CN111269215B

CN111269215B - Nitrogen-containing heterocyclic organic compound and preparation method and application thereof

Info

Publication number: CN111269215B
Application number: CN202010250340.5A
Authority: CN
Inventors: 徐登峰; 何创; 雷桥仕; 李孟心; 马洪艳; 涂田勇
Original assignee: Zhongkelijian Pharmaceutical Guangzhou Co ltd
Current assignee: Zhongkelijian Pharmaceutical Guangzhou Co ltd
Priority date: 2020-04-01
Filing date: 2020-04-01
Publication date: 2021-10-26
Anticipated expiration: 2040-04-01
Also published as: CN111269215A

Abstract

The invention relates to a nitrogen-containing heterocyclic organic compound, a preparation method and application thereof, wherein the nitrogen-containing heterocyclic organic compound has a structure shown in a general formula (I):

Description

Nitrogen-containing heterocyclic organic compound and preparation method and application thereof

Technical Field

The invention relates to the technical field of medicine preparation, in particular to a nitrogen heterocyclic ring organic compound and a preparation method and application thereof.

Background

In recent years, there have been many important advances in the field of cancer treatment, and cancer-targeted therapies, immunotherapy, cell transplantation therapies, and the like have made it hope that patients will survive and live at high quality. However, the current cancer therapies still have many disadvantages, such as the effective response rate of the PD-1/PD-L1 drug in cancer immunotherapy is very low, and the drug is only effective for 10-20% of patients. Thus, there is a need for improvements to existing therapies, particularly cancer treatment regimens, that increase patient remission and survival.

The tissue microenvironment provides information conduction and strengthens connection for cell individuals and cell-to-cell behaviors, and is vital to maintaining normal operation of all functions in organisms. Under stress, the tissue microenvironment changes and a large body of experimental evidence suggests that disturbances in the tissue microenvironment are closely related to many diseases. For example, deregulation of the transforming growth factor-beta (TGF- β) signaling pathway in the tissue microenvironment is directly associated with the development and metastasis of cancer, and its hyperactivity is also closely associated with the therapeutic resistance of cancer immunotherapy (e.g., the PD-1/PD-L1 monoclonal antibody drug). Intervention towards important factors/signaling pathways in the tissue microenvironment, such as the TGF- β signaling pathway, is an important direction in the field of pharmaceutical development in recent years.

Transforming growth factor-beta (TGF-beta) signaling pathways are involved in a variety of cellular processes, including cell growth, migration, differentiation, apoptosis, etc., and play important roles in embryonic development of organisms, formation of tissues and organs, repair, immune inspection, and homeostasis of adults. The generalized TGF-beta signaling pathway is relatively complex, with the TGF-beta/SMAD signaling pathway being relatively well studied. The TGF- β/SMAD signaling pathway activates receptor-specific SMAD proteins (R-SAMD) by binding three subtype (. beta.1,. beta.2,. beta.3) free ligands to cell surface TGF- β transmembrane serine/threonine kinase receptors type I and type II to form heterologous complexes, which bind to common SMAD (co-SMAD) and enter the nucleus where they interact with other cytokines to co-regulate gene transcription. Abnormalities in the TGF- β signaling pathway can lead to a number of diseases, such as cancer, tissue fibrosis, dysplasia of cartilage, pulmonary hypertension, and the like. In particular, at advanced stages of various cancers, tumor cells and intratumoral stromal cells often promote immune escape and metastasis of the cancer by over-activating TGF- β signaling pathways, leading to cancer progression and increased difficulty in cancer treatment. It has been shown that inhibition of the TGF- β signalling pathway is effective in enhancing the immune response of cancer, particularly solid tumors, and also in reducing tumor metastasis. Preliminary experiments also demonstrated that administration of a TGF- β signaling pathway inhibitor in combination with a PD-1/PD-L1 pathway monoclonal antibody increased the survival time of experimental animals in the laboratory. Therefore, the development of safe and effective drugs for TGF-beta signaling pathway has been a research hotspot in the field of biological medicine.

Based on the characteristics of a TGF-beta signal channel, a type I receptor (TGF-beta receptor I, also called active-like kinase 5, ALK5) of TGF-beta protein on the cell surface is an ideal target in the field of small molecule targeted drug development. The small molecule selectively and targetedly inhibits the ALK5 protein from phosphorylating the downstream signal transduction protein Smad2/3, reduces or blocks the transmission of TGF-beta signals to the cell nucleus, and regulates the TGF-beta signal path to return to normal, thereby treating various TGF-beta signal path mediated diseases. Currently, some small molecule ALK5 inhibitors have entered clinical trials, such as gallunertitib from gifts (LY2157299), vacsertib from medpaco co-development (TEW-7197), etc.

Disclosure of Invention

Therefore, the nitrogen-containing heterocyclic organic compound has an inhibitory effect on ALK5, and provides a brand-new small molecular compound for preparing medicines for TGF-beta signal channel mediated diseases.

A nitrogen-containing heterocyclic organic compound having a structure represented by the general formula (I):

wherein, Y₁Is O, NR₃₀Or (CH)₂)_nN is an integer of 1 to 6;

X₁～X₄each independently is N or CR₃₁；

A is selected from any one of the following structures:

Z₁～Z₄each independently is CR₁₄Or N; and Z is₁～Z₄At most two N;

Z₅～Z₈each independently is CR₁₄R₁₅N or O; z₅～Z₈At most two N;

R₁and R₄Each independently selected from: H. substituted or unsubstituted C_1-6An alkyl group, a substituted or unsubstituted 3-20 membered cycloalkyl group, a substituted or unsubstituted 3-20 membered heterocyclyl group, a substituted or unsubstituted 5-20 membered aryl group, or a substituted or unsubstituted 5-20 membered heteroaryl group;

R₂、R₃、R₅～R₁₅each independently selected from: H. substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted 3-20 membered cycloalkyl, substituted or unsubstituted 3-20 membered heterocyclic group, substituted or unsubstituted 5-20 membered aromatic group, substituted or unsubstituted 5-20 membered heteroaromatic group, keto, carbonyl, carboxyl, ester group, alkoxycarbonyl, aryloxycarbonyl, amino, cyano, carbamoyl, haloformyl, isocyano, isocyanate group, thiocyanate group, isothiocyanate group, hydroxyl, nitro or halogen; and said R is₁、R₂And R₃Not H at the same time;

Y₂is O, NR₁₆Or (CH)₂)_nN is an integer of 1 to 6; r₁₆Is H or an alkanoyl group;

b is selected from: 6-10 membered nitrogen-containing heterocycle, -CH₂-or-CO-;

l is a substituent on B, and is selected from: H. substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted 3-to 8-membered cycloalkyl, substituted or unsubstituted 3-to 8-membered heterocyclyl, -NR₂₁R₂₂、COR₂₁、-COOR₂₁、-CONR₂₁R₂₂or-NCOR₂₁；

m is an integer of 1 to 7, and when m is greater than 1, L's are the same as or different from each other;

R₂₁and R₂₀Each independently is: H. substituted or unsubstituted C_1-6An alkyl group, a substituted or unsubstituted acyl group, a substituted or unsubstituted 3-8 membered cycloalkyl group, a substituted or unsubstituted 3-8 membered heterocyclyl group, or a substituted or unsubstituted 5-10 membered aryl or a substituted or unsubstituted 5-10 membered heteroaryl group; and said R is₂₁And R₂₀Can be reacted with R₂₁、R₂₀The attached N together form a substituted or unsubstituted 3-6 membered heterocyclyl;

R₃₀selected from: H. c_1-6Alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl;

R₃₁selected from: H. c_1-6Alkyl radical, C_1-6Alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-10 membered aryl, 5-10 membered heteroaryl, hydroxy or halogen.

The preparation method of the nitrogen heterocyclic ring organic compound comprises the following steps:

m represents halogen;

providing a compound shown as a formula (II-1);

reacting a compound shown as a formula (II-1) with compounds shown as (II-2) and (II-3) to prepare a compound shown as a formula (II-4);

reacting the compound shown in the formula (II-4) with the compound shown in the formula (II-5) to prepare a compound shown in the formula (II); or

M represents halogen;

providing a compound represented by the formula (III-1);

reacting a compound represented by the formula (III-1) with a compound represented by the formula (III-2) to obtain a compound represented by the formula (III-3);

reacting a compound shown as a formula (III-3) with a compound shown as a formula (II-3) to prepare a compound shown as a formula (III-4);

reacting the compound shown in the formula (III-4) with the compound shown in the formula (III-5) to prepare a compound shown in the formula (III);

reacting the compound represented by the formula (III-4) with the compound represented by the formula (IV-1) to obtain the compound represented by the formula (IV).

A composition, which comprises the nitrogenous heterocyclic organic compound or pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials.

The application of the nitrogen heterocyclic organic compound or the composition in preparing a medicament for treating or preventing TGF-beta signal channel mediated diseases.

A method of treating or preventing a TGF- β signalling pathway mediated disease comprising the administration of an effective amount of a nitrogen-containing heterocyclic organic compound as described above.

A medicament for treating or preventing TGF-beta signal channel mediated diseases, which comprises the nitrogen heterocyclic organic compound or pharmaceutically acceptable salt thereof.

The nitrogen heterocyclic organic compound can be used as an ALK5 inhibitor, has the potential of restoring a TGF-beta signal pathway to normal by regulating ALK5so as to intervene various diseases, and provides a series of compounds with brand new structures for the development of TGF-beta signal pathway mediated disease drugs.

Detailed Description

In order that the invention may be more fully understood, a more particular description of the invention will now be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

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. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Definition of terms

Unless otherwise indicated, the terms used in the present invention have the following definitions:

when stereoisomers of the compounds exist in the invention, the compounds are understood to include R configuration, S configuration and racemate when not particularly indicated.

The term "substituted" as used herein means substituted with one or more groups. When a plurality of groups are selected from the same series of candidate substituents, they may be the same or different.

The term "optionally" as used herein means that the defined group may or may not be selected from a list of candidate groups.

