WO2022166482A1

WO2022166482A1 - Benzimidazole enl protein inhibitor, preparation method therefor, and use thereof

Info

Publication number: WO2022166482A1
Application number: PCT/CN2021/142675
Authority: WO
Inventors: 李海涛; 林木; 张士猛; 李小林; 吴亦钦
Original assignee: 清药同创(北京)药物研发中心有限公司
Priority date: 2021-02-04
Filing date: 2021-12-29
Publication date: 2022-08-11
Also published as: CN114853679A

Abstract

A benzimidazole ENL protein inhibitor compound as represented by formula I, and a racemate, stereoisomer, tautomer, isotopic marker, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug, or pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising same, a preparation method therefor, and a pharmaceutical use thereof. The structure of formula I is as follows.

Description

A kind of benzimidazole ENL protein inhibitor and preparation method and use thereof

This application claims to enjoy the patent application number 202110166935.7 submitted by the applicant to the State Intellectual Property Office of China on February 4, 2021, and the name of the invention is "a benzimidazole ENL protein inhibitor and its preparation method and use" in The priority of the first application. The entire contents of this prior application are incorporated herein by reference.

technical field

The invention belongs to the field of pharmaceutical compounds, in particular to a benzimidazole ENL protein inhibitor and a preparation method and application thereof.

Background technique

Epigenetic modification plays an important role in the occurrence and development of tumors, and a variety of epigenetic regulation mechanisms have been found, including DNA methylation, histone modification, and non-coding RNA regulation. Covalent modification of histones plays an important role in gene regulation. Common histone modifications include methylation, acetylation, phosphorylation, ubiquitination, and small ubiquitination (SUMO). Different types of histone-modifying proteins or domains are called "readers" in cells. The "reader" is one of the main ways of epigenetic regulation. The "readers" of histone acetylation that have been discovered so far mainly include the bromodomain, the plant zinc finger homology domain and the YEATS domain. Abnormal expression of these domains or proteins containing these domains can lead to the occurrence and development of a series of diseases, such as cancer, inflammatory diseases, cardiovascular diseases and neurological diseases.

AF9 protein is an important class of transcriptional regulators, which can form complexes with histone H3K79 methyltransferase DOT1L and superelongation complex (SEC), respectively, and play an important regulatory role in the transcription initiation-elongation transition. At the same time, clinical genetic studies have shown that the fusion of the AF9 gene and the MLL gene (MLL-AF9), as the most common type of MLL rearrangement, is the driver of diseases such as human acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). sexual factors. Further cell biology and functional genomics studies have found that human AF9 and H3K9ac modifications have strong co-localization at the genome-wide level, and regulate the expression of cell proliferation and differentiation genes including MYC, BMP2, and HOXA gene clusters; studies have shown that AF9 recruits DOTL1 to specific chromatin segments by recognizing histone H3K9 acetylation modifications, promoting histone H3K79 methylation co-deposition and gene activation, revealing a novel histone modification cross-session mechanism that embodies eukaryotic The complexity and sophistication of biological epigenetic regulation. At present, hundreds of proteins containing YEATS domains have been identified in nature, distributed in more than 70 different species, and conserved from yeast to humans. Protein factors containing YEATS domains, such as AF9, ENL, GAS41, etc., are not only closely related to transcriptional regulation, but their abnormal regulation usually leads to the occurrence of human diseases such as leukemia and cancer.

Among them, ENL proteins are a class of important histone acetylation "reader" epigenetic modification proteins, which play an important role in the progression of leukemia. Studies have shown that AML has a variety of cell molecular genetics and epigenetic abnormalities including abnormal expression of cell surface molecules, gene mutation and abnormal gene methylation in the process of occurrence and development, while ENL is involved in the process of leukemia. It plays an important role in maintaining the protein necessary for the occurrence and development of AML, so targeting ENL protein is a good drug target for the treatment of AML. In 2017, Nature simultaneously published the research results of two research teams on ENL protein. It was confirmed that ENL protein is a key factor affecting the survival ability of leukemia MLL-r cells. The ENL YEATS domain is important for the transcriptional control of leukemia-causing gene expression (Wan L et al, Nature, 2017, 543(7644):265.). Disrupting the interaction of the ENL YEATS domain with histone acetylation renders histone acetylation unrecognized and inhibits the growth of many types of leukemia cell lines, including MLL-r and non-MLL-r leukemia cell lines ( Wilkinson A W et al, Nature, 2017, 543(7644):186-188.). These studies all show that ENL protein is a good drug target. Acute myeloid leukemia (AML) is a complex blood disorder characterized by an increase in leukemic blasts in the blood or bone marrow. It is the most common type of acute leukemia in adult leukemia, accounting for about 70% of the incidence of adult leukemia. In recent years, with the continuous development of combined chemotherapy methods, the cure rate of AML has improved, but about 40% of patients still cannot achieve complete remission (CR), and the prognosis of AML is poor. Only 40%-50% have a 5-year survival period, while the average survival period of elderly patients is less than 1 year, and the overall 5-year survival rate of patients is only 26.9%. Recurrent episodes and chemoresistance of AML remain a therapeutic challenge in this field.

At present, only bromodomain inhibitors of the three histone acetylation "readers" - BET inhibitors are in active development, and no ENL inhibitors have entered the clinical stage. Therefore, there is an urgent need to develop a new class of ENL inhibitors with novel structures, good activity, and more safety and efficacy.

SUMMARY OF THE INVENTION

In order to solve the problems existing in the prior art, the present invention provides a compound represented by formula I and its racemate, stereoisomer, tautomer, isotopic label, nitrogen oxide, solvate, polyamide, etc. Crystal forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof:

Wherein, the Ar ₁ and Ar ₂ may be the same or different, and are each independently selected from the following groups optionally substituted by one, two or more R _a :

Said R _a is selected from H, =O, halogen, OH, CN, SH, NH ₂ , COOH, or optionally substituted with one, two or more R _b : C ₁ -C ₁₂ aliphatic Hydrocarbyl; the R _b is selected from halogen, OH, CN, SH, NH ₂ , COOH;

The L ₁ and L ₂ can be the same or different, each independently selected from the bond, or optionally substituted by one, two or more R _L

(*, ** represent the connection site with other group parts of the general formula, and * represents the connection with the corresponding left group of the general formula, then ** represents the connection with the corresponding right group of the general formula); R _L is selected from H, =O, halogen, OH, CN, SH, NH ₂ , COOH, or optionally substituted with one, two or more R _c : C ₁ -C ₁₂ aliphatic hydrocarbyl; The R _c is selected from halogen, OH, CN, SH, NH ₂ , COOH;

The W is selected from (C ₁ -C ₂₀ ) aliphatic hydrocarbon groups, optionally containing one, two or more heteroatoms (C ₁ -C ₂₀ ) aliphatic hydrocarbon groups; or more R _W substituted; said R _W is selected from H, =O, halogen, OH, CN, SH, NH ₂ , COOH, or optionally substituted with one, two or more R _d Group: C ₁ -C ₁₂ aliphatic hydrocarbon group; the R _d is selected from halogen, OH, CN, SH, NH ₂ , COOH;

The R ₁ and R ₂ can be the same or different, and are independently selected from -NH-C(=O)R ₃ , -(CH ₂ ) _p R ₃ , (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally (C ₁ -C ₁₂ )aliphatic hydrocarbon groups containing one, two or more heteroatoms;

Said _R is selected from the following groups optionally substituted by one, two or more R _e : C _6-20 aryl, 5-14 membered heteroaryl or 3-12 membered heterocyclyl;

The _Re is selected from H, -C(=O)OC ₁ -C ₁₂ aliphatic hydrocarbon group, -OC ₁ -C ₁₂ aliphatic hydrocarbon group, C ₁ -C ₁₂ aliphatic hydrocarbon group, =O, halogen, OH, CN, SH, NH ₂ , COOH; p is selected from 0, 1, 2, 3, 4, 5;

According to an embodiment of the present invention, the Ar ₁ and Ar ₂ may be the same or different, and are each independently selected from the following groups (*, ** represent the attachment site to other group parts of the general formula, and * represents the The general formula corresponds to the left group part of the connection, then ** represents the connection with the corresponding right group of the general formula):

According to an embodiment of the present invention, the W is selected from the following groups: -( _CH2 )n-, -O-( _CH2 )nO-, -O-( _CH2 )n-, _- ( _OCH2CH2 ) _m -O-, -(OCH ₂ CH ₂ )m-, -(CH ₂ CH ₂ O)m-CH ₂ CH ₂ -, -(CH ₂ )s-NR _s -(CH ₂ CH ₂ O)m -CH2CH2-NRs-( _CH2 ) _s- , -( _CH2 ) _s -NRs-( _CH2 ₎ n-NRs-( _CH2 ₎ _s- ;

The Rs is selected from H, C ₁ -C ₁₂ aliphatic hydrocarbon group;

The s is selected from 0, 1, 2; the n is selected from 1-16, for example, selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 , 15, 16; the m is selected from 1-8, such as selected from 1, 2, 3, 4, 5, 6, 7, 8;

According to an embodiment of the present invention, preferably, when the R ₁ and R ₂ are independently selected from (C ₁ -C ₁₂ ) aliphatic hydrocarbon groups optionally containing one, two or more heteroatoms, further is selected from -(CH ₂ ) _q N(CH ₃ ) ₂ , the q is selected from 0, 1, 2, 3;

According to an embodiment of the present invention, preferably, the R ₃ is selected from the following groups optionally substituted by one, two or more R _e : phenyl, indazolyl, pyrrolidinyl.

According to an embodiment of the present invention, preferably, the R ₁ and R ₂ may be the same or different, and each is independently selected from the following groups:

According to an embodiment of the present invention, the "halogen" is selected from F, Cl, Br, I; the (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, (C ₁ -C ₂₀ ) aliphatic hydrocarbon group may be selected from (C ₁ -C 20 ) aliphatic hydrocarbon group C ₁₂ ) alkyl, (C ₂ -C ₁₂ ) alkenyl, (C ₂ -C ₁₂ ) alkynyl;

According to an embodiment of the present invention, the structure of formula I is selected from the following formula Ia or formula Ib:

In the structure of formula Ia, R ₁ , R ₂ , W, L ₁ , and L ₂ are as defined in formula I above.

