CN114478568A

CN114478568A - Thienopyrimidine compound, pharmaceutical composition containing thienopyrimidine compound and application of thienopyrimidine compound

Info

Publication number: CN114478568A
Application number: CN202011166899.6A
Authority: CN
Inventors: 陈旭星; 李京; 陈艳红; 赵兆
Original assignee: Suzhou Youli Biomedical Technology Co ltd
Current assignee: Suzhou Youli Biomedical Technology Co ltd
Priority date: 2020-10-27
Filing date: 2020-10-27
Publication date: 2022-05-13
Also published as: WO2022089296A1; TW202216728A; TWI804016B; CN116438183A

Abstract

The invention provides a thienopyrimidine compound, a pharmaceutical composition containing the thienopyrimidine compound and application of the thienopyrimidine compound. The structural formula of the thienopyrimidine compound has a structure shown in a formula I. The compounds interfere with the interaction between menin protein and MLL1 or MLL2 or MLL-fusion oncoprotein, and are expected to be drugs for treating tumors, diabetes and other diseases that depend on MLL1, MLL2, MLL fusion protein, and/or the activity of menin protein.

Description

Thienopyrimidine compound, pharmaceutical composition containing thienopyrimidine compound and application of thienopyrimidine compound

Technical Field

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

Background

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

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

In particular hematological neoplasms, there are certain genetic abnormalities or mutations, such as nuclear porin 98(NUP98) gene fusion, nuclear phosphoprotein (NPM1) gene mutation, DNA methyltransferase 3A (DNMT3A) mutation, MLL gene amplification, etc., which are often accompanied by high levels of HOX gene expression. Abnormal overexpression of the posterior HOXD gene, particularly HOXD13, in ewing's sarcoma, was associated with high levels of meinin and MLL1 protein, whereas HOXD13 was a downstream gene regulated by menin and MLL 1. Thus, interfering with the interaction between menin and MLL proteins, especially by covalent binding, is a very promising strategy for treating tumors.

Therefore, there is still a strong need in the art to develop effective drugs that can interfere with the interaction between menin and MLL proteins.

Disclosure of Invention

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

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

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

wherein the content of the first and second substances,

R¹selected from H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 member heterocyclyl, substituted or unsubstituted C1-C6 alkylcarbonyl, substituted or unsubstituted

Substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted C1-C6 alkyl sulfone, substituted or unsubstituted C1-C6 alkyl sulfoxide, substituted or unsubstituted C,

Wherein R is^a、R^bEach independently selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclyl, or R^aAnd R^bA substituted or unsubstituted 4-8 membered heterocyclic ring formed by linking with N, wherein R^aAnd R^bThe heterocyclic ring formed by connecting with N contains 1-3 heteroatoms selected from N, O, S, P;

R²selected from H, halogen, substituted or notSubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C16 aryl, substituted or unsubstituted 5-16 membered heteroaryl, substituted or unsubstituted 4-8 membered heterocyclyl, substituted or unsubstituted C1-C6 alkylcarbonyl, substituted or unsubstituted

Wherein R is^c、R^dEach independently selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclyl, or R^cAnd R^dA substituted or unsubstituted 4-8 membered heterocyclic ring formed by linking with N, wherein R^cAnd R^dThe heterocyclic ring formed by connecting with N contains 1-3 heteroatoms selected from N, O, S, P;

R³each independently selected from H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, -NH₂、-NO₂COOH, -CN, -OH, substituted or unsubstituted C1-C6 alkyl sulfone group, substituted or unsubstituted C1-C6 alkyl sulfoxide group, substituted or unsubstituted C1-C6 alkylthio group,

Wherein R is^3'Selected from H, methyl or fluoro; r^3”Is selected from the group consisting of chlorine or bromine,

represents the attachment position of the group;

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

w is selected from N or C;

U¹、U²、U³、U⁴、U⁵、U⁶、U⁷、U⁸are each independently selected from

And U is¹、U²、U³、U⁴At most one of them is

U⁵、U⁶At most one of them is

U⁷、U⁸At most one of them is

Wherein the content of the first and second substances,

represents the position of attachment of the group,

each R' is independently selected from: H. halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

each R' "is independently selected from: H. halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

each R "" is independently selected from: H. halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano; (ii) a

A is selected from substituted or unsubstituted C6-C16 aryl, substituted or unsubstituted 5-16 membered heteroaryl, wherein said heteroaryl contains 1-3 heteroatoms selected from N, O, S, P;

L¹is absent,

Wherein, the first and the second end of the pipe are connected with each other,

represents the attachment position of a group, R^L1'、R^L1”Each independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, or R^L1'And R^L1”Form a substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclic ring with the attached carbon atom, wherein R^L1'And R^L1”The heterocyclic ring formed with the attached carbon atom contains 1 to 3 heteroatoms selected from N, O, S, P;

L²is selected from

Wherein the content of the first and second substances,

represents the attachment position of the group;

x is selected from carbon atoms or

R⁴Is selected from

Wherein R4' is selected from fluorine or chlorine, R4' is selected from H, methyl or fluorine, R4' is selected from H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, substituted or unsubstituted (C1-C4 alkyl)₂Amino, substituted or unsubstituted C1-C4 alkylthio, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclyl, substituted or unsubstituted C2-C4 acyl, said heterocyclyl containing 1-3 heteroatoms selected from N, O, S, P, wherein

Indicates the attachment position of the group.

Preferably, said R is¹Selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, and substituted or unsubstituted 4-8 membered heterocyclyl, wherein said heterocyclyl contains 1-3 heteroatoms selected from N, O, S, P.

Preferably, said R is¹Selected from substituted or unsubstituted C1-C6 alkyl groups, more preferably halogen-substituted C1-C6 alkyl groups, and still more preferably trifluoroethyl groups.

Preferably, said R is²Selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C16 aryl, substituted or unsubstituted 5-16 membered heteroaryl, substituted or unsubstituted 4-8 membered heterocyclyl, wherein said heteroaryl or heterocyclyl contains 1-3 heteroatoms selected from N, O, S, P, further preferably, said R is²Is H.

Preferably, said R is³Is H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, -NH₂or-CN.

Preferably, the Y, Z are each selected from N.

Preferably, W is C.

