CN110981803A - Anti-tumor compound and synthesis method and application thereof - Google Patents

Abstract

The invention belongs to the field of medicinal chemistry, and particularly relates to an anti-tumor compound and a synthesis method and application thereof. The compound has a structure shown in a general formula I or a general formula II, shows good inhibitory activity on proteasome regulation kinase DYRK2, and can further achieve anti-tumor activity by inhibiting proteasomes. The invention also relates to a synthetic method of the compound, which adopts a chemical total synthetic route, and the convergent synthetic route can be applied to the chemical synthesis of compounds with similar structures and related derivatives, thereby opening up a wide development space for novel antitumor drugs.

Description

Anti-tumor compound and synthesis method and application thereof

Technical Field

The invention relates to the field of medicinal chemistry, in particular to an anti-tumor compound and a synthesis method and application thereof.

Background

Malignant tumor is the first disease threatening human health, is the first of the three main causes of death of human diseases, accounts for one fourth of all causes of death of diseases, and the number of the diseases and the number of the deaths are increased year by year. The tumor is caused by the interaction of a plurality of factors such as genetic factors, the environment in the body, the environment in vitro and the like, the molecular mechanism of the tumor relates to abnormal changes such as genomics, epigenetics, gene expression regulation and the like, and the tumor is a multi-factor, multi-step and intricate process.

Among them, Multiple Myeloma (MM) is a common malignant tumor of the blood system, characterized by unlimited clonal proliferation of malignant plasma cells in the bone marrow and secretion of a single immunoglobulin strain, and is clinically accompanied by anemia, osteolytic bone disease, renal damage, and damage caused by extramedullary infiltration of neoplastic cells. Since the first proteasome inhibitor, "bortezomib", was approved by the FDA in 2003, proteasome inhibitors were widely used in the treatment of multiple myeloma, and the second-generation proteasome inhibitor, "carfilzomib", and the third-generation proteasome inhibitor, "ixazozomib", were also approved by the FDA in 2012 and 2015, respectively, for marketing. However, almost all patients with multiple myeloma will have severe drug resistance after being treated by proteasome inhibitors, which causes relapse, and the patients still can not be cured so far, so that the development of novel targeted drugs for overcoming the drug resistance is urgently needed in clinic. In addition, toxic and side effects such as cardiotoxicity, thrombocytopenia, neuropathy and the like caused by proteasome inhibitors are also attracting more and more attention, so that a practical and effective combined drug administration scheme is explored while a novel targeted drug is developed, the drug treatment effect is improved, the drug dosage is reduced, the toxic and side effects are reduced, and the occurrence of drug resistance is relieved.

Breast cancer is one of the most common malignant tumors of women, occurs in the malignant tumors of mammary gland epithelial tissues, and is mainly characterized by breast mass, skin change, axillary lymphadenectasis and the like in clinical features. The development of the medicines such as paclitaxel, trastuzumab and lapatinib solves the treatment dilemma of the breast cancer. However, triple negative breast cancer (the result of immunohistochemical examination of cancer tissues is breast cancer in which an Estrogen Receptor (ER), a Progestogen Receptor (PR) and a proto-oncogene Her-2 are negative and account for 10.0% -20.8% of all pathological types of breast cancer) is the worst type of prognosis in breast cancer, and currently, no targeted drug is on the market, and no specific treatment guide exists, so that the research and development of a novel targeted drug are urgently needed clinically, and a practical and effective combined medication scheme is explored.

The enzyme is a protein for catalyzing other molecules in a living body to be phosphorylated, and regulates a series of important physiological processes such as cell cycle, signal transduction, cell autophagy, growth and development and the like by regulating the phosphorylation degree of a downstream substrate.

There are over 500 kinases in humans, and one novel class of kinases is the DYRK family of bispecific protein kinases in the CMGC family. The DYRK family is conserved in evolution, contains 5 members in total, such as DYRK1A, DYRK1B, DYRK2, DYRK3 and DYRK4, has phosphorylation activity on tyrosine and serine/threonine fragments of exogenous substrates, and plays a key role in cell proliferation and apoptosis induction. Recent studies have shown that phosphorylation at Thr25, a critical site on the Rpt3 subunit of the human 26s proteasome, is a necessary condition for tumor cells to pass through their cell cycle, while DYRK2 is the major kinase to phosphorylate this site. Thus, DYRK2 is a potential novel target for tumor therapy.

Selective inhibitors of DYRK family members have been reported with a focus on DYRK1A, whereas a lesser number of reports have been reported for DYRK2, which are also less active and less selective. The reason for this is that the DYRK family members are highly homologous, especially near the ATP binding pocket, where only a few hydrophobic amino acids differ.

Disclosure of Invention

In order to solve the above technical problems, a first object of the present invention is to provide an anti-tumor compound or a pharmaceutically acceptable salt or stereoisomer thereof, which exhibits excellent inhibitory activity against protein kinase DYRK2 and has a desirable anti-tumor effect. In particular, the method comprises the following steps of,

the compound has a structure shown in a general formula I or a general formula II:

in the general formula I, R₁Represents hydrogen, acyl, sulfonyl, benzene which can be substituted, pyridine which can be substituted, furan which can be substituted, thiophene which can be substituted, or straight-chain or branched-chain alkyl of C1-C12, the tail end of which can be substituted by one or more of amino, cyano, amido or a ring containing hetero atoms; preferably R₁Represents a linear alkyl group of C1-C4 which may be substituted at the terminal by an amino group, cyano group, amido group or N-containing heterocycle; further preferred is R₁Represents any one of propylamino, Boc-propylamino, acetylapropylamino, acetylbutylamino, propionitrile group, butyronitrile group or side chain with the following terminal group as N-containing heterocycle: 4-N cyclohexylamine, 4-methylpiperidine, (S) -3-methylpyrrolidine, (R) -3-methylpyrrolidine, 3-methylazetidine, 1- (4-methylpiperidin-1-yl) propan-2-en-1-one, 4- ((4-methylpiperidin-1-yl) methyl) benzenesulfonyl fluoride, N- (4- (4-methylpiperazin-1-yl) -4-oxobutyl) acrylamide, N- (5- (4-methylpiperidin-1-yl) -5-oxopentyl) acrylamide, (2-formyl-5- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid, and mixtures thereof, (2-formyl-4- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid;

R₂、R₃independently of one another, represents hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, phenyl or benzyl; preferably, R₂、R₃Any one of which represents methyl or benzyl, the other represents methyl, isopropyl or benzyl; more preferably, R₂、R₃Any one of which represents methyl and the other represents methyl, isopropyl or benzyl; most preferably, R₂、R₃All represent methyl;

R₄represents hydrogen, carboxyl, ester group, amide group, or C1-C3 linear alkyl with the terminal substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl; preferably, R₄Represents hydrogen, carboxyl, ester group, amido or methyl which can be substituted by one or more of halogen, methyl, hydroxyl, amino, cyano and guanidyl; further preferably, R₄Represents hydrogen, carboxylA group, hydroxymethyl, aminomethyl, difluoromethyl, guanidinomethyl, dimethyl-substituted hydroxymethyl;

in the general formula II, R₅Represents hydrogen, acyl, sulfonyl, benzene which can be substituted, pyridine which can be substituted, furan which can be substituted, thiophene which can be substituted, or straight-chain or branched-chain alkyl of C1-C12, the tail end of which can be substituted by one or more of amino, cyano, amido and N-containing heterocyclic ring; preferably R₅Represents a linear alkyl of C1-C4, the tail end of the linear alkyl can be substituted by one or more of amino, cyano, amido and N-containing heterocycle; further preferred is R₅Represents any one of propylamino, Boc-propylamino, acetylapropylamino, acetylbutylamino, propionitrile group, butyronitrile group or side chain with the following terminal group as N-containing heterocycle: 4-N cyclohexylamine, 4-methylpiperidine, (S) -3-methylpyrrolidine, (R) -3-methylpyrrolidine, 3-methylazetidine; most preferably, R₅Represents 4-methylpiperidine; r₆Represents hydrogen, carboxyl, ester group, amide group or C1-C3 linear alkyl with the terminal substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl; preferably, R₆Represents hydrogen, carboxyl, ester group, amido or methyl which can be substituted by one or more of halogen, methyl, hydroxyl, amino, cyano and guanidyl; further preferably, R₆Represents hydrogen, carboxyl, hydroxymethyl, aminomethyl, difluoromethyl, guanidinomethyl, dimethyl-substituted hydroxymethyl; most preferably, R₆Represents a hydroxymethyl group.

As a preferred embodiment, in formula I, R₁R represents hydrogen, acyl, sulfonyl, aryl such as substituted benzene, pyridine, furan, thiophene, etc., straight chain or branched chain alkyl of C1-C12, the terminal of which may be substituted by amino, cyano, amido, or heteroatom-containing ring₂、R₃Independently of one another, represents hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, phenyl or benzyl, R₄Represents hydrogen, carboxyl, ester group, amide group, or a C1-C3 linear alkyl group which may be substituted at the terminal by halogen, methyl, ethyl, hydroxyl, amino, cyano, guanidino;

as a preferred embodiment, in formula I, R₁Represents a C1-C4 linear alkyl group optionally substituted at the end by an amino, cyano, amido or heteroatom-containing ring, R₂、R₃Independently of one another, represents methyl, isopropyl, benzyl; r₄Represents hydrogen, carboxyl, ester group or methyl with the terminal substituted by one or more of halogen, methyl, hydroxyl, amino and guanidyl. Preferably, the compound of formula I is selected from the compounds of the following structures:

as a preferred embodiment, in formula II, R₅Represents a linear alkyl of C1-C4, the tail end of the linear alkyl can be substituted by one or more of amino, cyano, amido or N-containing heterocyclic rings; r₆Represents hydrogen, carboxyl, ester group, amido or methyl which can be substituted by one or more of halogen, methyl, hydroxyl, amino, cyano and guanidyl;

as the most preferable embodiment, in the formula II, R₅Represents 4-methylpiperidine, R₆Represents hydrogen or hydroxymethyl.

Preferably, the compound of formula II is selected from the compounds of the following structures:

the inhibitor acridine based on DYRK2 is subjected to multiple rounds of chemical derivatization, two types of 32 analogs are finally obtained through optimization, and the currently best compound has higher activity and selectivity on DYRK2 and IC₅₀0.009 μ M, selective for other DYRK family members; the selectivity of the same family of DYRK1A was nearly 300-fold, and its tumor suppressor activity was preliminarily verified at the cellular level and the animal level. The acridine DYRK2 inhibitor small molecule can be used for preventing or treating proteasome-related tumorsOpens up a new way.

The invention also provides pharmaceutically acceptable salts of the compounds, wherein the pharmaceutically acceptable salts include, but are not limited to, inorganic acid salts selected from, for example, hydrochloride, phosphate, hydrobromide, sulfate; also included are organic salts selected from, for example, trifluoroacetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, benzoate, salicylate, stearate, alkanoates such as acetate and HOOC- (CH)₂) A salt of n-COOH, wherein n is selected from 0 to 4. Similarly, examples of pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium, and ammonium.

Preferably the pharmaceutically acceptable salt is trifluoroacetate; particularly preferably, when the compound has a structure shown in formula I-22, the pharmaceutically acceptable salt is trifluoroacetate.

Based on the structure of the compound, when the amino group of R1 in the general formula I and R5 in the general formula II is protected by an acyl protecting group (such as acetyl, methylsulfonyl, p-toluenesulfonyl, o-nitrobenzenesulfonyl, tert-butoxycarbonyl, N-benzyloxycarbonyl and the like), and/or when the hydroxyl group of R4 in the general formula I and R6 in the general formula II is protected by benzyl, p-methoxybenzyl, acyl protecting groups (the same as above), ether protecting groups (tetrahydropyran, methoxymethyl ether and the like), silyl ether protecting groups (trimethylsilyl, triisopropylsilyl, tert-butyldiphenylsilyl and the like), the compound also belongs to derivatives which are easily obtained based on the compound.

The second objective of the present invention is to provide a method for synthesizing the anti-tumor compound or the pharmaceutically acceptable salt or stereoisomer thereof, wherein the synthetic route of the compound of the general formula I is as follows:

when R is₄When the compound represents hydrogen, the compound 4 is subjected to at least one step of side chain introduction reaction to prepare a compound IA;

when R is₄When the compound represents carboxyl, ester group, amide group or C1-C3 linear alkyl of which the tail end can be substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl, the compound IB is prepared by at least one step of side chain introduction reaction of a compound 4'.

