CN114276328A

CN114276328A - Compound as small molecule immunosuppressant, preparation method and application thereof

Info

Publication number: CN114276328A
Application number: CN202011041992.4A
Authority: CN
Inventors: 孙效华; 尹大伟; 杨阳; 刘洋; 陈庆广; 王静; 王静然; 杨红振; 胡怀忠
Original assignee: Beijing Konruns Pharmaceutical Co Ltd
Current assignee: Beijing Konruns Pharmaceutical Co Ltd
Priority date: 2020-09-28
Filing date: 2020-09-28
Publication date: 2022-04-05
Anticipated expiration: 2040-09-28
Also published as: CN114276328B

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a compound serving as a small molecular immunosuppressant, and a preparation method and application thereof. The invention specifically discloses a compound shown as a formula I or pharmaceutically acceptable salt, prodrug, metabolite, isotope derivative and solvate thereof. The compound of the invention is a compound which acts on a PD-1/PD-L1 signal channel and is relevant to the treatment of tumor diseases and other immune diseases and contains a pyrimidine ring. The compound of the invention can improve the drug effect and reduce the side effect of the drug. The invention also provides a preparation method of the compound and application of the compound in treating diseases related to PD-1/PD-L1 signal pathways.

Description

Compound as small molecule immunosuppressant, preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a compound for treating diseases related to a PD-1/PD-L1 signal pathway. The invention also relates to a preparation method of the compound and application of the compound in treating cancer or immune diseases.

Background

Programmed Cell Death receptor 1 (PD-1), an immune co-suppressor molecule, a type I transmembrane Protein consisting of 268 amino acids, which is expressed specifically only on T cells. In normal organisms, PD-1 plays an important role in maintaining immune tolerance of the organism as a negative regulatory molecule of T cell proliferation. Programmed death receptor 1 ligand (PD-L1) is also a type I transmembrane protein, and is highly selectively expressed in tumor cells. When the T cell receptor is activated, PD-1 is combined with PD-L1 and is highly expressed in tumor infiltrating lymphocytes at an effector stage, and the activity of the T lymphocytes is inhibited. When a tumor develops, the PD-1/PD-L1 signal channel can inhibit the immune response of T cells so as to promote the development of tumor immune escape. Research shows that blocking the PD-1/PD-L1 signal channel can enhance the effect T lymphocyte to reach tumor part, reduce the inhibition of tumor part regulating T lymphocyte and enhance the endogenous tumor immunity effect, so that PD-1/PD-L1 becomes the important medicine target for tumor immunotherapy.

A plurality of monoclonal antibody PD-1/PD-L1 inhibitors are on the market, but small molecule inhibitors are not on the market at present. Compared with monoclonal antibody drugs, small molecule drugs have the advantages of flexible administration mode (oral administration), short half-life, stable property, high exposure, low price, no side effect related to immune rejection and the like, so the development of small molecule immunosuppressants is imminent.

Disclosure of Invention

The invention provides a novel pyrimidine ring-containing compound serving as a PD-1/PD-L1 inhibitor, which has a structure shown in a formula I. The compounds of the present invention, and pharmaceutically acceptable salts, prodrugs, isotopic derivatives and solvates thereof, and pharmaceutical compositions comprising the same, are useful for treating neoplastic diseases associated with the inhibition of the PD-1/PD-L1 signaling pathway.

The compounds of the invention have the structure shown in formula I:

wherein X is selected from Br, Cl and F;

y is selected from methyl, ethyl, isopropyl, cyclopropyl, Cl, F and H;

R₁and R₂Independently selected from H, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstitutedAryl, substituted or unsubstituted heterocyclic aryl, the substituents being selected from C1-C6 alkyl, C1-C6 alkoxy;

R₁and R₂Optionally taken together with the nitrogen atom to which they are attached to form a substituted or unsubstituted 4-7 membered heterocyclyl;

n＝1、2、3；

R₃selected from hydrogen, C1-C6 alkyl, substituted or unsubstituted 6-to 10-membered aryl, substituted or unsubstituted 6-to 10-membered cycloalkyl, substituted or unsubstituted 6-to 10-membered heterocycloalkyl, substituted or unsubstituted 5-to 7-membered heterocycloaryl;

R₄and R₅Independently selected from hydrogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted 3-6 membered cycloalkyl, and amido;

R₄and R₅Optionally taken together with the nitrogen atom to which they are attached to form a substituted or unsubstituted 4-7 membered heterocyclyl;

R₁、R₂、R₁and R₂Together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic ring, R₄、R₅、R₄And R₅A 4-7 membered heterocyclic ring which is formed by the heterocyclic ring and the connected nitrogen atom, wherein the substituent of the formed 4-7 membered heterocyclic ring is at least one of 1-4C 1-C6 alkyl groups, 3-6 membered cycloalkyl groups, 3-6 membered heterocycloalkyl groups, hydroxyl, carboxyl, amino, nitrile groups, halogen, trifluoromethyl and nitro; the 3-6 membered heterocycloalkyl or heteroaryl group contains 1-3 heteroatoms selected from N, O, S.

In the compound, a biphenyl substituent in the molecular structure of the compound can increase the conjugated system of the whole molecule, has a certain steric hindrance effect, and can increase the directionality of the molecule, so that the compound is better embedded with a dimer formed by two PD-L1. Meanwhile, the substituent extending out of the structure on the left side of the molecule can increase the solubility and the membrane permeability of the whole molecule, thereby increasing the druggability of the molecule. When the compound is embedded with a PD-L1 dimer, PD-L1 can not be combined with PD-1 any more, so that PD-L1 can be identified and killed by T cells, and the high expression of PD-L1 in tumor cells is inhibited, thereby achieving the purpose of treating tumor diseases.

More specifically, in the compounds of the present invention, Y is selected from the group consisting of methyl, ethyl, isopropyl, cyclopropyl, Cl, F, H, which are smaller in volume or shorter in chain length. Mainly in view of the research process, the inventors found that when Y is selected to be a larger or longer group, the activity decreases significantly. The main reason is that the activity of the compounds of the present invention on PD-L1 is related to Y and X in the molecule. The angle formed by the two benzene rings of the biphenyl group is related to the groups X and Y. When two benzene rings of a biphenyl group in the compound form an included angle of less than 45 degrees, the compound has high activity, and when Y is too large or too long, the included angle exceeds 45 degrees, and the directionality is reduced, so that the activity of the compound is reduced.

In the compounds of the formula I above, n preferably has a value of not more than 3. The carbon chain of n is not suitable for being too long or too short, and the value of n is preferably 3. The presence of this side chain is on the one hand related to the physicochemical properties (e.g. solubility, lipid solubility, etc.). When n is less than or equal to 3, the compound can be made to have good physicochemical properties. When N is greater than 3, the side chain is too long, resulting in a decrease in solubility and lipid solubility of the compound. On the other hand, the presence of this side chain is related to the ability of the compound to bind to the target. If the carbon chain is too long, the compound will not be in effective contact with the target.

In the compounds of the invention of the formula I, R₁A 4-to 7-membered heterocyclic ring which may be taken alone or together with the nitrogen atom to which it is attached, R₂Or a 4-to 7-membered heterocyclic ring which may be independently combined with the nitrogen atom to which it is attached. Further, R₁And R₂And a 4-7 membered heterocyclic ring which may be simultaneously formed with the nitrogen atom to which it is attached.

Furthermore, R₁And R₂The volume of (A) should not be too large. R₁And R₂Too large a volume or too long a chain results in an included angle of more than 45 degrees, resulting in a decrease in the activity of the compound.

Similarly, R₄、R₅、R₄And R₅Together with the nitrogen atom to which they are attached form a 4-to 7-membered heterocyclic ring R₄、R₅、R₄And R₅To the nitrogen attachedThe atoms together form a 4-7 membered heterocyclic ring.

In the compounds of the invention of the formula I, the radical R₁、R₂、R₄、R₅The cyclization can restrict the conformation of the molecule, and moderately increases the rigidity, which is beneficial to increasing the directionality of the molecule. The chain substituent has higher activity, good flexibility and stronger fat solubility, but the directionality is reduced, and meanwhile, the metabolic sites are increased. R is to be₁、R₂、R₄、R₅Together with the other groups in formula I, different compound molecules can be designed specifically.

The compounds according to the invention or pharmaceutically acceptable salts, prodrugs, metabolites, isotopic derivatives, solvates thereof,

wherein R is₁And R₂Each preferably selected from methyl, ethyl, isopropyl, cyclopropyl, phenyl, pyridyl;

or, R₁And R₂A 4-7 membered heterocyclic group taken together with the nitrogen atom to which it is attached, wherein R₁And R₂The 4-7 membered heterocyclic group, which is formed together with the attached nitrogen atom, is preferably selected from the following groups:

R₃preferably one selected from hydrogen, pyridyl and nitrile substituted phenyl.

In the above-mentioned 4-to 7-membered heterocyclic group, with respect to 4-hydroxypiperidine, the polarity as well as the solubility of the compound molecule are increased due to the presence of a hydroxyl group.

For the inclusion of R as described herein₁And R₂The group compound, the small molecule compound and two molecules of PD-L1 form dimerization, the small molecule compound is embedded in the dimer, so that the compound is more firmly embedded with PD-L1, the activity of the compound is improved, and simultaneously, R₁And R₂Forming chain or ring substituents, especially in R₁Or R₂With shorter chain lengths or with formation of nonaromatic cyclic groupsAnd the ability of forming hydrogen bonds with amino acids on a target is better, so that the activity of the compound is improved, and the druggability of molecules is improved.

wherein R is₄And R₅Identical or different, R₄And R₅Together with the nitrogen atom to which they are attached form a substituted or unsubstituted 4-7 membered heterocyclic group; wherein R is₄And R₅Together with the nitrogen atom to which they are attached form a substituted or unsubstituted 4-7 membered heterocyclyl group preferably selected from 1-piperidinyl, 1-pyrrolidinyl, 4-methyl-1-piperidinyl, 1-cyclobutylamino, 1-cyclopentylamino.

Preferably, R₄And R₅Independently selected from amide groups, wherein the amide group is selected from-R₆-C(＝O)-NH-R₇or-R₆-NH-C(＝O)R₇，R₆And R₇Each independently is a C1-C5 alkyl group.