Preferably, the U is¹、U²、U³、U⁴、U⁵、U⁶、U⁷、U⁸Are each independently selected from

And U is¹、U²、U³、U⁴At most one of them is

U⁵、U⁶At most one of them is

U⁷、U⁸At most one of them is

Wherein the content of the first and second substances,

represents the position of attachment of the group,

each R' is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

each R' "is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

each R "" is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano; .

Preferably, A is selected from substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, wherein said heteroaryl contains 1-3 heteroatoms selected from N, O, S, P.

Preferably, a is a substituted or unsubstituted benzene ring, a substituted or unsubstituted pyridine ring, a substituted or unsubstituted pyridazine ring, a substituted or unsubstituted pyrimidine ring, a substituted or unsubstituted triazazine ring, a substituted or unsubstituted thiophene ring, a substituted or unsubstituted thiazole ring, a substituted or unsubstituted imidazole ring, a substituted or unsubstituted pyrrole ring, a substituted or unsubstituted pyrazole ring, a substituted or unsubstituted oxazole ring, a substituted or unsubstituted isoxazole ring, a substituted or unsubstituted triazoizole ring.

Preferably, said L¹Is absentor-CH₂-, preferably-CH₂-。

Preferably, said L²Is selected from

Preferably, it is

Preferably, said X is selected from carbon atoms.

Preferably, said R is⁴Is selected from

Wherein R4' is selected from fluorine or chlorine, R4' is selected from H, methyl or fluorine, R4' is selected from H, substituted or unsubstituted C1-C4 alkyl,

indicates the attachment position of the group.

Preferably, the structural formula of the spiro moiety in the compound shown in the formula I is shown in the specification

Wherein

Represents the attachment position of a group selected from any one of the following groups:

Wherein

Represents the connecting position of the group and is selected from any one of the following groups:

preferably, the structural formula in the formula I is

Shown as a fused ring portion, wherein

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

Preferably, the structural formula in the formula I is

A ring-shaped part as shown, wherein

preferably, said R is⁴Is selected from-CH₂F、-CH₂Cl、

Preferably, the compound represented by the formula I is selected from any one of the following compounds:

preferably, the thienopyrimidine compound further includes any one of pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, cis-trans isomers, solvates, polymorphs, or deuterations of the compound represented by formula I.

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

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

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

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

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

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

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

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

Description of the terms

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

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

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

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

"pharmaceutically acceptable acid addition salts" refers to salts with inorganic or organic acids which retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, and the like; organic acid salts include, but are not limited to, formates, acetates, 2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, caprates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbates, salicylates, 4-aminosalicylates, napadisylates, and the like. These salts can be prepared by methods known in the art.

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

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

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

As used herein, a "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, solvent, or emulsifying agent that is approved by the relevant governmental regulatory agency for human or livestock use.

The "tumor" of the present invention includes, but is not limited to, glioma, sarcoma, melanoma, chondroma of joint, cholangioma, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, squamous carcinoma of lung, adenocarcinoma of lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, renal cancer, oral cancer, etc.

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

As used herein, the term "treatment" and other similar synonyms include the following meanings:

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

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

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

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

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein, refer to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

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

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

Radical definition

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

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

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

Certain chemical groups defined herein are preceded by a shorthand notation to indicate the total number of carbon atoms present in the group. For example, C1-C6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the shorthand notation excludes carbons that may be present in a substituent of the group.

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

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

"hydroxy" means an-OH group.

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

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

"nitro" means-NO₂。

"cyano" means-CN.

"amino" means-NH₂。

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

"carboxyl" means-COOH.

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

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

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

The term "cyclic hydrocarbon group" as used herein as a group or part of another group means a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting of only carbon and hydrogen atoms, which may include fused, bridged or spiro ring systems, having from 3 to 15 carbon atoms, preferably from 3 to 10 carbon atoms, more preferably from 3 to 8 carbon atoms, and which is saturated or unsaturated and may be attached to the rest of the molecule by a single bond via any suitable carbon atom. Unless otherwise specifically indicated in the specification, carbon atoms in the cyclic hydrocarbon group may be optionally oxidized. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2, 3-indanyl, 1,2,3, 4-tetrahydro-naphthyl, 5,6,7, 8-tetrahydro-naphthyl, 8, 9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8, 9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9, 10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo [2.2.1] heptyl, 7-dimethyl-bicyclo [2.2.1] heptyl, bicyclo [2.2.1] heptenyl, bicyclo [2.2.2] octyl, bicyclo [3.1.1] heptyl, bicyclo [3.2.1] octyl, bicyclo [2.2.2] octenyl, Bicyclo [3.2.1] octenyl, adamantyl, octahydro-4, 7-methylene-1H-indenyl, octahydro-2, 5-methylene-pentalenyl and the like.

The term "heterocyclyl" as used herein as a group or part of another group means a stable 3-to 20-membered non-aromatic cyclic group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen, and sulfur. Unless otherwise specifically indicated in the specification, a heterocyclic group may be a monocyclic, bicyclic, tricyclic or higher ring system, which may include fused ring systems, bridged ring systems or spiro ring systems; wherein the nitrogen, carbon or sulfur atom in the heterocyclic group may be optionally oxidized; the nitrogen atoms may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be attached to the rest of the molecule via a carbon atom or a heteroatom and by a single bond. In heterocyclic groups containing fused rings, one or more of the rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purposes of the present invention, heterocyclyl is preferably a stable 4-to 11-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 4-to 8-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclyl groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2, 7-diaza-spiro [3.5] nonan-7-yl, 2-oxa-6-aza-spiro [3.3] heptan-6-yl, 2, 5-diaza-bicyclo [2.2.1] heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxolanyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolyl, octahydroisoindolyl, pyrazolinyl, pyrazolidinyl, phthalimido, and the like.

The term "aryl" as used herein as a group or part of another group means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably having 6 to 10 carbon atoms. For the purposes of the present invention, an aryl group may be a monocyclic, bicyclic, tricyclic or higher polycyclic ring system and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the aryl group is attached to the remainder of the molecule by a single bond via an atom on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, fluorenyl, 2, 3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-benzoxazin-3 (4H) -one-7-yl, and the like.

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

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

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

The term "substituted or unsubstituted" herein means that a hydrogen in the structure is substituted with the substituent or that the hydrogen is unsubstituted. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group, or more than one (to the substitutable position on the substituted structure) position in the structure may be substituted.