As a preferred embodiment, when the compound of formula I has a structure as shown in IA1, the synthetic route is as follows:

wherein R is₁As defined above; preferably, the structure of the compound of the general formula IA1 is shown as I-1-I-6 or I-12-I-17;

the synthesis method comprises the following specific steps:

(i) after the compound 4 and sodium hydrosulfide are subjected to thiolation reaction, the obtained product is subjected to substitution reaction with a corresponding side chain compound to introduce a side chain, so that a compound IA1 is obtained;

preferably, when R is₁When the side chain compound represents 4-N cyclohexylamine or 4-methylpiperidine, the side chain compound is provided with a protecting group, and after a substitution reaction is carried out and a side chain is introduced, the obtained product is subjected to deprotection to obtain a compound IA 1;

in some embodiments, when the compounds of formula I have structures as shown in formulas I-12 to I-17, they can also be synthesized by the following synthetic routes:

wherein R is₁Represents 1- (4-methylpiperidin-1-yl) prop-2-en-1-one, 4- ((4-methylpiperidin-1-yl) methyl) benzenesulfonyl fluoride, N (4- (4-methylpiperazin-1-yl-4-oxobutyl) acrylamide, n- (5- (4-methylpiperidin-1-yl) -5-oxopentyl) acrylamide, (2-formyl-5- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid or (2-formyl-4- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid;

the synthesis method comprises the following specific steps:

(i) the compound I-6 is subjected to at least one-step amidation or alkylation reaction to obtain the compounds I-12 to I-17.

When the compound of the general formula I has a structure shown as a general formula IA2, the synthetic route is as follows:

wherein R is₁、R₃As defined above; preferably, the structure of the compound of the general formula IA2 is shown as I-7-I-10, I-18 or I-19;

the synthesis method comprises the following specific steps:

(i) carrying out deprotection reaction on the compound 4 by boron tribromide to obtain a compound 5;

(ii) performing alkylation reaction on the compound 5 to obtain a compound 6;

(iii) carrying out thiolation reaction on the compound 6 and sodium hydrosulfide, carrying out substitution reaction on the obtained product and a corresponding side chain compound with a protecting group, introducing a side chain, and carrying out deprotection reaction to obtain a compound IA 2;

in some embodiments, when the compound of formula I has a structure as shown in formula I-18 or I-19, it can also be synthesized using the following synthetic route:

wherein R is₁Represents 1- (4-methylpiperidin-1-yl) prop-2-en-1-one or 4- ((4-methylpiperidin-1-yl) methyl) benzenesulfonyl fluoride;

the synthesis method comprises the following specific steps:

(i) the compound I-9 is subjected to at least one-step amidation or alkylation reaction to obtain compounds I-18 and I-19.

When the compound of the general formula I has a structure shown in a formula I-11, the synthetic route is as follows: the synthesis method comprises the following steps

The method comprises the following specific steps:

(i) carrying out deprotection reaction on the compound 4 by boron tribromide to obtain a compound 7;

(ii) the compound 7 reacts with benzyl bromide to obtain a compound 8;

(iii) carrying out thiolation reaction on the compound 8 and sodium hydrosulfide, carrying out alkylation reaction on the obtained product and a corresponding side chain compound with a protecting group, and carrying out Boc protection removal in dichloromethane solution containing trifluoroacetic acid to obtain a compound I-11;

when the compound of the general formula I has a structure shown in a formula IB1, the synthetic route is as follows:

wherein R is₁As defined above; r₄Represents carboxyl, ester group, amide group or C1-C3 linear alkyl with the terminal substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl; preferably, the structure of the compound of the general formula IB1 is shown as formulas I-20-I-30;

the synthesis method comprises the following specific steps:

(i) treating the compound 4' with oxalic acid chloride, and performing methyl esterification in methanol to obtain a compound 12; and (3) carrying out at least one-step one-pot thiolation alkylation on the compound 12, and/or one-step functional group conversion, and/or one-step deprotection reaction to obtain a compound IB 1.

Preferably, the synthetic route of the compound 4 is as follows:

the method comprises the following specific steps:

(i) carrying out copper catalytic coupling on the compounds 1 and 2 to obtain a compound 3;

(ii) and refluxing, cyclizing and aromatizing the compound 3 in phosphorus oxychloride to obtain the compound 4.

Preferably, the synthetic route of the compound 4' is as follows:

the method comprises the following specific steps:

(i) carrying out coupling reaction on the compounds 1 ' and 2 ' under the catalysis of copper and cuprous oxide to obtain a compound 3 ';

(ii) cyclizing and aromatizing the compound 3 'in phosphorus oxychloride to obtain the compound 4'.

Preferably, when the compound of formula II has a structure represented by formula II-1, the synthetic route is as follows:

the synthesis method comprises the following specific steps:

(i) carrying out heterocyclic synthesis reaction on the compound 22 and the compound 23 under alkaline condition to obtain a compound 24;

(ii) cyclizing and aromatizing the compound 24 in phosphorus oxychloride to obtain a compound 25;

(iii) carrying out thiolation reaction on the compound 25 and sodium hydrosulfide, carrying out alkylation reaction on the obtained product and a corresponding side chain compound with a protecting group, and then carrying out deprotection to obtain a compound II-1;

when the compound of the general formula II has the structure shown in the formula II-2, the synthetic route is as follows:

the synthesis method comprises the following specific steps:

(i) reacting the compound 22 with a compound 23 'to obtain a compound 24';

(ii) cyclizing and aromatizing the compound 24 'in phosphorus oxychloride to obtain a compound 25';

(iii) thiolating the compound 25 'with sodium hydrosulfide, and alkylating the compound by a one-pot method to obtain a compound 26';

(iv) and (3) carrying out deprotection on the compound 26' after reduction by diisopropylaluminum hydride to obtain a compound II-2.

Preferably, the protecting group described in the present invention is a Boc-protecting group. In some embodiments, the resulting product is subjected to an alkylation reaction with the corresponding side chain compound bearing a Boc-protecting group followed by de-Boc protection in dichloromethane solution containing trifluoroacetic acid to provide the corresponding compound.

"pharmaceutically acceptable salts" as defined in the present invention include salts of the compounds, as well as salts of stereoisomers of the compounds, such as salts of enantiomers, and/or salts of diastereomers.

If the compounds described herein are obtained in the conventional salt-forming form, those skilled in the art will recognize a variety of synthetic procedures which may be employed without undue experimentation in preparing non-toxic pharmaceutically acceptable addition salts.

The compounds described herein can be asymmetric (e.g., have one or more stereogenic centers). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are contemplated. Compounds of the present invention containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods for how to prepare optically active forms from optically active starting materials are known in the art, for example by resolution of racemic mixtures or by stereoselective synthesis.

In some examples, 4-N cyclohexylamine refers to

In some examples, 4-methylpiperidinyl refers to

In some examples (S) -3-methylpyrrolidine refers to

In some examples, (R) -3-methylpyrrolidine refers to

In some examples, 3-methylazetidine refers to

In some examples, 1- (4-methylpiperidin-1-yl) prop-2-en-1-one refers to

In some examples, 4- ((4-methylpiperidin-1-yl) methyl) benzenesulfonyl fluoride refers to

In some examples, N- (4- (4-methylpiperazin-1-yl) -4-oxobutyl) acrylamide refers to

In some examples, N- (5- (4-methylpiperidin-1-yl) -5-oxopentyl) acrylamide refers to

In some examples, 2-formyl-5- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid refers to

In some examples, (2-formyl-4- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid refers to

The third purpose of the invention is to provide a pharmaceutical composition, which contains the compound or one or more of the pharmaceutically acceptable salts or stereoisomers thereof, and at least one medicinal diluent, carrier or excipient.

Preferably, the pharmaceutical composition further comprises a second therapeutic agent that can be used to treat a tumor;

the second therapeutic agent is preferably a proteasome inhibitor and/or a natural anti-cancer drug;

the proteasome inhibitor is preferably one or more of bortezomib, carfilzomib and ixazoib;

the natural anticancer drug is preferably paclitaxel.

The fourth purpose of the invention is to provide the application of the compound or the pharmaceutically acceptable salt or the stereoisomer thereof in preparing tumor treatment medicines.

Preferably, the compound or the pharmaceutically acceptable salt or the stereoisomer thereof is used as a DYRK2 inhibitor to inhibit the activity of the 26s proteasome of the human body;

preferably said compound or a pharmaceutically acceptable salt or stereoisomer thereof acts by blocking the cell cycle of the tumour cell;

the tumors comprise hematological tumors and solid tumors; preferably, the hematological tumor is one or more of multiple myeloma (including multiple myeloma resistant to proteasome inhibitor 'velcade'), leukemia, chronic lymphoma, mantle cell lymphoma and Fahrenheit megasphaemia; preferably, the solid tumor is breast cancer.

The combination effect of the compound and the proteasome inhibitor in the invention is better than that of the current clinical commonly used treatment scheme (such as combining the proteasome inhibitor and the adrenocortical hormone dexamethasone, combining the proteasome inhibitor and the immunomodulator thalidomide and the like).

When the compound, the derivative and the pharmaceutically acceptable salt thereof are used for treating and relieving tumors, ideal application effects are generally shown.

When used, the compound and its pharmaceutically acceptable salt or stereoisomer are administered to a patient in an effective amount to treat, alleviate or prevent the above diseases, and the specific form can adopt various known means, which is not particularly limited in the present invention.

Herein, "treating" or "alleviating" refers to administering at least one compound disclosed herein and/or at least one derivative and/or at least one pharmaceutically acceptable salt thereof to a subject in need thereof identified, e.g., the subject has a tumor.

Herein "effective amount" refers to an amount of at least one compound and/or at least one derivative thereof, and/or at least one pharmaceutically acceptable salt thereof disclosed herein that is effective to "treat" (as defined above) a disease or disorder in a subject.

The beneficial effects of the invention at least comprise:

(1) the compound provided by the invention shows good inhibitory activity on proteasome regulatory kinase DYRK2, and further achieves anti-tumor activity by inhibiting proteasomes.

(2) The synthetic method of the invention adopts a chemical total synthetic route, and the convergent synthetic route can be applied to the chemical synthesis of compounds with similar structures and related derivatives, thereby opening up a wide development space for novel antitumor drugs.

Drawings

FIG. 1 shows the proteasome inhibitory activity of compounds measured at the cellular level;

FIG. 2 shows the combined effect of a cell level detection compound and a first-line clinical anti-tumor drug.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

The compounds disclosed herein, or pharmaceutically acceptable salts thereof, can be synthesized from commercially available starting materials along with the disclosure herein. The following schemes describe some of the methods for the preparation of the compounds disclosed in the present invention.

The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products available from regular distributors, not indicated by the manufacturer.

EXAMPLE 1 Synthesis of Compounds I-1, I-2, I-3, I-4

I-1 is synthesized according to the following steps:

(1) synthesis of Compound 3:

to commercially available starting materials 1(9.26g, 40mmol), 2(6.90g, 56mmol) were added Cu (0..73g, 11mmol), Cu₂O (0.82g, 5.7mmol), potassium carbonate (7.74g, 56mmol) and DMF 100ml were stirred at 80 ℃ overnight. The reaction solution was cooled to room temperature, and 2M dilute hydrochloric acid was added thereto until the system became acidic, and a large amount of solid was precipitated. The precipitated solid was filtered by suction, washed with water, and dried to obtain compound 3(4.02g, 37%) which was directly used in the subsequent reaction without purification.

(2) Synthesis of Compound 4:

the compound 3(2.58g, 9.45mmol) is placed in a sealed tube, 30ml of phosphorus oxychloride is added under the protection of argon, and the reaction is carried out for 8h at 130 ℃. After the reaction was completed, the reaction solution was cooled slightly, poured onto ice and stirred sufficiently, and a large amount of yellow solid was precipitated. And (3) carrying out suction filtration, washing and drying on the precipitated solid to obtain the compound 4(1.65g, quant.) which can directly participate in the subsequent reaction without purification.

(3) Synthesis of Compound I-1:

to compound 4(50mg, 0.183mmol) and 67% sodium hydrosulfide hydrate (22.9mg, 0.274mmol) was added DMF5ml under argon protection, and the mixture was reacted at 50 ℃ for 2 h. Subsequently, N-Boc-3-aminopropylbromide (60.9mg, 0.274mmol) and potassium carbonate (50.5mg, 0.365mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Separation and purification by silica gel column (dichloromethane/ethyl acetate ═ 90/10) gave compound I-1(47.4mg, 61%).