The term "substituted or unsubstituted" as used herein means that the defined group may or may not be substituted. When a defined group is substituted, it is understood to be optionally substituted with art-acceptable groups including, but not limited to: c_1-30An alkyl group, a cycloalkyl group having 3 to 20 ring atoms, a heterocyclic group having 3 to 20 ring atoms, an aryl group having 5 to 20 ring atoms, a heteroaryl group having 5 to 20 ring atoms, a silane group, a carbonyl group, an alkoxycarbonyl group, an aryloxycarbonyl group, a carbamoyl group, a haloformyl group, a formyl group, -NRR', a cyano group, an isocyano group, an isocyanate group, a thiocyanate group, an isothiocyanate group, a hydroxyl group, a trifluoromethyl group, a nitro group or a halogen, and the above groups may be further substituted by a substituent acceptable in the art; it is understood that R and R 'in-NRR' are each independently substituted with art-acceptable groups including, but not limited to, H, C_1-6An alkyl group, a cycloalkyl group having 3 to 8 ring atoms, a heterocyclic group having 3 to 8 ring atoms, an aryl group having 5 to 20 ring atoms or a heteroaryl group having 5 to 10 ring atoms; said C is_1-6Alkyl, cycloalkyl having 3 to 8 ring atoms, heterocyclyl having 3 to 8 ring atoms, aryl having 5 to 20 ring atoms or heteroaryl having 5 to 10 ring atoms optionally further substituted by one or moreAnd (3) substitution: c_1-6Alkyl, cycloalkyl having 3 to 8 ring atoms, heterocyclyl having 3 to 8 ring atoms, halogen, hydroxy, nitro or amino.

The term "a" in the context of the present invention denotes a linking site, for example in formula (I) A is

When is, represents Y₁Attached to the N linked to the "X" on the imidazole ring.

The "alkyl group" in the present invention represents saturated straight-chain and branched-chain alkyl groups in a specific number of atoms, and specific examples thereof include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, and tert-pentyl groups. C₁-C₆Alkyl refers to an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl.

"cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbyl substituent. 3-20 membered cycloalkyl means a monocyclic cycloalkyl group comprising 3 to 20 carbon atoms, preferably 3-8 membered. In one embodiment, the 3-8 membered monocyclic cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like. Polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups. Cycloalkyl groups may be optionally substituted with one or more substituents.

"Heterocyclyl" means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent in which one or more ring atoms are selected from nitrogen, oxygen, or S (O)_m(wherein m is an integer of 0 to 2), preferably nitrogen or oxygenA heteroatom; but not the ring moiety of-O-, -O-S-or-S-, the remaining ring atoms being carbon. A 3-20 membered heterocyclyl group refers to a ring containing 3 to 20 ring atoms, at least one of which is a heteroatom; preferably, the heterocyclyl ring contains 5 to 6 ring atoms of which 1-2 are heteroatoms. In one embodiment, heterocyclyl is dihydrofuranyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, or homopiperazinyl, and the like.

"aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, more preferably phenyl and naphthyl, most preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, and the aryl group may be substituted or unsubstituted.

A 5-20 membered "heteroaryl" refers to a heteroaromatic system containing at least one heteroatom, including but not limited to oxygen, sulfur, and nitrogen, and containing 5 to 20 ring atoms. Heteroaryl is preferably 5 or 6 membered, including but not limited to: furan, benzofuran, thiophene, benzothiophene, pyrrole, pyrazole, triazole, imidazole, oxazole, oxadiazole, thiazole, tetrazole, indole, carbazole, pyrroloimidazole, pyrrolopyrrole, thienopyrrole, thienothiophene, furopyrrole, furofuran, thienofuran, benzisoxazole, benzisothiazole, benzimidazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine, quinoline, isoquinoline, phthalazine, quinoxaline, phenanthridine, primadine, quinazoline, quinazolinone, and derivatives thereof. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, the ring to which the parent structure is attached being a heteroaryl ring. Heteroaryl groups may be optionally substituted or unsubstituted.

The "alkoxy group" as used herein means all linear or branched isomers having a specific number of carbon atoms, and specific examples thereof include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and n-butoxy.

The "halogen" represents fluorine, chlorine, bromine, iodine.

The substituent "amino" in the present invention includes primary, secondary and tertiary amino groups, specificallyAmino radicals comprising-NR₂₁R₂₂Wherein R is₂₁And R₂₂Is a hydrogen atom or any optional group such as: H. substituted or unsubstituted straight-chain alkyl groups, substituted or unsubstituted branched-chain alkyl groups, substituted or unsubstituted cycloalkyl groups, substituted or unsubstituted heterocyclic groups, substituted or unsubstituted aryl groups, or substituted or unsubstituted heteroaryl groups, and the like.

In the present invention, the substitution on the alkyl group or the cycloalkyl group, if it is not specified to occur on a specific carbon atom, means that it may occur on any carbon atom for which the number of substituents has not yet reached saturation. When a plurality of substituents are selected from the same series, they may be the same or different.

In the present invention, the substitution on the benzene ring, the heteroaromatic ring or the heterocyclic ring, if it is not specified to occur on a specific atom, means that it may occur at any position not substituted by other atoms than hydrogen. When a plurality of substituents are selected from the same series, they may be the same or different.

It is to be understood that when a group of the present invention contains a plurality of substituents, the plurality of substituents may be the same or different.

By "pharmaceutically acceptable salt" is meant that the compound of formula (I) retains the desired biological activity with minimal toxic side effects. The pharmaceutically acceptable salts may be obtained directly during the preparation and purification of the compound or indirectly by reacting the free acid or free base of the compound with another suitable base or acid.

The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). The term "hydrate" is used when the solvent is water.

Application method

For use in therapy, the compounds within the invention are generally administered in the form of a standard pharmaceutical composition. The compound comprises one or more compounds shown in a general formula (I) with effective treatment dose and pharmaceutically acceptable auxiliary materials. The pharmaceutically acceptable auxiliary materials are pharmaceutically acceptable carriers, excipients or sustained-release agents and the like.

The compounds and pharmaceutical compositions provided herein may be in a variety of forms such as tablets, capsules, powders, syrups, solutions, suspensions, and aerosols, and may be presented in a suitable solid or liquid carrier or diluent. The pharmaceutical compositions of the present invention may also be stored in a suitable injection or drip sterilization device. The pharmaceutical composition may also comprise flavoring agent, etc.

In the present invention, the pharmaceutical composition comprises a safe and effective amount (e.g. 0.1-99.9 parts by weight, preferably 1-90 parts by weight) of a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof; and the balance of pharmaceutically acceptable auxiliary materials, wherein the total weight of the composition is 100 parts by weight. Alternatively, the pharmaceutical composition of the present invention comprises 0.1 to 99.9 wt%, preferably 1 to 90 wt%, of the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof; and the balance of pharmaceutically acceptable excipients, wherein the total weight of the composition is 100% by weight.

The compound represented by the general formula (I) and a pharmaceutically acceptable carrier, excipient or sustained-release agent are preferably used in such a ratio that the compound represented by the general formula (I) as an active ingredient accounts for 60% or more of the total weight, the remainder accounts for 0 to 40% of the total weight, and the amount of the remainder is preferably 1 to 20%, most preferably 1 to 10%.

The compound shown in the general formula (I) or the pharmaceutical composition containing the compound shown in the general formula (I) can be clinically used for mammals including human and animals, and the administration route can comprise oral administration, nasal cavity inhalation, transdermal absorption, pulmonary administration or gastrointestinal tract administration and the like. The preferred route of administration is oral. Preferably in unit dosage form, and each dose contains 0.01mg-200mg, preferably 0.5mg-100mg of the active ingredient, and is administered once or in divided portions. Regardless of the method of administration, the optimal dosage for an individual will depend on the particular treatment. Usually starting with a small dose and gradually increasing the dose until the most suitable dose is found.

The pharmaceutical composition of the present invention can be administered orally as well as intravenously, intramuscularly or subcutaneously. Preferred pharmaceutical compositions are solid compositions, especially tablets and solid-filled or liquid-filled capsules, from the standpoint of ease of preparation and administration. Oral administration of the pharmaceutical composition is preferred.

The solid support comprises: starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose, kaolin, and the like, and liquid carriers include: sterile water, polyethylene glycols, non-ionic surfactants, edible oils (e.g., corn, peanut and sesame oils), and the like, as appropriate to the nature of the active ingredient and the particular mode of administration desired. Adjuvants commonly used in the preparation of pharmaceutical compositions may also advantageously be included, for example flavouring agents, colouring agents, preservatives and antioxidants such as vitamin E, vitamin C, BHT and BHA.

Injectable formulations include, but are not limited to, sterile, injectable, aqueous, oleaginous solutions, suspensions, emulsions and the like. These formulations may also be formulated with parenterally suitable diluents, dispersing agents, wetting agents, suspending agents and the like. Such injectable formulations can be sterilized by filtration in a bacterial-retaining filter. These formulations may also be formulated with an antimicrobial agent dissolved or dispersed in an injectable medium or by other methods known in the art.

Detailed Description

The invention provides a nitrogen heterocyclic ring organic compound, which has a structure shown in a general formula (I):

wherein, Y₁Comprises the following steps: o, NR₃₀Or (CH)₂)_nN is an integer of 1 to 6; further, n is an integer of 1 to 3; further, n is 1.

X₁～X₄Each independently is N or CR₃₁(ii) a Further, X₁～X₄At least one of which is N; further, X₁And X₃Is CR₃₁(ii) a Further, X₂And X₄Is N, or X₂Is CR₃₁、X₄Is N;

a is selected from any one of the following structures:

Z₁～Z₄each independently is CR₁₄Or N; and Z is₁～Z₄At most two N; further, Z₁～Z₄Are all CR₁₄(ii) a Further, Z₂～Z₄Are all CR₁₄，Z₁Is N;

Z₅～Z₈each independently is CR₁₄R₁₅N or O; z₅～Z₈At most two N; further, Z₅～Z₈Are all CR₁₄R₁₅(ii) a Further, Z₆～Z₈Are all CR₁₄R₁₅，Z₅Is N;

further, R₁And R₄Each independently selected from: H. substituted or unsubstituted C_1-6An alkyl group, a substituted or unsubstituted 3-to 8-membered cycloalkyl group, a substituted or unsubstituted 3-to 8-membered heterocyclic group, a substituted or unsubstituted 5-to 10-membered aryl group, or a substituted or unsubstituted 5-to 10-membered heteroaryl group.

