane (30.0 mL. times.2), and the organic phases were combined, washed with saturated brine (30.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Crude preparation high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) isolated the formate salt of compound 21.

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

¹H 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

This example provides a compound 22 of formula I, wherein the structural formula of compound 22 is shown below:

the synthetic route of compound 22 is shown below

To a solution of the trifluoroacetate salt (100mg, 124. mu. mol) of compound 21-7 in example 21 in dichloromethane (5.0mL) was added triethylamine (12.5mg, 124. mu. mol). Compound 22-2(16.9mg, 186. mu. mol, 15.2. mu.L) was then added and the reaction stirred at-78 ℃ for 5 minutes under nitrogen. Dichloromethane (60mL) was added and the organic phase was washed with water (30mL) and saturated brine (60mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of compound 22.

MS-ESI[M+H]⁺Calculated 747, found 747

¹H 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

This example provides a compound of formula I, wherein the structural formula of compound 23 is shown below:

the synthetic route of compound 23 is shown below:

(1) to a solution of hydrochloride (440mg, 791. mu. mol) of intermediate compound 3-3 in example 3 in methanol (10.0mL) were added triethylamine (160mg, 1.58mmol), compound D (207mg, 791. mu. mol) and sodium cyanoborohydride (199mg, 3.17mmol), and the mixture was stirred at 25 ℃ for 12 hours. Water (100mL) and dichloromethane (200mL) were added to the reaction solution, the organic phase was washed with saturated brine (100mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 23-1.

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

(2) To a solution of compound 23-1(500mg, 726. mu. mol) in dichloromethane (12.0mL) was added trifluoroacetic acid (4.0mL), and the reaction 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, found 588.

(3) To a solution of the trifluoroacetate salt of compound 23-2 (500mg, 712. mu. mol) in dichloromethane (5.0mL) were added triethylamine (72.1mg, 712. mu. mol) and intermediate A (364mg, 1.43mmol), and the reaction mixture was stirred at 25 ℃ for 1 hour under nitrogen. Water (10.0mL) and methylene chloride (20.0mL) were added, and the organic phase was washed with saturated brine (10.0mL), 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 807, found 807.

(4) To a solution of compound 23-3(100mg, 123. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction 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 and measured value 707.

(5) To a solution of the hydrochloride of compound 23-5 (23.6mg, 182. mu. mol) in dichloromethane (3.0mL) were added triethylamine (12.3mg, 121. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (46.3mg, 121. mu. mol), to the reaction solution was added the trifluoroacetate salt of compound 23-4 (50mg, 60.9. mu. mol), and the reaction solution was stirred at 25 ℃ for 1 hour. Dichloromethane (200mL) was added and the organic phase was washed with water (100mL) and saturated brine (100mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 3% -33% for 10 minutes) to give the formate salt of compound 23.

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

¹H 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 compound 24 being shown below:

the synthetic route for compound 24 is shown below:

to a solution of the trifluoroacetate salt (50mg, 60.9. mu. mol) of compound 23-4 in example 23 in dichloromethane (2.0mL) was added triethylamine (6.16mg, 60.9. mu. mol). Compound 24-1(8.27mg, 91.3. mu. mol, 7.45. mu.L) was then added and the reaction stirred under nitrogen at-78 ℃ for 60 minutes. Dichloromethane (200mL) was added and the organic phase was washed with water (100mL) and saturated brine (100mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was isolated by preparative high performance liquid chromatography (Xtimate C18, 100mm x 30mm 10 μm, a: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) to give the formate salt of compound 24.

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

¹H NMR(400MHz，MeOD)δ8.37-8.40(m，1H)，8.28-8.30(m，1H)，7.81-7.84(m，1H)，7.66-7.70(m，1H)，7.12-7.16(m，2H)，6.86-6.91(m，1H)，6.19-6.39(m，2H)，5.73-5.82(m，1H)，4.54-4.60(m，4H)，4.39-4.44(m，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)。

Test example 1

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

1. the experimental principle is that MV-4-11 is a human leukemia cell strain with MLL translocation and expression of MLL fusion protein MLL-AF 4. The compounds of the present invention inhibit the proliferation of MV-4-11 by interfering with the menin/MLL protein/protein interaction.

2. Experimental materials Cell Counting Kit-8 was purchased from Shanghai Liji 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); iskoff's modified broth (IMDM) medium purchased from Invitrogen (cat # 12440046); the benchtop microplate reader SpectraMax i3X was purchased from Molecular Devices.

3. Experimental procedure cells in logarithmic growth phase were resuspended in complete medium (IMDM + 10% Fetal Bovine Serum (FBS)) and plated into 96-well plates (100. mu.L cell suspension per well, i.e., 15000 cells per well). Cells were incubated at 37 ℃ and 100% relative humidity, 5% CO₂Incubate in incubator for 24 hours.