The characterization information of the compound I-1 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.10(d,J＝9.3Hz,2H),7.94(s,2H),7.42(d,J＝9.3Hz,2H),4.04(s,6H),3.20(d,J＝5.9Hz,2H),2.95(t,J＝7.2Hz,2H),1.70–1.61(m,2H),1.39(s,9H)

this example also synthesizes Compound I-2, the specific procedures and reaction conditions being the same as those for the synthesis of I-1 above, except that the corresponding bromide is replaced, specifically: the bromide used for the synthesis of compound I-2 was N- (3-bromopropyl) acetamide, and the following is the characterization information for compound I-2:

the characterization information of the compound I-2 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.10(d,J＝9.4Hz,2H),7.94(d,J＝2.7Hz,2H),7.42(dd,J＝9.4,2.8Hz,2H),4.04(s,6H),3.30(dd,J＝13.2,6.7Hz,2H),2.96(t,J＝7.3Hz,2H),1.88(s,3H),1.69–1.59(m,2H).

this example also synthesizes Compound I-3, the specific procedures and reaction conditions being the same as those for the synthesis of I-1 above, except that the corresponding bromide is replaced, specifically: the bromide used for the synthesis of compound I-3 was N- (4-bromobutyl) acetamide, the following characterizing information for compound I-3:

the characterization information of the compound I-3 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.10(d,J＝9.3Hz,2H),7.95(d,J＝2.7Hz,2H),7.42(dd,J＝9.3,2.7Hz,2H),4.04(s,6H),3.15(dd,J＝13.1,6.7Hz,2H),2.94(t,J＝7.1Hz,2H),1.86(s,3H),1.59(m,2H),1.49(m,2H).

this example also synthesizes Compound I-4, the specific procedures and reaction conditions being the same as those for the synthesis of I-1 above, except that the corresponding bromide is replaced, specifically: the bromide used for the synthesis of compound I-4 is 4-bromobutyronitrile, and the following is the characterization information for compound I-4:

the characterization information of the compound I-4 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.11(d,J＝9.4Hz,2H),7.89(d,J＝2.7Hz,2H),7.43(dd,J＝9.4,2.8Hz,2H),4.04(s,6H),3.07(t,J＝7.0Hz,2H),2.47(t,J＝7.0Hz,2H),1.82–1.72(m,2H)。

EXAMPLE 2 Synthesis of Compounds I-5, I-6

I-5 was synthesized according to the following procedure:

to compound 4(50mg, 0.183mmol) and 67% sodium hydrosulfide hydrate (22.9mg, 0.274mmol) was added 5ml of dry DMF under an argon atmosphere, and the mixture was reacted at 50 ℃ for 2 hours. Thereafter, tert-butyl 4-bromopiperidine-1-carboxylate (72.4mg, 0.274mmol) and potassium carbonate (50.5mg, 0.365mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. The resulting crude product was put into 2ml of a dichloromethane solution containing 5% trifluoroacetic acid, reacted at room temperature for 2 hours until no starting material remained, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave Compound I-5(15.1mg, 23%)

The characterization information of the compound I-5 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.05(d,J＝9.4Hz,2H),8.00(d,J＝2.7Hz,2H),7.49(dd,J＝9.4,2.8Hz,2H),4.05(s,6H),3.36-3.33(m,1H),2.99(d,J＝6.8Hz,2H),2.85(td,J＝12.8,2.9Hz,2H),2.13(d,J＝16.5Hz,2H),1.70-1.65(m,2H).

this example also synthesizes Compound I-6, the specific procedures and reaction conditions being the same as those for the synthesis of I-5 above, except that the corresponding bromide is replaced, specifically: the bromide used for the synthesis of compound I-6 was 1-BOC-4-bromomethylpiperidine, and the following is the characterization information for compound I-6:

the characterization information of the compound I-6 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.24(d,J＝9.4Hz,2H),8.17(d,J＝2.3Hz,2H),7.85(dd,J＝9.4,2.4Hz,2H),4.14(s,6H),3.75-3.68(m,1H),3.39(d,J＝13.3Hz,2H),3.01(t,J＝10.9Hz,2H),2.18–2.09(m,2H),2.05–2.01(m,2H),1.37–1.32(m,2H).

EXAMPLE 3 Synthesis of Compounds I-7, I-8, I-9, I-10, I-11

I-10 was synthesized according to the following procedure:

(1) synthesis of compounds 5, 7:

to a suspension of compound 5(1.65g,6.05mmol) in 200ml of dried methylene chloride was slowly added 30ml of boron tribromide, and the mixture was reacted at 0 ℃ for 2 hours. Adding methanol into the reaction system to quench the reaction, and spin-drying the solvent under reduced pressure. The crude product was purified by silica gel column separation (dichloromethane/methanol-92/8%) to give product 5(0.22g, 14%) and reported compound 7(0.80g, 56%) with recovery of 0.70g of starting material.

The characterization information of compound 5 is specifically:

¹H NMR(400MHz,MeOD)δ8.08(dd,J＝9.4,4.8Hz,2H),7.64(dd,J＝7.6,4.4Hz,4H),4.07(s,3H).

(2) synthesis of Compound 6:

to compound 2(94.5mg, 0.364mmol), potassium carbonate (75.4mg,0.546mmol) and silver oxide (126.5mg,0.546mmol) under argon protection was added dried DMF10ml, then 64.5. mu.l benzyl bromide was added and the reaction was carried out at 40 ℃ for 2h until no starting material remained. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Separation and purification by silica gel column (petroleum ether/ethyl acetate 90/10) gave compound 6(126.9mg, quant.).

The characterization information of compound 6 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.08(dd,J＝9.3,7.6Hz,2H),7.60(d,J＝2.6Hz,1H),7.54(d,J＝7.1Hz,2H),7.51–7.35(m,6H),5.27(s,2H),4.02(s,3H).

(3) synthesis of compound 6 a:

to compound 8a (52.2mg, 0.364mmol) and 70% sodium hydrosulfide hydrate (13.15mg, 0.164mmol) was added DMF5ml under argon protection and reacted at 50 ℃ for 2 h. Subsequently, N-Boc-3-aminopropylbromide (53.3mg, 0.224mmol) and potassium carbonate (30.9mg, 0.224mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Purification by flash column chromatography on silica gel (dichloromethane/ethyl acetate-95/5) gave compound 6a, which was directly charged into the next reaction.

(4) Synthesis of Compound I-7:

the compound 6a was put into 2ml of a dichloromethane solution containing 5% trifluoroacetic acid, reacted at room temperature for 2 hours until no starting material remained, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave Compound I-7(8.6mg, 9.6%)

The characterization information of the compound I-7 is specifically as follows:

¹H NMR(400MHz,MeOD)δ8.13(dd,J＝12.1,9.4Hz,2H),8.01(dd,J＝8.5,2.6Hz,2H),7.76(ddd,J＝35.9,9.4,2.6Hz,2H),7.60-7.34(m,5H),5.42(s,2H),4.08(s,3H),3.00(t,J＝7.6Hz,2H),2.85(t,J＝7.6Hz,2H),1.70(quint,J＝7.6Hz,2H).

this example also synthesizes Compound I-8, the specific procedures and reaction conditions being the same as those for the synthesis of I-7 above, except that the corresponding bromide is replaced, specifically: the bromide used for the synthesis of compound I-8 was tert-butyl 4-bromopiperidine-1-carboxylate, and the following information characterizes compound I-8:

the characterization information of the compound I-8 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.01(dd,J＝14.3,9.4Hz,2H),7.90(d,J＝2.4Hz,2H),7.55(m,3H),7.49–7.42(m,3H),7.38(m,1H),5.38(s,2H),4.03(s,3H),3.28(d,J＝13.1Hz,2H),3.21–3.12(m,1H),2.83(t,J＝10.8Hz,2H),1.88(d,J＝11.5Hz,2H),1.72(td,J＝14.4,3.7Hz,2H).

this example also synthesizes Compound I-9, the specific procedures and reaction conditions being the same as those for the synthesis of I-7 above, except that the corresponding bromide is replaced, specifically: the bromide used for the synthesis of compound I-9 was 1-BOC-4-bromomethylpiperidine, the following characterizing information for compound I-9:

the characterization information of the compound I-9 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.04–7.91(m,2H),7.83(s,2H),7.53(dd,J＝20.5,8.0Hz,3H),7.47–7.32(m,4H),5.34(s,2H),4.00(s,3H),3.21(d,J＝12.5Hz,2H),2.71-2.69(m,3H),1.95(d,J＝12.5Hz,2H),1.35(dd,J＝30.8,18.6Hz,4H).

this example also synthesizes Compound I-10, using the same specific procedures and reaction conditions as those described above for the synthesis of I-7, except that the corresponding bromide was replaced during the alkylation of intermediate 5, specifically: the bromide used for the synthesis of compound I-10 was isopropyl bromide, and the following is the characterization information for compound I-10:

the characterization information of the compound I-10 is specifically as follows:

¹H NMR(400MHz,MeOD)δ8.15(dd,J＝9.4,3.6Hz,2H),8.06(t,J＝3.0Hz,2H),7.71(td,J＝9.3,2.6Hz,2H),4.98–4.92(m,1H),4.09(s,3H),3.36(d,J＝12.9Hz,2H),3.11(d,J＝6.8Hz,2H),2.88(t,J＝11.9Hz,2H),2.12(d,J＝14.0Hz,2H),1.79–1.69(m,1H),1.54-1.41(m,2H),1.51(s,3H),1.49(s,3H).

in this example, compound I-11 was synthesized, and the specific steps and reaction conditions were the same as those for the synthesis of I-9, except that the starting materials were different, specifically: the starting material for the synthesis of compound I-11 was compound 7, and the following was the characterization information of compound I-11:

the characterization information of the compound I-11 is specifically as follows:

¹H NMR(400MHz,MeOD)δ8.09(d,J＝9.4Hz,2H),7.85(d,J＝2.5Hz,2H),7.73(dd,J＝9.4,2.6Hz,2H),7.43(ddd,J＝28.6,28.0,7.3Hz,10H),5.37(s,4H),3.22(d,J＝12.8Hz,2H),2.73–2.62(m,4H),1.75(d,J＝13.6Hz,2H),1.41–1.31(m,1H),1.25(dt,J＝21.6,7.6Hz,2H).

EXAMPLE 4 Synthesis of Compounds I-12, I-13, I-18, I-19

I-12 was synthesized according to the following procedure:

(1) synthesis of Compound I-12:

to a solution of the hydrochloride of the compound I-6 (4.4mg,0.012mmol) in dry dichloromethane 1ml was slowly added 1.8. mu.l of acryloyl bromide and 4.5. mu.l of triethylamine, and reacted at room temperature for 1.5 h. The solvent was dried under reduced pressure, and the crude product was isolated and purified by silica gel column separation (dichloromethane/ethyl acetate 70/30) to give product I-12(2.2mg, 44%).

The characterization information of the compound I-12 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.10(d,J＝9.3Hz,2H),7.91(s,2H),7.42(d,J＝9.3Hz,2H),6.53(dd,J＝16.8,10.6Hz,1H),6.24(d,J＝16.8Hz,1H),5.65(d,J＝10.5Hz,1H),4.64(d,J＝12.3Hz,1H),4.02(s,6H),3.94(d,J＝12.7Hz,1H),2.91(t,J＝12.4Hz,1H),2.83(d,J＝6.7Hz,2H),2.52(t,J＝12.5Hz,1H),1.94(dd,J＝46.4,12.4Hz,2H),1.57(d,J＝3.7Hz,1H),1.33–1.12(m,3H).

(2) synthesis of Compound I-13:

to 10ml of a dry acetonitrile solution of the hydrochloride of the compound I-6 (86mg,0.213mmol) and 4- (bromomethyl) benzenesulfonyl fluoride (107.5mg, 0.425mmol) was added slowly 0.12ml of diisopropylamine, and the mixture was reacted at 80 ℃ for 5 hours. The solvent was dried under reduced pressure, and the crude product was separated and purified by silica gel column (dichloromethane/ethyl acetate 90/10-50/50) to obtain product I-13(22mg, 19%).