For example, as used herein, the term "substituted" or "in the absence of a specific list of substituents"By … … "is meant that one or more hydrogen atoms on a given atom or group are independently substituted with one or more, e.g., 1,2,3, or 4, substituents independently selected from: deuterium (D), halogen, OH, mercapto, cyano, -CD₃、-C₁-C₆Alkyl (preferably-C)_1-3Alkyl group), C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, cycloalkyl (preferably 3-to 8-membered cycloalkyl), aryl, heterocyclyl (preferably 3-to 8-membered heterocyclyl), heteroaryl, aryl C₁-C₆Alkyl, heteroaryl C₁-C₆Alkyl radical, C₁-C₆Haloalkyl-, OC₁-C₆Alkyl (preferably-OC)₁-C₃Alkyl), -OC₂-C₆Alkenyl, OC₁-C₆Alkylphenyl, -C₁-C₆Alkyl OH (preferably-C)₁-C₄Alkyl OH), -C₁-C₆Alkyl SH, -C₁-C₆Alkyl radical O-C₁-C₆Alkyl, OC₁-C₆Haloalkyl, NH₂、C₁-C₆Alkyl NH₂(preferably C)₁-C₃Alkyl NH₂)、N(C₁-C₆Alkyl radical)₂(preferably N (C)₁-C₃Alkyl radical)₂)、NH(C₁-C₆Alkyl) (preferably NH (C)₁-C₃Alkyl)), N (C)₁-C₆Alkyl) (C₁-C₆Alkylphenyl), NH (C)₁-C₆Alkylphenyl), nitro, C (O) -OH, C (O) OC₁-C₆Alkyl (preferably-C (O) OC₁-C₃Alkyl), -CONRiri (where Ri and Rii are H, D and C)_1-6Alkyl, preferably C_1-3Alkyl), -NHC (O) (C)₁-C₆Alkyl), NHC (O) (phenyl), N (C)₁-C₆Alkyl radical C (O) (C)₁-C₆Alkyl group), N (C)₁-C₆Alkyl group C (O) (phenyl), C (O) C₁-C₆Alkyl, C (O) heteroaryl (preferably C (O) -5-7 membered heteroaryl), C (O) C₁-C₆Alkylphenyl, C (O) C₁-C₆Haloalkyl, OC (O) C₁-C₆Alkyl (preferably O)C(O)C₁-C₃Alkyl), -S (O)₂-C₁-C₆Alkyl, -S (O) -C₁-C₆Alkyl, -S (O)₂-phenyl, -S (O)₂-C₁-C₆Haloalkyl, -S (O)₂NH₂、S(O)₂NH(C₁-C₆Alkyl), S (O)₂NH (phenyl), -NHS (O)₂(C₁-C₆Alkyl), -NHS (O)₂(phenyl) and NHS (O)₂(C₁-C₆Haloalkyl), wherein each of said alkyl, cycloalkyl, phenyl, aryl, heterocyclyl and heteroaryl is optionally further substituted with one or more substituents selected from the group consisting of: halogen, -OH, -NH₂Cycloalkyl, 3-8 membered heterocyclyl, C₁-C₄Alkyl radical, C₁-C₄Haloalkyl-, -OC₁-C₄Alkyl, -C₁-C₄Alkyl OH, -C₁-C₄Alkyl radical O-C₁-C₄Alkyl, OC₁-C₄Haloalkyl, cyano, nitro, -C (O) -OH, -C (O) OC₁-C₆Alkyl, CON (C)₁-C₆Alkyl radical)₂、CONH(C₁-C₆Alkyl), CONH₂、NHC(O)(C₁-C₆Alkyl), NH (C)₁-C₆Alkyl radical C (O) (C)₁-C₆Alkyl), -SO₂(C₁-C₆Alkyl), -SO₂(phenyl), -SO₂(C₁-C₆Haloalkyl), -SO₂NH₂、SO₂NH(C₁-C₆Alkyl), SO₂NH (phenyl), -NHSO₂(C₁-C₆Alkyl), -NHSO₂(phenyl) and-NHSO₂(C₁-C₆Haloalkyl). When an atom or group is substituted with a plurality of substituents, the substituents may be the same or different. The terms "moiety," "structural moiety," "chemical moiety," "group," "chemical group" as used herein refer to a specific fragment or functional group in a molecule. Chemical moieties are generally considered to be chemical entities that are embedded in or attached to a molecule.

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

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

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

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

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

In the present invention, in the case of the present invention,

represents a C1-C6 alkyl-substituted ester radical, and may be, for example

In the present invention, (C1-C4 alkyl)₂Amino, representing 2C 1-C4 alkyl-substituted aminesFor example, can be

And the like.

In the present invention, "each R '", "each R" "each R '", "each R" "refer to" each R ' "," each R "" and "each R" "which are present in the thienopyrimidine compound represented by formula I.

Conventional techniques for preparing/separating individual isomers include Chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, Chiral high performance liquid chromatography, see, for example, Gerald Gubitz and Martin G.Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol.243, 2004; m. Stalcup, Chiral Separations, Annu. Rev. anal. chem.3:341-63, 2010; fumiss et al (eds.), VOGEL' S ENCYCOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5. TH ED., Longman Scientific and Technical Ltd., Essex,1991, 809-816; heller, acc, chem, res, 1990,23,128.

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

Protecting groups may be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting Groups is described in detail in Greene, T.W. and P.G.M.Wuts, Protective Groups in organic Synthesis, (1999),4th Ed., Wiley. The protecting group may also be a polymeric resin.

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

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

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

Detailed Description

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

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

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

In each of the embodiments described herein, the first,¹h NMR was recorded by a BRUKER AVANCE NEO model 400MHz NMR spectrometer with chemical shifts expressed in δ (ppm); liquid chromatography-mass spectrometry (LCMS) was recorded by Shimadzu LC-20AD, SIL-20A, CTO-20AC, SPD-M20A, CBM-20A, LCMS-2020 type mass spectrometer; preparative HPLC separation was performed using a Gilson-281 model liquid chromatograph.