Further, R₁And R₄When the group (b) is further substituted, it is preferably substituted with: c_1-6Alkyl radical, C_1-6Alkoxy, 3-to 8-membered cycloalkyl, 3-to 8-membered heterocyclyl, 5-to 20-membered aryl, 5-to 20-membered heteroaryl, silyl, carbonyl, alkoxycarbonyl, aryloxycarbonyl, carbamoyl, haloformyl, formyl, -NR₂₁R₂₂Cyano, isocyano, isocyanate, thiocyanate, isothiocyanate, hydroxyTrifluoromethyl, nitro or halogen; wherein-NR₂₁R₂₂R in (1)₂₁And R₂₂Each independently substituted with art-acceptable groups, preferably R₂₁And R₂₂Each independently is H or C_1-4Alkyl, and R₂₁And R₂₂Can be reacted with R₂₁And R₂₂The attached N together form a 5-6 membered heterocyclyl or heteroaryl group, preferably a heterocyclyl or heteroaryl group containing 1-2 heteroatoms; more preferably a 6-membered heterocyclyl group containing 2 heteroatoms which may be one or more of N, O and S, preferably N and/or O, or a 6-membered heteroaryl group containing 1-2 heteroatoms.

Further, R₁And R₄Each independently selected from: H. c_1-6Alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl; said C is_1-6Alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl optionally further substituted with: c_1-4Alkyl radical, C_1-4Alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-6 membered aryl, 5-6 membered heteroaryl, hydroxy, trifluoromethyl, nitro or halogen.

R₂、R₃、R₅～R₁₅Each independently selected from: H. substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted 3-20 membered cycloalkyl, substituted or unsubstituted 3-20 membered heterocyclic group, substituted or unsubstituted 5-20 membered aromatic group, substituted or unsubstituted 5-20 membered heteroaromatic group, keto, carbonyl, carboxyl, ester group, alkoxycarbonyl, aryloxycarbonyl, amino, cyano, carbamoyl, haloformyl, isocyano, isocyanate group, thiocyanate group, isothiocyanate group, hydroxyl, nitro or halogen; and R is₁、R₂And R₃Not H at the same time;

further, R₂、R₃、R₅～R₁₅Each independently selected from: H. substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted 3-to 8-membered cycloalkyl, or a substituted or unsubstituted aryl, heteroaryl, and heteroarylA substituted or unsubstituted 3-to 8-membered heterocyclic group, a substituted or unsubstituted 5-to 10-membered aromatic group, a substituted or unsubstituted 5-to 10-membered heteroaromatic group, a ketone group, a carbonyl group, a carboxyl group, an ester group, an alkoxycarbonyl group, an aryloxycarbonyl group, an amino group, a cyano group, a carbamoyl group, a haloformyl group, an isocyano group, an isocyanate group, a thiocyanate group, an isothiocyanate group, a hydroxyl group, a nitro group or a halogen; when C is present_1-6Alkyl radical, C_1-6When the alkoxy group, the 3-to 8-membered cycloalkyl group, the 3-to 8-membered heterocyclic group, the 5-to 10-membered aromatic group or the 5-to 10-membered heteroaromatic group is further substituted, it is preferably substituted with: c_1-4Alkyl radical, C_1-4Alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-6 membered aryl, 5-6 membered heteroaryl, hydroxy, trifluoromethyl, nitro or halogen.

Further, R₁Selected from: 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or 5-6 membered aryl, said 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered aryl being optionally further substituted by: c_1-6Alkyl, hydroxy, halogen or halogen substituted C_1-4An alkyl group;

and R is₂And R₃Each independently selected from the group consisting of: H. 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or 5-6 membered aryl, said 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered aryl being optionally further substituted by: c_1-6Alkyl, halogen or halogen substituted C_1-4An alkyl group.

and R is₂And R₃One of which is H and one is selected from the following groups: 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or 5-6 membered aryl, said 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered aryl being optionally further substituted by: c_1-6Alkyl, halogen or halogen radicalsGeneration C_1-4An alkyl group.

Further, R₃Is H, R₂Selected from the following groups: 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or 5-6 membered aryl, said 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered aryl being optionally further substituted by: c_1-6Alkyl, halogen or halogen substituted C_1-4An alkyl group.

Further, R₄Selected from: H. c_1-6Alkyl, 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or 5-6 membered aryl, C_1-6Alkyl, 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered aryl are optionally further substituted with: c_1-6Alkyl, halogen, hydroxy or halogen substituted C_1-4An alkyl group; and Z is₁Is CH or N, Z₂～Z₄Is CH; z₅～Z₈Is CH₂。

Further, the above-mentioned C_1-6Alkyl, 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and 5-6 membered aryl are optionally substituted with: methyl, hydroxy, F or trifluoromethyl;

further, R₁And R₄Each independently selected from H, C_1-4Alkyl or the following group:

and the above groups are optionally further substituted by C_1-4Alkyl or halogen substitution; further, optionally further substituted with methyl or F.

Y₂Is O, NR₁₆Or (CH)₂)_nN is an integer of 1 to 6; r₁₆Is H or an alkanoyl group; further, Y₂O, NH or N-Ac; further, Y₂Is NH.

B is selected from: 6-10 membered nitrogen-containing heterocycle, -CH₂-or-CO-; further, B is-CO-, phenyl, pyridyl, pyrimidinyl, or quinoline; further, B is-CO-; further, in the present invention,b is pyrimidinyl.

L is a substituent on B, and L is selected from: H. substituted or unsubstituted C_1-6Alkyl, substituted or unsubstituted C_1-6Alkoxy, substituted or unsubstituted 3-to 8-membered cycloalkyl, substituted or unsubstituted 3-to 8-membered heterocyclyl, substituted or unsubstituted 5-to 10-membered aryl, substituted or unsubstituted 5-to 10-membered heteroaryl, -NR₂₁R₂₂、COR₂₂、-COOR₂₂、-CONR₂₁R₂₂or-NCOR₂₁；

Still further, L is selected from: H. substituted or unsubstituted C_1-4Alkyl, substituted or unsubstituted C_1-4Alkoxy, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted 3-6 membered heterocyclyl, NR₂₁R₂₂、COR₂₂、-COOR₂₂、-CONR₂₁R₂₂or-NCOR₂₁。

Further, when said C is_1-6Alkyl radical, C_1-6When alkoxy, 3-to 8-membered cycloalkyl, 3-to 8-membered heterocyclyl, 5-to 10-membered aryl, 5-to 10-membered heteroaryl are further substituted, it is preferably substituted with: H. c_1-4Alkyl, hydroxy substituted C_1-4Alkyl, halogen substituted C_1-4Alkyl, -NR₂₁R₂₂、COR₂₂、-COOR₂₂、-CONR₂₁R₂₂or-NCOR₂₁；

m is an integer of 1 to 7, and when m is greater than 1, L's are the same as or different from each other; further, m is 1,2 or 3. Further, m is 1 or 3.

R₂₁And R₂₂Each independently is: H. substituted or unsubstituted C_1-6An alkyl group, a substituted or unsubstituted acyl group, a substituted or unsubstituted 3-8 membered cycloalkyl group, a substituted or unsubstituted 3-8 membered heterocyclyl group, or a substituted or unsubstituted 5-10 membered aryl or a substituted or unsubstituted 5-10 membered heteroaryl group; and R is₂₁And R₂₀Can be reacted with R₂₁、R₂₀The attached N together form a substituted or unsubstituted 3-6 membered heterocyclyl; further, R₂₁And R₂₂Can be reacted with R₂₁And R₂₂The attached N together form a substituted or unsubstituted 5-6 membered heterocyclyl or heteroaryl group,preferably to form a heterocyclic or heteroaryl group containing 1-2 heteroatoms; more preferably a 6-membered heterocyclyl group containing 2 heteroatoms which may be one or more of N, O and S, preferably N and/or O, or a 6-membered heteroaryl group containing 1-2 heteroatoms.

Further, L is selected from: H. c_1-4Alkyl, hydroxy substituted C_1-4Alkyl (preferably)

)、C_1-4Alkoxy, -N (R)₂₂)₂、-NH-Ac、-COR₂₂、-COOR₂₂、-CONH₂Or the following groups

R₂₂Is H or C_1-4An alkyl group;

R₂₃is H, C_1-4Alkyl, aryl, heteroaryl, and heteroaryl,

Wherein p and q are each independently an integer from 1 to 5; further, p is 1, q is 1;

R₂₄、R₂₅and R₂₆Each independently selected from: H. c_1-6Alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl; further, R₂₅Is H; further, R₂₅Is H; further, R₂₄And R₂₆One of them is H and one is C_1-6Alkyl or₂₄And R₂₆Are all H; further, R₂₄Is C_1-6Alkyl radical, R₂₆Is H.

Further, B is-CH₂When, L is not hydrogen or alkyl.

Further, when B is-CO-, L is not hydrogen or-NR₂₁R₂₂。

R₃₀Selected from: H. c_1-6Alkyl, 3-8 membered cycloalkyl3-8 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl; further, R₃₀Is H or C_1-4An alkyl group.

R₃₁Selected from: H. c_1-6Alkyl radical, C_1-6Alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, 5-10 membered aryl, 5-10 membered heteroaryl, hydroxy or halogen; further, R₃₁Is H.

Further, when X is present₁～X₄Are all CR₃₁When A is

Further, when A is

X₁～X₄At least one of which is N.

Further, when A is

When, Y₁Is (CH)₂)_nOr NR₃₀(ii) a When A is

When, Y₁Is O.

Further, when A is

Y₁Is (CH)₂)_n，Y₂Is NH or N-Ac; further, Y₁Is CH₂，Y₂Is NH.

Further, when A is

Y₁Is O, Y₂Is NH or O.

Further, the nitrogen-containing heterocyclic organic compound has a structure represented by any one of the following general formulae (II) to (IV):

the definitions of the groups in the general formulae (II) to (IV) are as described above and are not described in detail here.

Further, in the structure represented by the formula (II), R₁And R₂Each independently selected from the group consisting of:

further, in the structure of formula (II), R₁、R₂And R₃In which one group is

Further, in the structure of formula (II), R₁Is composed of

R₂Is composed of

Further, in the structure shown in the formula (II), L is-NH₂Or

Further, in the structure shown in the formula (III), R₄Selected from: methyl or the following groups:

further, in the structure shown in the formula (III), L is-NH₂、-N(CH₃)₂、-NH-Ac、-COOR₂₂、-CONH₂A methyl group,

Further, the nitrogen-containing heterocyclic organic compound is selected from the following compounds:

the invention also provides a preparation method of the compound, which comprises the following steps:

providing compounds with structures shown in a formula (I-1), a formula (I-2) and a formula (I-3), and respectively carrying out reaction;

it is to be understood that the above reaction may be a condensation reaction, a substitution reaction, and the like, which are conventional in the art, and the reaction reagents and reaction conditions may be selected according to actual needs, and are not particularly limited herein.