R₄Or R₅Preferably with a shorter chain length or with cyclic groups. These groups have a good ability to form hydrogen bonds with amino acids on the target, which contributes to the activity of the compounds.

The compound of the invention is selected from any one of the following compounds:

(S) -1- ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) piperidine-2-carboxylic acid

((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-serine

(2S,4R) -1- ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid

((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-alanine

(S) -1- ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinyloxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) piperidine-2-carboxylic acid

((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-serine

(2S,4R) -1- ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid

((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-alanine

((2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-bromo-3 ' - (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-3 ' - (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-fluoro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-fluoro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-fluoro-3 ' - (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (3-piperidin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (3-piperidin-1-yl) ethoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-3 '- (3-piperidinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2,2' -dichloro-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy-4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-3 ' - (3- (dimethylamino) propoxy) -2' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy-4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -ethyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -isopropyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -cyclopropyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4- ((3-cyanobenzyl) oxy) -6-methoxypyrimidin-5-yl) methyl) -L-proline

((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4-methoxy-6- (pyridin-3-ylmethoxy) pyrimidin-5-yl) methyl) -L-proline

(2S,4R) -1- ((2- ((2-bromo- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid

((2- ((2-chloro-2 ' -butyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

The present invention also provides a process for the preparation of a compound according to formula I, wherein the process comprises the following reaction scheme:

the method comprises the following steps:

(1) dissolving SM1 and SM2 in a solvent 1, adding an alkaline reagent, and reacting at the temperature ranging from room temperature to 70 ℃ to obtain a compound C01;

(2) dissolving a compound C01 in a solvent 2, adding an alkaline reagent and a palladium metal catalyst, and reacting at the temperature of 70-120 ℃ to obtain a compound C02;

(3) dissolving compounds C02 and SM3 in a solvent 3, adding an alkaline reagent and a palladium metal catalyst, and reacting at the temperature of 70-120 ℃ to obtain a compound C03;

(4) dissolving C03 and SM4 in solvent 4, and carrying out reductive amination reaction to obtain the compound of formula I.

Preferably, in step (1), the solvent 1 is selected from one or more of tetrahydrofuran, DMSO, DMF, acetonitrile; the alkaline reagent is selected from one or more of cesium carbonate, potassium carbonate, sodium carbonate, potassium tert-butoxide and sodium tert-butoxide;

preferably, in step (2), the solvent 2 is selected from one or more of 1, 4-dioxane, toluene and benzene; the alkaline reagent is selected from one or more of potassium acetate, cesium carbonate, potassium carbonate and sodium carbonate; the palladium metal catalyst is selected from Pd (PPh)₃)₄、Pd(dppf)Cl₂Palladium acetate, Pd₂(dba)₃One or more of;

preferably, in step (3), the solvent 3 is selected from one or more of 1, 4-dioxane, toluene and water; the alkaline reagent is selected from one or more of cesium carbonate, potassium carbonate, sodium carbonate, potassium tert-butoxide and sodium tert-butoxide; the palladium metal catalyst is selected from Pd (PPh)₃)₄、Pd(dppf)Cl₂Palladium acetate, Pd₂(dba)₃One or more of;

preferably, in step (4), the solvent 4 is selected from one or more of tetrahydrofuran, DMSO, DMF, acetonitrile, methanol, dichloromethane; the reducing agent of the reductive amination reaction is selected from NaBH₃CN, sodium triacetoxyborohydride and/or sodium borohydride.

The invention also provides a pharmaceutical composition comprising a compound or a pharmaceutically salt of a compound or a solvate of a compound of the invention, and a pharmaceutically acceptable carrier.

In order to adapt to different administration modes, the pharmaceutical composition of the invention can be prepared into various dosage forms. Preferably, the pharmaceutical composition of the present invention is formulated in the form of an oral preparation or an injection.

The compound is a PD-1/PD-L1 signal channel inhibitor, and can be applied to treating diseases related to the PD-1/PD-L1 signal channel after being prepared into various preparations, wherein the diseases can be malignant tumors or cancers and can also be immunological diseases.

Optionally, the cancer is selected from skin cancer, lung cancer, hematological tumors, breast cancer, glioma, digestive system tumors, reproductive system tumors, lymphoma, nervous system tumors, head and neck cancer; the immune diseases are selected from diabetes, myasthenia gravis, rheumatoid arthritis, systemic lupus erythematosus and dermatitis.

Accordingly, the present invention also provides a method of treating cancer or immune disease, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition of the present invention.

Within the meaning of the present invention, the terms used are defined as follows:

"halogen" means F, Cl, Br, I.

"C1-C6 alkyl" refers to an alkyl chain having 1-6 carbon atoms and may be straight or branched. For example: methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and the like.

"C1-C8 alkoxy" refers to an alkyl chain of 1-8 carbon atoms which may be straight or branched, wherein at least one carbon atom in the middle is replaced by an oxygen atom.

"6-10 membered aryl" means an aryl group having 6-10 carbon atoms, such as phenyl, naphthyl. The hydrogen atoms on the carbon atoms in the ring may be substituted with specified substituents.

"3-6 membered cycloalkyl" and "6-10 membered cycloalkyl" refer to saturated carbocyclic rings having 3-6 and 6-10 carbon atoms, respectively, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like, wherein hydrogen atoms on carbon atoms in the ring may be substituted by specified substituents.

"6-10 membered heterocycloaryl" means an aryl group having 6-10 atoms, wherein 1-3 carbon atoms in the ring may be substituted with N, O, S. Such as quinolinyl, isoquinolinyl, pyridinyl, benzofuranyl, and the like.

"4-7 membered heterocyclic ring" means a heterocyclic ring having 4 to 7 atoms, such as a tetrahydropyrrole ring, a piperidine ring, a piperazine ring, and the like.

"3-6 membered heterocycloalkyl" means a carbocyclic ring of 3-6 carbon atoms containing at least one N, O, S heteroatom from the N, O, S heteroatoms forming a heterocyclic group.

The "prodrug" refers to a derivative which is converted into the compound of the present invention in vivo by an oxidation, reduction, hydrolysis or the like reaction catalyzed by an enzyme under physiological conditions.

"metabolite" refers to all molecules produced in a cell or organism (preferably a human) that are derived from a compound of the invention.

By "isotopic derivative" is meant a compound of the invention which has a structure comprising one or more isotopic atoms present in unnatural proportions. Such as deuterium (2H or D), carbon-13 (13C), nitrogen-15 (15N).

By "solvate" is meant a compound of the invention that forms a solvent complex with a solvent molecule by physical association. The physical bond comprises a hydrogen bond. Conventional solvents include water, methanol, ethanol, acetic acid, tetrahydrofuran, ethyl acetate, acetonitrile, and the like. The compounds of formula I may be prepared in crystalline form and may be in the form of solvates (including hydrates).

Pharmaceutically acceptable salts of the compounds of formula I, containing one or more basic or acidic groups, especially pharmaceutically usable salts thereof. Such as alkali metal salts, alkaline earth metal salts, ammonium salts. More precisely sodium, potassium, calcium, magnesium or organic amines such as ethylamine, ethanolamine, triethylamine or amino acid salts. The compounds of the present invention may form protonated compounds of formula I with inorganic or organic acids, examples of which include hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, methanesulfonic acid, lactic acid, malic acid, maleic acid, tartaric acid, and the like, as well as other acids known to those skilled in the art.

"pharmaceutical compositions" when used as a medicament means the compounds of formula I of the present invention and their salts, isotopic derivatives, metabolites, prodrugs, solvates and other compositions with or without biologically active substances, which are useful for the treatment or prevention of diseases associated with the PD1/PDL-1 signaling pathway, such as solid tumors, hematological tumors, and the like.

Drawings

FIG. 1 shows the general structure of the compound of formula I according to the present invention.

Detailed Description

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

Example 1

The structural formula of the compound:

the synthetic route is as follows:

the synthesis method comprises the following steps:

preparation of 4- (3- (3-bromo-2-methylphenoxy) propyl) morpholine:

to a reaction flask were added 3-bromo-2-methylphenol (1.61g,1eq), 4- (3-chloropropyl) morpholine (1.55g,1.1eq), acetonitrile (40mL), potassium carbonate (3.56g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (150mL) is added to quench the system, ethyl acetate (60mL x 3) is used for extraction, organic phases are combined and washed once by saturated salt solution, dried by anhydrous sodium sulfate, filtered by suction, concentrated, and crude silica gel column chromatography is carried out to obtain 4- (3- (3-bromo-2-methylphenoxy) propyl) morpholine (2.06g, yield: 76%). MS [ +1] + ] 314.1.

Preparation of 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine:

to a reaction flask was added 4- (3- (3-bromo-2-methylphenoxy) propyl) morpholine (2.06g,1eq), pinacol diboron (3.33g,2eq), potassium acetate (1.29g,2eq), pd (dppf) Cl₂(0.24g,0.05eq) and 1, 4-dioxane (50 mL). The reaction was stopped by heating to 90 ℃ under nitrogen, stirring for two hours and LCMS to characterize the reaction complete, adding water (350mL) to quench the system, extracting with ethyl acetate (100mL x 3), combining the organic phases and washing with saturated brine once, drying over anhydrous sodium sulfate, suction filtering, concentrating, and chromatography on crude silica gel column to give 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) morpholine (1.28g, yield: 54%). MS: [ M +1]+:362.2.

Preparation of methyl 2-bromo-3-iodobenzoate:

to the reaction flask was added methyl 3-amino-2-bromobenzoate (25.1g,1eq), acetonitrile (250mL) and water (400mL), and concentrated hydrochloric acid (40mL) was added to the above system with stirring. After stirring at room temperature for 10min, the temperature was reduced to 0 ℃ and a solution of sodium nitrite (15.1g,2eq) in water (200mL) was added dropwise with stirring. After stirring at 0 ℃ for 30min, an aqueous potassium iodide solution (21.8g,1.2eq,200mL of water) was added dropwise to the system, and the mixture was stirred at room temperature for 3 hours. LCMS characterization reaction complete. Extraction with ethyl acetate (300 mL. times.3), merging of the organic phases and washing with saturated brine once, drying over anhydrous sodium sulfate, suction filtration, concentration, and crude silica gel column chromatography gave methyl 2-bromo-3-iodobenzoate (34g, yield: 92%).