According to an embodiment of the present invention, the structure of formula I may be selected from the following formulae II-VII:

In the structures of formula II-VII, R ₁ , R ₂ , R _S , s, n, m are as defined in formula I above.

According to an embodiment of the present invention, the compounds represented by Formula I (including Formula II to Formula VII) and their racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, and solvates , polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, illustrative, non-limiting specific examples of compounds of formula I are shown below:

According to an embodiment of the present invention, the compound of formula I may further be selected from, for example, the following structures:

The present invention also provides the compounds represented by the formula I (including formula II to VII) and their racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs The preparation method of the form, metabolite, ester, prodrug or its pharmaceutically acceptable salt, but not limited to the method described below. All starting materials are based on the group characteristics of the target molecule conforming to the general formula, and are prepared through the schemes in these routes, methods well known to those of ordinary skill in the field of organic chemistry, or purchased directly. The compounds of the present invention can be synthesized using the methods described below in combination with synthetic methods known in the art of synthetic organic chemistry, or with relevant modifications recognized by those skilled in the art. Those skilled in the art know that, according to a specific target structure, one or more of the following schemes can be optionally combined, or any steps in one or more of the schemes can be combined to obtain a synthesis scheme.

The preparation method of the compound of the present invention comprises: under suitable conditions, reacting a raw material containing a benzimidazole structure with a raw material containing a R ₁ or R ₂ group in a suitable reagent; or connecting a material containing -W- The starting material of the moiety is reacted with the starting material containing the benzimidazole structure in a suitable reagent, or the two reaction steps described above are involved.

The present invention further provides a pharmaceutical composition comprising the compound of formula I described in the present invention and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof.

In some embodiments, the pharmaceutical composition of the present invention further comprises a therapeutically effective amount of the compound of formula I of the present invention and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides compounds, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers.

According to embodiments of the present invention, the compounds of the present invention may be used in combination with additional therapeutic agents. The pharmaceutical compositions of the present invention further comprise a second therapeutic agent.

A carrier in the pharmaceutical composition is "acceptable" which is compatible with (and preferably, is capable of stabilizing) the active ingredient of the composition and which is not deleterious to the subject being treated. One or more solubilizers can be used as pharmaceutical excipients for delivery of the active compounds.

The present invention further provides the compounds of formula I and their racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or Use of a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of an ENL inhibitor.

The present invention further provides the compounds of formula I and their racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or Use of a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a medicament for preventing, regulating or treating diseases or conditions mediated by ENL.

The present invention also provides a method of inhibiting ENL activity in a subject, the method comprising administering a compound of the present invention as a therapeutic agent.

According to an embodiment of the present invention, the ENL-mediated related disease or disorder is selected from cancer, in particular leukemia. The leukemia is selected from acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML); and some less common types. Preferably, the leukemia is AML.

The present invention further provides a method for treating the ENL-mediated disease or condition, the method comprising administering to a patient in need thereof a therapeutically effective amount of a first and a second therapeutic agent, wherein the first therapeutic agent is the compound of the present invention. In some embodiments, the present invention provides a combined formulation of a compound of the present invention and an additional therapeutic agent for simultaneous, separate or sequential use in therapy.

Terminology Explanation:

Unless otherwise stated, definitions of groups and terms set forth in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions set forth in tables, and definitions of specific compounds in the examples etc., can be arbitrarily combined and combined with each other. Such combinations, group definitions and compound structures after the combination should fall within the scope of the description of the present application.

The numerical range described in the specification and claims of the present application, when the numerical range is defined as an "integer" or is generally understood as an "integer" according to the conventional understanding in the art, it should be understood that the two endpoints of the range and the range are described. each integer within. For example, when "0-6" represents a carbon number, it should be understood that each integer of 0, 1, 2, 3, 4, 5 and 6 is described. "More" means three or more.

In the group definitions described in this article, *, ** represent the connection site of the group and other group parts of the general formula, and * represents the connection with the group on the left side corresponding to the general formula, then ** represents the corresponding group of the general formula. The right group is attached.

The term "halogen" refers to F, Cl, Br and I. In other words, F, Cl, Br and I may be described as "halogens" in this specification.

Optionally substituted with substituents described herein encompasses both unsubstituted as well as substituted with one or more substituents, eg "optionally substituted with one, two or more R" means that there may be no substitution Substituted with R (unsubstituted) or substituted with one, two or more Rs.

The term "aliphatic hydrocarbon group" includes saturated or unsaturated, straight or branched chain or cyclic hydrocarbon groups, the type of the aliphatic hydrocarbon group can be selected from alkyl, alkenyl, alkynyl, etc., the carbon atoms of the aliphatic hydrocarbon group The number is preferably 1-20 (specifically selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 ), the further preferred range can be 1-12, the more preferred range can be 1-6, specifically can include but not limited to the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, vinyl, 1- propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 1-ethylvinyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-Methyl-1-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butanyl Alkynyl, 1-methyl-2-propynyl, 3-butynyl, 1-pentynyl, 1-hexynyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl;

The "aliphatic hydrocarbon group" may optionally contain one, two or more heteroatoms (or be interpreted as optionally inserted into the aliphatic hydrocarbon group optionally with CC bonds and CH bonds). Suitable heteroatoms will be apparent to those skilled in the art and include, for example, sulfur, nitrogen, oxygen, phosphorous and silicon. The heteroatom-containing aliphatic hydrocarbyl group may be selected from the following groups: (C ₁ -C ₁₂ ) aliphatic hydrocarbyloxy, (C ₁ -C ₁₂ ) aliphatic hydrocarbyl mercapto, (C ₁ -C ₆ ) aliphatic hydrocarbyloxy base, (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto, (C ₁ -C ₆ ) aliphatic hydrocarbyloxy (C ₁ -C ₆ ) aliphatic hydrocarbyl, (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto (C ₁ -C ₆ ) ) aliphatic hydrocarbyl, (C ₁ -C ₆ ) aliphatic hydrocarbyloxy (C ₁ -C ₆ ) aliphatic hydrocarbyloxy, (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto, N- (C ₁ -C ₃ ) aliphatic hydrocarbylamino (C ₁ -C ₆ ) aliphatic hydrocarbyl, N,N-di-(C ₁ -C ₃ ) aliphatic hydrocarbylamine (C ₁ -C ₆ ) aliphatic hydrocarbyl; for example, it can be Methoxy, ethoxy, propoxy, butoxy, pentoxy, methoxymethyl, ethoxymethyl, propoxymethyl, methoxyethyl, ethoxyethyl , Propoxyethyl, Methoxypropyl, Ethoxypropyl, Propoxypropyl, N-methylaminomethyl, N-methylaminoethyl, N-ethylaminoethyl, N , N-dimethylaminomethyl, N,N-dimethylaminoethyl, N,N-diethylaminoethyl; the "aliphatic hydrocarbon" part contained in other groups is the same as the above explanation.

The term "3-12 membered heterocyclyl" means a saturated or unsaturated monovalent monocyclic or bicyclic ring containing 1-5 heteroatoms independently selected from N, O and S, and the heteroatom-containing group does not have For aromaticity, the 3-12-membered heterocyclic group is preferably a 3-10-membered heterocyclic group. The term 3-12 membered heterocyclyl means a saturated monovalent monocyclic or bicyclic ring containing 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclyl group can be attached to the remainder of the molecule through any of the carbon atoms or a nitrogen atom, if present. In particular, the heterocyclic group may include, but is not limited to: 4-membered ring, such as azetidinyl, oxetanyl; 5-membered ring, such as tetrahydrofuranyl, tetrahydrothienyl, dioxane Pentenyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholine group, piperazinyl, or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclyl group can be benzo-fused. The heterocyclyl group may be bicyclic, such as, but not limited to, a 5,5 membered ring, such as a hexahydrocyclopento[c]pyrrole-2(1H)-yl ring, or a 5,6 membered bicyclic ring, such as a hexahydropyrrole The [1,2-a]pyrazin-2(1H)-yl ring. The nitrogen-containing ring may be partially unsaturated, i.e. it may contain one or more double bonds such as, but not limited to, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiene oxazinyl, 4,5-dihydrooxazolyl, or 4H-[1,4]thiazinyl, alternatively, it may be benzo-fused such as, but not limited to, dihydroisoquinolinyl, 2,3- Dihydrobenzofuranyl, 3,4-dihydro-2H-1-benzopyranyl (chromanyl), 2,3-dihydrobenzo[b][1,4]dioxanyl. The 3-12 membered heterocyclic group can also be selected from, for example, the following groups:

The term "C _6-20 aryl" is to be understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6-20 carbon atoms, preferably a "C _6-10 aryl" . The term C _6-20 aryl is understood to preferably mean a monovalent aromaticity having 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon rings, especially those having 6 carbon atoms ("C ₆ aryl"), such as phenyl; or biphenyl, or those having 9 carbon atoms a ring ("C ₉ aryl") such as indanyl or indenyl, or a ring having 10 carbon atoms ("C ₁₀ aryl") such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, Either a ring with 13 carbon atoms (" _C13 aryl"), such as fluorenyl, or a ring with 14 carbon atoms (" _C14 aryl"), such as anthracenyl.

The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, especially 5 or 6 or 9 or 10 carbon atoms, and which contain 1-5, preferably 1-3 heteroatoms independently selected from N, O and S, and, in addition, in each may be benzo-fused. In particular, heteroaryl is selected from the group consisting of thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl oxazolyl, thi-4H-pyrazolyl, etc. and their benzo derivatives such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzene Triazolyl, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo derivatives, such as quinoline Or azinyl, indolizinyl, purinyl, etc. and their benzo derivatives; or cinnoline, phthalazinyl, quinazolinyl, quinoxaline olinyl, naphthyridinyl, pteridyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl and the like.

Unless otherwise specified, a heterocyclyl or heteroaryl group includes all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative non-limiting examples, pyridyl or pyridylene includes pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl and pyridin-4-yl; thienyl or thienylene includes thien-2-yl, thien-2-yl, thien-3-yl and thien-3-yl.