The characterization information of the compound I-13 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.10(d,J＝9.4Hz,2H),7.95(d,J＝2.8Hz,2H),7.93(d,J＝8.5Hz,2H),7.56(d,J＝8.2Hz,2H),7.42(dd,J＝9.3,2.8Hz,2H),4.02(s,6H),3.53(s,2H),2.84(d,J＝5.6Hz,2H),2.78(d,J＝11.3Hz,2H),1.94-1.86(m,4H),1.70(br s,1H),1.35-1.31(m,2H).

the compound 4- (bromomethyl) benzenesulfonyl fluoride can be synthesized by the following method:

commercially available p-toluenesulfonyl fluoride (250mg, 1.43mmol), N-bromosuccinimide (306mg, 1.72mmol), dibenzoyl peroxide (17.4mg, 0.072mmol) were dissolved in 4ml dry ethyl acetate and reacted at 80 ℃ for 1h under microwave. After the system was cooled, the solvent was dried by evaporation under reduced pressure, and the crude product was isolated and purified by silica gel column (petroleum ether/ethyl acetate ═ 5:2) to give 4- (bromomethyl) benzenesulfonyl fluoride (136mg, 37%).

The characterization information of the compound 4- (bromomethyl) benzenesulfonyl fluoride is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.00(d,J＝8.4Hz,2H),7.65(d,J＝8.3Hz,2H),4.52(s,2H).

in this example, compound I-18 was synthesized, the specific steps and reaction conditions were the same as those for the synthesis of I-12 above, except that the corresponding starting materials were replaced, specifically: the starting materials for the synthesis of compound I-18 were 1 to 9, and the following is the characterization information of compound I-1:

the characterization information of the compound I-18 is specifically:

1H NMR(400MHz,CDCl₃)δ8.11(dd,J＝12.1,9.4Hz,2H),7.93(dd,J＝13.2,2.7Hz,2H),7.56–7.49(m,3H),7.46–7.33(m,4H),6.52(dd,J＝16.8,10.6Hz,1H),6.24(dd,J＝16.8,1.9Hz,1H),5.65(dd,J＝10.6,1.9Hz,1H),5.32(s,2H),4.60(d,J＝12.7Hz,1H),4.02(s,3H),3.90(d,J＝12.8Hz,1H),2.87(t,J＝12.9Hz,1H),2.65(dd,J＝6.8,1.8Hz,2H),2.49(t,J＝12.1Hz,1H),1.85(dd,J＝36.0,12.1Hz,2H),1.52-1.46(m,1H),1.23–1.06(m,2H).

in this example, compound I-19 was synthesized, the specific steps and reaction conditions were the same as those for the synthesis of I-13 above, except that the corresponding raw materials were replaced, specifically: the starting materials for the synthesis of compound I-19 were 1-9, and the following is the characterization information for compound I-19:

the characterization information of the compound I-19 is specifically:

1H NMR(400MHz,CDCl₃)δ8.10(dd,J＝11.1,9.4Hz,2H),8.00–7.88(m,4H),7.58–7.47(m,5H),7.45–7.32(m,4H),5.30(s,2H),4.00(s,3H),3.51(s,2H),3.48(s,1H),2.75(d,J＝10.9Hz,2H),2.68(d,J＝5.2Hz,2H),2.16(s,1H),1.92–1.73(m,5H).

EXAMPLE 5 Synthesis of Compounds I-14, I-15

I-14 was synthesized according to the following procedure:

(1) synthesis of Compound I-14:

the known compound, 4-acrylamidobutyric acid (50mg, 0.318mmol), was dissolved in 5ml of dry DMF, HATU (132mg, 0.347mmol) and triethylamine (0.12 ml) were added thereto, and the reaction was carried out at room temperature for 40 min; then, hydrochloride salt of I-6 (117.2mg, 0.289mmol) was added and reacted at room temperature overnight. Diluting with dichloromethane, washing with saturated saline twice, mixing organic phases, drying with anhydrous sodium sulfate, filtering, and spin-drying the solvent under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC 1817. mu. m2.1X50mm column) gave Compound I-14(22.1mg, 15%) with the characterisation information of Compound I-14 specified as:

¹H NMR(400MHz,CDCl₃)δ8.09(d,J＝9.3Hz,2H),7.91(d,J＝2.2Hz,2H),7.41(dd,J＝9.3,2.4Hz,2H),6.54(br s,1H),6.22(d,J＝16.8Hz,1H),6.06(dd,J＝17.0,10.2Hz,1H),5.58(d,J＝10.1Hz,1H),4.57(d,J＝13.1Hz,1H),4.02(s,6H),3.77(t,J＝13.2Hz,1H),3.35(q,J＝5.6Hz,2H),2.85(t,J＝12.4Hz 1H),2.82(d,J＝6.7Hz,2H),2.43(t,J＝12.4Hz 1H),2.37(t,J＝6.6Hz,2H),1.89–1.81(m,4H),1.52-1.46(m,1H),1.22–1.07(m,2H).

this example also synthesizes Compound I-15, the specific procedures and reaction conditions being the same as those for the synthesis of I-14 above, except that the corresponding carboxylic acid is replaced, specifically: the carboxylic acid used for the synthesis of compound I-15 was 5-acrylamidopentanoic acid, and the following is the characterization information for compound I-15:

the characterization information of the compound I-15 is specifically as follows:

1H NMR(400MHz,CDCl3)δ8.09(d,J＝9.4Hz,2H),7.91(d,J＝2.7Hz,2H),7.42(dd,J＝9.4,2.8Hz,2H),6.30(br s,1H),6.25(dd,J＝17.0,1.5Hz,1H),6.09(dd,J＝17.0,10.2Hz,1H),5.60(dd,J＝10.2,1.5Hz,1H),4.58(d,J＝13.4Hz,1H),4.02(s,6H),3.78(d,J＝13.6Hz,1H),3.31(q,J＝6.3Hz,2H),2.85(t,J＝12.4Hz 1H),2.83(d,J＝6.8Hz,2H),2.43(td,J＝12.9,2.5Hz,1H),2.31(t,J＝7.0Hz,2H),1.98-1.86(m,4H),1.60–1.50(m,3H),1.23-1.08(m,2H).

EXAMPLE 6 Synthesis of Compounds I-16, I-17

I-16 was synthesized according to the following procedure:

(1) synthesis of compound 10:

2-bromo-5-hydroxybenzaldehyde (1.32g, 6.57mmol) was dissolved in 13ml of 0.5M sodium hydroxide solution and heated to boiling; to the boiling solution was added dropwise 13ml of a 0.5M sodium hydroxide solution of commercially available Compound 9(1.00g, 6.57mmol) and reacted for 3 hours under reflux. The reaction system is cooled to room temperature and neutralized to acidity with hydrochloric acid, and a large amount of solid is precipitated. Filtering to remove solid, diluting with n-pentane, washing with water for 2 times, combining organic phases, drying with anhydrous sodium sulfate, filtering, and spin-drying the solvent under reduced pressure. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate: 2/1) to obtain compound 10(0.21g, 12%)

The characterization information for compound 10 is specifically: 1H NMR (400MHz, CDCl3) δ 10.31(s,1H),7.54(d, J ═ 8.8Hz,1H),7.43(d, J ═ 3.2Hz,1H),7.04(dd, J ═ 8.8,3.2Hz,1H),4.29(t, J ═ 6.1Hz,2H),2.87(t, J ═ 6.1Hz,2H).

(2) Synthesis of compound 11:

compound 10(40.0mg, 0.147mmol) was dissolved in 2ml of dry dichloromethane and 18.6. mu.l of oxalyl chloride and one drop of DMF were added and reacted at room temperature for 2 h. The solvent was dried under reduced pressure, and the crude product was directly added to hydrochloride salt of 1-6 (58mg, 0.144mmol), 2ml of dried dichloromethane and 44. mu.l triethylamine were added, and reacted at room temperature for 3 hours. Diluting with dichloromethane, washing with saturated saline twice, mixing organic phases, drying with anhydrous sodium sulfate, filtering, and spin-drying the solvent under reduced pressure. Purification by flash silica gel column (dichloromethane/ethyl acetate 95/5) gave compound 11(69mg, 77%) which was directly charged to the next reaction.

(3) Synthesis of Compound I-16:

compound 11(31.4mg, 0.051mmol), pinacoldiboron ester (25.6mg, 0.102mmol), dppf palladium dichloride (3.7mg, 0.005mmol), potassium acetate (14.8mg, 0.153mmol) were dissolved in 1ml of dry 1,4 dioxane under argon protection and reacted at 100 ℃ for 2 h. The solvent was dried under reduced pressure and the crude product was taken up in 2M methanolic hydrochloric acid and reacted at 70 ℃ overnight. The solvent was spun dry under reduced pressure and purified by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) to give Compound I-16(2.8mg, 9.4%) the characterization information for Compound I-16 specifically:

1H NMR(400MHz,CDCl3)δ9.85(s,1H),8.63(d,J＝9.4Hz,2H),8.16(d,J＝8.4Hz,1H),7.97(d,J＝2.6Hz,2H),7.72(dd,J＝9.5,2.6Hz,2H),7.40(d,J＝2.6Hz,1H),7.16(dd,J＝8.3,2.6Hz,1H),4.68(d,J＝12.9Hz,1H),4.44–4.38(m,2H),4.08(s,6H),3.91(d,J＝15.2Hz,1H),3.00-2.95(m,3H),2.84(t,J＝6.6Hz,2H),2.51(t,J＝12.9Hz,1H),2.03–1.88(m,2H),1.70-1.60(m,1H),1.24-1.18(m,2H).

in this example, compound I-17 was synthesized, the specific steps and reaction conditions were the same as those for the synthesis of I-16 above, except that the corresponding benzaldehyde was replaced, specifically: synthesis of Compound I-17 was performed using 2-bromo-4-hydroxybenzaldehyde, the following characterizing information for Compound I-17:

the characterization information of the compound I-17 is specifically:

¹H NMR(400MHz,CDCl₃)δ9.74(d,J＝5.9Hz,1H),8.64(d,J＝9.4Hz,2H),7.97(d,J＝2.5Hz,2H),7.84(d,J＝8.5Hz,1H),7.74(d,J＝2.6Hz,1H),7.71(dd,J＝9.4,2.6Hz,2H),7.11(dd,J＝8.5,2.7Hz,1H),4.68(d,J＝13.3Hz,1H),4.44(dd,J＝12.2,6.2Hz,2H),4.07(s,6H),3.93(d,J＝12.6Hz,1H),3.00-2.96(m,3H),2.84(dd,J＝12.4,6.3Hz,2H),2.51(t,J＝12.9Hz,1H),2.02-1.88(d,J＝13.1Hz,1H),1.90(d,J＝12.4Hz,1H),1.70-1.60(m,1H),1.28-1.22(m,4H).

EXAMPLE 7 Synthesis of Compounds I-20, I-21, I-22, I-23, I-24

I-23 was synthesized according to the following procedure:

(1) synthesis of compound 3':

to commercially available raw materials 1 '(2.00 g, 8.66mmol), 2' (1.59g, 9.52mmol) were added Cu (0.11g, 1.73mmol), Cu₂O (0.12g, 0.87mmol), potassium carbonate (3.59g, 25.97mmol) and 15ml of DMF were stirred at 80 ℃ overnight. The reaction solution was cooled to room temperature, and 2M dilute hydrochloric acid was added thereto until the system became acidic, and a large amount of solid was precipitated. Will separateThe obtained solid was filtered, washed with water and dried to obtain compound 3(2.00g, 73%), which was used in the subsequent reaction without purification.

(2) Synthesis of compound 4':

the compound 3' (1.00g, 3.16mmol) is put into a sealed tube, 10ml of phosphorus oxychloride is added under the protection of argon, and the reaction is carried out for 8h at 130 ℃. After the reaction was completed, the reaction solution was cooled slightly, poured onto ice and stirred sufficiently, and a large amount of yellow solid was precipitated. The precipitated solid was filtered, washed with water and dried to obtain compound 4(1.00g, 95%) which was used in the subsequent reaction without purification.

(3) Synthesis of compound 12:

0.40ml of oxalyl chloride was added dropwise to a 10ml suspension of compound 4' (0.89g, 2.67mmol) in dry dichloromethane, and 1 drop of DMF was added thereto, to release a large amount of bubbles; reacting at room temperature for 0.5h until the system becomes a brown-black solution. The mixture was quenched for 2h by adding dry methanol and neutralized by adding triethylamine. Diluting with dichloromethane, washing with saturated saline twice, mixing organic phases, drying with anhydrous sodium sulfate, filtering, and spin-drying the solvent under reduced pressure. Separation and purification by silica gel column (dichloromethane/ethyl acetate 95/5) gave compound 12(0.74g, 80%).