Preparation of methyl-2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-carboxylate:

adding into a reaction flask4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine (2.56g,1.2eq), methyl 2-bromo-3-iodobenzoate (2.01g,1eq), potassium carbonate (2.44g,3eq), Pd (dppf) Cl₂(0.22g,0.05eq), toluene (100mL), water (10mL) and methanol (10 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) representation of complete reaction, stopping the reaction, adding water (700mL) to quench the system, extracting with ethyl acetate (200mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain methyl-2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl ] crude product]-3-formic ester (0.93g, yield: 36%). MS: [ M +1]+:448.1/450.1.

Preparation of (2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl ] -3-yl) methanol:

adding methyl-2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl into a reaction bottle]-3-formic ester (0.93g,1eq), freshly distilled tetrahydrofuran (50mL) and LiAlH added portionwise at 5 ℃₄(0.16g,2eq), stirring for 30min at room temperature, TLC (thin layer chromatography) for complete reaction, slowly dropping the reaction system into saturated NH4Cl aqueous solution for quenching reaction, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, filtering, and concentrating to obtain 2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl]-3-yl) methanol (0.74g, yield: 85%) crude product. The crude product was used directly in the next reaction without any purification treatment. MS: [ M +1]+:420.1/422.1.

Preparation of 2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethylpyrimidine-5-carbaldehyde:

to the reaction flask was added 2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl ] -3-yl) methanol (0.74, 1eq), 2-chloro-4, 6-dimethoxypyrimidine (0.54g,1.5eq), acetonitrile (50mL), potassium carbonate (0.73g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS indicated reaction was complete, reaction was stopped, the system was quenched with water (400mL), extracted with ethyl acetate (150mL x 3), the organic phases were combined and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered with suction, concentrated, and chromatographed on crude silica gel to give 2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethylpyrimidine-5-carbaldehyde (0.76g, yield: 74%). MS (M + 1) +:586.2/588.2

Preparation of (S) -1- ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) piperidine-2-carboxylic acid:

2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethylpyrimidine-5-carbaldehyde (86.8mg,1eq) and L-2-pipecolic acid (57.3mg,3eq) were dissolved in a mixed solvent of DMF (5mL) and methanol (2mL), and 1 drop of acetic acid was added thereto with stirring to react at room temperature for 30 minutes, and then sodium cyanoborohydride (27.9mg, 3eq) was added to the system. Stirring overnight at room temperature, LCMS to characterize the reaction was complete, stop the reaction, add water (50mL) to quench the system, extract with ethyl acetate (20mL x 3), combine the organic phases and wash with saturated brine once, dry over anhydrous sodium sulfate, pump filter, concentrate, and purify the crude product by preparative liquid phase to give (S) -1- ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) piperidine-2-carboxylic acid (29.0mg, yield: 28%). MS [ +1] + ] 699.2/701.2.

1H NMR(400MHz,d₆-DMSO)δ7.58(m,1H),7.47(t,J＝7.6Hz,1H),7.27–7.18(m,2H),6.99(d,J＝8.0Hz,1H),6.70(d,J＝7.2Hz,1H),5.59–5.46,2H),4.18–3.99(m,2H),3.85(s,6H),3.69–3.62(m,2H),3.61–3.48(m,5H),2.96–2.82(m,2H),2.50–2.43(m,2H),2.40–2.30(m,4H),2.20–2.10(m,1H),1.96–1.80(m,5H),1.68–1.60(m,2H),1.49–1.34(m,3H)。

Example 2

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

referring to the synthetic method for reductive amination in example 1, 2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]Reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde and L-proline to synthesize (2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:685.2/687.2.

¹H NMR(400MHz,d₆-DMSO)δ7.61(m,1H),7.47(t,J＝7.6Hz,1H),7.26–7.14(m,2H),6.98(d,J＝8.0Hz,1H),6.68(d,J＝7.4Hz,1H),5.56–5.42(m,2H),4.12–4.00(m,2H),3.94–3.75(m,8H),3.60–3.54(m,4H),3.25–3.07(m,4H),2.49–2.43(m,2H),2.42–2.30(m,4H),2.09–2.00(m,1H),1.95–1.87(m,2H),1.87(s,3H),1.83–1.75(m,1H),1.74–1.55(m,1H)。

Example 3

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-serine:

referring to the synthetic method of reductive amination in example 1, 2- ((2-Bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl]Synthesis of ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-serine]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-serine. MS: [ M +1]⁺:631.2.

¹H NMR(400MHz,d₆-DMSO)δ7.61(m,1H),7.48(t,J＝7.6Hz,1H),7.28–7.17(m,2H),6.99(d,J＝8.2Hz,1H),6.71(d,J＝7.4Hz,1H),5.61–5.43(m,2H),4.18–3.99(m,2H),3.85(s,6H),3.75–3.61(m,2H),3.69–3.62(m,2H),3.61–3.48(m,4H),2.94–2.82(m,1H),2.51–2.43(m,2H),2.42–2.31(m,4H),1.96–1.80(m,5H)。

Example 4

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of (2S,4R) -1- ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid:

referring to the synthetic method for reductive amination in example 1, 2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]Synthesis of (2S,4R) -1- ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde and (2S,4R) -4-hydroxypyrrole-2-carboxylic acid]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid. MS: [ M +1]⁺:657.3。

¹H NMR(400MHz,d₆-DMSO)δ7.56(d,J＝6.6Hz,1H),7.47(t,J＝7.6Hz,1H),7.26–7.14(m,2H),6.98(d,J＝8.2Hz,1H),6.68(d,J＝7.6Hz,1H),5.59–5.37(m,2H),4.20–3.99(m,4H),3.88(s,6H),3.73(s,2H),3.61–3.50(m,4H),3.27–3.15(m,2H),2.47–2.42(m,2H),2.39–2.30(m,4H),2.02–1.77(m,7H)。

Example 5

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-alanine:

referring to the synthetic method for reductive amination in example 1, 2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]Synthesis of ((2- ((2-bromo-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-alanine]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-alanine. MS: [ M +1]⁺:614.2.

¹H NMR(400MHz,d₆-DMSO)δ7.59(m,1H),7.49(t,J＝7.6Hz,1H),7.29–7.17(m,2H),7.01(d,J＝8.2Hz,1H),6.72(d,J＝7.4Hz,1H),5.60–5.43(m,2H),4.17–3.98(m,2H),3.86(s,6H),3.69–3.62(m,2H),3.61–3.34(m,5H),2.51–2.43(m,2H),2.42–2.31(m,4H),1.94–1.82(m,5H),1.24(d,J＝6.6HZ,3H)。

Example 6

The structural formula of the compound:

the synthetic route is as follows:

the synthesis method comprises the following steps:

preparation of 2-chloro-4, 6-dimethoxypyrimidine-5-carbaldehyde:

under the condition of ice-water bath, phosphorus oxychloride (229.2g,15eq) is dropwise added into a reaction bottle filled with DMF (109.2g,15eq), the mixture is stirred for 30min under the condition of ice-water bath, a DMF (200mL) solution of 2-chloro-4, 6-dimethoxypyrimidine (17.4g,1eq) is dropwise added into a reaction system, the temperature is raised to 50 ℃, and the mixture is stirred and reacted for two hours. LCMS indicated reaction was complete and stopped. While stirring, the reaction system was dropped into an ice-water bath, adjusted to neutral with sodium bicarbonate (pH 7), the solid was filtered with suction, washed with water twice, and the dried crude product was purified by silica gel column chromatography to obtain a product (11.4g, yield 56%) of 2-chloro-4, 6-dimethoxypyrimidine-5-carbaldehyde. MS: [ M +1]⁺:203.1.

Preparation of 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 3-bromo-2-chlorobenzyl alcohol (2.6g,1eq), 2-chloro-4, 6-dimethoxypyrimidine (3.6g,1.5eq), acetonitrile (100mL), potassium carbonate (4.8g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (700mL) is added to quench the system, ethyl acetate (200mL x 3) is used for extraction, organic phases are combined and washed once by saturated salt solution, anhydrous sodium sulfate is used for drying, suction filtration and concentration are carried out, and crude silica gel column chromatography is carried out to obtain 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-formaldehyde (3.4g, yield: 75%). MS [ +1] + ] 389.0.

Preparation of 2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine (1.28g,1.1eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (1.24g,1eq), potassium carbonate (1.32g,3eq), pd (dppf) Cl₂(0.12g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (350mL) to quench a system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl) crude product]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.78g, yield: 45%). MS: [ M +1]+:542.2.

Preparation of (S) -1- ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) piperidine-2-carboxylic acid:

2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (80.2mg,1eq) and L-2-pipecolic acid (57.3mg,3eq) were dissolved in a mixed solvent of DMF (5mL) and methanol (2mL), and 1 drop of acetic acid was added thereto with stirring to react at room temperature for 30 minutes, and then sodium cyanoborohydride (27.9mg, 3eq) was added to the system. Stirring overnight at room temperature, LCMS to characterize the reaction was complete, stop the reaction, add water (50mL) to quench the system, extract with ethyl acetate (20mL x 3), combine the organic phases and wash with saturated brine once, dry over anhydrous sodium sulfate, pump filter, concentrate, and purify the crude product by preparative liquid phase to give (S) -1- ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) piperidine-2-carboxylic acid (32mg, yield: 30%). MS [ M +1] +:655.2.

¹H NMR(400MHz,d₆-DMSO)δ7.60(dd,J＝7.6,1.3Hz,1H),7.43(t,J＝7.6Hz,1H),7.18-7.27(m,2H),6.99(d,J＝8.2Hz,1H),6.70(d,J＝7.4Hz,1H),5.59–5.46(m,2H),4.11–3.99(m,2H),3.85(s,6H),3.69–3.62(m,2H),3.61–3.48(m,5H),2.96–2.82(m,2H),2.50–2.43(m,2H),2.40–2.30(m,4H),2.20–2.10(m,1H),1.96–1.82(m,5H),1.68–1.60(m,2H),1.49–1.34(m,3H)。

Example 7

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

referring to the synthetic method for reductive amination in example 6, 2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]Reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde and L-proline to synthesize (2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:641.2.