Depending on their molecular structure, the compounds of the present invention may be chiral and thus may exist in various enantiomeric forms. Thus these compounds may exist in racemic or optically active forms. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids such as N- Benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulfonic acids. It is also possible to use optically active resolving agents such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel. Chromatographic enantiomeric resolution is advantageously performed. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, eg hexane/isopropanol/acetonitrile.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles can form N-oxides since nitrogen needs to have available lone pairs of electrons for oxidation to nitrogen oxides; those skilled in the art will recognize that N-oxides can be formed - Nitrogen-containing heterocycles of oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for the preparation of N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the use of peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA), peroxyacids Hydrogen oxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes such as dimethyldioxirane oxidize heterocycles and tertiary amines. These methods for preparing N-oxides have been extensively described and reviewed in the literature.

Pharmaceutically acceptable salts may be, for example, acid addition salts of sufficiently basic compounds of the invention having nitrogen atoms in the chain or ring, such as acid addition salts with inorganic acids such as hydrochloric acid, hydrofluoric acid acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfate, or acid addition salts with organic acids such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid , propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentane Propionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, niacin, pamoic acid, pectic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, Camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid , ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

In addition, another suitable pharmaceutically acceptable salt of the compounds of the present invention that is sufficiently acidic are alkali metal salts (eg sodium or potassium salts), alkaline earth metal salts (eg calcium or magnesium salts), ammonium salts, or salts with organic bases that provide physiologically acceptable cations, such as salts with sodium ions, potassium ions, N-methylglucamine, dimethylglucamine, ethylglucamine, Lysine, Dicyclohexylamine, 1,6-Hexanediamine, Ethanolamine, Glucosamine, Meglumine, Sarcosine, Serinol, Trishydroxymethylaminomethane, Aminopropanediol, 1-Amino-2 , 3,4-Butanetriol. By way of example, such pharmaceutically acceptable salts include salts of the group -COOH with sodium ions, potassium ions, calcium ions, magnesium ions, N-methylglucamine, dimethylglucamine, Ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropylene glycol , 1-amino-2,3,4-butanetriol.

In addition, basic nitrogen-containing groups can be quaternized with the following reagents: lower alkyl halides such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as sulfuric acid Dimethyl, diethyl, dibutyl, and dipentyl sulfates; long-chain halides such as decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; aralkyl groups Halides such as benzyl and phenethyl bromide etc. As examples, pharmaceutically acceptable salts include hydrochloride, sulfate, nitrate, bisulfate, hydrobromide, acetate, oxalate, citrate, mesylate, formate or Meglumine salts, etc.

Since the compounds of the present invention may have multiple salt-forming sites, the pharmaceutically acceptable salts include not only the salts formed on one of the salt-forming sites of the compounds of the present invention, but also 2, 3 or all of them. salts formed at the site. To this end, the molar ratio of the compound of formula (I) to the acid ion (anion) or base cation required for salt formation in the pharmaceutically acceptable salt can vary within a wide range, for example, it can be 4:1 ~1:4, such as 3:1, 2:1, 1:1, 1:2, 1:3, etc.

Depending on the position and nature of the various substituents, the compounds of the present invention may also contain one or more asymmetric centers. Asymmetric carbon atoms can exist in (R) or (S) configuration. When there is only one asymmetric center, a racemic mixture is produced, and when multiple asymmetric centers are contained, a diastereomeric mixture is obtained. In some cases, asymmetry may also exist due to hindered rotation about a particular bond, such as the central bond connecting two substituted aromatic rings of a particular compound. In addition, the substituents may also exist in the form of cis or trans isomers.

The compounds of the present invention also include all possible stereoisomers of each, either a single stereoisomer or said stereoisomer (eg, R-isomer or S-isomer, or E-isomer or Z-isomers) in any ratio in the form of any mixture. Single stereoisomers (eg single enantiomers or single diastereomers) of the compounds of the invention can be achieved by any suitable prior art method (eg chromatography, particularly eg chiral chromatography) separation.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist as two or more interconvertible species. Proton tautomers arise from the migration of covalently bonded hydrogen atoms between two atoms. Tautomers generally exist in equilibrium, and attempts to separate individual tautomers usually result in a mixture whose physicochemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form predominates; in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

In the present invention, referenced compounds also include isotopically-labeled compounds that are the same as those shown in formula I, but in which one or more atoms are assigned an atomic mass or mass number different from the usual Atomic substitution for naturally occurring atomic mass or mass number. Examples of isotopes that may be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F, and Cl, such as ² H, ³ H, ¹³ C, ¹¹ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, respectively , ¹⁷ O, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. Compounds of the invention, prodrugs thereof, or pharmaceutically acceptable salts of said compounds or said prodrugs containing the above isotopes and/or other isotopes of other atoms are within the scope of the invention. Certain isotopically-labeled compounds of the invention, eg, compounds incorporating radioactive isotopes such ^as3H ^and14C , are useful in drug and/or substrate tissue distribution assays. Tritium (ie ³ H) and carbon 14 (ie ¹⁴ C) isotopes are particularly preferred for ease of preparation and detectability. Furthermore, substitution with heavier isotopes (such as deuterium, i.e., 2H) may provide certain therapeutic advantages (eg, increased in vivo half ^- life or reduced dosage requirements) derived from greater metabolic stability, and may therefore be is preferred in some cases. The compounds of the invention as claimed in the claims may in particular be substituted with deuterium or tritium. Furthermore, the presence of hydrogen in a substituent group is not listed individually. The term deuterium or tritium does not imply the exclusion of deuterium or tritium, but may equally well include deuterium or tritium.

The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound of the present invention sufficient to achieve the intended application, including but not limited to disease treatment as defined below. A therapeutically effective amount may vary depending on the intended application (in vitro or in vivo), or the subject and disease condition being treated such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc., which It can be easily determined by one of ordinary skill in the art. The specific dose will vary depending on the particular compound chosen, the dosing regimen followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system on which it is carried.

The term "solvates" are those forms of the compounds of the present invention which, in solid or liquid state, form complexes by coordination with solvent molecules. Hydrates are a specific form of solvates in which the coordination is with water. In the present invention, preferred solvates are hydrates. Further, the pharmaceutically acceptable solvates (hydrates) of the compounds of the general formula I of the present invention refer to co-crystals and clathrates formed by the compound I with stoichiometric one or more molecules of water or other solvents. Useful solvents for solvates include, but are not limited to, water, methanol, ethanol, ethylene glycol, and acetic acid.

The term "prodrug" or "prodrug" refers to the in vivo conversion of a compound to a compound of the aforementioned general formula or specific compound. Such conversion is effected by hydrolysis of the prodrug in blood or enzymatic conversion to the parent structure in blood or tissue. The prodrugs of the present invention can be esters. In the present invention, esters can be used as prodrugs including phenyl esters, aliphatic esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, a compound of the present invention contains a hydroxyl/carboxyl group, which can be acylated to give the compound in the form of a prodrug. Other prodrug forms include phosphates, such as these phosphates are phosphorylated by the hydroxyl group on the parent.

beneficial effect

The present invention provides a compound of general formula I with novel structure, which has a good inhibitory effect on ENL.

Detailed ways

The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following examples are only for illustrating and explaining the present invention, and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the intended protection scope of the present invention.

Unless otherwise stated, the starting materials and reagents used in the following examples are commercially available or can be prepared by known methods.

Instrumentation and General Methods

¹ H and ¹⁹ F NMR were recorded on a Bruker Ascend 400 spectrometer using deuterated reagents (DMSO-d ₆ , CDCl ₃ , CD ₃ OD). Deuterated solvent or TMS was used as internal standard. The chemical shift is expressed in ppm, the coupling constant (J) is expressed in Hz, and the splitting modes are s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad peak). LC-MS adopts Agilent 1260-6120 system equipped with Waters Cortecs C18, 2.7 μm, 4.6 × 30 mm column; HPLC adopts Waters Acquity UPLC H-class instrument equipped with Acquity BEH C18, 1.7 μm, 50 × 2.1 mm column. Preparative HPLC (Kromasil-C18 (100×21.2mm, 5μm) column, Xtimate-C18 (250×30mm) column or Xbridge-C18 (150×19mm, 5μm) column), chiral preparative SFC (Daicel ChiralPak IG) (250mm×30mm, 10μm) column, (S,S) Whelk-O1 (100×4.6mm) column, Daicel Chiralpak IC-3 (50×4.6mm) column, Daicel Chiralpak AD-H (250×4.6mm) column and Chirex S-VAL R-NEA column (250 x 4.6 mm)) to purify or resolve the final compound.

Compound Abbreviations:

T ₃ P: 2,4,6-tripropyl-1,3,5,2,4,6-trioxytriphosphoric acid-2,4,6-trioxide or tripropylphosphoric anhydride

BOP: Benzotriazol-1-oxytris(dimethylamino)phosphorus hexafluorophosphate

For the series of compounds of the present invention, there are two forms of full numbering and abbreviated numbering, wherein, for example, BV021 is the abbreviation of PI1701-BV021-1, both of which point to the same compound structure. The rest of the compound numbers can be understood with reference to this example.

Synthesis Example

The synthesis of embodiment 1 compound BV021

Step 1: N,N-Dimethyl-1-(5-nitro-1H-benzo[d]imidazol-2-yl)dimethylamine

2-(Chloromethyl)-5-nitro-1H-1,3-benzodiazole (10 g, 47 mmol) was dissolved in a THF solution of dimethylamine (the concentration was 1 M, the volume was 0.10 L), and the system Stir at 70°C for 14 hours. Concentration and slurrying with acetonitrile gave the title compound (9.9 g, 95%) as a red solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 13.04 (brs, 1H), 8.40 (s, 1H), 8.09 (dd, J=8.8and 2.0Hz, 1H), 7.67 (s, 1H), 3.75 ( s, 2H), 2.27 (s, 6H). LC-MS: ESI m/z 221.2 [M+H] ⁺ ; calcd for C ₁₀ H ₁₂ N ₄ O ₂ 220.23.

Step 2: 2-((Dimethylamino)methyl)-1H-benzo[d]imidazol-5-amine

N,N-Dimethyl-1-(5-nitro-1H-benzo[d]imidazol-2-yl)dimethylamine (4.5 g, 0.02 mol) was dissolved in methanol (50 mL) and palladium was added Carbon (0.87 g, 80 mmol). The system was stirred at 25°C for 14 hours. The mixture was filtered, and the organic phase was concentrated to give the title compound (3.5 g, 90%) as a yellow oily liquid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 7.14 (d, J=8.4Hz, 1H), 6.60 (d, J=1.6Hz, 1H), 6.44 (dd, J=8.4and 2.0Hz, 1H) , 3.53 (s, 2H), 2.20 (s, 6H). LC-MS: ESI m/z 191.1 [M+H] ⁺ ; calcd for C ₁₀ H ₁₄ N ₄ 190.25.