Further, it is preferable to prepare compounds having structures represented by formulas (II) to (IV) by the following method:

s101: providing a compound shown as a formula (II-1);

it will be appreciated that the choice of suitable starting materials, not specifically limited herein, may be made depending on the desired end product, e.g. R₁In the case of pyridine, 2-amidinopyridine hydrochloride can be used.

S102: reacting a compound shown as a formula (II-1) with compounds shown as (II-2) and (II-3) to prepare a compound shown as a formula (II-4);

it is understood that, in S102, the compound represented by the formula (II-1) and the compound represented by the formula (II-2) may be reacted first and then reacted with the compound represented by the formula (II-3), or the compound represented by the formula (II-1), the compound represented by the formula (II-2) and the compound represented by the formula (II-3) may be simultaneously mixed and reacted, and the reaction is not particularly limited and is understood to be within the scope of the present invention. It is preferable that the compound represented by the formula (II-1) is reacted with the compound represented by the formula (II-2) and then reacted with the compound represented by the formula (II-3).

Further, it is preferable to carry out the reaction according to the following steps: mixing the compound shown in the formula (II-1) and the compound shown in the formula (II-2) with alkali and solvent, and respectively carrying out microwave reaction for 10min to 20min (preferably 15min) at 70 ℃ to 90 ℃ (preferably 80 ℃) and 140 ℃ to 160 ℃ (preferably 150 ℃). Then cooling to room temperature, adding the compound shown in the formula (II-3) and alkali, carrying out microwave reaction under the condition of 140-240 ℃ (preferably 150 ℃) until the reaction is finished (preferably 20min), and separating and purifying.

Further, the molar ratio of the compound represented by the formula (II-1), the compound represented by the formula (II-2) and the compound represented by the formula (II-3) is 1:1: 1.

Further, the base is potassium carbonate and the solvent is acetonitrile.

S103, reacting the compound represented by the formula (II-4) with the compound represented by the formula (II-5) to obtain the compound represented by the formula (II).

Further, step S103 includes the steps of:

mixing the compound shown in the formula (II-4) and the compound shown in the formula (II-5) in a catalyst, alkali, a ligand and a solvent, reacting, separating and purifying to obtain the required product.

Further, the reaction is carried out for 20min to 40min (preferably 30min) under the microwave condition at the temperature of 140 ℃ to 240 ℃ (preferably 150 ℃).

Further, the catalyst is palladium acetate, the ligand is Xantphos ligand, and the alkali is cesium carbonate solvent and is 1, 4-dioxane.

Further, the molar ratio of the compound represented by the formula (II-4) to the compound represented by the formula (II-5) is 1 (2-2.5).

S201: providing a compound represented by the formula (III-1);

the compound (1, 2-phenylenediamine dihydrochloride) represented by the formula (III-1) is a commercially available raw material.

S202: reacting a compound represented by the formula (III-1) with a compound represented by the formula (III-2) to obtain a compound represented by the formula (III-3);

further, step S202 includes the steps of: dissolving a compound shown as a formula (III-1), a compound shown as a formula (III-2) and polyphosphoric acid in a solvent, reacting at the temperature of 100-140 ℃, and separating and purifying after the reaction is finished.

Further, the molar ratio of the compound represented by the formula (III-1) to the compound represented by the formula (III-2) was 1: 1.

S203: reacting a compound shown as a formula (III-3) with a compound shown as a formula (II-3) to prepare a compound shown as a formula (III-4);

further, step S203 includes the steps of: mixing the compound shown in the formula (III-3) and the compound shown in the formula (II-3), alkali and a solvent, reacting for 10min to 20min (preferably 15min) at 110 ℃ to 130 ℃ (preferably 120 ℃) under the microwave condition, and separating and purifying after the reaction is finished.

Further, the molar ratio of the compound represented by the formula (III-3) to the compound represented by the formula (II-3) is 1: (1-1.2).

S204 a: reacting the compound shown in the formula (III-4) with the compound shown in the formula (III-5) to prepare a compound shown in the formula (III);

the step is as step S103, and is not described herein again.

S204 b: reacting the compound represented by the formula (III-4) with the compound represented by the formula (IV-1) to obtain the compound represented by the formula (IV).

The invention also provides a pharmaceutically acceptable salt which is prepared from the nitrogenous heterocyclic organic compound.

The invention also provides a composition which comprises the nitrogenous heterocyclic organic compound or pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials.

The invention also provides the application of the nitrogenous heterocyclic organic compound or the composition in preparing a medicament for treating or preventing TGF-beta signal channel mediated diseases.

Further, the drug is an ALK5 kinase inhibitor.

Further, the medicament is a medicament for treating cancer, infectious diseases or autoimmune diseases.

The invention also provides a method of treating or preventing TGF-beta signaling pathway mediated diseases comprising administering an effective amount of the above-described nitrogen-containing heterocyclic organic compound.

The present invention will be described below with reference to specific examples.

In the following examples, analytical data of samples were determined by the following instruments: the nuclear magnetic resonance is measured by a Bruker AMX-400 nuclear magnetic resonance instrument and a Bruker AMX-500 nuclear magnetic resonance instrument, TMS (tetramethylsilane) is an internal standard, the unit of chemical shift is ppm, and the unit of coupling constant is Hz; mass spectra were determined by an Agilent1200/MSD mass spectrometer.

Silica gel 200-300 mesh for column chromatography (produced by Qingdao ocean factory); the TLC silica gel plate is an HSGF-254 thin-layer chromatography prefabricated plate produced by a cigarette bench chemical plant; the boiling range of petroleum ether is 60-90 ℃; the color is developed by adopting an ultraviolet lamp, an iodine cylinder, phosphomolybdic acid and the like.

The starting materials, reaction reagents and the like used in the following examples are commercially available products unless otherwise specified. Reagents and solvents used in the experiment are all processed according to the specific conditions of the reaction.

The abbreviations used in the present invention are the same as those commonly used in the art and have the following meanings:

PE: petroleum ether, DCM: dichloromethane, EA: ethyl acetate, THF: tetrahydrofuran, MeOH: methanol, DMF: n, N-dimethylformamide, DME: ethylene glycol dimethyl ether, DMA: n, N-dimethylacetamide, DIEA: n, N-diisopropylethylamine, TEA: triethylamine, DCC: dicyclohexylcarbodiimide, DMAP: 4-N, N-dimethylaminopyridine, HOBt: 1-hydroxybenzotriazole, edc.hcl: 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride, Boc anhydride: di-tert-butyl dicarbonate.

Example 1

Synthetic route of compound 1:

synthesis of an intermediate A:

add 2-amidinopyridine hydrochloride (0.16g,1.0mmol), 2-bromo-1- (2-pyridyl) -1-ethanone hydrobromic acid (0.28g, 1.0mmol), sodium bicarbonate (0.30g,3.5mmol) and acetonitrile (15mL) to the microwave tube. After the addition, the reactant is in a micro stateStirring was carried out under a wave condition at 80 ℃ and 150 ℃ for 15 minutes each. Then cooled to room temperature, 2-chloro-4- (chloromethyl) pyridine (0.16g,1.0mmol), potassium carbonate (0.27g,2.0mmol) were added to the microwave tube and stirred at 150 ℃ for 20 minutes. Cooled to room temperature, concentrated to remove the solvent and the residue purified by column chromatography on silica gel to give intermediate product a (0.10g, 28.8%) as a brown solid. LC-MS (ESI) M/z 348.11[ M + H ]]⁺。

Target compound synthesis:

intermediate product A (0.10g,0.29mmol), 4-amino-6- (4-methyl-1-piperazinyl) pyrimidine (0.13g,0.67mmol), cesium carbonate (0.24g,0.74mmol), palladium acetate (0.010g,0.04mmol), Xantphos ligand (0.040g,0.07mmol) and 1, 4-dioxane (18mL) were added to the microwave tube. After the addition, the reaction was stirred at 150 ℃ for 30 minutes under microwave conditions. Cooled to room temperature, concentrated to remove the solvent and the residue was purified by silica gel column chromatography to give compound LEG001122(0.030g, 20.7%) as a brown solid.¹H NMR(500MHz,Chloroform-d)δ8.58(s,1H),8.53(s,1H),8.38(d,J＝8.0Hz,1H),8.27(d,J＝7.9Hz,1H),8.20(d,J＝5.2Hz,1H),8.12(d,J＝7.9Hz,1H),7.79(dd,J＝14.9,7.6Hz,2H),7.75(s,1H),7.55(s,1H),7.24(t,J＝6.3Hz,1H),7.18(t,J＝6.3Hz,1H),7.13(s,1H),7.03(d,J＝13.3Hz,1H),6.74(d,J＝5.2Hz,1H),6.00(s,2H),3.65(s,3H),2.55–2.44(m,4H),2.38-2.34((m,4H).LC-MS(ESI):m/z＝503.18[M-H]^-.

Examples 2 to 10 were prepared according to the procedure of example 1

Example 2

Substantially the same as in example 1 except that 1-cyclopropyl-2-bromoethanone was used in place of 2-bromo-1- (2-pyridyl) -1-ethanone hydrobromic acid.

¹H NMR(500MHz,Chloroform-d)δ8.44(d,J＝4.8Hz,1H),8.27(s,1H),8.22–8.12(m,2H),7.71(t,J＝7.9Hz,1H),7.19–7.11(m,1H),7.04(s,1H),6.95(s,1H),6.70(s,1H),6.65(d,J＝5.3Hz,1H),5.84(s,2H),3.64(s,3H),3.49(s,1H),2.48(t,J＝5.2Hz,4H),2.34(t,J＝5.2Hz,4H),1.95–1.86(m,1H),0.89(t,J＝7.9Hz,2H),0.77(t,J＝5.2Hz,2H).LC-MS(ESI):m/z＝468.21[M+H]⁺

Example 3

Substantially the same as in example 1, except that tetrahydropyran-4-formamidine hydrochloride was used in place of 2-amidinopyridine hydrochloride and 2-bromo-1- (tetrahydro-2H-pyran-4-yl) ethanone was used in place of 2-bromo-1- (2-pyridyl) -1-ethanone hydrobromide

¹H NMR(500MHz,Chloroform-d)δ8.28(s,1H),8.22(d,J＝5.3Hz,1H),7.12(s,1H),6.91(s,1H),6.51(s,1H),6.48(d,J＝5.3Hz,1H),5.05(s,2H),4.01(dd,J＝11.4,5.6Hz,5H),3.67(d,J＝5.7Hz,4H),3.57–3.47(m,3H),3.47–3.37(m,3H),2.89–2.80(m,1H),2.80–2.72(m,1H),2.50(t,J＝5.2Hz,5H),2.35(s,3H),2.11–1.94(m,4H).LC-MS(ESI):m/z＝517.31[M-H]^-.