¹H NMR(400MHz,d₆-DMSO)δ7.60(d,J＝7.2Hz,1H),7.44(t,J＝7.6Hz,1H),7.31–7.18(m,2H),6.99(d,J＝8.0Hz,1H),6.70(d,J＝7.2Hz,1H),5.52(s,2H),4.10–4.00(m,2H),3.95–3.78(m,8H),3.60–3.50(m,4H),3.20–3.05(m,4H),2.50–2.43(m,2H),2.40–2.30(m,4H),2.04–1.96(m,1H),1.97–1.75(m,6H),1.73–1.55(m,1H)。

Example 8

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-serine:

referring to the synthetic method for reductive amination in example 6, 2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]Synthesis of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-serine]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-serine. MS: [ M +1]⁺:631.2.

¹H NMR(400MHz,d₆-DMSO)δ7.60(dd,J＝7.6,1.3Hz,1H),7.44(t,J＝7.6Hz,1H),7.28–7.17(m,2H),6.99(d,J＝8.2Hz,1H),6.70(d,J＝7.4Hz,1H),5.59-5.47(m,2H),4.12–3.99(m,2H),3.85(s,6H),3.72–3.64(m,2H),3.61–3.47(m,2H),3.61–3.48(m,4H),2.94–2.82(m,1H),2.51–2.43(m,2H),2.42–2.31(m,4H),1.96–1.80(m,5H)。

Example 9

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of (2S,4R) -1- ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid:

referring to the synthetic method for reductive amination in example 6, 2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]Synthesis of (2S,4R) -1- ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinylpropoxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde and (2S,4R) -4-hydroxypyrrole-2-carboxylic acid]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid. MS: [ M +1]⁺:657.3.

¹H NMR(400MHz,d₆-DMSO)δ7.60(d,J＝7.6Hz,1H),7.43(t,J＝7.6Hz,1H),7.29–7.16(m,2H),6.98(d,J＝8.2Hz,1H),6.70(d,J＝7.6Hz,1H),5.55–5.45(m,2H),4.17–3.99(m,3H),3.87(s,6H),3.75–3.65(m,2H),3.60–3.51(m,4H),3.24–3.10(m,2H),2.46(t,J＝7.1Hz,2H),2.40–2.26(m,5H),1.97–1.70(m,7H)。

Example 10

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-alanine:

referring to the synthetic method for reductive amination in example 6, 2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehydeAnd L-alanine reaction to synthesize ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl group)]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-alanine. MS: [ M +1]⁺:614.2.

¹H NMR(400MHz,d₆-DMSO)δ7.60(d,J＝7.6Hz,1H),7.43(t,J＝7.6Hz,1H),7.29–7.17(m,2H),6.99(d,J＝8.2Hz,1H),6.70(d,J＝7.4Hz,1H),5.55–5.43(m,2H),4.17–3.98(m,2H),3.89(s,6H),3.69–3.62(m,2H),3.61–3.34(m,5H),2.51–2.43(m,2H),2.42–2.31(m,4H),1.94–1.82(m,5H),1.20(d,J＝6.6HZ,3H)。

Example 11

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of 1- (3- (3-bromo-2-methylphenoxy) propyl) -4-methylpiperazine:

to the reaction flask was added 3-bromo-2-methylphenol (2.82g,1eq), 1- (3-chloropropyloxy) -4-methylpiperazine (2.91g,1.1eq), acetonitrile (60mL), potassium carbonate (6.24g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (250mL) is added to quench the system, ethyl acetate (100mL x 3) is extracted, organic phases are combined and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered by suction, concentrated, and subjected to crude silica gel column chromatography to obtain 1- (3- (3-bromo-2-methylphenoxy) propyl) -4-methylpiperazine (3.69g, yield: 76%). MS [ M +1] +:327.1/329.1.

Preparation of 1-methyl-4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperazine:

to a reaction flask was added 1- (3- (3-bromo-2-methylphenoxy) propyl) -4-methylpiperazine (3.69g,1eq), pinacol diboride (5.73g,2eq), potassium acetate (2.22g,2eq), pd (dppf) Cl₂(0.42g,0.05eq) and 1, 4-dioxane (150 mL). Heating to 90 ℃ under the protection of nitrogen, stirring the reaction for two hours, LCMS (liquid chromatography) for characterization of complete reaction, stopping the reaction, adding water (850mL) to quench the system, extracting with ethyl acetate (200mL x 3), combining organic phases, washing with saturated saline once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography on a crude product to obtain 1-methyl-4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) piperazine (2.25g, yield: 53%). MS: [ M +1]+:374.3/376.3.

Preparation of methyl-2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenyl ] -3-carboxylic acid ester:

to a reaction flask was added 1-methyl-4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperazine (0.75g,1.2eq), methyl 2-bromo-3-iodobenzoate (0.56g,1eq), potassium carbonate (0.68g,3eq), pd (dppf) Cl₂(0.06g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (400mL) to quench a system, extracting by ethyl acetate (100mL x 3), combining organic phases, washing by saturated salt water once, drying by anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography on a crude product to obtain methyl-2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenyl]-3-formic ester (0.41g, yield: 54%). MS: [ M +1]+:461.1/463.1

Preparation of (2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperidin-1-yl) propyl) - [1,1' -biphenylyl ] -3-yl) methanol:

adding methyl-2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenyl into a reaction bottle]-3-formic ester (0.41g,1eq), freshly distilled tetrahydrofuran (50mL) and LiAlH added portionwise at 5 ℃₄(67.4mg,2eq), stirring at room temperature for 30min, TLC characterization reaction complete, slowly dropping the reaction system into saturated NH₄The reaction was quenched with aqueous Cl, extracted with ethyl acetate (100 mL. times.3), and the organic phases were combined and washed with saturated brineWashing once, drying with anhydrous sodium sulfate, filtering, concentrating, and performing silica gel column chromatography to obtain (2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperidin-1-yl) propyl) - [1,1' -biphenyl]-3-yl) methanol (0.25g, yield: 65%). MS: [ M +1]+:433.1/435.1.

Preparation of 2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added (2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperidin-1-yl) propyl) - [1,1' -biphenyl ] -3-yl) methanol (0.25g, 1eq), 2-chloro-4, 6-dimethoxypyrimidine (0.17g,1.5eq), acetonitrile (30mL), potassium carbonate (0.24g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS indicated reaction was complete, reaction was stopped, water (200mL) was added to quench the system, ethyl acetate (50mL x 3) was extracted, organic phases were combined and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered with suction, concentrated, and crude silica gel column chromatographed to give 2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (287.3mg, yield: 83%). MS [ +1] + ] 599.2/602.2

Preparation of ((2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (60.1mg,1eq) and L-proline (34.5mg,3eq) were dissolved in a mixed solvent of DMF (5mL) and methanol (2mL), and 1 drop of acetic acid was added thereto with stirring, followed by reaction at room temperature for 30 minutes, and then sodium cyanoborohydride (20.9mg, 3eq) was added to the system. Stirring overnight at room temperature, LCMS to characterize reaction completion, stop reaction, quench the system with water (50mL), extract with ethyl acetate (20mL x 3), combine the organic phases and wash once with saturated brine, dry over anhydrous sodium sulfate, pump filter, concentrate, and purify the crude by preparative liquid phase to give (2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline (23.4mg, yield: 33%). MS [ +1] + ] 698.2/700.2.

¹H NMR(400MHz,d₆-DMSO)δ7.66(d,J＝7.2Hz,1H),7.48(t,J＝7.6Hz,1H),7.31–7.22(m,2H),7.01(d,J＝8.0Hz,1H),6.68(d,J＝7.2Hz,1H),5.58(s,2H),4.10–4.03(m,2H),3.95(s,6H),3.37–3.33(m,2H),3.23–3.17(m,1H),2.74–2.63(m,1H),2.47–2.24(m,9H),2.17(s,3H),2.12–2.05(m,1H),1.97(s,3H),1.90–1.64(m,6H)。

Example 12

The structural formula of the compound:

the synthesis method comprises the following steps:

with reference to the synthesis procedure of example 11, ((2- ((2-bromo-3 '- (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1 '-biphenylyl) propoxy) -2' -methyl ] is synthesized starting with tert-butyldimethylchlorosilane-protected 1- (3-chloropropyl) piperidin-4-ol]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:699.2/701.2.

¹H NMR(400MHz,d₆-DMSO)δ7.62(d,J＝7.2Hz,1H),7.48(t,J＝7.6Hz,1H),7.15-7.26(m,2H),6.99(d,J＝8.0Hz,1H),6.70(d,J＝7.4Hz,1H),5.59-5.43(m,2H),4.12–4.06(m,2H),3.95(s,6H),3.68–3.57(m,2H),3.25–3.18(m,1H),2.72–2.64(m,1H),2.47–2.27(m,8H),2.09–2.04(m,1H),1.95(s,3H),1.84–1.64(m,9H)。

Example 13

The structural formula of the compound:

the synthesis method comprises the following steps:

preparation of 2- ((2-chloro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-methoxypyrimidine-5-carbaldehyde:

to the reaction flask was added 1-methyl-4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperazine (0.75g,1.2eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.65g,1eq), potassium carbonate (0.68g,3eq), pd (dppf) Cl₂(0.06g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) representation of complete reaction, stopping the reaction, adding water (400mL) to quench a system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated common salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 2- ((2-chloro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenyl ] crude product]-3-yl) methoxy) -4, 6-methoxypyrimidine-5-carbaldehyde (0.57g, yield: 51%). MS: [ M +1]+:555.2/557.2。

Preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

2- ((2-chloro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-methoxypyrimidine-5-carbaldehyde (55.5mg,1eq) and L-proline (34.5mg,3eq) were dissolved in a mixed solvent of DMF (5mL) and methanol (2mL), and 1 drop of acetic acid was added thereto with stirring, followed by reaction at room temperature for 30 minutes, and then sodium cyanoborohydride (20.9mg, 3eq) was added to the system. Stirring overnight at room temperature, LCMS to characterize reaction completion, stop reaction, quench the system with water (50mL), extract with ethyl acetate (20mL x 3), combine the organic phases and wash once with saturated brine, dry over anhydrous sodium sulfate, pump filter, concentrate, and purify the crude by preparative liquid phase to give (2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline (20.9mg, yield: 32%). MS [ M +1] +:654.3/656.3.