Preparation of intermediates

1. Preparation of intermediate A

The synthetic route of the intermediate A is shown as follows:

(1) triethylamine (5.84g,57.7mmol) and compound A-2(7.27g,63.5mmol) were added to a solution of compound A-1(5.0g,28.9mmol) in dichloromethane (25.0mL), and the reaction mixture was stirred at 0 ℃ for 1 hour. Adding saturated sodium bicarbonate water solution (40.0mL) to quench the reaction, extracting with dichloromethane (40.0mL × 3), combining the organic phases, washing with saturated brine (30.0mL × 1), drying over anhydrous sodium sulfate, filtering, and concentrating the organic phase under reduced pressure to obtain compound A-3;

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

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

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

(3) compound A-5(5.0g,28.9mmol) was dissolved in acetic acid (20.0mL) and water (2.0mL), N-chlorosuccinimide (865.9mg,6.48mmol) was added, and the reaction solution was stirred at 25 ℃ for 0.5 hour. Quenching the reaction with water (10.0mL), extracting with dichloromethane (10.0mL × 2), combining the organic phases, washing with saturated brine (10.0mL × 1), drying over anhydrous sodium sulfate, filtering, and concentrating the organic phase under reduced pressure to give intermediate A;

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

2. preparation of intermediate B

The synthetic route of intermediate B is shown below:

(1) dimethyl sulfoxide (6.10g,78.1mmol) was slowly added dropwise over 30 minutes to a solution of oxalyl chloride (5.45g,42.9mmol) in dichloromethane (50.0mL) and the reaction was held at-78 ℃. A solution of Compound B-1(5.0g,39.0mmol) in methylene chloride (50.0mL) was then added and the reaction stirred at-78 deg.C for 1 hour. Triethylamine (19.8g,195mmol) was added and the mixture was stirred at-78 ℃ for 0.5 hour, and the reaction was heated to 0 ℃ for 1 hour. Quenching the reaction with water (40.0mL), extracting with dichloromethane (40.0 mL. times.2), combining the organic phases, washing with saturated brine (40.0 mL. times.1), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure to give compound B-2;

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

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

MS-ESI[M+H]⁺calculated value 225, found value 225.

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

MS-ESI[M+H]⁺Calculated 235, found 235.

(4) Compound B-6(800mg,3.42mmol) was dissolved in phosphorus oxychloride (3.0mL), and the reaction was stirred at 115 ℃ for 3 hours. The reaction solution is directly concentrated under reduced pressure to remove phosphorus oxychloride, ice and saturated sodium bicarbonate solution are added to adjust the pH value to be more than 7, ethyl acetate (20.0mL multiplied by 2) is used for extraction, the combined organic phases are washed by saturated saline solution (20.0mL multiplied by 2), dried by anhydrous sodium sulfate, filtered, and the organic phase is concentrated under reduced pressure. Separating the crude product by silica gel column chromatography (petroleum ether/ethyl acetate 1:0 to 0:1) to obtain an intermediate B;

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

example 1

This example provides compound 1 of formula I, wherein compound 1 has the following structural formula:

the synthetic route of compound 1 is shown below:

(1) to a solution of intermediate B (400mg,1.58mmol) and compound 1-1(394mg,1.74mmol) in N, N-dimethylformamide (8.0mL) was added potassium carbonate (437mg,3.16mmol), and the reaction solution was stirred at 50 ℃ for 3 hours. Adding ethyl acetate (20.0mL), washing with saturated brine (20.0mL × 5), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and separating the crude product by silica gel column chromatography (petroleum ether/ethyl acetate 10:1 to 1:1) to obtain compound 1-2;

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

(2) to a solution of compound 1-2(576mg,1.30mmol) in dichloromethane (5.0mL) was added a dioxane solution of hydrogen chloride (4mol/L,2.0mL), and the reaction solution was stirred at 20 ℃ for 2 hours. Concentrating the reaction solution under reduced pressure to obtain hydrochloride of the compound 1-3;

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

(3) to a solution of the hydrochloride salt of compound 1-3 (370mg,0.98mmol) in methanol (15.0mL) was added triethylamine to adjust the pH to 7. Compound 1-4(255mg,0.98mmol) was added, and the reaction solution was stirred at 25 ℃ for 15 minutes. Sodium cyanoborohydride (245mg,3.91mmol) was then added and stirring continued at 25 ℃ for 12 h. Quenching the reaction with water (20.0mL), extracting with ethyl acetate (10.0mL × 1), washing the organic phase with saturated brine (10.0mL × 3), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and separating the crude product by silica gel column chromatography (petroleum ether/ethyl acetate ═ 100:1 to 1:1) to obtain compounds 1-5;

MS-ESI[M+H]⁺calculated 588, found 588.

(4) To a solution of compound 1-5(347mg,0.59mmol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was filtered and concentrated under reduced pressure to give trifluoroacetate salts of compounds 1 to 6.

The crude product is directly used for the next reaction;

(5) to a solution of the trifluoroacetate salt of compounds 1 to 6 (340mg, 565. mu. mol) in N, N-dimethylformamide (8.0mL) was added triethylamine to adjust the pH to 8, followed by addition of intermediate A (289mg,1.13mmol), and the reaction solution was stirred at 25 ℃ for 0.5 hour. The reaction was quenched by the addition of water (15.0mL), extracted with ethyl acetate (10.0 mL. times.3), and the combined organic phases were washed with saturated brine (10.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 1-7;

MS-ESI[M+H]⁺value 707 is calculated as an actual value 707.

(6) To a solution of compounds 1 to 7(100mg,0.14mmol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.5mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was filtered and concentrated under reduced pressure to give trifluoroacetate salts of compounds 1 to 8. The crude product was used directly in the next reaction.

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

MS-ESI[M+H]⁺Calculated 718, found 718.

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

Example 2

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

the synthetic route of compound 2 is shown below:

to a solution of the trifluoroacetate salts of intermediates 1-8 in example 1 (100mg, 139. mu. mol) in dichloromethane (5.0mL) was added triethylamine to adjust the pH to 8. Compound 2-1(18.8mg, 208. mu. mol, 17.0. mu.L) was then added, and the reaction was stirred at-78 ℃ for 0.5 hour. The reaction was quenched by the addition of water (10.0mL), extracted with dichloromethane (10.0 mL. times.3), and the combined organic phases were washed with saturated brine (10.0 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparation of crude product by high performance liquid chromatography (Kromasil C18, 150mm. times.30 mm 5 μm, A: water (0.2% trifluoroacetic acid), B: acetonitrile, 5% -65%: 20 min; 100%: 5 min) to give Compound 2.