Example 4

Substantially the same as in example 2, except that tetrahydropyran-4-formamidine hydrochloride was used in place of 2-amidinopyridine hydrochloride and 4, 6-diaminopyrimidine was used in place of 4-amino-6- (4-methyl-1-piperazinyl) pyrimidine

¹H NMR(500MHz,Chloroform-d)δ8.90(s,1H),8.18(d,J＝6.3Hz,2H),7.01(s,1H),6.96(s,1H),6.49(s,1H),6.45(s,1H),5.25(s,2H),5.00(s,2H),3.99(t,J＝11.9,4.1Hz,2H),3.39(t,J＝11.8Hz,2H),2.77-2.69(m,1H),2.08–1.99(m,2H),1.90–1.86(m,1H),1.61(t,J＝13.6Hz,2H),0.83(d,J＝7.6Hz,2H),0.63(d,J＝5.2Hz,2H).LC-MS(ESI):m/z＝392.21[M+H]⁺.

Example 5

Substantially the same as in example 1, except that 2-bromo-1- (tetrahydro-2H-pyran-4-yl) ethanone was used in place of 2-bromo-1- (2-pyridyl) -1-ethanone hydrobromic acid, and 4, 6-diaminopyrimidine was used in place of 4-amino-6- (4-methyl-1-piperazinyl) pyrimidine

¹H NMR(500MHz,Chloroform-d)δ8.45(d,J＝4.8Hz,1H),8.23(s,1H),8.21–8.12(m,2H),7.72(t,J＝7.9Hz,1H),7.17(t,J＝6.2Hz,2H),6.94(s,1H),6.85(s,1H),6.73(s,1H),6.68(d,J＝5.3Hz,1H),5.87(s,2H),4.76(s,2H),4.05(dd,J＝12.5,4.0Hz,2H),3.55(t,J＝11.6Hz,2H),2.89(t,J＝11.9Hz,1H),2.03–1.97(m,2H),1.77-1.74(m,2H).LC-MS(ESI):m/z＝427.16[M-H]^-.

Example 6

Substantially the same as in example 4, except that 2-amidinobenzene was used in place of tetrahydropyran-4-formamidine hydrochloride

¹H NMR(500MHz,Chloroform-d)δ8.26(s,1H),8.18(d,J＝5.2Hz,1H),7.51(td,J＝7.5,1.8Hz,1H),7.42-7.37(m,1H),7.22–7.17(m,1H),7.15–7.09(m,1H),6.91(s,1H),6.86(s,1H),6.70(s,1H),6.56(d,J＝5.2Hz,1H),4.96(s,2H),4.83(s,2H),3.74(s,1H),1.93-1.87(m,1H),0.89-0.85(m,2H),0.82–0.77(m,2H).LC-MS(ESI):m/z＝400.14[M-H]^-.

Example 7

Substantially the same as example 5, except that cyclopropanecarboxamidine was used in place of 2-amidinopyridine hydrochloride and 3,4, 5-trimethoxyaniline was used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)δ8.12(s,1H),7.02(s,1H),6.58(s,2H),6.49(s,1H),6.44(d,J＝5.0Hz,1H),6.39(s,1H),5.03(s,2H),3.96(d,J＝10.0Hz,2H),3.80(s,3H),3.79(s,6H),3.45(t,J＝10.0Hz,2H),2.71(t,J＝10.0Hz,2H),1.60(t,J＝10.0Hz,2H),1.56(m,3H),0.94(m,2H),0.0.84(m,2H).MS(MM-ES+APCI)m/z:467.24[M+H]⁺.HPLC purity:94.58％.

Example 8

Substantially the same as in example 7 except that 4, 6-diaminopyrimidine was used in place of 3,4, 5-trimethoxyaniline

¹H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.18(d,J＝5.2Hz,1H),8.04(s,1H),7.26(s,1H),6.91(s,1H),6.82(s,1H),6.60(d,J＝4.8Hz,1H),6.50(s,2H),5.22(s,2H),3.88-3.84(m,2H),3.41-3.32(m,2H),2.65-2.59(m,1H),1.87-1.76(m,3H),1.56-1.50(m,2H),0.84-0.78(m,4H).MS(MM-ES+APCI)m/z:392.29[M+H]⁺.HPLC purity:98.95％.

Example 9

Essentially the same as in example 8, except that 2- (4-aminopyridine) -2-propanol was used in place of 4, 6-diaminopyrimidine

¹H NMR(400MHz,CDCl3)δ9.26(s,1H),8.28(d,J＝5.2Hz,1H),8.23(s,1H),7.61(s,1H),7.48(s,1H),6.67(d,J＝4.8Hz,1H),6.52(s,1H),6.41(s,1H),5.13(s,2H),3.90(m,2H),3.36(t,J＝8.0Hz,2H),2.66-2.61(m,1H),1.82(m,2H),1.58-1.51(m,3H),1.48(s,6H),0.86(m,2H),0.80(m,2H).MS(MM-ES+APCI)m/z:434.27[M+H]⁺.HPLC purity:91.47％.

Example 10

Substantially the same as in example 8, except that 1-cyclopropyl-2-bromoethanone was used in place of 2-bromo-1- (tetrahydro-2H-pyran-4-yl) ethanone

¹H NMR(500MHz,CH₃OD)δ8.09(s,1H),7.1(s,1H),7.01(s,1H),6.69-6.67(m,2H),5.22(s,2H),1.78-1.73(m,2H),0.91-0.89(m,4H),0.84-0.80(m,2H),0.62-0.59(m,2H).MS(MM-ES+APCI)m/z:348.21[M+H]⁺.HPLC purity:86.55％.

Example 11

The synthesis route of the compound is as follows:

the synthesis steps of the compound are as follows:

intermediate 11-a synthesis:

to the microwave tube were added 1, 2-phenylenediamine dihydrochloride (0.18g,1.0mmol), 2-tetrahydropyranoic acid (0.13g, 1.0mmol) and polyphosphoric acid (4 mL). After the addition, the reaction was stirred at 120 ℃ for 3 hours in an oil bath. Then cooled to room temperature. The reaction mass was made basic with sodium bicarbonate solution, extracted with ethyl acetate, concentrated to remove the solvent and the residue was purified by silica gel column chromatography to give the product intermediate 11-a (0.070g, 35%) as a brown solid. LC-MS (ESI) M/z 201.1[ M-H ]]^-。

Intermediate 11-b synthesis:

intermediate 11-a (0.070g,0.35mmol), 2-chloro-4- (chloromethyl) pyridine (0.070g,0.41mmol), potassium carbonate (0.10g,0.70mmol) and acetonitrile (10mL) were added to a microwave tube. The mixture was stirred at 120 ℃ for 15min under microwave conditions, concentrated to remove the solvent, and the residue was purified by silica gel column chromatography to give intermediate 11-b (0.10g, 23%) as a brown solid. LC-MS (ESI) M/z 326.12[ M-H ═]^-。

Target compound synthesis:

add intermediate 11-b (0.10g,0.0.30mmol), 4, 6-diaminopyrimidine (0.10g,0.90mmol), cesium carbonate (0.20g,0.65mmol), palladium acetate (0.010g,0.03mmol), Xantphos ligand (0.035g,0.06mmol) and 1, 4-dioxane (18mL) to the microwave tube. After the addition, the reaction was stirred at 150 ℃ for 15 minutes under microwave conditions. Cooled to room temperature, concentrated to remove the solvent and the residue was purified by column chromatography on silica gel to give the desired product (0.020g, 16.3%) as a brown solid. LC-MS (ESI) m/z ═ m/z400.18[M-H]^-。¹H NMR(500MHz,Chloroform-d)δ8.20(d,J＝5.3Hz,2H),7.83(d,J＝8.0Hz,1H),7.32–7.28(m,1H),7.24–7.22(m,1H),7.17(d,J＝7.9Hz,1H),6.94(s,1H),6.62(d,J＝1.1Hz,1H),6.54(d,J＝5.4Hz,1H),5.37(s,2H),4.73(s,2H),4.11–4.05(m,2H),3.49(d,J＝4.1Hz,3H),2.24-2.17(m,1H),2.04(d,J＝5.4Hz,1H),1.79(d,J＝13.6Hz,2H).

Examples 12 to 45 were prepared according to the preparation scheme of example 11.

Example 12

Essentially the same as in example 11, except that 2-methyl-3H-imidazopyridine was used in place of intermediate 11-a

¹H NMR(500MHz,DMSO-d6)δ9.52(s,1H),8.28(d,J＝5.0Hz,1H),8.16(d,J＝5.0Hz,1H),8.01(d,J＝5.0Hz,1H),7.97(s,1H),7.27(dd,J＝5.0,10.0Hz,1H),7.18(s,1H),6.92(s,1H),6.67(d,J＝5.0Hz,1H),6.50(s,2H),5.49(s,2H),2.55(s,3H).MS(MM-ES+APCI)m/z:355.2[M+Na]⁺.HPLC purity:98.24％.

Example 13

Substantially the same as in example 11 except that 2-pyrazinecarboxylic acid was used in place of 2-tetrahydropyranoic acid

¹H NMR(500MHz,DMSO-d6)δ9.58(s,1H),9.45(s,1H),8.74-8.72(m,2H),8.11(d,J＝5.5Hz,1H),7.93(s,1H),7.86-7.84(dd,J＝5.1,10.3Hz,1H),7.64-7.62(dd,J＝5.2,10.0Hz,1H),7.37-7.33(m,2H)，δ7.19(s,1H),6.87(s,1H),6.67(d,J＝5.1Hz,1H),6.47(s,2H),6.12(s,2H).MS(MM-ES+APCI)m/z:418.17[M+Na]⁺.HPLC purity:98.55％.