¹H NMR(400MHz,d₆-DMSO)δ7.61(d,J＝7.2Hz,1H),7.44(t,J＝7.6Hz,1H),7.28–7.20(m,2H),6.98(d,J＝8.0Hz,1H),6.70(d,J＝7.2Hz,1H),5.53(s,2H),4.08–4.03(m,2H),3.91(s,6H),3.33–3.31(m,2H),3.23–3.19(m,1H),2.70–2.63(m,1H),2.47–2.27(m,9H),2.14(s,3H),2.09–2.04(m,1H),1.91–1.64(m,9H)。

Example 14

The synthesis method comprises the following steps:

preparation of 2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-chloro-2 ' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 4- ((tert-butyldimethyl) oxy) -1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine (1.07g,1.3eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.65g,1eq), potassium carbonate (0.68g,3eq), pd (dppf) Cl₂(0.06g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, LCMS (liquid chromatography-liquid chromatography) for completely reacting, stopping the reaction, adding water (400mL) to quench the system, extracting with ethyl acetate (100mL x 3), and synthesizingWashing the organic phase with saturated salt water, drying with anhydrous sodium sulfate, vacuum filtering, concentrating, and performing silica gel column chromatography to obtain 2- ((3' - (3- (4- ((tert-butyl dimethyl silicon) oxygen) piperidine-1-yl) propyl) -2-chloro-2 ' -methyl- [1,1' -biphenyl ] e]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.51g, yield: 45%). MS: [ M +1]+:670.3.

Preparation of (S) -1- ((2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-chloro-2 ' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) pyrrolidinyl-2-carboxylic acid:

2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-chloro-2 ' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (67.0mg,1eq) and L-proline (34.5mg,3eq) were dissolved in a mixed solvent of DMF (5mL) and methanol (2mL), and after 1 drop of acetic acid was added with stirring and reacted at room temperature for 30min, sodium cyanoborohydride (20.9mg, 3eq) was added to the system. Stirring was carried out overnight at room temperature, LCMS indicated that the reaction was complete, the reaction was stopped, a quenching system was added with water (50mL), ethyl acetate (20mL × 3) was extracted, the organic phases were combined and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by preparative liquid phase to give (S) -1- ((2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-chloro-2 ' -methyl- [1,1' -dibiphenyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) pyrrolidinyl-2-carboxylic acid (34.5mg, yield: 35%). MS [ +1] + ] 769.4.

Preparation of ((2- ((2-chloro-3 ' - (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

(S) -1- ((2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-chloro-2 ' -methyl- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) pyrrolidinyl-2-carboxylic acid (34.5mg,1eq) was dissolved in ultra-dry tetrahydrofuran (2mL), and a tetrahydrofuran solution of tetra-n-butylammonium fluoride (TBAF,1Min THF,0.13mL,3eq) was added dropwise to the system with stirring at room temperature. After stirring at room temperature for 2 hours, LCMS indicated reaction completion, reaction stopped, quench system with water (50mL), extract with ethyl acetate (20mL x 3), combine organic phases and wash with saturated brine once, dry over anhydrous sodium sulfate, pump filter, concentrate, and purify the crude by preparative liquid phase to give ((2- ((2-chloro-3 ' - (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline (10.5mg, yield: 35%). MS [ +1] + ] 655.3.

¹H NMR(400MHz,d₆-DMSO)δ7.61(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.27-7.16(m,2H),6.98(d,J＝8.0Hz,1H),6.68(d,J＝7.4Hz,1H),5.56-5.41(m,2H),4.09–4.03(m,2H),3.91(s,6H),3.68–3.57(m,2H),3.25–3.17(m,1H),2.72–2.64(m,1H),2.47–2.27(m,8H),2.09–2.04(m,1H),1.95(s,3H),1.84–1.64(m,9H)。

Example 15

The synthetic route is as follows:

Preparation of 2- ((3-bromo-2-fluorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 3-bromo-2-fluorobenzyl alcohol (2.4g,1eq), 2-chloro-4, 6-dimethoxypyrimidine (3.6g,1.5eq), acetonitrile (100mL), potassium carbonate (4.8g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (700mL) is added to quench the system, ethyl acetate (200mL x 3) is used for extraction, organic phases are combined and washed once by saturated salt solution, anhydrous sodium sulfate is used for drying, suction filtration and concentration are carried out, and crude silica gel column chromatography is carried out to obtain 2- ((3-bromo-2-fluorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-formaldehyde (3.0g, yield: 70%). MS is [ M +1] +:371.0/373.0.

Preparation of 2- ((2-fluoro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine (1.28g,1.1eq), 2- ((3-bromo-2-fluorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (1.18g,1eq), potassium carbonate (1.32g,3eq), pd (dppf) Cl₂(0.12g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (350mL) to quench a system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 2- ((2-fluoro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl) crude product]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.69g, yield: 41%). MS: [ M +1]+:526.2.

Preparation of ((2- ((2-fluoro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

2- ((2-fluoro-2 ' -methyl-3 ' - (3-morpholinopropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (52.5mg,1eq) and L-proline (34.5mg,3eq) were dissolved in a mixed solvent of DMF (5mL) and methanol (2mL), and 1 drop of acetic acid was added thereto with stirring to react at room temperature for 30 minutes, and then sodium cyanoborohydride (20.9mg, 3eq) was added to the system. Stirring overnight at room temperature, LCMS to characterize reaction completion, stop reaction, quench the system with water (50mL), extract with ethyl acetate (20mL x 3), combine the organic phases and wash once with saturated brine, dry over anhydrous sodium sulfate, pump filter, concentrate, and purify the crude by preparative liquid phase to give (2- ((2-bromo-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline (26.8mg, yield: 43%). MS [ +1] + ] 625.3.

¹H NMR(400MHz,d₆-DMSO)δ7.68(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.33–7.18(m,2H),7.01(d,J＝8.0Hz,1H),6.68(d,J＝7.2Hz,1H),5.59–5.52(m,2H),4.12–4.02(m,2H),3.95–3.78(m,8H),3.61–3.51(m,4H),3.23–3.05(m,4H),2.52–2.43(m,2H),2.40–2.20(m,4H),2.04–1.94(m,1H),1.97–1.63(m,7H)。

Example 16

The synthetic route is as follows:

referring to the synthesis of example 15, 1-methyl-4- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperazine was reacted with 2- ((3-bromo-2-fluorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde to produce 2- ((2-chloro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propyl) - [1,1' -biphenylyl) biphenyl]-3-yl) methoxy-4, 6-methoxypyrimidine-5-carbaldehyde, which is then reductively aminated with L-proline to form ((2- ((2-fluoro-2 ' -methyl-3 ' - (3- (4-methylpiperazin-1-yl) propoxy) - [1,1' -biphenylyl)]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:638.3.

¹H NMR(400MHz,d₆-DMSO)δ7.65(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.28–7.21(m,2H),6.99(d,J＝8.0Hz,1H),6.69(d,J＝7.2Hz,1H),5.56–5.49(m,2H),4.08–4.03(m,2H),3.95(s,6H),3.30–3.28(m,2H),3.24–3.19(m,1H),2.74–2.63(m,1H),2.46–2.27(m,9H),2.15(s,3H),2.10–2.04(m,1H),1.91–1.64(m,9H)。

Example 17

The synthetic route is as follows:

referring to the synthesis of example 14, 4- ((tert-butyldimethylsilyl) oxy) -1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine was reacted with 2- ((3-bromo-2-fluorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde to give 2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-fluoro-2 ' -methyl- [1,1' -biphenylyl) propyl) -2]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde, and then the aldehyde and L-proline are subjected to reductive amination reaction to generate (S) -1- ((2- ((3' - (3- (4- ((tert-butyldimethylsilyl) oxy) piperidin-1-yl) propyl) -2-fluoro-2 ' -methyl- [1,1' -biphenyl]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) pyrrolidinyl-2-carboxylic acid, and final TBAF deprotection of the TBS protecting group to yield ((2- ((2-fluoro-3 '- (3- (4-hydroxypiperidin-1-yl) propoxy) -2' -methyl- [1,1 '-biphenylyl) propoxy) -2' -methyl]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:639.3.

¹H NMR(400MHz,d₆-DMSO)δ7.66(d,J＝7.2Hz,1H),7.47(t,J＝7.6Hz,1H),7.26-7.17(m,2H),7.01(d,J＝8.0Hz,1H),6.70(d,J＝7.4Hz,1H),5.55-5.48(m,2H),4.12–4.04(m,2H),3.95(s,6H),3.71–3.57(m,2H),3.25–3.14(m,1H),2.73–2.64(m,1H),2.48–2.26(m,8H),2.10–2.04(m,1H),1.95(s,3H),1.84–1.65(m,9H)。

Example 18:

The synthetic route is as follows:

preparation of 4- (2- (3-bromo-2-methylphenoxy) ethyl) morpholine:

to the reaction flask was added 3-bromo-2-methylphenol (1.87g,1eq), 4- (2-chloroethyl) morpholine (1.64g,1.1eq), acetonitrile (50mL), potassium carbonate (4.14g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (150mL) is added to quench the system, ethyl acetate (60mL x 3) is used for extraction, organic phases are combined and washed once by saturated salt solution, dried by anhydrous sodium sulfate, filtered by suction, concentrated, and crude silica gel column chromatography is carried out to obtain 4- (2- (3-bromo-2-methylphenoxy) ethyl) morpholine (2.16g, yield: 72%). MS is [ M +1] +:300.1.

Preparation of 4- (2- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) ethyl) morpholine:

to a reaction flask was added 4- (2- (3-bromo-2-methylphenoxy) ethyl) morpholine (2.16g,1eq), pinacol diboron (3.65g,2eq), potassium acetate (1.41g,2eq), pd (dppf) Cl₂(0.26g,0.05eq) and 1, 4-dioxane (60 mL). The reaction was stopped by heating to 90 ℃ under nitrogen, stirring for two hours and LCMS to characterize the reaction was complete, water (350mL) was added to quench the system, ethyl acetate (100mL x 3) was extracted, the organic phases were combined and washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and chromatographed on crude silica gel to give 4- (2- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) ethyl) morpholine (1.27g, yield: 51%). MS: [ M +1]+:348.2.