Dissolving a compound to be detected in dimethyl sulfoxide (DMSO) to prepare stock solution with the concentration of 10mmol/L, and gradually diluting the stock solution with DMSO according to a 4-fold gradient for 8 times. Then diluted 20-fold with the medium. The final concentration of the compound was 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 this order, and the amount of the compound was added to the 96-well plate seeded with cells at 25. mu.L/well.

Cells to which test compounds were added were placed at 37 ℃ and 100% relative humidity, 5% CO₂Respectively incubating for 72 hours in the incubator; cell viability was measured using the CCK-8 method by adding 10. mu.L of CCK-8 detection reagent to each well and incubating in an incubator for an additional 4 hours. The reading wavelength was 450nM (reference wavelength 650nM) using a benchtop microplate reader.

4. Data processing:

the inhibition rate of the drug on the growth of tumor cells was calculated according to the following formula:

the tumor cell growth inhibition rate is [ (ODc-ODs)/(ODc-ODb) ]. times.100%

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

And calculating IC of Compounds with Graphpad software₅₀。

The specific test results are shown in table 1:

TABLE 1

Test compounds	MV 4-11IC₅₀(nM)
		Example 1	30.46
Example 2	374.9
		Example 3	6.03
Example 4	12.31
		Example 6	362.3
Example 8	142.0
		Example 9	67.18
Example 10	196.9
		Example 11	102.5

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

Test example 2

Determination of the antiproliferative effect of 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 Excell (cat # FND 500); dimethyl sulfoxide (DMSO) was purchased from Sigma (cat # D2650); 384 well cell culture plates were purchased from Corning (cat # 3756); automatic cell counters were purchased from Life technologies (model number Countess II); the microplate Reader was purchased from PerkinElmer (model EnVision Multilabel Reader).

3. Fruit of Chinese wolfberryAssay method cells in logarithmic growth phase were resuspended in growth medium (IMDM + 10% FBS) and diluted to target density (50000/mL). The cell suspension was seeded into 384-well plates at 50. mu.L per well; at 37 ℃ 5% CO₂Incubate overnight in the incubator.

Test compounds were dissolved in DMSO to prepare stock solutions at a concentration of 10 mM. Stock was first diluted to 2mM with DMSO and further diluted in 3-fold gradients for 10 concentrations. mu.L of each concentration of the above solution was diluted with 94.5. mu.L of growth medium. Then 5. mu.L/well was added to the 384 well plates seeded with cells.

Cells containing test compound were incubated at 37 ℃ with 5% CO₂Incubate in incubator for 72 hours. Equilibrating 384 well plates at room temperature, adding 15. mu.L of CellTiter-Glo reagent to each well, vortexing for 2 minutes, incubating for 60 minutes at room temperature, EnVision Multilabel Reader reading luminescence, calculating IC of compound using GraphPad Prism 5.0software₅₀。

4. Experimental data:

the specific test results are shown in table 2:

TABLE 2

Test compounds	MV 4-11IC₅₀(nM)
		Example 2	237.2
Example 3	8.36
		Example 5	575.1
Example 6	422.5
		Example 7	81.22
Example 12	124.2
		Example 13	317.6
Example 14	485.7
		Example 15	47.09
Example 16	91.37
		Example 17	90.38
Example 18	167.2
		Example 19	8.83
Example 20	19.36
		Example 21	95.84
Example 22	78.54
		Example 23	12.06
Example 24	29.70

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

The applicant states that the present invention is illustrated by the above examples, including the spirocyclic compound, the pharmaceutical composition containing the same and the application thereof, but the present invention is not limited to the above examples, i.e. it does not mean that the present invention must rely on the above examples to be implemented. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. A spiro compound is characterized in that the structural formula of the spiro compound is shown as the following formula I:

wherein the content of the first and second substances,

R¹selected from the group consisting of-C (O) (NR)^aR^b) (ii) a Wherein R is^a、R^bEach independently selected from H, optionally substituted C1-C6 alkyl, optionally substituted 3-8 membered cycloalkyl and optionally substituted 4-8 membered heterocyclyl, or R^aAnd R^bIs linked to N to form an optionally substituted 4-8 membered heterocyclic ring; wherein the heterocyclic ring contains 1-3 groups selected from N,O, S, P;

R²selected from H, halogen, methyl and trifluoromethyl;

R³selected from H and halogen;

R⁴selected from H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, halogen, -NH₂、-NO₂-COOH, -CN, -OH, optionally substituted C1-C6 alkyl sulfone group, optionally substituted C1-C6 alkyl sulfoxide group, optionally substituted C1-C6 alkylthio group, -NHCORR^4'＝CH₂、-NHCOCHR^4'R^4”、-SO₂C(R^4')＝CH₂、-NHSO₂CR^4'＝CH₂and-NHSO₂CHR^4'R^4”(ii) a Wherein R is^4'Selected from H, methyl and fluoro; r^4”Selected from chlorine and bromine atoms;

w is selected from N and C;