The characterization information of compound 12 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.12(d,J＝9.3Hz,1H),7.70(s,1H),7.64(s,1H),7.46(s,1H),7.42(d,J＝9.4Hz,1H),4.09(s,3H),4.03(s,6H).

(4) synthesis of compound 13:

to compound 12(0.38g, 1.16mmol) and 70% sodium hydrosulfide hydrate (0.10g, 1.21mmol) was added dry DMF10ml and reacted at 50 ℃ for 2 h. Then, 1-BOC-4-bromomethylpiperidine (0.64g, 2.31mmol) and potassium carbonate (0.40g, 2.89mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate 95/5) gave compound 13(0.38g, 63%).

The characterization information of compound 13 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.13(d,J＝9.4Hz,1H),8.08(d,J＝2.0Hz,1H),7.89(s,1H),7.68(d,J＝1.9Hz,1H),7.41(dd,J＝9.4,1.9Hz,1H),4.09(s,3H),4.02(s,6H),2.78(d,J＝6.7Hz,2H),2.55(t,J＝12.2Hz,2H),1.82(br s,2H),1.45-1.35(m,3H),1.43(s,9H),1.20-1.09(m,2H)

(5) synthesis of compound 14:

compound 13(348mg, 0.661mmol) was dissolved in 10ml of dry dichloromethane, and 3.1ml of 1.5M diisopropylaluminum hydride was added thereto and reacted at room temperature for 2 hours. Adding saturated potassium hydrogen tartrate solution to quench reaction, adding dichloromethane for dilution, separating by water/dichloromethane, separating organic phase, drying with anhydrous sodium sulfate, and spin-drying solvent under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate 80/20) gave compound 14(180mg, 55%).

The characterization information for compound 14 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.03(d,J＝9.3Hz,1H),7.88(s,1H),7.81(s,1H),7.39(d,J＝9.3Hz,1H),7.27(s,1H),5.41(s,1H),5.21(s,2H),4.02(s,3H),4.00(s,3H)2.79(d,J＝6.7Hz,2H),2.57(t,J＝11.7Hz,2H),1.84(br s,2H),1.47-1.25(m,3H),1.43(s,9H),1.20-1.09(m,2H).

(6) synthesis of Compound I-20:

compound 14(24.2mg, 0.048mmol) was dissolved in 5ml of 5% trifluoroacetic acid in dichloromethane, reacted at room temperature for 2h, and the solvent was dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound I-20 (11.2mg, 45%) characterization information for compound I-20 specifically:

¹H NMR(400MHz,MeOD)δ8.09(d,J＝9.4Hz,1H),7.76(d,J＝2.2Hz,1H),7.71(d,J＝2.2Hz,1H),7.54(d,J＝1.1Hz,1H),7.50(dd,J＝9.4,2.5Hz,1H),5.23(s,2H),4.04(s,3H),4.03(s,3H),3.33–3.30(m,2H),2.95(d,J＝6.7Hz,2H),2.83(td,J＝12.8,1.6Hz,2H),2.07-1.99(m,2H),1.66–1.55(m,1H),1.51–1.40(m,2H).

this example also synthesizes Compound I-21, the specific procedures and reaction conditions being the same as those for the synthesis of I-20 above, except that the corresponding bromide is replaced, specifically: synthesis of Compound I-21 was performed using N-Boc-3-aminopropylbromide, and the following is the characterization information for Compound I-21:

the characterization information of the compound I-21 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.13(d,J＝9.4Hz,1H),7.94(d,J＝26.6Hz,2H),7.58–7.54(m,1H),7.50(dd,J＝9.4,2.7Hz,1H),5.31(s,2H),4.05(s,3H),4.05(s,3H),3.09(t,J＝7.4Hz,2H),2.95–2.88(t,J＝7.4Hz,2H),1.79–1.70(m,2H).

this example also synthesizes Compound I-22, the specific procedures and reaction conditions being the same as those described above for the synthesis of I-20, except that the corresponding bromide is replaced, specifically: synthesis of Compound I-22 was performed using N-Boc-3(S) -bromomethylpyrrolidine, the following characterization information for Compound I-22:

the characterization information of the compound I-22 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.11(d,J＝9.4Hz,1H),7.81(s,1H),7.75(s,1H),7.57(s,1H),7.51(d,J＝9.4Hz,1H),5.27(s,2H),4.07(s,3H),4.07(s,3H),3.36–3.27(m,2H),3.19–3.05(m,3H),2.96(dd,J＝11.4,8.3Hz,1H),2.25(hept,J＝7.6Hz,1H),2.17–2.05(m,1H),1.80–1.70(m,1H).

this example also synthesizes Compound I-23, the specific procedures and reaction conditions being the same as those for the synthesis of I-20 above, except that the corresponding bromide is replaced, specifically: synthesis of Compound I-23 was performed using N-Boc-3(R) -bromomethylpyrrolidine, the following characterization information for Compound I-23:

the characterization information of the compound I-23 is specifically as follows:

¹H NMR(400MHz,MeOD)δ8.14(d,J＝9.3Hz,1H),7.96(s,1H),7.89(s,1H),7.58(s,1H),7.53–7.48(m,1H),5.32(s,2H),4.06(s,6H),3.35–3.26(m,2H),3.20–3.08(m,3H),2.97–2.92(m,1H),2.26(hept,J＝7.6Hz,1H),2.17–2.07(m,1H),1.80–1.71(m,1H).

this example also synthesizes Compound I-24, the specific procedures and reaction conditions being the same as those for the synthesis of I-20 above, except that the corresponding bromide is replaced, specifically: 1-BOC-3- (bromomethyl) azetidine was used for the synthesis of compound I-24, and the following is the characterization information for compound I-24:

the characterization information of the compound I-24 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.07(d,J＝9.4Hz,1H),7.80(d,J＝2.2Hz,1H),7.73(d,J＝2.1Hz,1H),7.53(s,1H),7.46(dd,J＝9.3,2.2Hz,1H),5.28(s,2H),4.04(s,3H),4.03(s,3H),3.82(t,J＝9.7Hz,2H),3.69–3.62(m,2H),3.26(d,J＝7.9Hz,2H),2.71–2.58(m,1H).

EXAMPLE 8 Synthesis of Compound I-25

I-28 was synthesized according to the following procedure:

compound 13(42mg, 0.080mmol) was dissolved in 2ml of tetrahydrofuran, and 0.4ml of an aqueous solution of 1M lithium hydroxide was added thereto to conduct a reaction at 40 ℃ for 20 hours. After cooling to room temperature, 5ml of a 10% trifluoroacetic acid solution in methylene chloride was added to the system, and the reaction was carried out at room temperature for 6 hours. The solvent was spin dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound I-25 (21.6mg, 51%).

The characterization information of the compound I-25 is specifically as follows:

1H NMR(600MHz,MeOD)δ8.39(d,J＝2.8Hz,1H),8.23(d,J＝2.7Hz,1H),8.12(d,J＝9.3Hz,1H),7.97(d,J＝2.5Hz,1H),7.62(dd,J＝9.3,2.6Hz,1H),4.09(s,3H),4.07(s,3H),3.36-3.33(m,2H),3.02(d,J＝6.8Hz,2H),2.89–2.82(m,2H),2.11(d,J＝14.0Hz,2H),1.70–1.64(m,1H),1.50–1.41(m,2H).

EXAMPLE 9 Synthesis of Compound I-26

I-29 was synthesized according to the following procedure:

(1) synthesis of compound 15:

mu.l of oxalyl chloride was dissolved in 1ml of dry tetrahydrofuran, cooled to-78 deg.C, 25.6. mu.l of dimethyl sulfoxide was slowly added to the system, and the reaction was carried out at-78 deg.C for 30min, then 1ml of tetrahydrofuran solution of compound 14(60mg, 0.12mmol) was added to the system, and the reaction was continued at-78 deg.C for 1h, after which 0.10ml of triethylamine was added, and the reaction was slowly returned to room temperature for 2 h. The solvent was dried under reduced pressure, and the residue was separated and purified by silica gel column chromatography (dichloromethane/ethyl acetate 95/5) to obtain compound 15(56mg, 94%).

The characterization information of compound 15 is specifically:

¹H NMR(400MHz,CDCl₃)δ11.54(s,1H),8.19(d,J＝2.9Hz,1H),8.09(d,J＝9.3Hz,1H),7.98(d,J＝2.9Hz,1H),7.86(d,J＝2.6Hz,1H),7.44(dd,J＝9.4,2.7Hz,1H),4.10-3.98(m,2H),4.04(s,3H),4.03(s,3H),2.80(d,J＝6.8Hz,2H),2.58(t,J＝12.2Hz,2H),1.83(m,2H),1.52–1.44(m,1H),1.43(s,9H),1.23-1.08(m,2H).

(2) synthesis of compound 16:

to 60mg of a 4A molecular sieve activated with compound 15(55mg, 0.11mmol), ammonium acetate (85mg, 1.11mmol) and sodium cyanoborohydride (6.9mg, 0.11mmol) was added 3ml of dry methanol under an argon atmosphere, and the mixture was reacted at-5 ℃ overnight. The solid was removed by filtration, diluted with dichloromethane, washed twice with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the solvent was spin-dried under reduced pressure. Separation and purification on a silica gel column (dichloromethane/methanol-95/5) gave compound 16(25mg, 46%).

The characterization information of compound 16 is specifically:

¹H NMR(400MHz,CDCl₃)δ7.54(d,J＝2.6Hz,1H),7.52(d,J＝2.5Hz,1H),7.30(d,J＝9.3Hz,1H),7.08(d,J＝2.3Hz,1H),6.90(dd,J＝9.3,2.6Hz,1H),4.86(s,2H),4.06–3.95(m,2H),4.04(s,3H),3.96(s,3H),2.62(d,J＝6.7Hz,2H),2.52(t,J＝12.0Hz,2H),1.85-1.70(m,2H),1.41(s,9H),1.35-1.30(m,1H),1.18-1.04(m,2H).

(3) synthesis of Compound I-26:

compound 16(13.5mg, 0.027mmol) was dissolved in 2ml of 5% trifluoroacetic acid in dichloromethane, reacted at room temperature for 2h, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound I-26 (6.3mg, 45%).

The characterization information of the compound I-26 is specifically as follows:

1H NMR(400MHz,MeOD)δ7.55(dd,J＝18.2,2.6Hz,2H),7.47(d,J＝9.3Hz,1H),7.17(d,J＝2.5Hz,1H),6.95(dd,J＝9.3,2.7Hz,1H),4.14–4.07(m,3H),4.02(s,3H),2.86–2.72(m,4H),2.07–1.98(m,3H),1.57–1.31(m,4H).

EXAMPLE 10 Synthesis of Compounds I-27

I-27 was synthesized according to the following procedure:

(1) synthesis of compound 17:

compound 12(40mg, 0.12mmol) was dissolved in 5ml of dry dichloromethane, cooled to-70 ℃ and 0.28ml of 1M diisopropylaluminum hydride, and reacted at-70 ℃ overnight. Adding saturated potassium hydrogen tartrate solution to quench reaction, adding dichloromethane for dilution, separating by water/dichloromethane, separating organic phase, drying with anhydrous sodium sulfate, and spin-drying solvent under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate 90/10) gave compound 17(31mg, 84%).

The characterization information of compound 17 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.00(d,J＝9.3Hz,1H),7.44–7.34(m,3H),7.27(s,1H),5.28(br s,1H),5.19(s,2H),4.02(s,3H),4.00(s,3H).

(2) synthesis of compound 18:

to compound 17(30mg, 0.10mmol), dess-martin oxidizer (60mg, 0.14mmol) was dissolved in 3ml of dry dichloromethane and reacted at room temperature for 2 h. Dilute with dichloromethane, separate the water/dichloromethane layer, separate the organic layer, dry over anhydrous sodium sulfate, and spin dry the solvent under reduced pressure. The crude product was dissolved in 2ml of dry dichloromethane in a plastic container under argon, 0.1ml of diethylaminosulfur trifluoride was added and the reaction was carried out overnight at room temperature. Sodium bicarbonate was added to dissolve in the quench reaction, diluted with dichloromethane, water/dichloromethane separated, the organic phase separated, dried over anhydrous sodium sulfate, and the solvent was spin dried under reduced pressure. Separation and purification by silica gel column (petroleum ether/ethyl acetate: 5/1) gave compound 18(7.4mg, 23%).