Preparation of 2- ((2-chloro-2 ' -methyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to the reaction flask was added 4- (2- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) ethyl) morpholine (1.27g,1.1eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-methylAldehyde (1.29g,1eq), potassium carbonate (1.39g,3eq), Pd (dppf) Cl₂(0.12g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (350mL) to quench a system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 2- ((2-chloro-2 ' -methyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenyl) crude product]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.93g, yield: 53%). MS: [ M +1]+:528.2.

Preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

referring to the synthetic method for reductive amination in example 6, 2- ((2-chloro-2 ' -methyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenylyl)](2- ((2-chloro-2 ' -methyl-3 ' - (2-morpholinoethoxy) - [1,1' -biphenylyl) ether) synthesized by reacting (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-proline]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:627.2.

¹H NMR(400MHz,d₆-DMSO)δ7.61(dd,J＝7.6,1.5Hz,1H),7.44(t,J＝7.6Hz,1H),7.28–7.18(m,2H),7.02(d,J＝8.1Hz,1H),6.71(d,J＝7.4Hz,1H),5.61–5.46(m,2H),4.21–4.02(m,2H),3.95–3.81(m,8H),3.63–3.56(m,4H),3.27(dd,J＝9.1,5.3Hz,1H),3.19–3.10(m,1H),2.75(t,J＝5.7Hz,2H),2.66–2.57(m,1H),2.52–2.47(m,3H),2.12–2.00(m,1H),1.87(s,3H),1.84–1.55(m,4H)。

Example 19:

The synthetic route is as follows:

preparation of 1- (3- (3-bromo-2-methylphenoxy) propyl) piperidine:

to the reaction flask was added 3-bromo-2-methylphenol (1.87g,1eq), 1- (3-chloropropyl) piperidine (1.78g,1.1eq), acetonitrile (50mL), potassium carbonate (4.14g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (150mL) is added to quench the system, ethyl acetate (60mL x 3) is used for extraction, organic phases are combined and washed once by saturated salt solution, dried by anhydrous sodium sulfate, filtered by suction, concentrated, and the crude product is subjected to silica gel column chromatography to obtain 1- (3- (3-bromo-2-methylphenoxy) propyl) piperidine (2.15g, yield: 69%). MS [ +1] + ] 312.1.

Preparation of 1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine:

to a reaction flask was added 1- (3- (3-bromo-2-methylphenoxy) propyl) piperidine (2.15g,1eq), pinacol diboron (3.51g,2eq), potassium acetate (1.35g,2eq), pd (dppf) Cl₂(0.26g,0.05eq) and 1, 4-dioxane (60 mL). Under the protection of nitrogen, the temperature is raised to 90 ℃, the reaction is stirred for two hours, LCMS (liquid crystal display system) indicates that the reaction is complete, the reaction is stopped, water (350mL) is added for quenching the system, ethyl acetate (100mL x 3) is used for extraction, organic phases are combined and washed once by saturated saline, anhydrous sodium sulfate is used for drying, suction filtration and concentration are carried out, and crude silica gel column chromatography is carried out to obtain 1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) piperidine (1.21g, yield: 49%). MS: [ M +1]+:360.2.

Preparation of 2- ((2-chloro-2 ' -methyl-3 ' - (3- (piperidin-1-yl) propyl) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 1- (3- (2-methyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine (1.21g,1.2eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (1.09g,1eq), potassium carbonate (1.16g,3eq), pd (dppf) Cl₂(0.11g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for reaction for 16 hours, and displaying an LCMS (liquid Crystal display System)After the reaction is finished, stopping the reaction, adding water (350mL) to quench a system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated common salt water once, drying with anhydrous sodium sulfate, filtering, concentrating, and performing silica gel column chromatography to obtain 2- ((2-chloro-2 ' -methyl-3 ' - (3- (piperidin-1-yl) propyl) - [1,1' -biphenyl) of crude product]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.81g, yield: 53%). MS: [ M +1]+:540.2.

Preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-piperidin-1-yl) propoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

referring to the synthetic method for reductive amination in example 6, 2- ((2-chloro-2 ' -methyl-3 ' - (3- (piperidin-1-yl) propyl) - [1,1' -biphenylyl ] is synthesized]Synthesis of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-piperidin-1-yl) propoxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-proline]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:639.2.

¹H NMR(400MHz,d₆-DMSO)δ7.61(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.30–7.18(m,2H),6.98(d,J＝8.0Hz,1H),6.68(d,J＝7.3Hz,1H),5.55(s,2H),4.11–4.03(m,2H),3.95–3.79(m,8H),3.21–3.05(m,4H),2.46–2.39(m,2H),2.35–2.28(m,4H),2.04–1.96(m,1H),1.97–1.45(m,13H)。

Example 20:

((2- ((2-chloro-2 ' -methyl-3 ' - (2- (piperidin-1-yl) ethoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline

The synthetic route is as follows:

referring to the synthesis of example 19, 1- (2-chloroethyl) piperidine was used as a starting material,(2- ((2-chloro-2 ' -methyl-3 ' - (2- (piperidin-1-yl) ethoxy) - [1,1' -biphenylyl ] was synthesized according to the synthesis method of example 19]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:624.3.

¹H NMR(400MHz,d₆-DMSO)δ7.61(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.30–7.18(m,2H),6.98(d,J＝8.0Hz,1H),6.68(d,J＝7.3Hz,1H),5.55(s,2H),4.11–4.03(m,2H),3.95–3.79(m,8H),3.32–3.15(m,4H),2.48–2.39(m,2H),2.38–2.28(m,4H),2.04–1.96(m,1H),1.97–1.45(m,11H)。

Example 21:

The synthetic route is as follows:

with reference to the synthesis method of example 19, 3-bromo-phenol was used as a starting material, and ((2- ((2-chloro-3 '- (3-piperidinopropoxy) - [1,1' -biphenylyl) was synthesized according to the synthesis method of example 19]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:627.3.

¹H-NMR(400MHz,d₆-DMSO)δ7.62(dd,J＝7.6,1.8Hz,1H),7.45(t,J＝7.6Hz,1H),7.42–7.33(m,2H),7.00–6.91(m,3H),5.54(s,2H),4.05(t,J＝6.4Hz,2H),3.91(s,6H),3.87(s,2H),3.59–3.51(m,4H),3.33–3.25(m,2H),3.23–3.14(m,2H),2.64(dd,J＝17.2,9.8Hz,1H),2.42(t,J＝7.1Hz,2H),2.39–2.31(m,4H),2.14–2.00(m,1H),1.96–1.51(m,3H)。

Example 22:

The synthetic route is as follows:

((2- ((2-chloro-3 '- (3- (dimethylamino) propoxy) -2' -methyl- [1,1 '-biphenylyl) propoxy) -2' -methyl ] amine was synthesized according to the synthesis method of example 19, starting with 3-chloro-N, N-dimethylpropyl-1-amine as a starting material in accordance with the synthesis method of example 19]-3-yl) methoxy-4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:599.2.

¹H NMR(400MHz,d₆-DMSO)δ7.60(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.29–7.18(m,2H),6.99(d,J＝8.0Hz,1H),6.70(d,J＝7.2Hz,1H),5.55(s,2H),4.11–4.03(m,2H),3.95(s,6H),3.91–3.79(m,2H),3.21–3.05(m,4H),2.46–2.39(m,4H),2.35–2.28(m,4H),2.04–1.96(m,1H),1.95–1.45(m,7H)。

Example 23:

The synthetic route is as follows:

preparation of 4- (3- (3-bromo-2-chlorophenoxy) propyl) piperidine:

to the reaction flask was added 3-bromo-2-chlorophenol (2.07g,1eq), 1- (3-chloropropyl) piperidine (1.78g,1.1eq), acetonitrile (50mL), potassium carbonate (4.14g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (150mL) is added to quench the system, ethyl acetate (60mL x 3) is extracted, organic phases are combined and washed once with saturated saline, dried over anhydrous sodium sulfate, filtered by suction, concentrated, and subjected to crude silica gel column chromatography to obtain 4- (3- (3-bromo-2-chlorophenoxy) propyl) piperidine (2.31g, yield: 69%). MS [ +1] + ] 334.0.

Preparation of 4- (3- (2-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine:

to a reaction flask was added 4- (3- (3-bromo-2-chlorophenoxy) propyl) piperidine (2.31g,1eq), pinacol diboride (3.51g,2eq), potassium acetate (1.35g,2eq), Pd (dppf) Cl₂(0.26g,0.05eq) and 1, 4-dioxane (60 mL). Under the protection of nitrogen, the temperature is raised to 90 ℃, the reaction is stirred for two hours, LCMS (liquid crystal display system) indicates that the reaction is complete, the reaction is stopped, water (350mL) is added for quenching the system, ethyl acetate (100mL x 3) is used for extraction, organic phases are combined and washed once by saturated saline, anhydrous sodium sulfate is used for drying, suction filtration and concentration are carried out, and crude silica gel column chromatography is carried out to obtain 4- (3- (2-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) piperidine (1.29g, yield: 49%). MS: [ M +1]+:382.2.

Preparation of 2- ((2,2' -chloro-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 4- (3- (2-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine (1.29g,1.2eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (1.09g,1eq), potassium carbonate (1.16g,3eq), pd (dppf) Cl₂(0.11g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (350mL) to a quenching system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 2- ((2,2' -chloro-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl ] crude product]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (1.00g, yield: 53%). MS: [ M +1]+:562.2.

Preparation of ((2- ((2,2' -dichloro-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy-4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

referring to the synthetic method for reductive amination in example 6, 2- ((2,2' -chloro-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)](2- ((2,2 '-dichloro-3' - (3-morpholinepropoxy) - [1,1 '-biphenylyl) oxy) - [1,1' -dimethoxypyrimidine-5-carbaldehyde was synthesized by reacting (E) -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-proline]-3-yl) methoxy-4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:661.2.