U¹、U²、U³、U⁴、U⁵、U⁶、U⁷、U⁸each 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 ') (NH) —)₂) -, and U¹、U²、U³、U⁴At most one of which is-C (═ O) -, -C (R ') (R ") -O-or-C (R ') (R") -NR ' "-, U⁵、U⁶At most one of which is-C (═ O) -, -C (R ') (R ") -O-, -C (R ') (R") -NR ' "-or-N ═ C (NH)₂)-，U⁷、U⁸At most one of which 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;

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

L¹is absent, -CR^L1'R^L1”-、-CO-、-SO₂-, -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^L1”Form an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the carbon atom to which it is attached; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P;

L²selected from: -SO₂-、-SO-、-CO-、-CF₂-and-C (N ═ N) -;

L³selected from: oxygen atom, sulfur atom, -SO₂-、-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^L3”Form an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the carbon atom to which it is attached; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P; r^L3”'Selected from: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkane, and optionallyAn optionally substituted 4-8 membered saturated or unsaturated heterocyclic ring, wherein said heterocyclic ring comprises 1-3 heteroatoms selected from N, O, S, P;

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

R⁵selected from: -CH₂R^5'、

indicates the attachment position of the group.

2. The spirocyclic compound of claim 1, wherein said R is²Selected from H and halogen; further preferably, said R²Is fluorine;

preferably, said R is³Is H or a fluorine atom; further preferably, said R³Is H;

preferably, said R is⁴Is H, optionally substituted C1-C6 alkyl, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 alkylamino, -NH₂or-CN;

preferably, said Y, Z are each N;

preferably, said W is C;

preferably, said V is N;

preferably, the U is¹、U²、U³、U⁴、U⁵、U⁶、U⁷、U⁸Each independently selected from: -C (R ') (R ") -, -C (R') (R") -C (R '") (R") -, -C (═ O) -, and-C (R') (R ") -C (═ O) -, and U¹、U²、U³、U⁴Wherein at most one is-C (═ O) -or-C (R') (R ") -C (═ O) -, U⁵、U⁶Wherein at most one is-C (═ O) -or-C (R') (R ") -C (═ O) -, U⁷、U⁸Wherein at most one 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 said heteroaromatic ring contains 1-3 heteroatoms selected from N, O, S, P; further preferably, said 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 triazazole ring;

preferably, said L¹Is absent or-CH₂-; further preferably, said L¹is-CH₂-；

Preferably, said L²Selected from: -SO₂-, -SO-and-CO-; further preferably, said L²is-SO₂-；

Preferably, said L³Selected from: oxygen atom, sulfur atom, -CR^L3'R^L3”-and-NR^L3”'-; wherein R is^L3'、R^L3”Are independently selected respectivelyFrom: H. optionally substituted C1-C4 alkyl and halogen, or R^L3'And R^L3”Form an optionally substituted 3-8 membered saturated or unsaturated cycloalkane, an optionally substituted 4-8 membered saturated or unsaturated heterocycle with the carbon atom to which it is attached; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P; r is^L3”'Selected from the group consisting of: H. optionally substituted C1-C4 alkyl, optionally substituted 3-8 membered saturated or unsaturated cycloalkane, and optionally substituted 4-8 membered saturated or unsaturated heterocycle; wherein said heterocycle contains 1-3 heteroatoms selected from N, O, S, P; further preferably, said L³Is an oxygen atom or a sulfur atom;

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

preferably, said R is⁵Selected from: -CH₂R^5'、

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

3. The spirocyclic compound of claim 1 or 2, wherein in said formula I

The spiro ring is selected from any one of the following groups:

preferably, in said formula I

The spiro ring is selected from any one of the following groups:

4. the spirocyclic compound of any one of claims 1-3, wherein in said formula I

The cyclic moiety is selected from any one of the following groups:

5. The spiro compound according to any one of claims 1 to 4, wherein the formula in formula I is

The cyclic part is selected from any one of the following groups:

6. the spirocyclic compound of any one of claims 1-5, wherein said R is⁵Selected from: -CH₂F、-CH₂F、-CH₂Cl、

7. The spirocyclic compound of any one of claims 1-6, wherein said compound of formula I is selected from any one of the following compounds:

8. the spiro compound according to any of claims 1 to 7, further comprising any of the pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, cis-trans-isomers, solvates or polymorphs, or deuterates of the compound of formula I.

9. A pharmaceutical composition comprising a spirocyclic compound according to any one of claims 1-8 and a pharmaceutically acceptable carrier.

10. Use of a spirocyclic compound according to any one of claims 1-8 or a pharmaceutical composition according to claim 9, selected from any one of the following (a) - (c):

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