Step 3: tert-Butyl 4-((2-((dimethylamino)methyl)-1H-benzo[d]imidazol-5-yl)carbamoyl)benzoate

2-((Dimethylamino)methyl)-1H-benzo[d]imidazol-5-amine (0.95 g, 5.0 mmol) was dissolved in dichloromethane (20 mL), followed by the addition of 4-((tert. Butoxy)carbonyl)benzoic acid (1.1 g, 5.0 mmol), _T3P (2.4 g, 7.5 mmol) and triethylamine (1.5 g, 15 mmol). After the system was stirred at 25°C for 3 hours, it was dissolved in dichloromethane (30 mL) and washed with saturated brine (30 mL*3 times). The organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound (2.5 g, crude) as a red solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.43(brs,1H), 8.11-7.99(m,7H), 7.47(s,1H), 3.71(s,2H), 2.28(s,6H), 1.58 (s, 9H). LC-MS: ESI m/z 395.2 [M ₊ H] ⁺ _; calcd for _C22H26N4O3 _394.48 .

Step 4: 4-((2-((Dimethylamino)methyl)-1H-benzo[d]imidazol-5-yl)carbamoyl)benzoic acid

Dissolve tert-butyl 4-((2-((dimethylamino)methyl)-1H-benzo[d]imidazol-5-yl)carbamoyl)benzoate (2.5 g, 60 mmol) in dichloro To methane (30 mL) was added trifluoroacetic acid (30 mL). The system was stirred at 25°C for 14 hours, and concentrated to obtain the title compound (5 g, crude product) as a red oily liquid.

LC-MS: ESI m/z 339.1 [ _M ₊ H] ⁺ ; calcd for _C18H18N4O3 _338.37 .

Step ⁵ : N1,N1 ^' -(ethane-1,2-diyl)bis(N4-( ² -((dimethylamino)methyl)-1H-benzo[d]imidazole-5 - base) terephthalamide)

4-((2-((dimethylamino)methyl)-1H-benzo[d]imidazol-5-yl)carbamoyl)benzoic acid was dissolved in DMF (5 mL), followed by the addition of decane-1 , 10-diamine (51 mg, 0.29 mmol), BOP (0.52 g, 1.2 mmol) and triethylamine (0.23 g, 1.8 mmol). The system was reacted and stirred at 25°C for 14 hours, concentrated and purified by preparative HPLC under the following conditions [column: Kromasil-C18 100×21.2mm 5um; mobile phase: acetonitrile in water (containing 0.1% trifluoroacetic acid); gradient: 25-35% acetonitrile; flow rate 20 mL/min time: 14 min] to give the title compound (2.6 mg, 1.0%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.30(brs, 2H), 8.60(brs, 2H), 8.29(brs, 2H), 8.03(d, J=7.2Hz, 6H), 7.98-7.88( m, 4H), 7.44 (s, 4H), 3.66–3.60 (m, 4H), 3.30–3.23 (m, 4H), 2.22 (s, 12H), 1.63–1.50 (m, 4H), 1.30–1.22 ( m, 12H). LC-MS: (ESI) m/z _813.5 [ _M +H] ⁺ ; calcd for _C46H56N10O4 _812.45 . HPLC purity: 98.82% (214 nm), 98.83% (254 nm).

Example 2: The following compounds were further synthesized with reference to the synthetic method of Example 1:

BV022:

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.41 (brs, 2H), 8.62 (brs, 2H), 8.24 (brs, 2H), 8.12–7.94 (m, 10H), 7.64–7.54 (m, 4H) ),4.47(s,4H),3.28–3.25(m,4H),2.85(s,12H),1.63–1.50(s,4H),1.44–1.28(s,8H).LC-MS:(ESI) m/z _813.5 [M ₊ H] ⁺ ; calcd for _C44H52N10O4 _784.42 . HPLC purity: 98.73% (214 nm), 96.69% (254 nm).

BV023:

¹ H NMR (400MHz, DMSO) δ 10.33 (brs, 2H), 8.64 (brs, 2H), 8.26 (brs, 2H), 8.10–8.02 (m, 6H), 8.01–7.96 (m, 4H), 7.46 (s, 4H), 3.70–3.64 (m, 4H), 3.31 (d, J=5.6Hz, 4H), 2.25 (s, 12H), 1.66–1.57 (m, 4H), 1.44–1.39 (m, 2H) ).LC-MS: ESI m/z 743.3 [M+H] ⁺ ; calcd for C ₄₁ H ₄₆ N ₁₀ O ₄ 742.37. HPLC purity: 98.44% (214 nm), 98.45% (254 nm).

BV024:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.32(brs,2H), 10.31(brs,2H), 8.24(brs,2H), 8.08-8.02(m, 6H), 7.99-7.94(m,4H) ), 7.55–7.25 (m, 4H), 3.63 (s, 4H), 3.23–3.20 (m, 4H), 2.23 (s, 12H), 1.65–1.56 (m, 4H). LC-MS: (ESI) m/z 729.3 [M ₊ H] ⁺ _; calcd for _C40H44N10O4 _728.35 . HPLC purity: 97.39% (214 nm), 95.74% (254 nm).

BV025:

¹ H NMR (400 MHz, MeOD-d ₄ ) δ 8.27 (s, 2H), 8.05 (d, J=8.4 Hz, 4H), 7.99 (d, J=8.4 Hz, 4H), 7.66 (d, J= 8.8Hz, 2H), 7.55(dd, J=8.8and 1.6Hz, 2H), 4.60(s, 4H), 3.54(t, J=6.8Hz, 4H), 3.01(s, 12H), 2.00–1.94( m, 2H). LC-MS: ESI m/z _715.3 [M ₊ H] ⁺ ; calcd for _C39H42N10O4 _714.34 . HPLC purity: 98.22% (214 nm), 98.34% (254 nm).

BV026:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.32(brs, 2H), 10.33(brs, 2H), 8.81(brs, 2H), 8.07(d, J=8.4Hz, 4H), 8.00(d, J=8.4Hz, 4H), 7.52–7.44(m, 4H), 3.65(s, 4H), 3.51(s, 4H), 2.25(s, 12H). LC-MS: ESI m/z 701.3[M+ H] ⁺ _; calcd for _C38H40N10O4 _700.32 _. HPLC purity: 91.04% (214 nm), 92.14% (254 nm).

BV027:

¹ H NMR (400MHz, DMSO) δ 12.32 (brs, 2H), 10.33 (brs, 2H), 8.71 (brs, 2H), 8.05 (d, J=8.4Hz, 6H), 7.98 (d, J=8.4 Hz,4H),7.52-741(m,4H),3.64(s,4H),3.58(d,J=4.4Hz,8H),3.46(s,4H),2.23(s,12H).LC-MS : ESI m/z 789.3 [M+H] ⁺ ; calcd for C ₄₂ H ₄₈ N ₁₀ O ₆ 788.38. HPLC purity: 90.45% (214 nm), 93.00% (254 nm).

BV028:

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.30 (brs, 2H), 10.33 (brs, 2H), 8.71 (brs, 2H), 8.05 (d, J=8.4 Hz, 6H), 7.98 (d, J=8.4Hz, 4H), 7.46(s, 4H), 3.66(s, 4H), 3.62(d, J=5.6Hz, 4H), 3.49(s, 4H), 2.25(s, 12H).LC- MS: ESI m/z 745.7 [M+H] ⁺ _; calcd for _C40H44N10O5 _744.35 . HPLC purity: _96.54 % (214 nm), 96.49% (254 nm).

BV032:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.40 (brs, 2H), 10.31 (brs, 2H), 8.62 (brs, 2H), 8.47-8.39 (m, 4H), 8.04 (d, J=8.4 Hz, 4H), 7.96(d, J=8.4Hz, 2H), 7.45(s, 2H), 3.64(s, 4H), 3.30–3.26(m, 4H), 2.54(m, 6H), 2.24(s ,6H),2.03–1.96(m,2H),1.60–1.52(m,2H),1.39–1.32(m,4H),1.26–1.22(m,4H).LC-MS:(ESI)m/z _771.3 [M ₊ H] ⁺ ; calcd for _C43H50N10O4 _770.40 . HPLC purity: 97.21% (214 nm), 98.03% (254 nm).

BV033:

¹ H NMR (400MHz, DMSO-d ₆ )δ12.33(brs,2H), 10.31(brs,2H), 8.63(brs,2H), 8.26(brs,1H), 8.12-7.90(m,10H), 7.46(s, 4H), 3.64(s, 4H), 3.30(d, J=6.0Hz, 4H), 2.24(s, 12H), 1.57(s, 4H), 1.38(s, 4H).LC-MS : (ESI) m/z 757.3 [M+H] ⁺ ; calcd for C ₄₂ H ₄₈ N ₁₀ O ₄ 756.39. HPLC purity: 100% (214 nm), 99.27% (254 nm).

The synthesis of embodiment 3 compound BV005

Step 1: Di-tert-butyl O,O'-(pentane-1,5-diyl)diterephthalate

4-(tert-Butoxycarbonyl)benzoic acid (1.2 g, 5.2 mmol) and 4-dimethylaminopyridine (0.64 g, 5.2 mmol) were dissolved in dichloromethane (10 mL), followed by 1-(3-dimethylene) Aminopropyl)-3-ethylcarbodiimide hydrochloride (0.99 g, 5.2 mmol). The system was stirred at 0°C for 30 minutes and then 1,5-pentanediol (0.27 g, 2.6 mmol) was added. The system was stirred at 25°C for 4 hours, poured into 20 mL of water, extracted three times with dichloromethane (15 mL), and the organic phase was dried over anhydrous sodium sulfate. After concentration, column chromatography (petroleum ether/ethyl acetate=100%-95%) gave the title compound (0.53 g, 20%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ -d ₃ ) δ 8.05 (q, J=8.8 Hz, 8H), 4.38 (t, J=6.8 Hz, 4H), 1.93-1.83 (m, 4H), 1.66-1.58 (m,20H).