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

¹H NMR(400MHz,MeOD)δ8.49(s,1H),7.74(s,1H),7.38(d,J＝7.8Hz,1H),7.34-7.26(m,2H),6.35-6.18(m,2H),5.76(d,J＝9.6Hz,1H),4.61-4.48(m,4H),4.44-4.31(m,3H),4.20(d,J＝6.8Hz,2H),4.13-3.93(m,6H),3.67(t,J＝6.0Hz,4H),3.48(s,1H),3.12-2.89(m,3H),2.30(s,4H)。

Example 3

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

the synthetic route of compound 3 is shown below:

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

MS-ESI[M+H]⁺calculated 443, found 443.

¹H NMR(400MHz,CDCl₃)δ8.44(s,1H),7.13(s,1H),4.13(br s,4H),3.64(q,J＝10.0Hz,2H),3.45(br s,4H),1.84(br t,J＝5.6Hz,4H),1.48(s,9H)。

(2) To a solution of compound 3-2(270mg,1.30mmol) in ethyl acetate (5.0mL) was added a dioxane solution of hydrogen chloride (4mol/L,5.0mL), and the reaction mixture was stirred at 25 ℃ for 3 hours. Concentrating the reaction solution under reduced pressure to obtain hydrochloride of the compound 3-3;

MS-ESI[M+H]⁺calculated value 343, found value 343.

(3) To a solution of hydrochloride of compound 3-3 (200mg, 584. mu. mol) in methanol (10.0mL) were added triethylamine (177mg,1.75mmol), compound 3-4(153mg, 584. mu. mol) and sodium cyanoborohydride (110mg,1.75mmol), and the mixture was stirred at 25 ℃ for 12 hours. Adding an aqueous ammonium chloride solution (50.0mL) to quench the reaction, extracting with ethyl acetate (50.0 mL. times.1), washing the organic phase with a saturated saline solution (50.0 mL. times.2), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and separating the crude product by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to obtain compound 3-5;

MS-ESI[M+H]⁺calculated 588, found 588.

(4) To a solution of compound 3-5(150mg, 584. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (3.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 3-6. The crude product was used directly in the next reaction.

(5) To a solution of the trifluoroacetate salt of compound 3-6 (130mg, 267. mu. mol) in dichloromethane (3.0mL) were added triethylamine (81mg, 800. mu. mol) and intermediate A (82mg, 320. mu. mol), and the reaction solution was stirred at 25 ℃ for 1 hour. The reaction was quenched by the addition of water (10.0mL), extracted with ethyl acetate (20.0 mL. times.1), and the combined organic phases were washed with saturated brine (10.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 93:7) to afford compound 3-7.

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

¹H NMR(400MHz,CDCl₃)δ8.43(s,1H),7.08-7.18(m,3H),7.01-7.07(m,1H),4.48-4.57(m,2H),4.20-4.30(m,2H),4.08-4.12(m,4H),3.95(s,1H),3.62-3.69(m,4H),3.43-3.51(m,2H),2.89-2.98(m,2H),2.33-2.48(m,2H),1.82-1.98(m,4H),1.41-1.46(m,9H),1.28-1.35(m,4H)。

(6) To a solution of compound 3-7(143mg, 202. mu. mol) in dichloromethane (8.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 3-8. The crude product is directly used for the next reaction;

MS-ESI[M+H]⁺calculated 607, found 607.

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

MS-ESI[M+H]⁺Calculated 718, found 718.

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

Example 4

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

the synthetic route of compound 4 is shown below:

to a solution of the trifluoroacetate salt of intermediate 3-8 in example 3 (60mg, 99. mu. mol) in dichloromethane (5.0mL) was added triethylamine (10.0mg, 99. mu. mol). Compound 4-1(9mg, 99. mu. mol, 8.1. mu.L) was then added, and the reaction was stirred at-78 ℃ for 20 minutes. The reaction was quenched by the addition of brine (10.0mL), extracted with dichloromethane (20.0 mL. times.1), and the combined organic phases were washed with brine (10.0 mL. times.3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparation of crude product by high performance liquid chromatography (ACQUITY C18,4.6 mm. times.30 mm. 1.7 μm, A: water (0.01% trifluoroacetic acid), B: acetonitrile (0.01% trifluoroacetic acid), 5% -95% for 1.5 min, 95% for 1 min, 5% for 0.01 min) to isolate the trifluoroacetate salt of Compound 4;

MS-ESI[M+H]⁺calculated value 661, found value 661.

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

Example 5

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

the synthetic route of compound 5 is shown below:

(1) to a solution of hydrochloride of compound 1-3 (200mg, 438. mu. mol) in example 1 in methanol (5.0mL) were added triethylamine (44.3mg, 438. mu. mol), compound 5-1(115mg, 438. mu. mol) and sodium cyanoborohydride (82.6mg,1.31mmol), and the mixture was stirred at 25 ℃ for 2 hours. Quenching the reaction with saturated common salt solution (50.0mL), extracting with dichloromethane (50.0mL × 1), washing the organic phase with saturated common salt solution (50.0mL × 2), drying over anhydrous sodium sulfate, filtering, concentrating the organic phase under reduced pressure, and separating the crude product by silica gel column chromatography (petroleum ether/ethyl acetate 10:1 to 0:1) to obtain compound 5-2;

MS-ESI[M+H]⁺calculated value 589, found value 589.

(2) To a solution of Compound 5-2(72mg, 122. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 5-3. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculated 489, found 489.

(3) To a solution of the trifluoroacetate salt of compound 5-3 (70mg, 116. mu. mol) in N, N-dimethylformamide (8.0mL) was added triethylamine (35mg, 348. mu. mol), followed by intermediate A (30mg, 116. mu. mol), and the reaction solution was stirred at 25 ℃ for 5 minutes. The reaction was quenched by the addition of brine (20.0mL), extracted with dichloromethane (20.0 mL. times.1), and the combined organic phases were washed with brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 100:1 to 10:1) to give compound 5-4.

MS-ESI[M+H]⁺Calculated value 708 found value 708.

(4) To a solution of compound 5-4(53mg, 74.9. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 20 minutes. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 5-5. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculated 608 is measured 608.