The characterization information for compound 18 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.08(d,J＝9.3Hz,1H),7.91(t,J＝55.4Hz,1H),7.76–7.73(m,1H),7.59(d,J＝2.6Hz,1H),7.48–7.41(m,2H),4.05(s,3H),4.03(s,3H).

(3) synthesis of compound 19:

to compound 18(7.4mg, 0.023mmol) and 70% sodium hydrosulfide hydrate (1.9mg, 0.034mmol) was added dry DMF2ml and reacted at 60 ℃ overnight. Subsequently, 1-BOC-4-bromomethylpiperidine (12.7mg, 0.0461mmol) and potassium carbonate (15.8mg, 0.114mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Separation and purification by silica gel column (petroleum ether/ethyl acetate: 3/1) gave compound 19(5.3mg, 45%).

The characterization information for compound 19 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.11(d,J＝9.4Hz,1H),8.06(d,J＝2.7Hz,1H),7.95(t,J＝55.2Hz,1H),7.91(d,J＝2.7Hz,1H),7.77–7.74(m,1H),7.43(dd,J＝9.4,2.8Hz,1H),4.07-4.00(m,2H),4.05(s,3H),4.03(s,3H),2.81(d,J＝6.8Hz,2H),2.58(t,J＝12.4Hz,2H),1.82(d,J＝11.7Hz,2H),1.44(s,9H),1.42–1.41(m,1H),1.20-1.14(m,2H).

(4) synthesis of Compound I-27:

compound 19(5.3mg, 0.010mmol) was dissolved in 2ml of dichloromethane containing 5% trifluoroacetic acid, reacted at room temperature for 2h, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound I-27 (3.8mg, 70%).

The characterization information of the compound I-27 is specifically:

1H NMR(400MHz,MeOD)δ8.08(d,J＝6.3Hz,1H),7.93(d,J＝2.8Hz,1H),7.89(t,J＝59.6Hz,1H)7.89(d,J＝4.1Hz,1H),7.68(dd,J＝2.8,1.4Hz,1H),7.47(dd,J＝9.4,2.8Hz,1H),4.09-4.05(m,2H),4.06(s,3H),4.04(s,3H),2.96(d,J＝6.7Hz,2H),2.90–2.78(m,2H),2.11(d,J＝14.4Hz,2H),1.68–58(m,1H)，1.52–1.43(m,2H).

EXAMPLE 11 Synthesis of Compound I-28

I-28 was synthesized according to the following procedure:

(1) synthesis of compound 13':

to compound 12(0.10g, 0.30mmol) and 70% sodium hydrosulfide hydrate (0.02g, 0.45mmol) was added dry DMF5ml and reacted at 50 ℃ for 1 h. Subsequently, 70. mu.l of N-Boc-3(fS) -bromomethylpyrrolidine and potassium carbonate (0.12g, 0.90mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate 95/5) gave compound 13' (0.12g, 77%).

The characterization information of compound 13' is specifically:

¹H NMR(400MHz,CDCl₃)δ8.13(d,J＝9.4Hz,1H),8.08(s,1H),7.88(s,1H),7.68(s,1H),7.42(d,J＝8.8Hz,1H),4.10(s,3H),4.04(s,6H),3.63-3.29(m,2H),3.27–2.80(m,4H),2.06(br s,1H),1.91(br s,1H),1.61(br s,1H),1.43(s,9H)。

(2) synthesis of compound 14':

compound 13' (118mg, 0.23mmol) was dissolved in 5ml of dry dichloromethane, and 0.53ml of 1M diisopropylaluminum hydride was added thereto, followed by reaction at room temperature overnight. Adding saturated potassium hydrogen tartrate solution to quench reaction, adding dichloromethane for dilution, separating by water/dichloromethane, separating organic phase, drying with anhydrous sodium sulfate, and spin-drying solvent under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate 80/20) gave compound 14' (76mg, 68%).

The characterization information for compound 14' is specifically:

¹H NMR(400MHz,CDCl₃)δ8.06(d,J＝9.3Hz,1H),7.92(d,J＝2.6Hz,1H),7.85(d,J＝2.3Hz,1H),7.43(dd,J＝9.3,2.5Hz,1H),7.29(s,1H),5.37(s,1H),5.22(s,2H),4.05(s,3H),4.03(s,3H),3.63-3.34(m,2H),3.27–2.84(m,4H),2.15-2.06(m,1H),2.01–1.92(m,1H),1.69-1.61(m,1H),1.43(s,9H).

(3) synthesis of compound 20:

to compound 14' (13mg, 0.027mmol), triphenylphosphine (10.6mg, 0.040mmol), and N-bromosuccinimide (7.2mg, 0.040mmol), 1ml of dried tetrahydrofuran was added, reacted at room temperature for 30min, and the solvent was spin-dried under reduced pressure. Purification by silica gel column separation (pure dichloromethane) gave compound 20(9.6mg, 65%).

The characterization information for compound 20 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.16(d,J＝9.3Hz,1H),7.94(s,1H),7.91(s,1H),7.59(s,1H),7.43(d,J＝9.2Hz,1H),5.31(d,J＝8.9Hz,2H),4.05(s,3H),4.03(s,3H),3.64–3.33(m,2H),3.30–2.84(m,4H),2.12(br s,1H),2.01–1.92(m,1H),1.67-1.60(m,1H),1.43(s,9H).

(4) synthesis of Compound I-28:

to compound 20(5.5mg, 10.0. mu. mol), N' -di-Boc-guanidine (3.1mg, 12.1. mu. mol), potassium carbonate (1.7mg, 12.1. mu. mol) was added 1ml of dry DMF under argon protection and reacted at 50 ℃ for 2.5 h. The solvent was dried under reduced pressure, the crude product was dissolved in 4ml of dichloromethane containing 5% trifluoroacetic acid, reacted at room temperature for 2h, and the solvent was dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound I-28 (2.0mg, 27%).

The characterization information of the compound I-28 is specifically as follows:

¹H NMR(400MHz,MeOD)δ8.11(d,J＝9.4Hz,1H),7.96(d,J＝2.7Hz,1H),7.94(d,J＝2.7Hz,1H),7.52–7.48(m,2H),5.04(s,2H),4.07(s,3H),4.06(s,3H),3.39–3.32(m,2H),3.19–2.98(m,4H),2.28(hept,J＝7.6Hz,1H),2.20–2.09(m,1H),1.83–1.73(m,1H).

EXAMPLE 12 Synthesis of Compounds I-29, I-30

I-29 was synthesized according to the following procedure:

(1) synthesis of compound 21:

compound 12(0.20g, 0.60mmol) was dissolved in 10ml of dry tetrahydrofuran under argon and cooled to-78 ℃.1ml of 3M methyl Grignard reagent was then added dropwise thereto, and the reaction was carried out at-78 ℃ for 2 hours. The reaction was quenched by addition of ammonium chloride, diluted with dichloromethane, separated by water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate and the solvent was spin dried under reduced pressure. Separation and purification by silica gel column (dichloromethane/ethyl acetate ═ 95/5) gave compound 21(0.16g, 81%).

The characterization information of compound 21 is specifically:

¹H NMR(400MHz,CDCl₃)δ8.28(s,1H),8.03(d,J＝9.3Hz,1H),7.48(d,J＝2.6Hz,1H),7.45–7.40(m,2H),7.37(d,J＝2.7Hz,1H),4.03(s,3H),4.02(s,3H),1.81(s,6H).

(2) synthesis of Compound I-29:

to compound 21(0.022g, 0.066mmol) and 67% sodium hydrosulfide hydrate (8.3mg, 0.099mmol) were dissolved in 5ml dry DMF under argon protection and reacted at 50 ℃ for 2 h. Subsequently, 1-BOC-4-bromomethylpiperidine (27.7mg, 0.099mmol) and potassium carbonate (18.3mg, 0.133mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. The crude product was dissolved in 4ml of dichloromethane containing 5% trifluoroacetic acid, reacted at room temperature for 2h, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound I-29 (18.4mg, 51%).

The characterization information of the compound I-29 is specifically as follows:

1H NMR(400MHz,MeOD)δ8.08(m,1H),7.95–7.88(m,2H),7.61–7.55(m,2H),4.06(s,3H),4.05(s,3H),3.36(d,J＝12.8Hz,2H),3.03(d,J＝6.6Hz,2H),2.87(t,J＝11.9Hz,2H),2.10(d,J＝13.9Hz,2H),1.83(s,6H),1.73-1.63(m,1H),1.55–1.42(m,2H).

this example also synthesizes Compound I-30, the specific procedures and reaction conditions being the same as those for the synthesis of I-29 above, except that the corresponding bromide is replaced, specifically: synthesis of Compound I-30 was performed using N-Boc-3(S) -bromomethylpyrrolidine, the following characterization information for Compound I-30:

the characterization information of the compound I-30 is specifically as follows:

¹H NMR(400MHz,MeOD)δ8.06(d,J＝9.4Hz,1H),7.90(dd,J＝4.0,2.8Hz,2H),7.57–7.53(m,2H),4.07(s,3H),4.06(s,3H),3.40–3.27(m,2H),3.22–3.09(m,3H),3.01-2.94(m,1H),2.35–2.25(m,1H),2.20–2.09(m,1H),1.80–1.70(m,1H).

EXAMPLE 13 Synthesis of Compound II-1

II-1 was synthesized according to the following procedure:

(1) synthesis of compound 24:

under argon protection, 4ml of a tetrahydrofuran solution of known compound 22(1.04g, 5.02mmol) was added dropwise to a solution of 50ml of potassium tert-butoxide (0.79g, 7.02mmol) in THF at-78 deg.C, followed by slow addition of a solution of commercially available compound 23(0.83g, 5.02mmol) in 4ml of tetrahydrofuran to the system. After the addition, the reaction was carried out at-78 ℃ for 0.5h, followed by slow warming overnight. The reaction was quenched by addition of ammonium chloride, diluted with dichloromethane, separated by water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate and the solvent was spin dried under reduced pressure. Purification by silica gel column separation (pure dichloromethane) gave compound 24(0.29g, 18%).

The characterization information for compound 24 is specifically:

¹H NMR(400MHz,CDCl₃)δ9.39(s,1H),7.17–7.11(m,2H),6.91–6.83(m,2H),4.38(q,J＝7.1Hz,2H),3.77(s,3H),2.57(s,3H).

(2) synthesis of compound 25:

to compound 24(198mg, 0.61mmol) was added 5ml of phosphorus oxychloride in a sealed tube and reacted at 130 ℃ overnight. After the reaction is finished, slightly cooling the reaction liquid, pouring the reaction liquid on ice, and fully stirring the mixture to ensure that the phosphorus oxychloride is completely hydrolyzed. Diluting with dichloromethane, separating with water/dichloromethane, washing with saturated salt for 2 times, separating the organic phase, drying with anhydrous sodium sulfate, and spin-drying the solvent under reduced pressure. Purification by silica gel column separation (pure dichloromethane) afforded compound 25(74mg, 41%).

The characterization information for compound 25 is specifically:

¹H NMR(400MHz,CDCl₃)δ7.97(d,J＝9.2Hz,1H),7.55(d,J＝2.8Hz,1H),7.40(dd,J＝9.2,2.8Hz,1H),4.01(s,3H),2.88(s,3H).

(3) synthesis of compound 26:

to compound 25(0.020g, 0.067mmol) and 70% sodium hydrosulfide hydrate (10.8mg, 0.135mmol) were dissolved in 5ml dry DMF under argon protection and reacted at 50 ℃ for 2 h. Subsequently, 1-BOC-4-bromomethylpiperidine (37.5mg, 0.135mmol) and potassium carbonate (27.9mg, 0.202mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate 95/5) gave compound 26(14.9mg, 45%).

The characterization information for compound 26 is specifically:

¹H NMR(400MHz,CDCl₃)δ7.94(d,J＝9.2Hz,1H),7.85(d,J＝2.8Hz,1H),7.38(dd,J＝9.2,2.8Hz,1H),4.12–4.01(m,2H),4.00(s,3H),3.47(d,J＝6.9Hz,2H),2.84(s,3H),2.61(t,J＝12.7Hz,2H),1.86(d,J＝13.1Hz,2H),1.65–1.57(m,1H),1.44(s,9H),1.23–1.14(m,2H).