Step 2: 4,4'-((pentane-1,5-diylbis(oxy))bis(carbonyl))dibenzoic acid

Di-tert-butyl O,O'-(pentane-1,5-diyl)diterephthalate (0.53 g, 0.91 mmol) and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (3 mL) in After stirring at 25°C for 30 minutes, the system was concentrated to give the title compound (0.44 g, crude) which was used directly in the next step.

LC-MS: ESI m/z 401.1 [ _M +H] ⁺ ; calcd for _C21H20O8 _400.12 .

Step 3: Pentane-1,5-diylbis(4-((2-(((S)-2-methylpyrrolidin-1-yl)methyl)-1H-benzo[d]imidazole- 5-yl)carbamoyl)benzoate))

4,4'-((pentane-1,5-diylbis(oxy))bis(carbonyl))dibenzoic acid (0.12 g, 0.30 mmol), (S)-2-((2-methyl) pyrrolidin-1-yl)methyl)-1H-benzo[d]imidazol-5-amine (0.15 g, 0.66 mmol) (Angew.Chem.Int.Ed.2018,57,16302) and 2-( 7-Azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.29g, 0.75mmol), N,N-diisopropylethylamine (0.16g , 1.2 mmol) was dissolved in DMF (6 mL), and the reaction system was stirred at 25 °C for 14 hours. The system mixture was poured into 10 mL of water, and extracted three times with 10 mL of ethyl acetate. The organic phase was washed with saturated ammonium chloride solution (20 mL), then dried over anhydrous sodium sulfate, concentrated and purified by preparative HPLC under the following conditions [column: Kromasil-C18 100×21.2 mm 5um; mobile phase: acetonitrile in water ( with 0.1% trifluoroacetic acid); gradient: 25-35% acetonitrile; flow rate 20 mL/min time: 14 min] to give the title compound (37 mg, 13%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.74 (brs, 2H), 10.50 (brs, 2H), 8.24 (s, 2H), 8.17-8.05 (m, 8H), 7.66-7.50 (m, 4H) ), 4.81-4.69(m, 2H), 4.54-4.43(m, 2H), 4.37(t, J=6.4Hz, 4H), 3.71-3.53(m, 4H), 3.39-3.36(m, 2H), 2.27-2.17(m, 2H), 2.02-1.90(m, 4H), 1.89-1.79(m, 4H), 1.69-1.56(m, 4H), 1.38(d, J=5.6Hz, 6H).LC- MS: ESI m/z _825.3 [M+H] ⁺ _; calcd for _C47H52N8O6 _824.40 . HPLC purity: 98.5% (214 nm), 98.1% (254 nm).

Example 4 The following compounds were synthesized with reference to the synthetic method of Example 3:

BV006:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(brs, 2H), 10.39(brs, 2H), 8.29(s, 2H), 8.12-8.02(m, 8H), 7.45(s, 4H), 4.32(t, J＝6.4Hz, 4H), 4.05(d, J＝14.4Hz, 2H), 3.52(s, 2H), 2.95-2.89(m, 2H), 2.47-2.41(m, 2H), 2.31 –2.23(m,2H),1.97–1.88(m,2H),1.82–1.57(m,8H),1.40(s,10H),1.08(d,J=6.0Hz,6H).LC-MS:ESI m/z _867.4 [M+H] ⁺ _; calcd for _C50H58N8O6 _866.45 . HPLC purity: 98.0% (214 nm), 98.1% (254 nm).

BV007:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.51(brs, 2H), 8.24(brs, 2H), 8.10(s, 8H), 7.63(d, J=8.8Hz, 2H), 7.56(d, J=8.8Hz, 2H), 4.78(d, J=13.2Hz, 2H), 4.51(d, J=14.4Hz, 2H), 4.31(t, J=6.4Hz, 4H), 3.71-3.58(m, 4H), 3.38-3.28(m, 2H), 2.26-2.19(m, 2H), 2.02-1.89(m, 4H), 1.78-1.61(m, 6H), 1.49-1.27(m, 18H).LC- MS: ESI m/z _895.4 [M+H] ⁺ _; calcd for _C52H62N8O6 _894.48 . HPLC purity: 98.4% (214 nm), 97.3% (254 nm).

BV008:

¹ H NMR (400MHz, DMSO-d ₆ )δ12.24(brs,2H), 10.39(brs,2H), 8.15(s,2H), 8.09(s,6H), 8.05(s,2H), 7.46( s, 4H), 4.47–4.39 (m, 4H), 4.08 (d, J=14.2Hz, 2H), 3.81–3.78 (m, 4H), 3.65 (s, 2H), 3.61–3.54 (m, 2H) ,3.17(s,2H),2.95(dd,J＝12.0and 4.8Hz,2H),2.57-2.53(m,2H),2.33(t,J＝8.8Hz,2H),1.94(dd,J＝10.0 and 4.0Hz,2H),1.74–1.59(m,4H),1.38(d,J=8.8Hz,2H),1.09(d,J=6.0Hz,6H).LC-MS: ESI m/z 436.3[ M/ ₂ +H] ⁺ ; _calcd for _C48H54N8O8 _870.41 . HPLC purity: 99.2% (214 nm), 99.4% (254 nm).

BV009:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.15(brs, 2H), 10.31(brs, 2H), 8.63(t, J=5.6Hz, 2H), 8.32(s, 4H), 8.07-8.03( m, 6H), 7.97 (d, J=8.4Hz, 4H), 7.45 (s, 4H), 4.05 (d, J=14.0Hz, 2H), 3.56–3.50 (m, 2H), 3.33–3.29 (m ,2H),2.98–2.88(m,2H),2.49–2.41(m,4H),2.32–2.26(m,2H),1.99–1.89(m,2H),1.68–1.56(m,8H),1.46 -1.30 (m, 4H), 1.09 (d, J=6.0 Hz, 4H). LC-MS: ESI m/z 412.2 [M/2+H] ⁺ ; calcd for C ₄₇ H ₅₄ N ₁₀ O ₄ 823.43. HPLC purity: 99.74% (214 nm), 99.41% (254 nm).

BV010:

¹ H NMR (400MHz, DMSO) δ 12.20(brs, 2H), 10.30(s, 2H), 8.60(t, J=5.6Hz, 2H), 8.16(s, 2H), 8.04(d, J=8.1 Hz, 6H), 7.97(d, J＝8.3Hz, 4H), 7.45(s, 4H), 4.05(d, J＝14.1Hz, 2H), 3.54(d, J＝14.2Hz, 2H), 3.27( s, 2H), 2.95–2.89 (m, 2H), 2.54 (s, 4H), 2.31–2.24 (m, 2H), 1.93 (dd, J=12.0 and 6.5Hz, 2H), 1.69–1.51 (m, 8H), 1.33(s, 10H), 1.09(d, J=6.0Hz, 6H). LC-MS: ESI m/z 433.2[M/2+H] ⁺ ; Calculated for C ₅₀ H ₆₀ N ₁₀ O ₄ 433.24.HPLC: 98.70% (254nm), 98.39% (214nm).

BV011:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.18(brs, 2H), 10.30(s, 2H), 8.60(t, J=5.2Hz, 2H), 8.18(s, 2H), 8.06-8.00( m, 6H), 7.98–7.94 (m, 4H), 7.45 (s, 4H), 4.05 (d, J=14.0Hz, 2H), 3.54 (d, J=14.0Hz, 2H), 3.30–3.24 (m ,2H),2.95–2.87(m,2H),2.56–2.52(m,4H),2.31–2.26(m,2H),1.97–1.89(m,2H),1.70–1.61(m,4H),1.58 -1.50 (m, 4H), 1.41 - 1.28 (m, 14H), 1.08 (d, J=6.0Hz, 6H). LC-MS: ESI m/z 893.4 [M+H] ⁺ ; C ₅₂ H ₆₄ N Calculated for ₁₀ O _892.15 . HPLC purity: 99.17% (214 nm), 98.49% (254 nm).

BV012:

¹ H NMR (400MHz, DMSO-d ₆ )δ10.31(s, 2H), 8.71(t, J=5.2Hz, 2H), 8.34(brs, 4H), 8.06-7.97(m, 8H), 7.45( s, 2H), 4.04 (d, J=14.2Hz, 2H), 3.60–3.51 (m, 10H), 3.47–3.43 (m, 4H), 2.95–2.89 (m, 2H), 2.54 (s, 2H) ,2.31–2.24(m,2H),1.97–1.90(m,2H),1.70–1.58(m,4H),1.40–1.32(m,2H),1.08(d,J=6.0Hz,6H).LC - MS: ESI m/z 869.4 [M+H] ⁺ ; calcd for C ₄₈ H ₅₆ N ₁₀ O ₆ 868.44. HPLC purity: 99.30% (214 nm), 99.50% (254 nm).

BV031:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.15(brs, 2H), 10.43(brs, 2H), 8.09(s, 10H), 7.49(s, 4H), 4.47–4.40(m, 4H), 3.87(s,4H),3.81-3.78(m,4H),3.65(s,4H),2.40(s,12H).LC-MS:(ESI)m/z 396.3[M/2+H] ⁺ ; _Calcd . for _C42H46N8O8 _790.34 . HPLC purity: _97.9 % (214 nm), 97.7% (254 nm).

Example 5: Synthesis of compound BV013:

Step ¹ : N1, ^N7 -dibenzyl-N1,N7 ^- dimethylheptane- ^1,7 -diamine

1,7-Dibromoheptane (5.0 g, 19 mmol), N-methylbenzylamine (5.2 g, 42 mmol) and cesium carbonate (13 g, 39 mmol) were dissolved in DMF (50 mL). After the system was stirred at 80°C for 16 hours, the mixture was dissolved in ethyl acetate (50 mL) and washed with 20 mL of water 3 times. The organic phase was dried over anhydrous sodium sulfate and concentrated, followed by column chromatography (40 g silica gel, 10% methanol in dichloromethane) to give the title compound (2.3 g, 35% yield) as a yellow oily liquid.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.33-7.19(m, 10H), 3.41(s, 4H), 2.31-2.24(m, 4H), 2.08(s, 6H), 1.49-1.37(m , 4H), 1.29–1.19 (m, 6H). LC-MS: ESI m/z 339.4 [M+H] ⁺ ; calcd for C ₂₃ H ₃₄ N ₂ 338.27.