(5) To a solution of the compound 5-6(50mg, 69.3. mu. mol) in dichloromethane (5.0mL) was added triethylamine (7.0mg, 69.3. mu. mol), the trifluoroacetate salt of the compound 5-5 (9.0mg, 69.3. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (26.3mg, 69.3. mu. mol), and the reaction solution was stirred at 25 ℃ for 0.5 hour. The reaction was quenched by addition of brine (20.0mL), extracted with dichloromethane (20.0 mL. times.1), and the organic phase was washed with brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Preparation of crude product by high performance liquid chromatography (Phenomenexlunna C18, 100mm. times.40 mm 3 μm, A: water (0.225% formic acid), B: acetonitrile, 0% -35% for 10 min) to obtain the formate of Compound 5.

MS-ESI[M+H]⁺Calculated 719 and found 719.

¹H NMR(400MHz,MeOD)δ8.40-8.33(m,1H),8.31-8.22(m,1H),7.70-7.58(m,2H),6.87-6.70(m,1H),6.33-6.20(m,1H),4.61-4.51(m,4H),4.44-4.37(m,1H),4.34-4.23(m,2H),3.95-3.79(m,5H),3.74(br s,5H),3.38-3.33(m,2H),3.01-2.94(m,2H),2.85-2.79(m,1H),2.76-2.67(m,2H),2.62-2.54(m,1H),2.47-2.37(m,6H),2.16-2.02(m,2H),1.99-1.90(m,2H)。

Example 6

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

the synthetic route of compound 6 is shown below:

(1) to a solution of the trifluoroacetate salt of compound 3-3 in example 3 (360mg, 789. mu. mol) in methanol (15.0mL) was added triethylamine (160mg,1.58mmol), compound 6-1(247mg, 947. mu. mol), and the mixture was stirred at 25 ℃ for 15 minutes, added sodium cyanoborohydride (198mg,3.16mmol), and stirred at 25 ℃ for 1 hour. The reaction was quenched by addition of saturated brine (50.0mL), extracted with dichloromethane (50.0mL × 1), the organic phase was washed with saturated brine (25.0mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 20:1 to 0:1) to give compound 6-2.

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

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

(2) To a solution of Compound 6-2(220mg, 220. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 15 minutes. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 6-3. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculated 488, found 488.

(3) To a solution of the trifluoroacetate salt of compound 6-3 (220mg, 366. mu. mol) in dichloromethane (5.0mL) was added triethylamine to adjust the pH to 8. Intermediate A (140mg, 549. mu. mol) was added and the reaction was stirred at 25 ℃ for 0.5 hour. The reaction was quenched by the addition of water (20.0mL), extracted with dichloromethane (20.0 mL. times.3), and the combined organic phases were washed with saturated brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 6-4.

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

(4) To a solution of Compound 6-4(125mg, 177. mu. mol) in dichloromethane (6.0mL) was added trifluoroacetic acid (2.0mL), and the reaction was stirred at 25 ℃ for 30 minutes. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 6-5. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculated 607, found 607.

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

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

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

Example 7

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

the synthetic route of compound 7 is shown below:

to a solution of the trifluoroacetate salt (60mg, 83.3. mu. mol) of compound 6-5 of example 6 in dichloromethane (3.0mL) was added triethylamine (8.4mg, 83.3. mu. mol), compound 7-1(27.6mg, 167. mu. mol) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (63.3mg, 167. mu. mol), and the reaction mixture was stirred at 25 ℃ for 0.5 hour. The reaction was quenched by the addition of water (20.0mL), extracted with dichloromethane (20.0 mL. times.3), and the organic phase was washed with saturated brine (20.0 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Crude preparation by HPLC (1_ Welch Xtimate,70 mm. times.40 mm 3 μm, A: water (0.225% formic acid) and B: acetonitrile, 10% to 40% for 10 minutes) the formate salt of compound 7 was isolated.

MS-ESI[M+H]⁺Calculated 719 and found 719.

¹H NMR(400MHz,MeOD)δ8.32-8.25(m,1H),7.63-7.56(m,1H),7.36-7.25(m,2H),7.23-7.16(m,1H),6.82-6.71(m,1H),6.42-6.31(m,1H),4.62-4.50(m,4H),4.40-4.32(m,1H),4.28-4.21(m,2H),4.18-4.08(m,2H),3.99-3.78(m,6H),3.73-3.59(m,4H),3.28-3.05(m,4H),3.00-2.91(m,2H),2.73-2.60(m,6H),2.26-2.04(m,4H)。

Example 8

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

the synthetic route of compound 8 is shown below:

(1) to a solution of intermediate B (200mg, 792. mu. mol) and compound 8-1(179mg, 792. mu. mol) in N, N-dimethylformamide (8.0mL) was added potassium carbonate (219mg,1.58mmol), and the reaction was stirred at 80 ℃ for 2 hours. Saturated aqueous ammonium chloride (20.0mL) was added, extraction was performed with dichloromethane (50.0mL), and then the organic phase was washed with saturated brine (20.0mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 50:1 to 1:1) to give compound 8-2.

MS-ESI[M+H]⁺Calculated 443, found 443.

(2) To a solution of Compound 8-2(218mg, 493. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 8-3.

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

(3) To a solution of the trifluoroacetate salt of compound 8-3 (224mg, 491. mu. mol) in methanol (10.0mL) was added triethylamine (149mg,1.47 mmol). Subsequently, compound 8-4(129mg, 494. mu. mol) and sodium cyanoborohydride (308mg,4.90mmol) were added, and the reaction solution was stirred at 25 ℃ for 2 hours. Ethyl acetate (20.0mL) was added, the organic phase was washed with water (20.0mL) and saturated brine (20.0mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 1:0 to 10:1) to give compound 8-5.

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

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

(4) To a solution of compound 8-5(94.3mg, 160. mu. mol) in dichloromethane (3.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 1 hour. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 8-6. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculated 488, found 488.

(5) To a solution of the trifluoroacetate salt of compound 8-6 (96.5mg, 160. mu. mol) in dichloromethane (5.0mL) was added triethylamine (81.1mg, 801. mu. mol), followed by intermediate A (61.7mg, 241. mu. mol), and the reaction solution was stirred at 25 ℃ for 2 hours. The reaction was quenched by the addition of water (15.0mL), extracted with dichloromethane (20.0mL), and the organic phase was washed with water (20.0mL) and saturated brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 10:1) to afford compounds 8-7.

MS-ESI[M+H]⁺Value 707 is calculated as an actual value 707.