¹H NMR(400MHz,d₆-DMSO)δ7.63(dd,J＝7.6,1.4Hz,1H),7.46(t,J＝7.6Hz,1H),7.37(t,J＝8.0Hz,1H),7.30(dd,J＝7.6,1.5Hz,1H),7.23–7.19(m,1H),6.88(dd,J＝7.6,1.1Hz,1H),5.60–5.47(m,2H),4.22–4.09(m,2H),3.88(s,6H),3.80(d,J＝13.0Hz,1H),3.63(d,J＝12.8Hz,1H),3.60–3.50(m,4H),3.02–2.94(m,2H),2.46(t,J＝7.1Hz,2H),2.42–2.30(m,5H),2.01–1.86(m,3H),1.81–1.67(m,1H),1.66–1.56(m,2H)。

Example 24:

The synthetic route is as follows:

preparation of 4- (3- (3-bromo-2-ethylphenoxy) propyl) morpholine:

to the reaction flask was added 3-bromo-2-ethylphenol (2.01g,1eq), 1- (3-chloropropyl) piperidine (1.78g,1.1eq), acetonitrile (50mL), potassium carbonate (4.14g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents that the reaction is complete, the reaction is stopped, water (150mL) is added to quench the system, ethyl acetate (60mL x 3) is used for extraction, organic phases are combined and washed once by saturated salt solution, dried by anhydrous sodium sulfate, filtered by suction, concentrated, and crude silica gel column chromatography is carried out to obtain 4- (3- (3-bromo-2-ethylphenoxy) propyl) morpholine (2.36g, yield: 72%). MS [ +1] + ] 328.1.

Preparation of 4- (3- (2-ethyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine:

to a reaction flask was added 4- (3- (3-bromo-2-ethylphenoxy) propyl) morpholine (2.36g,1eq), pinacol diboron (3.65g,2eq), potassium acetate (1.41g,2eq), Pd (dppf) Cl₂(0.26g,0.05eq) and 1, 4-dioxane (60 mL). The reaction was stopped by heating to 90 ℃ under nitrogen, stirring for two hours and LCMS to characterize the reaction complete, adding water (350mL) to quench the system, extracting with ethyl acetate (100mL x 3), combining the organic phases and washing with saturated brine once, drying over anhydrous sodium sulfate, suction filtering, concentrating, and chromatography on crude silica gel column to give 4- (3- (2-ethyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenoxy) propyl) morpholine (1.43g, yield: 53%). MS: [ M +1]+:376.2.

Preparation of 2- ((2-chloro-2 ' -ethyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde:

to a reaction flask was added 4- (3- (2-ethyl-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) morpholine (1.43g,1.2eq), 2- ((3-bromo-2-chlorobenzyl) oxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (1.23g,1eq), potassium carbonate (1.31g,3eq), pd (dppf) Cl₂(0.12g,0.05eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (350mL) to quench a system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 2- ((2-chloro-2 ' -ethyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl) crude product]-3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde (0.87g, yield: 49%). MS: [ M +1]+:556.2.

Preparation of ((2- ((2-chloro-2 ' -ethyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline:

reference example 6 Synthesis Process for reductive amination2- ((2-chloro-2 ' -ethyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl)](2- ((2-chloro-2 ' -ethyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) amino acid) synthesized by reacting (3-yl) methoxy) -4, 6-dimethoxypyrimidine-5-carbaldehyde with L-proline]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:655.3.

¹H NMR(400MHz,d₆-DMSO)δ7.61(d,J＝7.2Hz,1H),7.43(t,J＝7.6Hz,1H),7.28–7.18(m,2H),6.98(d,J＝8.0Hz,1H),6.68(d,J＝7.2Hz,1H),5.52(s,2H),4.12–4.02(m,2H),3.95(s,6H),3.91–3.78(m,2H),3.61–3.50(m,4H),3.23–3.08(m,4H),2.51–2.43(m,2H),2.40–2.30(m,4H),2.04–1.97(m,3H),1.95–1.55(m,4H),1.35–1.25(m,3H)。

Example 25:

The synthetic route is as follows:

with reference to the synthesis method of example 23, 3-bromo-2-isopropyl-phenol as a starting material was synthesized according to the synthesis method of example 23 ((2- ((2-chloro-2 '-isopropyl-3' - (3-morpholinepropoxy) - [1,1 '-biphenylyl ] oxy) - [1,1' -biphenylyl ] phenyl]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:669.3.

¹H NMR(400MHz,d₆-DMSO)δ7.60(d,J＝7.2Hz,1H),7.44(t,J＝7.6Hz,1H),7.30–7.17(m,2H),6.99(d,J＝7.6Hz,1H),6.70(d,J＝7.2Hz,1H),5.60–5.49(m,2H),4.10–4.02(m,2H),3.95(s,6H),3.91–3.78(m,2H),3.61–3.50(m,4H),3.23–3.08(m,4H),2.51–2.43(m,2H),2.40–2.30(m,4H),2.04–1.97(m,2H),1.95–1.55(m,4H),1.35–1.24(m,6H)。

Example 26:

The synthetic route is as follows:

with reference to the synthesis method of example 23, 3-bromo-2-cyclopropyl-phenol was used as a starting material, and ((2- ((2-chloro-2 ' -cyclopropyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) was synthesized according to the synthesis method of example 23]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:667.3.

¹H NMR(400MHz,d₆-DMSO)δ7.60(d,J＝7.2Hz,1H),7.44(t,J＝7.6Hz,1H),7.30–7.17(m,2H),6.99(d,J＝7.6Hz,1H),6.70(d,J＝7.2Hz,1H),5.60–5.49(m,2H),4.10–4.02(m,2H),3.95–3.78(m,8H),3.60–3.50(m,4H),3.23–3.08(m,4H),2.51–2.43(m,2H),2.40–2.30(m,4H),2.04–1.97(m,1H),1.95–1.55(m,5H),1.24–0.99(m,4H)。

Example 27:

The synthetic route is as follows:

preparation of 3- (((2-chloro-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile:

to a solution of 3-hydroxymethylbenzonitrile (2.66g,1eq) in DMF (50mL) was added sodium hydride (1.21g,1.5eq, 60% purity) with stirring at 0 ℃. After stirring at 0 ℃ for 0.5 hour, dropwise adding a DMF (dimethyl formamide) solution of 2, 4-dichloro-6-methoxypyrimidine (3.58g,1eq) into the reaction system, heating to room temperature under the protection of nitrogen, stirring for reaction for two hours, LCMS (liquid crystal display system) characterization of complete reaction, stopping reaction, adding water (500mL) to quench the system, extracting with ethyl acetate (200mL 3), combining organic phases, washing with saturated common salt water once, drying with anhydrous sodium sulfate, filtering, concentrating, and performing silica gel column chromatography on a crude product to obtain 3- (((2-chloro-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (2.97g, yield: 54%). MS [ +1] + ] 276.0.

Preparation of 3- (((2- ((3-bromo-2-chlorobenzyloxy) oxy) -6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile:

to the reaction flask was added 3- (((2-chloro-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (2.97g,1eq), 3-bromo-2-chlorobenzyl alcohol (2.62g,1.1eq), acetonitrile (50mL), potassium carbonate (4.46g,3 eq). The temperature is raised to 70 ℃, and the reaction is stirred for two hours. LCMS represents the reaction is complete, the reaction is stopped, water (250mL) is added to quench the system, ethyl acetate (90mL x 3) is extracted, organic phases are combined and washed once with saturated common salt, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to crude silica gel column chromatography to obtain 3- (((2- ((3-bromo-2-chlorobenzyloxy) oxy) -6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (1.78g, yield: 36%). MS [ +1] + ] 460.0.

Preparation of 3- (((2- ((3-bromo-2-chlorobenzyloxy) oxy) -5-formyl-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile:

under the condition of ice-water bath, phosphorus oxychloride (8.89g,15eq) is dropwise added into a reaction bottle filled with DMF (4.23g,15eq), the mixture is stirred for 30min under the condition of ice-water bath, a solution of 3- (((2- ((3-bromo-2-chlorobenzyloxy) oxy) -6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (1.78g,1eq) in DMF (20mL) is dropwise added into a reaction system, the temperature is increased to 50 ℃, and the mixture is stirred and reacted for two hours. LCMS indicated reaction was complete and stopped. Dropwise adding the reaction system into an ice water bath with stirring, adjusting the pH value to be neutral (7) by using sodium bicarbonate, carrying out suction filtration on the solid, washing twice, and purifying the dried crude product by silica gel column chromatography to obtain 3- (((2- ((3-bromine)-2-chlorobenzyloxy) oxy) -5-formyl-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (0.87g, yield 46%). MS: [ M +1]⁺:488.0.

Preparation of 3- (((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -5-formyl-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile:

to the reaction flask was added 4- (3- (2-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) propyl) piperidine (0.96g,1.5eq), 3- (((2- ((3-bromo-2-chlorobenzyloxy) oxy) -5-formyl-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (0.87g,1eq), potassium carbonate (0.74g,3eq), pd (dppf) Cl₂(0.13g,0.1eq), toluene (50mL), water (5mL) and methanol (5 mL). Heating to 90 ℃ under the protection of nitrogen, stirring for 16 hours, performing LCMS (liquid Crystal display System) to characterize that the reaction is complete, stopping the reaction, adding water (350mL) to a quenching system, extracting with ethyl acetate (100mL x 3), combining organic phases, washing with saturated common salt water once, drying with anhydrous sodium sulfate, performing suction filtration, concentrating, and performing silica gel column chromatography to obtain 3- (((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenyl) crude product]-3-yl) methoxy) -5-formyl-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile (0.57g, yield: 50%). MS: [ M +1]+:643.2.

Preparation of ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4- ((3-cyanobenzyl) oxy) -6-methoxypyrimidin-5-yl) methyl) -L-proline:

referring to the synthetic method for reductive amination in example 6, 3- (((2- ((2-chloro-2 '-methyl-3' - (3-morpholinepropoxy) - [1,1 '-biphenylyl) oxy) - [1,1' -biphenylyl ] amine was added]Synthesis of ((2- ((2-chloro-2 '-methyl-3' - (3-morpholinyloxy) - [1,1 '-biphenylyl) oxy) - [1,1' -biphenylyl) by reaction of (3-yl) methoxy) -5-formyl-6-methoxypyrimidin-4-yl) oxy) methyl) benzonitrile with L-proline]-3-yl) methoxy) -4- ((3-cyanobenzyl) oxy) -6-methoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:743.3.