Step ² : N1,N7 ^- dimethylheptane-1,7-diamine

N1, ^N7 -dibenzyl ^- N1,N7 ^- dimethylheptane- ^1,7 -diamine (2.3 g, 0.01 mol) was dissolved in 100 mL methanol and palladium on carbon (0.30 g) was added. After the system was stirred at 25°C for 14 hours under a hydrogen atmosphere, the mixture was filtered through celite to obtain 1.2 g of the crude title compound as a brown gummy solid.

¹ H NMR (400MHz, CDCl ₃ )δ2.60-2.53(m,4H), 2.43(s,6H), 1.55-1.43(m,4H), 1.35-1.28(m,6H).LC-MS:ESI m/z 159.3 [M+H] ⁺ ; calcd for C ₉ H ₂₂ N ₂ 158.18.

Step ³ : N1,N7-Dimethyl-N1, ^N7 -bis(( ⁵ -nitro-1H-benzo[d]imidazol- ² -yl)methyl)heptane-1,7- Diamine

2-(Chloromethyl)-5-nitro-1H-benzo[d]imidazole (5.3 g, 25 mmol) was dissolved in 15 mL of acetonitrile, N ¹ , N ⁷ -dimethylheptane-1,7 was added - Diamine and sodium carbonate (1.2 g, 10 mmol) were dissolved in a solution of 5 mL DMF. After the system was stirred at 40° C. for 14 hours, the solvent was evaporated to dryness, 15 mL of water was added, and the mixture was extracted three times with 10 mL of ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated, and then subjected to column chromatography to obtain the title compound (0.40 g, 79%) as a brown gummy substance.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.80 (brs, 2H), 8.39 (s, 2H), 8.14 (s, 2H), 8.08 (dd, J=8.8, 2.0 Hz, 2H), 7.66 ( d, J=8.8Hz, 2H), 3.77(s, 4H), 2.40-2.32(m, 4H), 2.22(s, 6H), 1.47-1.38(m, 4H), 1.26-1.16(m, 6H) _.LC -MS: ESI m/z 509.3 [M ₊ H] ⁺ ; calcd for _C25H32N8O4 _508.25 .

Step ⁴ : N1,N7-bis(( ⁵ -amino-1H-benzo[d]imidazol-2-yl)methyl) ^-N1 ,N7-dimethylheptane ^- 1,7-di amine

N ¹ ,N ⁷ -dimethyl-N ¹ ,N ⁷ -bis((5-nitro-1H-benzo[d]imidazol-2-yl)methyl)heptane-1,7-diamine (0.62 g, 1.2 mmol) was dissolved in 10 mL of methanol and palladium on carbon (78 mg, 0.73 mmol) was added. Under a hydrogen atmosphere, the system was stirred at 25°C for 16 hours. The mixture was filtered through celite and concentrated to give the title compound (0.40 g, 73%) as a red liquid.

LC-MS: ESI m/z _449.3 [M+H] ⁺ ; calcd for _C25H36N8 _448.31 .

Step 5: 4,4'-(((((heptane-1,7-diylbis(methylazacyclobi))bis(methylene))bis(1H-benzo[d]imidazole- Dimethyl 2,5-diyl))bis(azadienyl))bis(carbonyl)dibenzoate

The N ¹ ,N ⁷ -bis((5-amino-1H-benzo[d]imidazol-2-yl)methyl)-N ¹ ,N ⁷ -dimethylheptane-1,7-diamine ( 0.10g, 0.20mmol), monomethyl terephthalate (0.11g, 0.60mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea Hexafluorophosphate (0.23 g, 0.60 mmol), N,N-diisopropylethylamine (0.10 g, 0.80 mmol) were dissolved in DMF (5 mL), and the reaction system was stirred at 25°C for 14 hours. The system mixture was poured into 10 mL of water, and extracted three times with 10 mL of ethyl acetate. The organic phase was washed with saturated ammonium chloride solution (20 mL), then dried over anhydrous sodium sulfate, concentrated and purified by preparative HPLC under the following conditions [column: Kromasil-C18 100×21.2 mm 5um; mobile phase: acetonitrile in water ( with 0.1% trifluoroacetic acid); gradient: 25-35% acetonitrile; flow rate 20 mL/min time: 14 min] to give the title compound (5 mg, 5%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.17(brs, 2H), 10.41(s, 2H), 8.26(s, 2H), 8.10-8.06(m, 8H), 7.45(s, 4H), 3.90(s,6H), 3.68(s,4H), 2.38–2.33(m,4H), 2.20(s,6H), 1.49–1.30(m,4H), 1.28–1.21(m,6H).LC- MS: ESI m/z _773.5 [M+H] ⁺ _; calcd for _C43H48N8O6 _772.37 . HPLC purity: 95.03% (214 nm), 95.51% (254 nm).

Example 6 The following compounds were synthesized with reference to the synthetic method of Example 5:

BV014:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.18(brs, 2H), 10.42(brs, 2H), 8.20-7.95(m, 10H), 7.52-7.37(m, 4H), 3.90(s, 6H) ),3.67(s,4H),2.39-2.32(m,4H),2.20(s,6H),1.49-1.39(m,4H),1.30-1.13(m,12H).LC-MS:ESI m/ z _815.4 [ _M +H] ⁺ _; calcd for C46H54N8O6 _814.42 . HPLC purity: 99.65% (214 nm), 99.66% (254 nm).

BV015:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.20(brs, 2H), 10.42(brs, 2H), 8.09(s, 10H), 7.50-7.41(m, 4H), 3.90(s, 6H), 3.67(s,4H), 2.38–2.31(m,4H), 2.20(s,6H), 1.48–1.41(m,4H), 1.24–1.05(m,16H).LC-MS: ESI m/z 422.3 [M/ ₂ +H] ⁺ _; calcd for _C48H58N8O6 _843.45 . HPLC purity: 95.16% (214 nm) 92.24% (254 nm).

BV016:

¹ H NMR (400 MHz, Me ₃ OD-d ₄ ) δ 8.30 (d, J=1.6 Hz, 2H), 8.14 (s, 2H), 8.11 (s, 2H), 8.01 (s, 2H), 7.98 ( s, 2H), 7.63(s, 1H), 7.61(s, 1H), 7.50(d, J=2.0Hz, 1H), 7.48(d, J=2.0Hz, 1H), 4.51(s, 4H), 3.94(s, 6H), 3.83–3.80(m, 4H), 3.68(d, J=2.0Hz, 12H), 2.87(s, 6H). LC-MS: ESI m/z 418.3[M/2+H ] ⁺ ; _calcd for _C44H50N8O9 _834.37 . HPLC purity: _95.03 % (214 nm), 95.43% (254 nm).

BV017:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.12 (brs, 2H), 10.24 (s, 2H), 8.47 (d, J=0.8Hz, 2H), 8.23 (d, J=0.8Hz, 2H) ,8.18(s,2H),8.08(s,2H),8.01(dd,J＝8.8and 1.6Hz,2H),7.75(d,J＝8.8Hz,2H),7.46(s,4H),4.10( s,6H),3.69(s,4H),2.39–2.34(m,4H),2.21(s,6H),1.47–1.43(m,4H),1.23(s,6H).LC-MS:ESI m /z 765.4 [M+H] ⁺ ; calcd for C ₄₃ H ₄₈ N ₁₂ O ₂ 764.40. HPLC purity: 95.03% (214 nm), 95.51% (254 nm).

BV018:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.17(brs, 2H), 10.24(brs, 2H), 8.48(s, 2H), 8.24(d, J=0.8Hz, 2H), 8.09(s, 2H), 8.02(dd, J=8.8and 1.6Hz, 2H), 7.75(d, J=8.8Hz, 2H), 7.46(s, 4H), 4.10(s, 6H), 3.67(s, 4H), 2.39–2.33(m,4H), 2.20(s,6H), 1.49–1.39(m,4H), 1.31–1.13(m,12H).LC-MS: ESI m/z 807.5[M+H] ⁺ ; Calcd. for _C46H54N12O2 _806.45 . HPLC purity _: _97.69 % (214 nm), 98.49% (254 nm).

BV019:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.16(brs, 2H), 10.23(brs, 2H), 8.48(s, 2H), 8.24(s, 2H), 8.10(d, J=10.0Hz, 2H), 8.02(d, J=7.2Hz, 2H), 7.75(d, J=8.8Hz, 2H), 7.50–7.39(m, 4H), 4.10(s, 6H), 3.67(s, 4H), 2.35–2.24(m,4H), 2.23–2.20(m,6H), 1.45–1.41(m,4H), 1.24–1.17(m,16H).LC-MS: ESI m/z 836.3[M+H] ⁺ ; _calcd for _C48H58N12O2 _835.48 . HPLC purity: _93.43 % (214 nm), 95.68% (254 nm).

BV020:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.19(brs, 2H), 10.26(brs, 2H), 8.47(s, 2H), 8.24(s, 2H), 8.11(s, 2H), 8.04– 7.99(m,2H),7.75(d,J=8.8Hz,2H),7.52-7.41(m,4H),4.10(s,6H),3.75(s,4H),3.55-3.48(m,12H) , 2.61-2.58 (m, 4H), 2.25 (s, 6H). LC-MS: (ESI) m/z 849.3 [M+Na] ⁺ ; calcd for C ₄₄ H ₅₀ N ₁₂ O ₅ 826.40. HPLC purity: 92.25% (214nm), 93.74% (254nm).

BV037:

¹ H NMR (400MHz, DMSO-d ₆ )δ10.21(brs,2H), 8.44(brs,2H), 8.20(s,2H), 8.10(brs,1H), 8.06(s,2H), 7.98( d, J＝8.8Hz, 2H), 7.71(d, J＝8.8Hz, 2H), 7.42(d, J＝9.2Hz, 4H), 4.07(s, 6H), 3.75(s, 4H), 3.54( t,J＝5.6Hz,4H),3.47(s,4H),2.60(t,J＝5.6Hz,4H),2.25(s,6H).LC-MS:(ESI)m/z 783.7[M+ H] ⁺ _; calcd for _C42H46N12O4 _782.38 . HPLC purity: 98.50% (214 nm), _97.23 % (254 nm).