Example 14

Substantially the same as in example 11 except that 2-pyridinecarboxylic acid is used in place of 2-tetrahydropyranoic acid and N, N-dimethylpyrimidine-4, 6-diamine is used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)δ8.58(d,J＝4.5Hz,1H),8.47(d,J＝8.0Hz,1H),8.23(s,1H),8.13(d,J＝5.2Hz,1H),7.88-7.51(m,2H),7.34-7.21(m,4H),7.11(s,1H),6.67(s,1H),6.64(d,J＝5.1Hz,1H),6.17(s,2H),3.03(s,6H).MS(MM-ES+APCI)m/z:445.2[M+Na]⁺.HPLC purity:99.94％.

Example 15

Substantially the same as in example 14, except that 4-amino-2, 6-dimethylpyrimidine was used in place of N, N-dimethylpyrimidine-4, 6-diamine

¹H NMR(500MHz,CDCl3)δ8.51(s,1H),8.44(d,J＝8.0Hz,1H),8.37(s,1H),8.11(d,J＝5.2Hz,1H),7.85(d,J＝5.2Hz,1H),7.77(dd,J＝1.6,9.3Hz,1H),7.36(s,1H),7.29-7.22(m,4H),6.91(s,1H),6.72(d,J＝5.1Hz,1H),6.14(s,2H),2.33(s,3H),2.27(s,3H).MS(MM-ES+APCI)m/z:430.16[M+Na]⁺.HPLC purity:99.11％.

Example 16

Substantially the same as in example 13, except that 6-methyl-2-pyridinecarboxylic acid was used instead of 2-pyrazinecarboxylic acid

¹H NMR(500MHz,CDCl3)δ9.48(s,1H),8.19(d,J＝5.1Hz,1H),8.08(d,J＝5.0Hz,1H),7.94(s,1H),7.86(t,J＝8.3Hz,1H),7.79-7.78(dd,J＝4.1,8.3Hz,1H),7.63-7.61(dd,J＝4.1,8.3Hz,1H),7.34-7.29(m,3H),7.23(s,1H),6.87(s,1H),6.59(d,J＝5.1Hz,1H),6.46(s,2H),6.18(s,2H),2.47(s,3H).MS(MM-ES+APCI)m/z:355.2[M+Na]⁺.HPLC purity:98.24％.

Example 17

Intermediate 17-a synthesis:

add 5-hydroxypyrimidine (0.1g,1.0mmol), 2, 4-dichloropyridine (0.15g,1.0mmol), cesium carbonate (0.35g,1.0mmol) and DMF (4mL) to the microwave tube. After the addition, the reaction was stirred for 3 hours at 110 ℃ in an oil bath. Then cooled to room temperature. The reaction mass was diluted with water, extracted with ethyl acetate, concentrated to remove the solvent and the residue was purified by silica gel column chromatography to give the product intermediate 17-a (0.12g, 60%) as a brown solid. LC-MS (ESI) 208.1[ M + H ]]⁺。

Target compound synthesis:

¹H NMR(400MHz,CDCl3)δ9.06(s,1H),8.59(s,2H),8.27(d,J＝6.0Hz,1H),8.23(d,J＝6.4Hz,1H),7.79(s,1H),7.44(s,1H),7.33-7.31(d,J＝5.6Hz,1H),6.49-6.47(d,J＝5.6Hz,1H),6.45(s,1H),3.69(s,1H),1.48(s,6H).MS(MM-ES+APCI)m/z:324.1[M+H]⁺.HPLC purity:98.24％.

Example 18

Essentially the same as in example 17, except that 5-hydroxypyrimidine was used in place of 2- (4-aminopyridine) -2-propanol

¹H NMR(400MHz,CDCl3)δ8.97(s,1H),8.92(s,1H),8.56(s,2H),8.51(s,2H),8.35(d,J＝5.6Hz,1H),6.79-6.77(d,J＝5.6Hz,1H).MS(MM-ES+APCI)m/z:269.0[M+H]⁺.HPLC purity:98.24％.

Example 19

Substantially the same as in example 17 except that 3-hydroxyquinoline is used in place of 5-hydroxypyrimidine

¹H NMR(400MHz,CDCl3)δ8.77(d,J＝2.8Hz,1H),8.27(d,J＝5.6Hz,1H),8.23(d,J＝6.0Hz,1H),8.12(d,J＝8.4Hz,1H),7.82(d,J＝1.6Hz,1H),7.77-7.69(m,2H),7.60-7.57(dd,J＝1.2,8.0Hz,1H),7.38(d,J＝2Hz,1H),7.32(s,1H),7.25-7.24(dd,J＝2.0,5.6Hz,1H),6.60-6.58(dd,J＝2.0,5.6Hz,1H),6.4(d,J＝2.0Hz,1H),1.45(s,6H).MS(MM-ES+APCI)m/z:395.1[M+Na]⁺.HPLC purity:96.30％.

Example 20

Substantially the same as in example 19, except that 4, 6-diaminopyrimidine was used in place of 2- (4-aminopyridine) -2-propanol

¹H NMR(400MHz,MeOD)δ8.73(d,J＝2.8Hz,1H),8.15(d,J＝6.0Hz,1H),8.06-8.04(m,2H),7.97(s,1H),7.89(d,J＝8.0Hz,1H),7.75-7.71(m,1H),7.63-7.61(t,J＝7.2Hz,1H),7.04(s,1H),6.98(s,1H),6.62-6.60(dd,J＝2.4,6.0Hz,1H),3.34(s,1H),1.25(s,2H).MS(MM-ES+APCI)m/z:353.12[M+Na]⁺.HPLC purity:99.22％.

Example 21

Substantially the same as in example 9, except that 2- (trifluoromethyl) benzimidazole was used instead of intermediate 11-a

¹H NMR(500MHz,CDCl3)δ8.32(d,J＝5.0Hz,1H),8.22(d,J＝5.0Hz,1H),7.77(d,J＝10.0Hz,1H),7.51(s,1H),7.43-7.38(m,2H),7.36-7.33(m,1H),7.27-7.25(m,2H),6.73(d,J＝5.0Hz,1H),6.32(s,1H),5.51(s,2H),1.46(s,6H).MS(MM-ES+APCI)m/z:450.19[M+Na]⁺.HPLC purity:88.03％.

Example 22

Substantially the same as in example 21, except that 2- (2-pyridine) benzimidazole was used instead of intermediate 11-a

¹H NMR(500MHz,DMSO-d6)δ9.47(s,1H),8.64(d,J＝5.0Hz,1H),8.39(d,J＝9.0Hz,1H),8.05(d,J＝5.0Hz,1H),7.97(t,1H),7.92(s,1H),7.77(dd,J＝2.4,6.0Hz,1H),7.55(dd,J＝2.4,6.0Hz,12H),7.48(dd,J＝2.4,6.0Hz,1H),7.29(dd,J＝2.4,6.0Hz,1H),7.20(s,1H),6.84(s,1H),6.59(d,J＝5.0Hz,1H),6.43(s,2H),6.19(s,2H).MS(MM-ES+APCI)m/z:395.21[M+H]⁺.HPLC purity:95.44％.

Example 23

Substantially the same as in example 22 except that 2- (4-aminopyridine) -2-propanol was used instead of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)δ8.54(d,J＝5.0Hz,1H),8.28(d,J＝5.0Hz,1H),8.25(s,1H),8.12(d,J＝5.0Hz,1H),8.05(d,J＝10.0Hz,1H),7.62-7.58(m,2H),7.42(s,1H),7.23(m,2H),7.18-7.14(m,2H),6.87(d,J＝5.0Hz,1H),6.49(s,1H),6.20(s,2H),3.62(s,1H),1.39(s,6H).MS(MM-ES+APCI)m/z:437.25[M+H]⁺.HPLC purity:92.33％.

Example 24

Substantially the same as in example 22 except that 4-amino-2-methoxypyridine was used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)δ8.81(s,1H),8.50(d,J＝5.0Hz,1H),8.28(d,J＝5.0Hz,1H),7.82(d,J＝10.0Hz,1H),7.77(d,J＝5.0Hz,1H),7.48-7.42(m,2H),7.24-7.18(m,3H),7.07-7.05(m,2H),6.83(d,J＝5.0Hz,1H),6.75(dd,J＝2.4,6.0Hz,1H),6.56(s,1H),6.20(s,1H),3.83(s,3H).MS(MM-ES+APCI)m/z:431.17[M+H]⁺.HPLC purity:98.56％.

Example 25

Substantially the same as in example 22 except that 1-amino-2-methyl-2-propanol was used in place of 4, 6-diaminopyrimidine

¹H NMR(400MHz,CDCl3)δ8.57(d,J＝5.2Hz,1H),8.39(d,J＝8.0Hz,1H),7.90(d,J＝5.2Hz,1H),7.84-7.78(m,2H),7.32-7.27(m,4H),6.43(d,J＝5.2Hz,1H),6.08(s,2H),6.02(s,1H),4.80(s,1H),4.81(d,J＝6.0Hz,1H),1.16(s,6H).MS(MM-ES+APCI)m/z:396.2[M+Na]⁺.HPLC purity:98.86％.

Example 26

Substantially the same as in example 22 except that 3,4, 5-trimethoxyaniline was used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)δ8.54(d,J＝2.0Hz,1H),8.39(d,J＝5.0Hz,1H),8.09(d,J＝5.0Hz,1H),8.82-7.78(m,2H),7.32-7.27(m,4H),6.76(s,1H),6.63(d,J＝5.0Hz,1H),6.47(s,1H),6.42(s,2H),6.11(s,2H),3.77(s,3H),3.65(s,6H).MS(MM-ES+APCI)m/z:490.2[M+Na]⁺.HPLC purity:94.58％.

Example 27

Essentially the same as in example 22, except that 2-morpholin-4-aminopyridine was used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)δ8.56(d,J＝5.0Hz,1H),8.32(d,J＝5.0Hz,1H),8.22(d,J＝5.0Hz,1H),7.88(d,J＝5.0Hz,1H),7.78-7.77(m,2H),7.31-7.28(m,4H),6.81-6.78(m,2H),6.49(s,1H),6.43-6.42(dd,J＝2.4,5.0Hz,1H),6.16(s,2H),3.76(t,J＝5.0Hz,4H),3.36(t,J＝5.0Hz,4H).MS(MM-ES+APCI)m/z:487.2[M+Na]⁺.HPLC purity:99.79％.