(4) synthesis of Compound II-1:

compound 26(14.9mg, 0.030mmol) was dissolved in 4ml of dichloromethane containing 5% trifluoroacetic acid, reacted at room temperature for 2h, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound II-1 (14.7mg, 96%).

The characterization information of the compound II-1 is specifically as follows:

1H NMR(400MHz,MeOD)δ7.85(d,J＝9.2Hz,1H),7.81(d,J＝2.7Hz,1H),7.40(dd,J＝9.2,2.8Hz,1H),3.97(s,3H),3.62(d,J＝6.9Hz,2H),3.37–3.32(m,2H),2.87(s,3H),2.91–2.80(m,2H),2.12(d,J＝13.8Hz,2H),1.82-1.70(m,1H),1.54–1.42(m,2H).

EXAMPLE 14 Synthesis of Compound II-2

II-2 was synthesized according to the following procedure:

(1) synthesis of compound 24':

under argon protection, 1ml of a tetrahydrofuran solution of known compound 22(150mg, 0.73mmol) was added dropwise to a THF solution of 8ml of potassium tert-butoxide (112mg, 0.92mmol) at-60 ℃ and then a 1ml of a tetrahydrofuran solution of known compound 23' (150mg, 0.66mmol) was slowly added dropwise to the system. After the addition, the reaction was carried out at-60 ℃ for 1.5h, followed by slow warming overnight. The reaction was quenched by addition of ammonium chloride, diluted with dichloromethane, separated by water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate and the solvent was spin dried under reduced pressure. Purification by silica gel column separation (pure dichloromethane) gave compound 24' (69.2mg, 28%).

The characterization information for compound 24' is specifically:

¹H NMR(400MHz,CDCl₃)δ11.61(s,1H),7.58–7.50(m,2H),7.11(dd,J＝9.0,3.2Hz,1H),4.49(q,J＝7.1Hz,2H),3.97(s,3H),3.83(s,3H),2.64(s,3H),1.42(t,J＝7.1Hz,3H).

(2) synthesis of compound 25':

to compound 24' (50mg, 0.14mmol) was added 1ml of phosphorus oxychloride in a sealed tube and reacted at 130 ℃ overnight. After the reaction is finished, slightly cooling the reaction liquid, pouring the reaction liquid on ice, and fully stirring the mixture to ensure that the phosphorus oxychloride is completely hydrolyzed. Diluting with dichloromethane, separating with water/dichloromethane, washing with saturated salt for 2 times, separating the organic phase, drying with anhydrous sodium sulfate, and spin-drying the solvent under reduced pressure. Purification by flash column chromatography on silica gel (dichloromethane/ethyl acetate-95/5) gave compound 25' (15mg, 32%).

The characterization information for compound 25' is specifically:

¹H NMR(400MHz,CDCl₃)δ7.69(s,2H),4.05(s,3H),4.02(s,3H),2.88(s,3H).

(3) synthesis of compound 26':

to compound 25' (15mg, 0.042mmol) and 70% sodium hydrosulfide hydrate (6.7mg, 0.084mmol) were dissolved in 2ml of dry DMF under argon protection and reacted at 50 ℃ for 2 h. Subsequently, 1-BOC-4-bromomethylpiperidine (29.3mg, 0.105mmol) and potassium carbonate (17.4mg, 0.126mmol) were added thereto, and reacted at room temperature overnight. The reaction mixture was spin-dried under reduced pressure, separated with water/dichloromethane, the organic phase was separated, dried over anhydrous sodium sulfate, and the solvent was spin-dried under reduced pressure. Purification by silica gel column separation (dichloromethane/ethyl acetate ═ 95/5) gave compound 26' (16.2mg, 70%).

The characterization information for compound 26' is specifically:

¹H NMR(400MHz,CDCl₃)δ8.02(d,J＝2.9Hz,1H),7.65(d,J＝2.8Hz,1H),4.04(s,3H),4.04-4.01(m,2H),4.00(s,3H),3.44(d,J＝6.9Hz,2H),2.84(s,3H),2.58(t,J＝12.6Hz,2H),1.82(d,J＝12.9Hz,2H),1.61–1.52(m,1H),1.44(s,9H),1.23-1.13(m,2H).

¹H NMR(400MHz,CDCl₃)δ8.02(d,J＝2.9Hz,1H),7.65(d,J＝2.9Hz,1H),4.05(s,3H),4.14-3.96(m,2H),4.04(s,3H),4.00(s,3H),3.45(d,J＝6.9Hz,2H),2.84(s,3H),2.58(t,J＝12.4Hz,2H),1.82(d,J＝12.3Hz,2H),1.57–1.50(m,1H),1.44(s,9H),1.24–1.11(m,2H).

(4) synthesis of Compound II-2:

compound 26' (16mg, 0.029mmol) was dissolved in 1ml of dry dichloromethane, cooled to-60 ℃ and 87. mu.l of 1M diisopropylaluminium hydride was added, after which the reaction was slowly returned to room temperature for 4 h. Adding saturated potassium hydrogen tartrate solution to quench reaction, adding dichloromethane for dilution, separating by water/dichloromethane, separating organic phase, drying with anhydrous sodium sulfate, and spin-drying solvent under reduced pressure. The crude product was dissolved in 4ml of dichloromethane containing 5% trifluoroacetic acid, reacted at room temperature for 2h, and the solvent was spin-dried under reduced pressure. Purification by HPLC/MS on a Waters Autopurification LC/MS system (ACQUITY UPLC BEHC1817 μm2.1X50mm column) gave the trifluoroacetate salt of compound II-2 (5.3mg, 34%).

The characterization information of the compound II-2 is specifically as follows:

1H NMR(400MHz,MeOD)δ7.78(d,J＝2.8Hz,1H),7.53(d,J＝2.7Hz,1H),5.20(s,2H),3.98(s,3H),3.57(d,J＝6.9Hz,2H),3.34(s,2H),2.88(s,3H),2.86–2.80(m,2H),2.15–2.08(m,2H),1.78–1.69(m,1H),1.55–1.41(m,2H).

experimental example I: cell level assay for anti-tumor activity of compounds

1640 complete medium: 500mL of 1640 liquid medium (Gibco), 55mL of fetal bovine serum (Gibco) and 5mL of streptomycin were thoroughly mixed, and filtered and sterilized through a sterile filter with a pore size of 0.22. mu.m.

DMEM complete medium: 500mL of DMEM liquid medium (Gibco), 55mL of fetal bovine serum (Gibco) and 5mL of streptomycin were thoroughly mixed, and then filtered through a sterile filter with a pore size of 0.22. mu.m and sterilized.

Tumor cell culture

1. Resuscitated multiple myeloma and leukemia cells were inoculated into 10mL 1640 complete medium, breast cancer cells into 10mL DMEM complete medium in 5% CO₂Cultured in an incubator at 37 ℃.

2. And carrying out cell passage when the cell density reaches 80-90%.

Second, antitumor Activity test

1. Inoculating the cultured tumor cells to a 96-well plate, wherein the density is 1000 cells/well, each well is 90 mu L of DMEM/1640 complete culture medium, and culturing adherent cells for 12h to allow the cells to grow in an adherent manner;

2. the compound was prepared as a 50mM stock solution in sterile DMSO, and then sequentially diluted with DMEM/1640 complete medium to give concentrations of 10mM, 100. mu.M, 10. mu.M, 1. mu.M, …,10^-2μ M dilution. The following compounds were used respectively: compounds I-1, I-2, …, I-30, II-1, II-2.

3. Adding compounds with different concentrations into a 96-well plate according to a one-tenth ratio, uniformly mixing, and putting into an incubator for culturing for 48 hours;

4. finally, by

The kit detects the cell activity: add 100. mu.L to each well

Reagent, then placing the microplate on a shaker and gently shakingAnd incubating for 15min at room temperature to generate stable luminescence signals. Luminescence signal values were recorded and IC of compounds determined by fitting curves using Graphpad Prism software₅₀The values, results are shown in tables 1 and 2.

IC₅₀The value, i.e., the concentration of a certain compound that induces apoptosis of tumor cells by 50%, is referred to as the 50% inhibitory concentration.

Test results show that the potential analogues of the compounds have stronger inhibitory activity on blood tumor cells and solid tumor cells.

Table 1: activity assay results (IC) of Compounds on human multiple myeloma cell lines and human breast cancer cell lines₅₀Value, μ M)

Table 2: results of Activity assays (IC) of Compounds on different tumor cell lines₅₀Value, μ M)

Experimental example II: antitumor mechanisms of Compounds

The DYRK (Dual-specificity transcription-phosphorylation-Regulated Kinase) family is a serine/threonine protein Kinase belonging to the CMGC family, comprising a total of 5 family members: DYRK1A,1B,2,3, 4. The DYRK family is closely related to cell growth, proliferation and differentiation. Among them, DYRK2 is a major kinase that phosphorylates threonine 25 (Rpt3-Thr25) of Rpt3 subunit of 26S proteasome to promote cell progression in S/G2 phase, and this lack of phosphorylation process will significantly inhibit tumor development. DYRK2 is therefore an important novel anti-tumor target.

1. Molecular level assay of compounds for DYRK2 kinase inhibitory activity and selectivity for similar kinases

Compounds were tested for inhibitory activity against DYRK2 and selectivity for similar kinases by mimicking, at the molecular level, the process by which DYRK2 catalyzes phosphorylation of substrates by isotopic quantitation assays.

Experimental group test Compound 2.5. mu.L (0.00001-100. mu.M final concentration) was mixed with 22.5. mu.L of 50mM Tris-HCl, pH 7.5,5mM MgCl2, 10. mu.M ATP (DYRK 2-10. mu.M ATP, DYRK 3-5. mu.M ATP, DYRK 1A-25. mu.M ATP, DYRKIB-50. mu.M ATP, Haspin-150. mu.M ATP), trace amounts of radiolabel³²P-ATP (50nM, PerkinElmer), 10nM DYRK2(DYRK3, DYRK1A, DYRK1B and Haspin are all 10nM), DYRK2, DYRK3, DYRK1A and DYRK1B with 150. mu.M biotin-Woodotide peptide substrate (biotin-KKISGRLSPIMTEQ-NH2) at 37 ℃ for 10 minutes and then stopped by adding 30mM EDTA, and Haspin with 200. mu. M H3(1-21) peptide substrate (biotin-ARTKQTARKSTGGKAPRKQLA-NH2) at 37 ℃ for 10 minutes and then stopped by adding 30mM EDTA. Then 15. mu.L of the reaction mixture was aspirated and added to P81 phosphocellulose filter paper (Whatman). The P81 phosphocellulose filter paper was washed three times in 0.75% phosphoric acid solution for 10 minutes to remove unreacted³²P-ATP, then P81 phosphocellulose filter paper was washed in 95% ethanol solution to air dry. The dried P81 phosphocellulose filter paper was placed in a special tube and quantified using a Microbeta liquid scintillation counter (Perkinelmer)³²P content, 2.5. mu.L buffer was added to the positive control group instead of the compound, and no peptide substrate was added to the negative control group. Data were processed with GraphPad Prism 7 software, and inhibition curves were fitted and IC was obtained₅₀The value is obtained. The results are shown in tables 3 and 4.

IC₅₀The value, i.e., the concentration of a compound that inhibits kinase activity by 50%, is referred to as the 50% inhibitory concentration.

Test results show that the compounds I-6, I-20, I-22 and I-29 and potential analogues thereof have strong inhibitory activity on DYRK2 kinase and good selectivity in similar kinases, and further influence a plurality of biological processes such as proteasome activity, cell cycle and the like, thereby playing an anti-tumor role.

Table 3: inhibitory Activity of different Compounds on DYRK2 and DYRK1A

TABLE 4 results of selective assays (IC) of highly active compounds in similar kinases₅₀Value, nM)

2. Cell level assay for proteasome inhibitory activity of compounds

An experiment system for reading the proteasome activity is constructed at a cellular level, the activity of proteasome hydrolysis chymotrypsin-like protease is utilized, and the inhibitory activity of the compound on proteasome is detected by using LLVY-R110 substrate as a fluorescent indicator of proteasome activity (LLVY-R110 is cleaved by proteasome to generate strong green fluorescence R110).