¹H NMR(400MHz,MeOD)δ8.33(s,1H),7.51(s,1H),7.15(dt,J＝16.0,8.0Hz,3H),4.49(s,2H),4.19-4.33(m,1H),4.09-4.16(m,4H),3.90-3.94(m,1H),3.85-3.89(m,5H),3.70(s,2H),3.62(t,J＝6.0Hz,2H),3.24(s,4H),2.94(t,J＝5.6Hz,2H),1.89-1.93(m,4H),1.43(s,9H)。

(6) Trifluoroacetic acid (1.0mL) was added to a solution of compound 8-7(56.0mg, 79.2. mu. mol) in dichloromethane (3.0mL), and the reaction was stirred at 25 ℃ for 1 hour. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 8-8. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculated value 607 found value 607.

(7) To a solution of the trifluoroacetate salt of compound 8-8 (57.1mg, 79.2. mu. mol) in dichloromethane (3.0mL) were added compound 8-9(10.7mg, 118. mu. mol) and triethylamine (40.0mg, 395. mu. mol), and the reaction solution was stirred at-78 ℃ for 1 hour. Dichloromethane (20.0mL) was added, and the organic phase was washed with water (20.0mL) and saturated brine (20.0 mL. times.2) successively, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Crude preparation high performance liquid chromatography (xtime C18, 100mm. times.30 mm 10 μm, A: water (0.225% formic acid), B: acetonitrile, 15% -45% for 10 min) isolated the formate salt of compound 8.

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

¹H NMR(400MHz,MeOD)δ8.35(s,1H),7.50(s,1H),7.24-7.30(m,2H),7.18-7.22(m,1H),6.21-6.36(m,2H),5.74-5.80(m,1H),4.49-4.56(m,4H),4.31-4.37(m,1H),4.24(d,J＝7.2Hz,2H),4.09(s,2H),3.85-3.93(m,6H),3.70(s,4H),3.65(td,J＝6.0,2.4Hz,2H),2.98(br t,J＝6.0Hz,2H),1.94-2.02(m,4H)。

Example 9

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

the synthetic route of compound 9 is shown below:

(1) to a solution of intermediate B (200mg, 792. mu. mol) and compound 9-1(188mg, 950. mu. mol) in N, N-dimethylformamide (10.0mL) was added potassium carbonate (219mg,1.58mmol), and the reaction was stirred at 25 ℃ for 2 hours. Saturated brine (50.0mL) was added, extraction was performed with dichloromethane (50.0mL), and then the organic phase was washed with saturated brine (50.0mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate ═ 50:1 to 1:1) to give compound 9-2.

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

(2) To a solution of Compound 9-2(274mg, 661. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction mixture was stirred at 25 ℃ for 0.5 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 9-3.

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

(3) To a solution of the trifluoroacetate salt of compound 9-3 (280mg, 891. mu. mol) in methanol (10.0mL) was added triethylamine (90.1mg, 891. mu. mol). Compound 9-4(349mg,1.34mmol) and sodium cyanoborohydride (55.9mg, 891. mu. mol) were then added, and the reaction was stirred at 25 ℃ for 1 hour. Ethyl acetate (40.0mL) was added, the organic phase was washed with water (20.0mL) and saturated brine (20.0mL × 2) in this order, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure from the organic phase, and the crude product was separated by silica gel column chromatography (dichloromethane/methanol ═ 100:1 to 10:1) to give compound 9-5.

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

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

(4) To a solution of compound 9-5(100mg, 179. mu. mol) in dichloromethane (10.0mL) was added trifluoroacetic acid (2.5mL), and the reaction was stirred at 25 ℃ for 0.5 hour. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 9-6. The crude product was used directly in the next reaction.

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

(5) To a solution of the trifluoroacetate salt of compound 9-6 (100mg, 174. mu. mol) in dichloromethane (5.0mL) was added triethylamine (70.6mg, 697. mu. mol), followed by intermediate A (66.9mg, 262. mu. mol), and the reaction solution was stirred at 25 ℃ for 5 minutes. The reaction was quenched by the addition of water (30.0mL), extracted with dichloromethane (40.0mL), and the organic phase was washed with saturated brine (20.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was isolated by silica gel column chromatography (dichloromethane/methanol ═ 100:1 to 10:1) to afford compound 9-7.

MS-ESI[M+H]⁺Calculated value 679 found value 679.

(6) To a solution of compound 9-7(77.0mg, 113. mu. mol) in dichloromethane (4.0mL) was added trifluoroacetic acid (1.0mL), and the reaction was stirred at 25 ℃ for 1 hour. The reaction solution was filtered and concentrated under reduced pressure to give the trifluoroacetate salt of compound 9-8. The crude product was used directly in the next reaction.

MS-ESI[M+H]⁺Calculating a value579 found 579.

(7) To a solution of the trifluoroacetate salt of compound 9-8 (75.0mg, 108. mu. mol) in dichloromethane (5.0mL) were added compound 9-9(9.8mg, 108. mu. mol) and triethylamine (32.9mg, 325. mu. mol), and the reaction solution was stirred at-78 ℃ for 5 minutes. Dichloromethane (50.0mL) was added, and the organic phase was washed with saturated brine (50.0 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Crude preparation high performance liquid chromatography (xtime C18, 100mm. times.30 mm 10 μm, A: water (0.225% formic acid), B: acetonitrile, 10% -40% for 10 min) isolated the formate salt of compound 9.

MS-ESI[M+H]⁺Calculated 633 and measured 633.

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

Test examples

Determination of the antiproliferative Effect of Compounds on MV-4-11 cells:

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

2. Experimental materials: cell Counting Kit-8 was purchased from Shanghai Liji Biotechnology Ltd (cat # D3100L 4057); 96-well clear-bottomed white cell culture plates were purchased from Corning Costar (cat No. 3610); fetal bovine serum was purchased from GIBCO (cat # 10099-141); eiskov modified broth (IMDM) medium was purchased from Invitrogen (cat # 12440046); desktop microplate reader SpectraMax i3X was purchased from Molecular Devices;

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

The test compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution with a concentration of 10mM, and diluted with DMSO in 4-fold gradient step by step for 8 times. Then diluted 20-fold with medium. The final concentration of the compound was 100. mu.M, 25. mu.M, 6.25. mu.M, 1.56. mu.M, 0.39. mu.M, 0.098. mu.M, 0.024. mu.M, 0.006. mu.M, 0.0015. mu.M (4-fold dilution, 9 concentrations) in this order, and the amount of the compound was added to the 96-well plate seeded with cells at 25. mu.L/well.