Example 28

The synthesis route of the compound is as follows:

target compound synthesis step:

triethylamine (70.0mg,0.68mmol) and LEC-005-073(180.0mg,0.46mmol) were dissolved in 5ml of anhydrous tetrahydrofuran under nitrogen protection, and then the reaction solution was cooled to 0 ℃ and acetyl chloride (45.0mg,0.50mmol) was slowly added dropwise. Reacting for 5 hours at 0 ℃ after the dripping is finished; after the reaction is finished, quenching the reaction by using a small amount of saturated sodium bicarbonate solution; adding ethyl acetate to extract and stratify; drying ethyl acetate by anhydrous sodium sulfate, and then spin-drying; the residue was subjected to silica gel column chromatography to give the objective product (49.0mg, 50.1%).¹H NMR(500MHz,CDCl3)δ8.63(s,1H),8.55(d,J＝5.0Hz,1H),8.45(d,J＝8.0Hz,1H),8.20(d,J＝5.0Hz,1H),8.03(s,1H),7.85-7.80(m,2H),7.75(s,1H),7.34-7.29(m,4H),7.05(s,1H),6.81(d,J＝5.0Hz,1H),6.20(s,2H),2.32(s,6H).MS(MM-ES+APCI)m/z:501.2[M+Na]⁺.HPLC purity:99.24％.

Example 29

Essentially the same as in example 22, except that 4-aminopyridine-2-carboxylic acid methyl ester was used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,CDCl3)8.55(d,J＝5.0Hz,1H),8.48(d,J＝8.0Hz,1H),8.40(d,J＝5.0Hz,1H),8.31(d,J＝5.0Hz,1H),8.08(s,1H),7.88(d,J＝10.0Hz,1H),7.84(s,1H),7.82(s,1H),7.77(m,2H),7.62(d,J＝5.0Hz,1H),7.33-7.28(m,2H),6.85(t,J＝5.0Hz,1H),6.48(s,1H),6.20(s,2H),3.96(s,3H).MS(MM-ES+APCI)m/z:459.2[M+Na]⁺.HPLC purity:96.30％.

Example 30

Substantially the same as in example 21, except that thiabendazole was used instead of intermediate 11-a

¹H NMR(500MHz,DMSO-d6)9.48(s,1H),9.29(d,J＝5.0Hz,1H),8.58(d,J＝5.0Hz,1H),8.08(d,J＝5.0Hz,1H),7.95(s,1H),7.74(d,J＝10.0Hz,1H),7.53(d,J＝10.0Hz,1H),7.28(m,2H),7.22(s,1H),6.86(s,1H),6.58(d,J＝5.0Hz,1H),6.46(s,2H),6.10(s,2H),3.17(d,J＝5.0Hz,1H).MS(MM-ES+APCI)m/z:423.1[M+Na]⁺.HPLC purity:99.47％.

Example 31

Intermediate 31-a synthetic route:

intermediate 31-a synthesis:

triethylamine (606.0mg,0.6mmol) and 4, 6-diaminopyrimidine (440.0mg,4.0mmol) were dissolved in 5ml of anhydrous tetrahydrofuran under nitrogen protection, and then the reaction mixture was cooled to 0 ℃ and tetrahydropyran-4-carbonyl chloride (300.0mg,2.0mmol) was slowly added dropwise. Reacting for 5 hours at 0 ℃ after the dripping is finished; after the reaction is finished, quenching the reaction by using a small amount of saturated sodium bicarbonate solution; adding ethyl acetate, extracting, layering, drying ethyl acetate with anhydrous sodium sulfate, spin-drying, and separating with silica gel column chromatography to obtain intermediate 31-a224.6mg, 51.1%. MS (MM-ES + APCI) M/z 245.10[ M + Na [ ]]⁺.

The synthesis of the target compound was carried out according to the preparation scheme of example 11.

¹H NMR(500MHz,DMSO-d6)10.47(s,1H),10.01(s,1H),8.66(d,J＝5.0Hz,1H),8.41(d,J＝10.0Hz,1H),8.33(s,1H),8.28(s,1H),8.15(d,J＝5.0Hz,1H),8.01(t,J＝5.0Hz,1H),7.80(d,J＝5.0Hz,1H),7.59(s,1H),7.58-7.46(m,2H),7.31(m,2H),6.71(d,J＝5.0Hz,1H),6.25(s,2H),3.90(d,J＝5.0Hz,2H),3.31(m,2H),2.75(m,1H),1.67-1.62(m,4H).MS(MM-ES+APCI)m/z:529.17[M+Na]⁺.HPLC purity:98.52％.

Example 32

¹H NMR(500MHz,CDCl3)10.30(s,1H),8.33(d,J＝5.0Hz,1H),8.28(d,J＝2.0Hz,1H),8.19(d,J＝5.5Hz,1H),7.55(d,J＝2.0Hz,1H),7.34(d,J＝8.0Hz,1H),7.19-7.10(m,4H),6.94(d,J＝8.0Hz,1H),6.90-6.88(m,2H),6.54(s,1H),6.14(s,2H),3.80(s,3H),2.38(s,3H).

Example 33

Essentially the same as in example 32, except that 2- (4- (6-amino-2-methylpyrimidin-4-yl) piperazin-1-yl) ethanol was used instead of methyl 4-aminopyridine-2-carboxylate

¹H NMR(500MHz,CDCl3)8.25(d,J＝5.0Hz,1H),8.15(s,1H),7.88(d,J＝5.5Hz,1H),7.72(m,1H),7.25(m,3H),7.15(d,J＝5.5Hz,1H),6.94(m,1H),6.80(m,1H),6.65(s,1H),6.14(s,2H),3.83(m,6H),2.58(m,6H),2.46(m,3H),2.35(m,3H).

Example 34

Can be prepared according to the preparation process of example 44.

Substantially the same as in example 22 except that 3-aminobenzamide was used in place of 4, 6-diaminopyrimidine

¹H NMR(500MHz,DMSO-d6)12.77(s,1H),9.13(d,J＝5.0Hz,1H),8.66(d,J＝10.0Hz,1H),8.48(d,J＝10.0Hz,1H),8.19(d,J＝10.0Hz,1H),8.10(d,J＝5.0Hz,1H),8.01(d,J＝5.0Hz,1H),7.82(m,2H),7.50-7.21(m,6H),6.62(d,J＝2.0Hz,1H),6.35(s,1H),6.20(s,2H),3.18(s,1H).

Example 35

Substantially the same as in example 21, except that 2- (4-pyridyl) benzimidazole was used instead of intermediate 11-a

MS(MM-ES+APCI)m/z:395.21[M+H].

Example 36

Substantially the same as in example 22, except that 4-amino-6- (4-methyl-1-piperazinyl) pyrimidine is used in place of 4, 6-diaminopyrimidine

¹H NMR(400MHz,CDCl3)8.51(d,J＝8.0Hz,1H),8.46(d,J＝8.0.0Hz,1H),8.21(s,1H),8.13(d,J＝5.2Hz,1H),7.88-7.81(m,2H),7.53(m,1H),7.35-7.30(m,4H),7.05(s,1H),6.93(s,1H),6.63(d,J＝8.0Hz,1H),6.17(s,2H),3.60(d,J＝8.0Hz,4H),2.46(d,J＝8.0Hz,4H),2.33(s,3H).MS(MM-ES+APCI)m/z:478.2[M+H]⁺.

Example 37

Essentially the same as in example 22, except that 4-aminoquinoline is used in place of 4, 6-diaminopyrimidine

H NMR(400MHz,CDCl3)8.55(m,2H),8.34(d,J＝8.0Hz,1H),8.28(d,J＝5.2Hz,1H),8.00(d,J＝8.4Hz,1H),7.84-7.75(m,3H),7.64-7.59(m,3H),7.36(t,J＝8.0Hz,1H),7.31-7.27(m,4H),6.84(d,J＝5.2Hz,1H),6.81(s,1H),6.22(s,2H).MS(MM-ES+APCI)m/z:429.2[M+H]⁺.HPLC purity:97.95％.

Example 38

Essentially the same as example 21 except that 2- (1H-imidazol-2-yl) pyridine was used instead of intermediate 11-a

¹H NMR(500MHz,DMSO)δ9.53(s,1H),8.54–8.51(m,1H),8.11(dd,J＝12.2,6.6Hz,2H),8.01(s,1H),7.86(td,J＝7.8,1.8Hz,1H),7.44(d,J＝0.9Hz,1H),7.32(ddd,J＝7.5,4.9,1.1Hz,1H),7.24(s,1H),7.14(d,J＝0.9Hz,1H),6.89(s,1H),6.56(d,J＝5.2Hz,1H),6.48(s,2H),5.91(s,2H).MS(MM-ES+APCI)m/z:367.07[M+Na]⁺.HPLC purity:99.48％.

Example 39

¹H NMR(500MHz,CDCl₃)δ8.48(d,J＝4.1Hz,1H),8.29(s,1H),8.24(d,J＝8.1Hz,1H),8.19(d,J＝5.3Hz,1H),7.76(td,J＝7.8,1.8Hz,1H),7.52(s,1H),7.23(d,J＝0.9Hz,1H),7.20(ddd,J＝7.5,4.9,1.0Hz,1H),7.05(d,J＝0.9Hz,1H),7.02(s,1H),7.00(s,1H),6.65(d,J＝5.1Hz,1H),5.96(s,2H),3.69–3.61(m,4H),2.54–2.45(m,4H),2.36(s,3H).MS(MM-ES+APCI)m/z:450.16[M+Na]⁺.HPLC purity:98.94％.

Example 40

Essentially the same as example 21, except that 4,5,6, 7-tetrahydro-1H-benzimidazole was substituted for intermediate 11-a

¹H NMR(500MHz,DMSO)δ9.56(s,1H),8.13(d,J＝5.2Hz,1H),8.00(d,J＝0.6Hz,1H),7.52(s,1H),7.17(s,1H),6.89(s,1H),6.54(dd,J＝5.2,1.1Hz,1H),6.48(s,2H),5.05(s,2H),2.42(s,2H),2.31(s,2H),1.66(s,4H).MS(MM-ES+APCI)m/z:344.52[M+Na]⁺.HPLC purity:98.52％.