1. Inoculating cultured human multiple myeloma cells and human breast cancer cells to a 384-well plate, wherein the density is 100000 cells/well, 20 mu L of DMEM/1640 complete culture medium is added to each well, and the adherent cells are cultured for 12h to allow the cells to grow in an adherent manner;

2. preparing a compound into 50mM mother solution by taking sterile DMSO as a solvent, and then diluting the mother solution by using a DMEM/1640 complete culture medium to obtain a diluent with the concentration of 50 mu M;

3. adding the compound into a 384-well plate according to the proportion of one fifth, then uniformly mixing and putting into an incubator for incubation for 1 h;

4. finally, the activity of the proteasome is detected by an Amplite fluorescence method proteasome 20S activity detection kit: add 25. mu.L of the protease assay loading solution to each well, incubate overnight in an incubator to allow for a steady fluorescent signal to be generated, and record the signal value. The results are shown in FIG. 1.

Test results show that the potential analogs of the compounds have obvious inhibition effect on proteasomes of tumor cells, thereby playing the role of resisting tumors.

Experimental example III: combined curative effect of cell level detection compound and clinical first-line antitumor drug

1. Inoculating the cultured tumor cells to a 96-well plate, wherein the density is 1000 cells/well, each well is 90 mu L of DMEM/1640 complete culture medium, and culturing adherent cells for 12h to allow the cells to grow in an adherent manner;

2. preparing a compound into 50mM mother solution by taking sterile DMSO as a solvent, and then diluting the mother solution by using a DMEM/1640 complete culture medium to obtain a diluent with a corresponding concentration;

2. preparing 50mM mother solution from bortezomib, carfilzomib, ixazoib, dexamethasone, thalidomide and paclitaxel which are first-line clinical antitumor drugs by taking sterile DMSO as a solvent, and then diluting the mother solution by using a DMEM/1640 complete culture medium to obtain a diluent with a corresponding concentration;

3. sequentially adding compounds with different concentrations and clinical first-line antitumor drugs into a 96-well plate according to the concentration combination listed in figure 2 by one tenth, uniformly mixing, and putting into an incubator for culturing for 48 h;

4. finally, by

The kit detects the cell activity: add 100. mu.L to each well

And (3) placing the microplate on an oscillator, gently oscillating, and incubating for 15min at room temperature to generate a stable luminescent signal. The luminescence signal values were recorded and fitted using Compuyn software to obtain CI values for the compound in combination with the first line clinical antineoplastic agent, the results are shown in FIG. 2.

CI value, i.e. union index. The CI value is less than 1, which indicates that the drug combination has synergistic effect; the CI value is equal to 1, which indicates that the drug combination has an additive effect; a CI value greater than 1 indicates that the drug combination is antagonistic.

The test result shows that the potential analogue of the compound has obvious synergistic effect when being combined with a first-line clinical antitumor drug, and the combined effect is better than the current clinical commonly used treatment scheme (combining a proteasome inhibitor and adrenocortical hormone dexamethasone, combining the proteasome inhibitor and an immunomodulator thalidomide and the like).

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Claims (10)

1. A compound or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the compound has a structure according to formula I or formula II:

in the general formula I, R₁Represents hydrogen, acyl, sulfonyl, benzene which can be substituted, pyridine which can be substituted, furan which can be substituted, thiophene which can be substituted, or straight-chain or branched-chain alkyl of C1-C12, the tail end of which can be substituted by one or more of amino, cyano, amido or a ring containing hetero atoms; preferably R₁Represents a linear alkyl group of C1-C4 which may be substituted at the terminal by an amino group, cyano group, amido group or N-containing heterocycle; further preferred is R₁Represents propylamino, Boc-propylamino, acetylapropylamino, acetylbutylamino, propionitrile, butyronitrile, 4-N cyclohexylamine, 4-methylpiperidine, (S) -3-methylpyrrolidine, (R) -3-methylpyrrolidine, 3-methylazetidine, 1- (4-methylpiperidin-1-yl) prop-2-en-1-one, 4- ((4-methylpiperidin-1-yl) methyl) benzenesulfonyl fluoride, N- (4- (4-methylpiperazin-1-yl)) -4-oxobutyl) acrylamide, N- (5- (4-methylpiperidin-1-yl) -5-oxopentyl) acrylamide, (2-formyl-5- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid or (2-formyl-4- (3- (4-methylpiperidin-1-yl) -3-oxopropoxy) phenyl) boronic acid;

R₂、R₃independently of one another, represents hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, phenyl or benzyl; preferably, R₂、R₃Any one of which represents methyl or benzyl, the other represents methyl, isopropyl or benzyl; more preferably, R₂、R₃Any one of which represents methyl and the other represents methyl, isopropyl or benzyl;

R₄represents hydrogen, carboxyl, ester group, amide group or C1-C3 linear alkyl with the terminal substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl; preferably, R₄Represents hydrogen, carboxyl, ester group, amido or methyl which can be substituted by one or more of halogen, methyl, hydroxyl, amino, cyano and guanidyl; further preferably, R₄Represents hydrogen, carboxyl, hydroxymethyl, aminomethyl, difluoromethyl, guanidinomethyl, dimethyl-substituted hydroxymethyl;

in the general formula II, R₅Represents hydrogen, acyl, sulfonyl, benzene which can be substituted, pyridine which can be substituted, furan which can be substituted, thiophene which can be substituted, or straight-chain or branched-chain alkyl of C1-C12, the tail end of which can be substituted by one or more of amino, cyano, amido and N-containing heterocyclic ring; preferably R₅Represents a linear alkyl of C1-C4, the tail end of the linear alkyl can be substituted by one or more of amino, cyano, amido or N-containing heterocyclic rings; further preferred is R₅Represents propylamino, Boc-propylamino, acetylamino, acetylbutylamino, propionitrile, butyronitrile, 4-N cyclohexylamine, 4-methylpiperidine, (S) -3-methylpyrrolidine, (R) -3-methylpyrrolidine or 3-methylazetidine;

R₆represents hydrogen, carboxyl, ester group, amide group or the tail end of which can be one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidylSeveral substituted C1-C3 linear alkyl groups; preferably, R₆Represents hydrogen, carboxyl, ester group, amido or methyl which can be substituted by one or more of halogen, methyl, hydroxyl, amino, cyano and guanidyl; further preferably, R₆Represents hydrogen, carboxyl, hydroxymethyl, aminomethyl, difluoromethyl, guanidinomethyl, dimethyl-substituted hydroxymethyl.

2. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the compound of formula I is selected from the group consisting of compounds of the following structures:

preferably the pharmaceutically acceptable salt is trifluoroacetate; particularly preferably, when the compound has a structure shown in formula I-22, the pharmaceutically acceptable salt is trifluoroacetate.

3. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the compound of formula II is selected from compounds of the following structures:

4. a method for synthesizing the compound of claim 1 or 2, or a pharmaceutically acceptable salt or a stereoisomer thereof, wherein the compound of the general formula I is synthesized by the following steps:

when R is₄When the compound represents hydrogen, the compound 4 is subjected to at least one step of side chain introduction reaction to prepare a compound IA;

when R is₄When the compound represents carboxyl, ester group, amide group or C1-C3 linear alkyl of which the tail end can be substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl, the compound IB is prepared by at least one step of side chain introduction reaction of a compound 4'.

5. The method of claim 4, wherein when the compound of formula I has the structure of IA1, the synthetic route is as follows:

wherein R is₁As defined in claim 1 or 2;

the synthesis method comprises the following specific steps:

(i) after the compound 4 and sodium hydrosulfide are subjected to thiolation reaction, the obtained product is subjected to substitution reaction with a corresponding side chain compound to introduce a side chain, so that a compound IA1 is obtained;

when the compound of the general formula I has a structure shown as a general formula IA2, the synthetic route is as follows:

wherein R is₁、R₃As defined in claim 1 or 2;

the synthesis method comprises the following specific steps:

(i) carrying out deprotection reaction on the compound 4 by boron tribromide to obtain a compound 5;

(ii) performing alkylation reaction on the compound 5 to obtain a compound 6;

(iii) carrying out thiolation reaction on the compound 6 and sodium hydrosulfide, carrying out substitution reaction on the obtained product and a corresponding side chain compound with a protecting group, introducing a side chain, and then carrying out deprotection to obtain a compound IA 2;

when the compound of the general formula I has a structure shown in a formula I-11, the synthetic route is as follows:

the synthesis method comprises the following specific steps:

(i) carrying out deprotection reaction on the compound 4 by boron tribromide to obtain a compound 7;

(ii) the compound 7 reacts with benzyl bromide to obtain a compound 8;

(iii) carrying out thiolation reaction on the compound 8 and sodium hydrosulfide, carrying out alkylation reaction on the obtained product and a corresponding side chain compound with a protecting group, and then carrying out deprotection to obtain a compound I-11;

when the compound of the general formula I has a structure shown in a formula IB1, the synthetic route is as follows:

wherein R is₁As defined in claim 1 or 2; r₄Represents carboxyl, ester group, amide group or C1-C3 linear alkyl with the terminal substituted by one or more of halogen, methyl, ethyl, hydroxyl, amino, cyano and guanidyl;

the synthesis method comprises the following specific steps:

(i) treating the compound 4' with oxalic acid chloride, and performing methyl esterification in methanol to obtain a compound 12; and (3) carrying out at least one-step one-pot thiolation alkylation on the compound 12, and/or one-step functional group conversion, and/or one-step deprotection reaction to obtain a compound IB 1.

6. The synthesis process according to claim 4 or 5, characterized in that the synthesis route of compound 4 is as follows:

the method comprises the following specific steps:

(i) carrying out copper catalytic coupling on the compounds 1 and 2 to obtain a compound 3;

(ii) refluxing, cyclizing and aromatizing the compound 3 in phosphorus oxychloride to obtain a compound 4;

and/or, the synthetic route of the compound 4' is as follows:

the method comprises the following specific steps:

(i) carrying out coupling reaction on the compounds 1 ' and 2 ' under the catalysis of copper and cuprous oxide to obtain a compound 3 ';

(ii) cyclizing and aromatizing the compound 3 'in phosphorus oxychloride to obtain the compound 4'.

7. The method for synthesizing an anti-tumor compound or a pharmaceutically acceptable salt or a stereoisomer thereof according to claim 3, wherein when the compound of the general formula II has a structure shown in formula II-1, the synthetic route is as follows:

the synthesis method comprises the following specific steps:

(i) carrying out heterocyclic synthesis reaction on the compound 22 and the compound 23 under alkaline condition to obtain a compound 24;

(ii) cyclizing and aromatizing the compound 24 in phosphorus oxychloride to obtain a compound 25;

(iii) carrying out thiolation reaction on the compound 25 and sodium hydrosulfide, carrying out alkylation reaction on the obtained product and a corresponding side chain compound with a protecting group, and then carrying out deprotection to obtain a compound II-1;

when the compound of the general formula II has the structure shown in the formula II-2, the synthetic route is as follows:

the synthesis method comprises the following specific steps:

(i) reacting the compound 22 with a compound 23 'to obtain a compound 24';

(ii) cyclizing and aromatizing the compound 24 'in phosphorus oxychloride to obtain a compound 25';

(iii) thiolating the compound 25 'with sodium hydrosulfide, and alkylating the compound by a one-pot method to obtain a compound 26';

(iv) and (3) carrying out deprotection on the compound 26' after reduction by diisopropylaluminum hydride to obtain a compound II-2.

8. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or stereoisomer thereof, in combination with at least one pharmaceutically acceptable diluent, carrier or excipient.

9. The pharmaceutical composition of claim 8, further comprising a second therapeutic agent useful for treating a tumor;

the second therapeutic agent is preferably a proteasome inhibitor and/or a natural anti-cancer drug;

the proteasome inhibitor is preferably one or more of bortezomib, carfilzomib and ixazoib;

the natural anticancer drug is preferably paclitaxel.

10. The use of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or stereoisomer thereof, for the manufacture of a medicament for the treatment of a tumour,

preferably, the compound or the pharmaceutically acceptable salt or the stereoisomer thereof is used as a DYRK2 inhibitor to inhibit the activity of the 26s proteasome of the human body;

preferably said compound or a pharmaceutically acceptable salt or stereoisomer thereof acts by blocking the cell cycle of the tumour cell;

preferably, the tumor comprises a hematological tumor and a solid tumor; the blood tumor is preferably one or more of multiple myeloma, leukemia, chronic lymphoma, mantle cell lymphoma and Fahrenheit megasphaemia; the solid tumor is preferably breast cancer.

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