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

4. Data processing:

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

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

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

And calculating IC of Compounds with Graphpad software₅₀。

The specific test method is shown in table 1:

TABLE 1

Test compounds	MV 4-11IC₅₀(nM)
		Example 1	1819
Example 2	6193
		Practice ofExample 3	1087
Example 4	>10000
		Example 5	409.9
Example 6	336.4
		Example 7	41.68
Example 8	2406
		Example 9	6062

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

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

Claims

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

wherein the content of the first and second substances,

Wherein R is^a、R^bEach independently selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclyl, or R^aAnd R^bA substituted or unsubstituted 4-8 membered heterocyclic ring formed by linking with N, wherein R^aAnd R^bThe heterocyclic ring formed by connecting with N contains 1-3 heteroatoms selected from N, O, S, P,

represents the attachment position of the group;

R²selected from H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C16 aryl, substituted or unsubstituted 5-16 membered heteroaryl, substituted or unsubstituted 4-8 membered heterocyclyl, substituted or unsubstituted C1-C6 alkylcarbonyl, substituted or unsubstituted

Substituted or notSubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted C1-C6 alkyl sulfone, substituted or unsubstituted C1-C6 alkyl sulfoxide,

Wherein R is^c、R^dEach independently selected from H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-8 membered heterocyclyl, or R^cAnd R^dA substituted or unsubstituted 4-8 membered heterocyclic ring formed by linking with N, wherein R^cAnd R^dThe heterocyclic ring formed by connecting with N contains 1-3 heteroatoms selected from N, O, S, P,

represents the attachment position of the group;

represents the attachment position of the group;

w is selected from N or C;

And U is¹、U²、U³、U⁴At most one of them is

U⁵、U⁶At most one of them is

U⁷、U⁸At most one of them is

Wherein the content of the first and second substances,

represents the attachment position of the group,

each R "" is independently selected from: H. halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

L¹to be out of stockAt the position of,

Wherein the content of the first and second substances,

L²is selected from

represents the attachment position of the group;

x is selected from carbon atoms or

Wherein the content of the first and second substances,

represents the attachment position of the group;

R⁴is selected from

Wherein R4 'is selected from fluorine or chlorine, R4' is selected from H, methyl or fluorine, R⁴"' is selected from H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, substituted or unsubstituted (C1-C4 alkyl)₂Amino, substituted or unsubstituted C1-C4 alkylthio, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 4-to 8-membered heterocyclyl, substituted or unsubstituted heteroalkylthioOr an unsubstituted C2-C4 acyl group, said heterocyclyl group containing 1-3 heteroatoms selected from N, O, S, P, wherein,

indicates the attachment position of the group.

2. The thienopyrimidine compound according to claim 1, wherein R is¹Selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, and substituted or unsubstituted 4-8 membered heterocyclyl, wherein said heterocyclyl contains 1-3 heteroatoms selected from N, O, S, P;

preferably, said R is¹Selected from substituted or unsubstituted C1-C6 alkyl, more preferably halogen substituted C1-C6 alkyl, still more preferably trifluoroethyl;

preferably, said R is²Selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C16 aryl, substituted or unsubstituted 5-16 membered heteroaryl, substituted or unsubstituted 4-8 membered heterocyclyl, wherein said heteroaryl or heterocyclyl contains 1-3 heteroatoms selected from N, O, S, P, further preferably, said R is²Is H;

preferably, said R is³Is H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C1-C4 alkylamino, -NH₂or-CN;

preferably, said Y, Z are each selected from N;

preferably, said W is C;

And U is¹、U²、U³、U⁴At most one of them is

U⁵、U⁶At most one of them is

U⁷、U⁸At most one of them is

Wherein the content of the first and second substances,

represents the attachment position of the group,

each R "" is independently selected from H, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, cyano;

preferably, A is selected from substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heteroaryl, wherein said heteroaryl contains 1-3 heteroatoms selected from N, O, S, P;

preferably, a is a substituted or unsubstituted benzene ring, a substituted or unsubstituted pyridine ring, a substituted or unsubstituted pyridazine ring, a substituted or unsubstituted pyrimidine ring, a substituted or unsubstituted triazazine ring, a substituted or unsubstituted thiophene ring, a substituted or unsubstituted thiazole ring, a substituted or unsubstituted imidazole ring, a substituted or unsubstituted pyrrole ring, a substituted or unsubstituted pyrazole ring, a substituted or unsubstituted oxazole ring, a substituted or unsubstituted isoxazole ring, a substituted or unsubstituted triazoizole ring;

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

Preferably, said L²Is selected from

Preferably, it is

Wherein the content of the first and second substances,

represents the attachment position of the group;

preferably, said X is selected from carbon atoms;

preferably, said R is⁴Is selected from

indicates the attachment position of the group.

3. The thienopyrimidine compound according to claim 1 or 2, wherein the spiro moiety in the compound represented by formula I has a structural formula of

Wherein

Wherein the content of the first and second substances,

4. the thienopyrimidine compound of any one of claims 1 to 3, wherein the formula in formula I is

Shown as a fused ring portion, wherein

wherein R is^e、R^fEach independently selected from H, methyl, trifluoromethyl, difluoromethyl, methoxy, ethoxy, trifluoromethoxy, halogen, hydroxy, amino, cyano, methylamino, dimethylamino, ethylamino, methylethylamino, diethylaminoA group, a trifluoroethylamino group, a carboxyl group, a methoxycarbonyl group, an ethoxycarbonyl group, a carbamoyl group, a methylcarbamoyl group, a dimethylcarbamoyl group, a methylethylcarbamoyl group or a diethylcarbamoyl group.

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

The ring-shaped portion is shown in which, among other things,

6. the thienopyrimidine compound according to any one of claims 1 to 5, wherein R is⁴Is selected from-CH₂F、-CH₂Cl、

7. The thienopyrimidine compound according to any one of claims 1 to 6, wherein the compound represented by the formula I is selected from any one of the following compounds:

8. the thienopyrimidine compound of any one of claims 1 to 7, further comprising any one of a pharmaceutically acceptable salt, enantiomer, diastereomer, tautomer, cis-trans isomer, solvate or polymorph or deuteron of the compound represented by formula I.

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

10. The thienopyrimidine compound according to any one of claims 1 to 8 or the pharmaceutical composition according to claim 9, wherein the use is selected from any one of the following (a) to (c):

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