EXAMPLE 41

Substantially the same as in example 40, except that 4-amino-6- (4-methyl-1-piperazinyl) pyrimidine was used in place of 46-diaminopyrimidine

¹H NMR(500MHz,CDCl₃)δ8.28(s,1H),8.19(d,J＝5.2Hz,1H),8.10(s,1H),7.40(s,1H),7.03(s,1H),6.98(s,1H),6.51(d,J＝5.0Hz,1H),4.96(s,2H),2.60(s,3H),2.54–2.42(m,6H),2.32(m,6H).MS(MM-ES+APCI)m/z:405.16[M+H]⁺.HPLC purity:96.95％.

Example 42

Substantially the same as in example 21, except that 2-phenylbenzimidazole was used instead of intermediate 11-a

¹H NMR(500MHz,DMSO)δ9.53(s,1H),8.14(d,J＝5.3Hz,1H),7.96(s,1H),7.79–7.68(m,3H),7.57–7.50(m,3H),7.45(dd,J＝6.8,1.7Hz,1H),7.27(dq,J＝7.3,5.8Hz,2H),7.15(s,1H),6.90(s,1H),6.56(d,J＝5.3Hz,1H),6.50(s,2H),5.55(s,2H).MS(MM-ES+APCI)m/z:416.09[M+H]⁺.HPLC purity:98.40％.

Example 43

Substantially the same as in example 22 except that 2-methyl-4-aminopyridine was used instead of 4, 6-diaminopyrimidine

MS(MM-ES+APCI)m/z:393.21[M+H]⁺.HPLC purity:96.30％.

Example 44

Reaction scheme of the target compound:

target compound synthesis reaction:

under the protection of nitrogen, adding raw material LEC-005-96(109.1mg,0.25mmol) and 5ml of concentrated ammonia water into a sealed tube, slowly heating to 100 ℃ and reacting for 16 hours; after completion of the reaction, the solvent was removed by concentration, and the residue was purified by silica gel column chromatography to give a compound (43.0mg, 40.5%) as a white solid. MS (MM-ES + APCI) M/z 444.10[ M + Na [ ]]⁺.HPLC purity:96.30％.

Example 45

Essentially the same as in example 13, except that acetamide was used in place of 4, 6-diaminopyrimidine

MS(MM-ES+APCI)m/z:345.15[M+H]⁺.HPLC purity:83.95％.

Example 46

Intermediate 46-a reaction step:

under the protection of nitrogen, dissolving raw material LEC-005-96(109.1mg,0.25mmol) in a mixed solvent composed of tetrahydrofuran, methanol and water, adding lithium hydroxide (7.2mg,0.30mmol), slowly heating to 50 ℃ for reaction for 2 hours; after the reaction was completed, the pH was adjusted to 3 with dilute hydrochloric acid, the solvent was removed by concentration, and the residue was used as it was in the next reaction.

A target compound reaction step:

under the protection of nitrogen, intermediate 46-a is dissolved in DMF (5ml), HATU (142.6mg,0.375mmol) and DIPEA (64.7mg,0.5mmol) are added at room temperature, and the mixture is stirred for reaction for 1 hour; then 1- (2-hydroxyethyl) piperazine (50.0mg,0.375mmol) is added and the reaction is carried out for 8 hours at 30 ℃; after the reaction is finished, adding saturated salt solution and ethyl acetate into the reaction solution, extracting and layering, and extracting the ethyl acetate phase twice with the saturated salt solution; the ethyl acetate was removed by concentration, and the residue was purified by silica gel column chromatography to give the objective compound (43.0mg, 40.5%) as a white solid.

¹H NMR(500MHz,DMSO)δ9.04(s,1H),8.65(d,J＝4.0Hz,1H),8.43(d,J＝8.0Hz,1H),8.32(d,J＝5.5Hz,1H),8.25(s,1H),8.22(d,J＝5.5Hz,1H),8.02(td,J＝8.0,1.7Hz,1H),7.87(d,J＝4.5Hz,1H),7.82(dt,J＝5.5,3.0Hz,1H),7.56(dt,J＝7.5,3.0Hz,1H),7.50(ddd,J＝7.5,5.0,1.0Hz,1H),7.36–7.27(m,2H),6.83(d,J＝5.0Hz,1H),6.50(s,1H),6.23(s,2H),3.66–3.59(m,2H),3.26(t,J＝6.0Hz,2H),3.11–3.03(m,4H),2.82–2.74(m,5H).

Example 47

Substantially the same as in example 46, except that N, N-dimethylethylenediamine was used instead of 1- (2-hydroxyethyl) piperazine

MS(MM-ES+APCI)m/z:493.2[M+H]⁺.HPLC purity:99.86％.

Example 48

Substantially the same as in example 46, except that 1-methyl-4- (6-aminopyridin-3-yl) piperazine was used instead of 1- (2-hydroxyethyl) piperazine

MS(MM-ES+APCI)m/z:477.2[M+H]⁺.HPLC purity:98.42％.

Example 49

Substantially the same as in example 2, except that 3-chloro-4-fluorobenzyl chloride was used in place of 2-chloro-4- (chloromethyl) pyridine

MS(MM-ES+APCI)m/z:483.3[M-H]-.HPLC purity:95.46％.

Biological evaluation test:

test example 1: test of inhibitory Effect of Compounds of the present invention on TGF-. beta.R 1(ALK5) kinase Activity

The invention adopts the following experimental method to determine the inhibition effect of the compound on the kinase activity of TGF-beta R1(ALK 5):

by LanthaScreen^TMThe Eu Kinase Binding Assay method detects the inhibitory activity of a series of compounds on ALK5 Kinase, and the specific test flow is as follows: the compounds were prepared in 10mM stock solution in DMSO, diluted 1:3 gradient, and loaded in 384-PP (from LABCYTE, P-05525) for use; preparing 1X Kinase Buffer A to dilute the following reaction reagents; 15nL of compound was added to a reaction plate (from Perkin Elmer, model ProxiPlate-384Plus) using a micropipette system (from LABCYTE, model Echo 520); add 5. mu.L ALK5solution per well (available from Thermo Scientific Forma Co., PV 5837); adding 5 μ L Eu-Anti-GST Antibody solution (from Thermo Scientific Forma, PV5594) into each well, and shaking for 15 min; mu.L of Kinase Tracer 178 solution (available from Thermo Scientific Forma, PV5593) was added to each well and mixed by shaking. The total reaction system contained 50mM Tris (pH 7.5),150mM NaCl,0.5mM EDTA, 0.02%

X-100, 2mM DTT, 50% Glycerol,50mM HEPES (pH 7.5),10mM MgCl2,1mM EGTA, 0.01% Brij-35,5nM ALK5,2nM Eu-Anti-GST Antibody,20nM Kinase Tracer 178, 0.1% DMSO. The reaction plate was incubated at 30 ℃ for 60 min. The test was performed using a multi-label microplate tester (available from PE corporation, model Envision multilabel reader). Using GraphData analysis was performed on Pad Prism 5 Demo software. Calculation of related Compound IC Using a non-Linear regression model₅₀Values, repeated three times, averaged.

Compound IC determined as described above₅₀The values are given in table 1 below.

TABLE 1 IC inhibition of ALK5 kinase Activity by Compounds of the invention₅₀Value of

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A nitrogen-containing heterocyclic organic compound having a structure represented by general formula (I):

wherein, Y₁Is CH₂；X₁And X₃Is CH, X₄Is N, X₂Is CH;

a is selected from any one of the following structures:

Z₂～Z₄each independently is CH; and Z is₁Is CH or N;

R₁is selected from

And said

Optionally further coated with C_1-4Alkyl or halogen substitution;

R₂selected from H or the following groups:

R₃is H;

R₄selected from: c_1-6Alkyl, 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered aryl or 5-6 membered heteroaryl; said C is_1-6Alkyl, 3-8 membered cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered aryl, 5-6 membered heteroaryl optionally further substituted with: c₁-₆Alkyl, hydroxy, halogen substituted C_1-4Alkyl or halogen;

Y₂is NH;

b is selected from: pyridyl or pyrimidinyl;

l is selected from: H. c_1-4Alkyl, hydroxy substituted C_1-4Alkyl radical, C_1-4Alkoxy, -N (R)₂₂)₂、-NH-Ac、-COR₂₂、-COOR₂₂、-CONH₂Or the following groups

R₂₂Is H or C_1-4An alkyl group;

R₂₃is H, C_1-4Alkyl or

p and q are each independently an integer from 1 to 5;

m is 1,2 or 3.

2. The nitrogen-containing heterocyclic organic compound according to claim 1, wherein R is₄Each independently selected from C_1-4Alkyl or the following group:

and the above groups are optionally further substituted by C_1-4Alkyl or halogen substitution.

3. The nitrogen-containing heterocyclic organic compound according to claim 1, wherein hydroxyl group in L is substituted for C_1-4Alkyl is

4. The nitrogen-containing heterocyclic organic compound according to any one of claims 1 to 3, which has a structure represented by any one of the following general formulae (II) to (III):

wherein Z₉Is N or-CH.

5. The nitrogen-containing heterocyclic organic compound according to claim 4, wherein the nitrogen-containing heterocyclic organic compound isSelected from the structures shown in formula (II), wherein R₁Is composed of

R₂Is composed of

R₃Is H, L is H, -NH₂Or

6. The nitrogen-containing heterocyclic organic compound according to claim 4, which is selected from the group consisting of the structures represented by the formula (III) wherein L is H, C_1-4Alkyl radical, C_1-4Alkoxy, -COOR₂₂、-CONH₂、

R₂₂Is H or C_1-4An alkyl group.

7. A nitrogen-containing heterocyclic organic compound selected from the group consisting of:

8. the method for producing a nitrogen-containing heterocyclic organic compound according to claim 4, characterized by comprising the steps of:

m represents halogen;

providing a compound shown as a formula (II-1);

reacting the compounds shown in the formulas (II-1), (II-2) and (II-3) to obtain a compound shown in a formula (II-4);

M represents halogen;

providing a compound represented by the formula (III-1);

reacting the compound represented by the formula (III-4) with the compound represented by the formula (III-5) to obtain the compound represented by the formula (III).

9. A composition comprising the nitrogen-containing heterocyclic organic compound of any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

10. Use of a nitrogen-containing heterocyclic organic compound according to any one of claims 1 to 7 or a composition according to claim 9 for the manufacture of a medicament for the treatment or prevention of a TGF- β signalling pathway mediated disease.

11. The use according to claim 10, wherein the medicament is an ALK5 kinase inhibitor.