Example 6: Monomer synthesis

LY120: methyl 4-((2-((dimethylamino)methyl)-1H-benzo[d]imidazol-5-yl)carbamoyl)benzoate

2-((dimethylamino)methyl)-1H-benzo[d]imidazol-5-amine (200 mg, 1.05 mmol), monomethyl terephthalate (189 mg, 1.05 mmol), 2-(7 -Azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.23g, 0.60mmol), N,N-diisopropylethylamine (0.10g, 0.80 mmol) was dissolved in DMF (5 mL) and the reaction was stirred at 25°C for 16 hours. The system mixture was poured into 10 mL of water, and extracted three times with 10 mL of ethyl acetate. The organic phase was washed with saturated ammonium chloride solution (20 mL), then dried over anhydrous sodium sulfate, concentrated and purified by preparative HPLC under the following conditions [column: Kromasil-C18 100×21.2 mm 5um; mobile phase: acetonitrile in water ( with 0.1% trifluoroacetic acid); gradient: 25-35% acetonitrile; flow rate 20 mL/min time: 14 min] to give the title compound (98.8 mg, 26%) as a white solid.

¹ H NMR (400MHz, DMSO) δ 10.53(s, 1H), 8.25(s, 1H), 8.11(s, 4H), 7.64(d, J=8.7Hz, 1H), 7.58(d, J=8.7 Hz,1H),4.60(s,2H),3.91(s,3H),2.91(d,J=12.6Hz,6H).LC-MS:ESI m/z353.2[M+H] ⁺ ; Calculated 352.15. HPLC purity: 97.44% (214 nm), 98.55% (254 nm).

The following compounds were synthesized with reference to the synthesis method of methyl 4-((2-((dimethylamino)methyl)-1H-benzo[d]imidazol-5-yl)carbamoyl)benzoate:

LY108:

¹ H NMR (400MHz, DMSO): δ12.49-12.12(brs,1H), 10.28-10.20(brs,1H), 8.48(d,J=0.7Hz,1H), 8.24(d,J=0.8Hz, 1H),8.20-8.15(brs,1H),8.07(s,1H),8.02(dd,J=8.9and 1.5Hz,1H),7.76(d,J=8.9Hz,1H),7.53-7.38(m , 2H), 4.10 (s, 3H), 3.65 (s, 2H), 2.24 (s, 6H). LC-MS: (ESI) m/z (M+H), 349.2. Calculated 348.17. HPLC purity: 98.78% (214nm), 99.46% (254nm).

BV002:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.18(brs,1H), 10.41(brs,1H), 8.28(s,1H), 8.12–8.02(m,4H), 7.45(s,2H), 4.05(d,J＝14.2Hz,1H),3.90(s,3H),3.53(d,J＝14.2Hz,1H),2.95-2.88(m,1H),2.51-2.43(m,1H),2.31 –2.22(m,1H),1.97-1.90(m,1H),1.71-1.59(m,2H),1.40-1.33(m,1H),1.08(d,J=6.0Hz,3H).LC-MS : ESI m/z 393.2 [M+H] ⁺ ; calcd for C ₂₂ H ₂₄ N ₄ O _392.18 .

BV003:

¹ H NMR (400MHz, DMSO-d ₆ )δ12.18(brs,1H),10.32(brs,1H),8.60(d,J=4.6Hz,1H),8.05-7.84(m,5H),7.50- 7.38(m, 2H), 4.04(d, J=13.6Hz, 1H), 3.53(d, J=14.2Hz, 1H), 3.30(s, 1H), 2.93–2.86(m, 1H), 2.81(t , J=4.8Hz, 3H), 2.29–2.24 (m, 1H), 1.95–1.89 (m, 1H), 1.72–1.63 (m, 2H), 1.39–1.32 (m, 1H), 1.08 (d, J = 6.0 Hz, 3H). LC-MS: ESI m/z 392.1 [M+H] ⁺ ; calcd for C ₂₂ H ₂₅ N ₅ O ₂ 391.20.

LY160:

¹ H NMR (400MHz, DMSO-d ₆ )δ12.53(brs,1H), 10.33(brs,1H), 8.62(brs,1H), 8.04(s,2H), 7.97(s,2H), 7.47( s, 2H), 6.56(s, 1H), 3.43(s, 2H), 2.82(s, 3H), 2.30(s, 6H). LC-MS: ESI m/z352.2[M+H]+; _Calcd . for _C19H21N5O2 _351.41 . HPLC purity _: 100% (214 nm), 98.66% (254 nm).

Biological Activity Evaluation Experiment

Biological experiment example 1: Inhibitory ability of two biochemical test compounds on ENL: AlphaScreen, Homogeneous Time-Resolved Fluorescence Technology (HTRF)

1.1AlphaScreen

Experimental materials: His-tagged human ENL protein was purchased from ACTIVE MOTIF (Cat#81098). Proteins were dissolved in sterile water and stored at -80°C after aliquoting. Biotin-labeled peptides were synthesized by GenScript, dissolved in DMSO and stored in aliquots at -20°C.

Hisdidine Detection Kit (Nickel Chelate) (6760619M) was purchased from PerkinElmer. The Source plate (PP-0200) was purchased from Labcyte, and the Proxiplate-384plus target plate was purchased from Perkin Elmer (Cat#6008280). The reaction buffer was prepared in-house: 50 mM HEPES, 0.005% Brij35, 1 mM TCEP, pH 7.4.

Experimental method: The candidate compound was dissolved in DMSO, added to the source plate according to a certain concentration gradient, and then the compound was transferred to the target plate with ECHO550 to form 11 concentrations. Proteins and peptides were diluted with reaction buffer, mixed gently, and 10 μl per well was added to the target plate with Multidrop (the working concentration of protein was 5nM, and the working concentration of peptide was 30nM), and incubated at room temperature for 1 hour. Add 10 μl Donor/Acceptor beads (working concentration 15 μg/mL) to each well and incubate at room temperature for one hour. Finally read with Envision Plate Reader. The _IC50 for each compound was obtained after fitting with XLfit.

1.2 Homogeneous Time-Resolved Fluorescence (HTRF)

Experimental materials: His-tagged human ENL protein was purchased from ACTIVE MOTIF (Cat#81098). Proteins were dissolved in sterile water and stored at -80°C after aliquoting. Biotin-labeled peptides were synthesized by GenScript, dissolved in DMSO and stored in aliquots at -20°C. Streptavidin-XL665 (Cat#610SAXLA) and Tb3+cryptate labeled anti-6His antibody (Cat#61HI2TLA) were purchased from Cisbio. Source plate (PP-0200) was purchased from Labcyte Company, Proxiplate-384plus target plate was purchased from Perkin Elmer Company. The reaction buffer was prepared by itself: PBS, 0.1% BSA, pH 7.2.

Experimental method: The candidate compound was dissolved in DMSO, added to the source plate according to a certain concentration gradient, and then the compound was transferred to the target plate with ECHO550 to form 11 concentrations. Proteins and peptides were diluted with reaction buffer, mixed gently, and 10 μl per well was added to the target plate with Multidrop (the working concentration of protein was 5nM and the working concentration of peptide was 100nM), and incubated at room temperature for 1 hour. Add 10 μl of the diluted mixture of Streptavidin-XL665 and Tb3+cryptate labeled anti-6His antibody to each well and incubate for one hour at room temperature. Finally read with Envision Plate Reader. The _IC50 for each compound was obtained after fitting with XLfit.

1.3 Experimental results

(1) The compounds of the present invention all have a good inhibitory effect on ENL, and the specific test results are shown in the following table:

Table 1 HTRF and AS test results of the compounds of the present invention

	AlfaScreenAlfaScreen	HTRFHTRF
BV013BV013	AA	BB
BV014BV014	BB	CC
BV015BV015	BB	BB
BV016BV016	AA	CC
BV017BV017	AA	BB
BV018BV018	AA	BB
BV020BV020	BB	CC
BV037BV037	BB	CC
BV005BV005	AA	BB
BV006BV006	AA	BB
BV007BV007	AA	BB
BV008BV008	AA	AA
BV009BV009	AA	AA
BV011BV011	CC	BB
BV012BV012	AA	BB
BV031BV031	AA	BB
BV023BV023	AA	--
BV024BV024	AA	--

BV025BV025	AA	--
BV026BV026	AA	--
BV027BV027	AA	--
BV028BV028	AA	--

Note: A: <100nM; B: 100-1000nM; C: >1000nM

(2) The structure of the dimer form of the compound of the present invention (shown as A-Linker-A) is composed of the corresponding monomer structure part (shown as A) (refer to Table 2 below), the inventors found that the dimer structure is directed to The activity of ENL is further improved compared to the monomer. According to the AS IC ₅₀ (nM) test results, some dimers have a better improvement in activity relative to monomers, and the Monomer/Dimer (AS) ratio is more than 10 times, such as BV013, BV018; some dimers are relative to The activity of the monomer has been particularly improved, and the Monomer/Dimer(AS) ratio has reached more than 100 times; for example, BV008, BV012, BV024, BV025, BV026, BV028; some dimers have a great degree of activity relative to the monomer. Improvement, the Monomer/Dimer(AS) ratio can even reach more than 500 times, such as BV009, BV023, BV027, BV031.

Table 2 dimer compound of the present invention and corresponding monomer structure

A(Monomer)A (Monomer)	LinkerLinker	A-Linker-A(Dimer)A-Linker-A(Dimer)
PI1701-LY120-1PI1701-LY120-1	─(CH ₂) ₇─ ─(CH ₂ ) ₇ ─	PI1701-BV013-1PI1701-BV013-1
PI1701-LY120-1PI1701-LY120-1	─(CH ₂) ₁₀─ ─(CH ₂ ) ₁₀ ─	PI1701-BV014-1PI1701-BV014-1
PI1701-LY120-1PI1701-LY120-1	─(CH ₂) ₁₂─ ─(CH ₂ ) ₁₂ ─	PI1701-BV015-1PI1701-BV015-1
PI1701-LY120-1PI1701-LY120-1	─CH ₂(CH ₂OCH ₂) ₃CH ₂─ ─CH ₂ (CH ₂ OCH ₂ ) ₃ CH ₂ ─	PI1701-BV016-1PI1701-BV016-1
PI1701-LY108-1PI1701-LY108-1	─(CH ₂) ₇─ ─(CH ₂ ) ₇ ─	PI1701-BV017-1PI1701-BV017-1
PI1701-LY108-1PI1701-LY108-1	─(CH ₂) ₁₀─ ─(CH ₂ ) ₁₀ ─	PI1701-BV018-1PI1701-BV018-1
PI1701-LY108-1PI1701-LY108-1	─(CH ₂) ₁₂─ ─(CH ₂ ) ₁₂ ─	PI1701-BV019-1PI1701-BV019-1
PI1701-LY108-1PI1701-LY108-1	─CH ₂(CH ₂OCH ₂) ₃CH ₂─ ─CH ₂ (CH ₂ OCH ₂ ) ₃ CH ₂ ─	PI1701-BV020-1PI1701-BV020-1
PI1701-BV002-1PI1701-BV002-1	─O(CH ₂) ₅O─ ─O(CH ₂ ) ₅ O─	PI1701-BV005-1PI1701-BV005-1
PI1701-BV002-1PI1701-BV002-1	─O(CH ₂) ₈O─ ─O(CH ₂ ) ₈ O─	PI1701-BV006-1PI1701-BV006-1
PI1701-BV002-1PI1701-BV002-1	─O(CH ₂) ₁₀O─ ─O(CH ₂ ) ₁₀ O─	PI1701-BV007-1PI1701-BV007-1

PI1701-BV002-1PI1701-BV002-1	─OCH ₂CH ₂OCH ₂CH ₂OCH ₂CH ₂O─ ─OCH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ O─	PI1701-BV008-1PI1701-BV008-1
PI1701-BV003-1PI1701-BV003-1	─NH(CH ₂) ₅NH─ ─NH(CH ₂ ) ₅ NH─	PI1701-BV009-1PI1701-BV009-1
PI1701-BV003-1PI1701-BV003-1	─NHCH ₂CH ₂OCH ₂CH ₂OCH ₂CH ₂NH─ ─NHCH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ NH─	PI1701-BV012-1PI1701-BV012-1
PI1701-LY160-1PI1701-LY160-1	─(CH ₂) ₅─ ─(CH ₂ ) ₅ ─	PI1701-BV023-1PI1701-BV023-1
PI1701-LY160-1PI1701-LY160-1	─(CH ₂) ₄─ ─(CH ₂ ) ₄ ─	PI1701-BV024-1PI1701-BV024-1
PI1701-LY160-1PI1701-LY160-1	─(CH ₂) ₃─ ─(CH ₂ ) ₃ ─	PI1701-BV025-1PI1701-BV025-1
PI1701-LY160-1PI1701-LY160-1	─(CH ₂) ₂─ ─(CH ₂ ) ₂ ─	PI1701-BV026-1PI1701-BV026-1
PI1701-LY160-1PI1701-LY160-1	─CH ₂CH ₂OCH ₂CH ₂OCH ₂CH ₂─ ─CH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ ─	PI1701-BV027-1PI1701-BV027-1
PI1701-LY160-1PI1701-LY160-1	─CH ₂CH ₂OCH ₂CH ₂─ ─CH ₂ CH ₂ OCH ₂ CH ₂ ─	PI1701-BV028-1PI1701-BV028-1
PI1701-LY120-1PI1701-LY120-1	─OCH ₂CH ₂OCH ₂CH ₂OCH ₂CH ₂O─ ─OCH ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ O─	PI1701-BV031-1PI1701-BV031-1

Biological Example 2: Surface Plasmon Resonance (SPR) Experiment Materials: ENL protein was purchased from ACTIVE MOTIF Company. Proteins were dissolved in sterile water and stored at -80°C after aliquoting. CM5 chip was purchased from GE Company, and DMSO was purchased from Sigma Company. Source plate (PP-0200) was purchased from Labcyte Company, Proxiplate-384plus target plate was purchased from Perkin Elmer Company.

Experimental method: The candidate compound was dissolved in DMSO, added to the source plate according to a certain concentration gradient, and then the compound was transferred to the target plate with ECHO550 to form 11 concentrations. The above-mentioned ENL recombinant protein was coupled to the CM5 chip by the method of amino coupling, and the coupling level was 5000RU. Each compound was tested by Biacore T200 of the State Key Laboratory of Bei Medicine.

Table 3 SPR test results of the compounds of the present invention.

	SPRSPR
BV002BV002	BB
BV003BV003	BB
BV008BV008	AA
BV009BV009	AA

Note: A: <100nM; B: 100-1000nM; C: >1000nM

The embodiments of the present invention have been described above. However, the present invention is not limited to the above-described embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

A compound of formula I and its racemate, stereoisomer, tautomer, isotope label, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or its Pharmaceutically acceptable salts:

Wherein, the Ar 1 and Ar 2 may be the same or different, and are each independently selected from the following groups optionally substituted by one, two or more R a :

Said R a is selected from H, =O, halogen, OH, CN, SH, NH 2 , COOH, or optionally substituted with one, two or more R b : C 1 -C 12 aliphatic Hydrocarbyl; the R b is selected from halogen, OH, CN, SH, NH 2 , COOH;

The L 1 and L 2 can be the same or different, each independently selected from the bond, or optionally substituted by one, two or more R L
(*, ** represent the connection site with other group parts of the general formula, and * represents the connection with the corresponding left group of the general formula, then ** represents the connection with the corresponding right group of the general formula); R L is selected from H, =O, halogen, OH, CN, SH, NH 2 , COOH, or optionally substituted with one, two or more R c : C 1 -C 12 aliphatic hydrocarbyl; The R c is selected from halogen, OH, CN, SH, NH 2 , COOH;

The W is selected from (C 1 -C 20 ) aliphatic hydrocarbon groups, optionally containing one, two or more heteroatoms (C 1 -C 20 ) aliphatic hydrocarbon groups; or more R W substituted; said R W is selected from H, =O, halogen, OH, CN, SH, NH 2 , COOH, or optionally substituted with one, two or more R d Group: C 1 -C 12 aliphatic hydrocarbon group; the R d is selected from halogen, OH, CN, SH, NH 2 , COOH;

The R 1 and R 2 can be the same or different, and are independently selected from -NH-C(=O)R 3 , -(CH 2 ) p R 3 , (C 1 -C 12 ) aliphatic hydrocarbon group, optionally (C 1 -C 12 )aliphatic hydrocarbon groups containing one, two or more heteroatoms;

Said R is selected from the following groups optionally substituted by one, two or more R e : C 6-20 aryl, 5-14 membered heteroaryl or 3-12 membered heterocyclyl;

The Re is selected from H, -C(=O)OC 1 -C 12 aliphatic hydrocarbon group, -OC 1 -C 12 aliphatic hydrocarbon group, C 1 -C 12 aliphatic hydrocarbon group, =O, halogen, OH, CN, SH, NH 2 , COOH; p is selected from 0, 1, 2, 3, 4, 5.
A compound of formula I according to claim 1 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites , ester, prodrug or a pharmaceutically acceptable salt thereof, characterized in that:

The W is selected from the following groups: -( CH2 )n-, -O-( CH2 )nO-, -O-( CH2 )n-, -( OCH2CH2 ) m -O-,- (OCH 2 CH 2 )m-, -(CH 2 CH 2 O)m-CH 2 CH 2 -, -(CH 2 )s-NR s -(CH 2 CH 2 O)m-CH 2 CH 2 -NR s- ( CH2 )s-,-( CH2 ) s -NRs-( CH2 )n-NRs-( CH2 ) s- ;

The Rs is selected from H, C 1 -C 12 aliphatic hydrocarbon group;

The s is selected from 0, 1, 2; the n is selected from 1-16, for example, selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 , 15, 16; the m is selected from 1-8, for example, selected from 1, 2, 3, 4, 5, 6, 7, 8.
A compound of formula I according to any one of claims 1-2 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, characterized by:

When the R 1 and R 2 are independently selected from (C 1 -C 12 ) aliphatic hydrocarbon groups optionally containing one, two or more heteroatoms, they may be further selected from -(CH 2 ) q N(CH 3 ) 2 , the q is selected from 0, 1, 2, 3.
A compound of formula I according to any one of claims 1-3 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, wherein the R is selected from the following groups optionally substituted by one, two or more Re : phenyl , indazole, pyrrolidine.
A compound of formula I according to any one of claims 1-4 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, characterized in that, the R 1 and R 2 can be the same or different, and are independently selected from the following groups:
A compound of formula I according to any one of claims 1-5 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, wherein the structure of formula I is selected from the following formula Ia or formula Ib:

In the structure of formula Ia, R 1 , R 2 , W, L 1 , and L 2 are as defined in formula I above.
A compound of formula I according to any one of claims 1-6 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, wherein the structure of formula I can be selected from the following formulas II-VII:

In the structures of formula II-VII, R 1 , R 2 , R S , s, n, m are as defined in formula I above.
A compound of formula I according to any one of claims 1-7 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, characterized in that Formula I is selected from the following specific compounds:
A compound of formula I according to any one of claims 1-7 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, characterized in that Formula I is selected from the following specific compounds:
The method for preparing the compound according to any one of claims 1-9, characterized in that it comprises the steps of: under suitable conditions, mixing the raw material containing the benzimidazole structure with the raw material containing the R 1 or R 2 group in a The reaction is carried out in a suitable reagent; or the raw material containing the -W- linking moiety and the raw material containing a benzimidazole structure are reacted in a suitable reagent, or the two reaction steps described above are included.
A pharmaceutical composition comprising the compound of formula I according to any one of claims 1-9 and its racemate, stereoisomer, tautomer, isotopic label, nitrogen oxide, solvate, Polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof; preferably, the pharmaceutical composition comprises a therapeutically effective amount of a compound of formula I and its racemates, stereoisomers, interconversions Isomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers.
The compound of formula I according to any one of claims 1-9 and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites , ester, prodrug or a pharmaceutically acceptable salt thereof or the pharmaceutical composition for the preparation of a medicament for preventing, regulating or treating a disease or condition mediated by ENL.
The use according to claim 12, wherein the ENL-mediated related diseases or conditions are selected from cancer, in particular leukemia. The leukemia is selected from acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML); and some less common types. Preferably, the leukemia is AML.