¹H NMR(400MHz,d₆-DMSO)δ8.19(s,1H),8.03–7.99(m,1H),7.69–7.60(m,2H),7.53–7.43(m,2H),7.32–7.20(m,2H),7.00(d,J＝8.0Hz,1H),6.68(d,J＝7.2Hz,1H),5.60–5.40(m,4H),4.08–4.00(m,2H),3.95–3.78(m,5H),3.60–3.50(m,4H),3.20–3.05(m,4H),2.50–2.43(m,2H),2.40–2.30(m,4H),2.04–1.96(m,1H),1.97–1.75(m,6H),1.73–1.65(m,1H)。

Example 28:

The synthetic route is as follows:

referring to the synthesis method of example 26, ((2- ((2-chloro-2 ' -methyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) was synthesized using 3-hydroxymethylpyridine as a starting material]-3-yl) methoxy) -4-methoxy-6- (pyridin-3-ylmethoxy) pyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:718.3.

¹H NMR(400MHz,d₆-DMSO)δ8.78(d,J＝1.6Hz,1H),8.58(dd,J＝4.8,1.6Hz,1H),7.98(d,J＝8.0Hz,1H),7.69–7.60(m,2H),7.48–7.43(m,1H),7.32–7.20(m,2H),7.00(d,J＝8.0Hz,1H),6.68(d,J＝7.2Hz,1H),5.60–5.40(m,4H),4.12–4.04(m,2H),4.01–3.78(m,5H),3.60–3.50(m,4H),3.20–3.05(m,4H),2.50–2.43(m,2H),2.40–2.30(m,4H),2.04–1.96(m,1H),1.97–1.55(m,7H)。

Example 29

The synthetic route is as follows:

preparation of (2S,4R) -1- ((2- ((2-bromo- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid:

(2S,4R) -1- ((2- ((2-bromo- [1,1 '-biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -4-hydroxypyrrole-2-carboxylic acid was synthesized by reacting 2- ((2-bromo- [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) formaldehyde with L-hydroxyproline.

MS:[M+1]⁺:544.1/546.1。

¹H NMR(400MHz,d₆-DMSO)δ7.60(dd,J＝7.6,1.6Hz,1H),7.54-7.31(m,7H),5.53(s,2H),4.94(s,1H),4.22-4.11(m,1H),3.96-3.74(m,8H),3.37(t,J＝8.0Hz,2H),3.28-3.24(m,1H),1.91-1.80(m,2H)。

Example 30:

The synthetic route is as follows:

with reference to the synthesis method of example 23, 3-bromo-2-butyl-phenol was used as a starting material, and ((2- ((2-chloro-2 ' -butyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl) was synthesized according to the synthesis method of example 23]-3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline. MS: [ M +1]⁺:683.3.

¹H NMR(400MHz,d₆-DMSO)δ7.61(d,J＝7.2Hz,1H),7.45(t,J＝7.6Hz,1H),7.31–7.18(m,2H),7.00(d,J＝7.6Hz,1H),6.71(d,J＝7.2Hz,1H),5.62–5.50(m,2H),4.12–4.05(m,2H),3.96(s,6H),3.92–3.80(m,2H),3.61–3.50(m,4H),3.24–3.11(m,4H),2.61–2.47(m,3H),2.40–2.30(m,4H),2.07–1.55(m,8H),1.37–1.24(m,2H),0.91–0.85(m,3H)。

Detecting inhibitory effects of compounds on the binding of PD-1/PD-L1 proteins to each other

In vitro kinase level was measured using the PD1/PD-L1 binding assay kit available from Cisbio.

The principle and the method for screening the PD-1/PD-L1 small molecule inhibitor comprise the following steps:

1) the principle is as follows: PD-L1 protein has Tag1 (Tag1 is a common Tag for purified proteins), PD-1 protein has Tag2 (Tag2 is a common Tag for purified proteins), and anti-Tag1 antibody labeled by Eu and anti-Tag2 antibody labeled by XL665 are combined with the two Tag proteins respectively. When excited by laser light, energy is transferred from the donor Eu to the acceptor XL665, causing XL665 to emit light. After the inhibitor is added, the combination of PD-1 and PD-L1 is blocked, so that Eu is far away from XL665, the occurrence of fluorescence energy transfer is influenced, and the signal is weakened. The blocking of the binding of PD-L1 and PD-1 by the inhibitor is judged by the change of the intensity of the signal.

2) The method comprises the following steps: the specific experimental procedures were performed with reference to PD1/PD-L1 from Cisbio, Inc., in combination with kit instructions 62ICP01PEG and 64ICP01 PEH. Briefly described as follows: and taking a new 384-well enzyme label plate, adding 2 mu L of diluent or target compounds diluted by the diluent and with different concentrations into each well, then adding 4 mu L of PD-1 protein and 4 mu L of PD-L1 protein, and incubating for 15min at normal temperature. 10 mu L of mixed solution of anti-Tag1-Eu and anti-Tag2-XL665 is added into each well, and after incubation for 2h at normal temperature, fluorescent signals of 620nM and 665nM are detected by a microplate reader. The signal ratio is calculated by the formula: (665nM signal)/(620 nM signal) × 10⁴. 8-10 concentration gradients are selected for each compound, three multiple holes are arranged for each concentration, and the obtained result is subjected to nonlinear fitting by GraphPad Prism software to obtain the IC50 value of the compound.

TABLE 1 IC50 values for the compounds of the invention

Note: letter a represents IC50 less than 10 nM;

letter B represents an IC50 of between 10-100 nM;

letter C indicates IC50 greater than 100 nM.

The results in Table 1 show that the compounds of the present invention are effective in inhibiting the binding of PD-1/PD-L1 at various concentrations and are therefore useful in the treatment of diseases resulting from binding to PD-1/PD-L1.

The invention provides a novel PD-1/PD-L1 small molecule inhibitor, and the results of the drug activity test show that the compound has excellent activity and is hopeful to become a novel PD-1/PD-L1 small molecule inhibitor.

In addition, as can be seen from Table 1, with respect to R₁And R₂Is a long chain group, the compound of the invention is due to R₁And R₂Forms a ring structure (or a short chain structure) together with the nitrogen atom, limits partial conformation, has better directionality, and has stronger binding force with PD-L1, thereby showing higher activity.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, prodrug, metabolite, isotopic derivative, solvate thereof:

wherein X is selected from Br, Cl and F;

y is selected from methyl, ethyl, isopropyl, cyclopropyl, Cl, F and H;

R₁and R₂Independently selected from H, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted 3-6 membered cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclic aryl, the substituent is selected from C1-C6 alkyl, C1-C6 alkoxy;

n＝1、2、3；

R₁、R₂、R₁and R₂Together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic ring, R₄、R₅、R₄And R₅A 4-7 membered heterocyclic ring together with the nitrogen atom to which it is attached; wherein the substituent in the 4-7 membered heterocycle is at least one of 1-4C 1-C6 alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, hydroxyl, carboxyl, amino, nitrile, halogen, trifluoromethyl and nitro; the 3-6 membered heterocycloalkyl or heteroaryl group contains 1-3 heteroatoms selected from N, O, S.

2. A compound according to claim 1, or a pharmaceutically acceptable salt, prodrug, metabolite, isotopic derivative, solvate thereof,

wherein R is₁And R₂Each independently selected from methyl, ethyl, isopropyl, cyclopropyl, phenyl, pyridyl;

or, R₁And R₂A 4-7 membered heterocyclic group taken together with the nitrogen atom to which it is attached, wherein R₁And R₂The 4-7 membered heterocyclic group, which is formed together with the attached nitrogen atom, is selected from the following groups:

R₃one selected from hydrogen, pyridyl and nitrile substituted phenyl.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, prodrug, metabolite, isotopic derivative, solvate thereof,

wherein R is₄And R₅Identical or different, R₄And R₅Together with the nitrogen atom to which they are attached form a 4-7 membered heterocyclic group, wherein R₄And R₅Together with the nitrogen atom to which they are attached form a substituted or unsubstituted 4-7 membered heterocyclyl selected from 1-piperidinyl, 1-pyrrolidinyl, 4-methyl-1-piperidinyl, 1-cyclobutylamino, 1-cyclopentylamino;

4. A compound or pharmaceutically acceptable salt according to claim 1, wherein the compound is selected from any one of the following compounds:

((2- ((2-chloro-2 ' -butyl-3 ' - (3-morpholinepropoxy) - [1,1' -biphenylyl ] -3-yl) methoxy) -4, 6-dimethoxypyrimidin-5-yl) methyl) -L-proline.

5. A process for the preparation of a compound according to claim 1, wherein the process comprises the following reaction scheme:

the method comprises the following steps:

(3) dissolving compounds C02 and SM3 in a solvent 3, adding an alkaline reagent and palladium metal, and reacting the catalyst at the temperature of 70-120 ℃ to obtain a compound C03;

(4) dissolving C03 and SM4 in a solvent 4 to perform reductive amination reaction to obtain a compound shown in the formula I;

preferably, in step (ii)(3) Wherein the solvent 3 is selected from one or more of 1, 4-dioxane, toluene, methanol and water; the alkaline reagent is selected from one or more of cesium carbonate, potassium carbonate, sodium carbonate, potassium tert-butoxide and sodium tert-butoxide; the palladium metal catalyst is selected from Pd (PPh)₃)₄、Pd(dppf)Cl₂Palladium acetate, Pd₂(dba)₃One or more of;

6. A pharmaceutical composition comprising a compound or a pharmaceutically salt of a compound or a solvate of a compound according to any one of claims 1 to 4 as an active ingredient, and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is formulated in the form of an oral formulation or an injection.

8. Use of a compound or a pharmaceutically salt of a compound or a solvate of a compound according to any one of claims 1 to 4, or a pharmaceutical composition according to claim 6 or 7, for the manufacture of a medicament for the treatment of a disease associated with the PD-1/PD-L1 signalling pathway.

9. The use of claim 8, wherein the disease is cancer or an immune disease.

10. The use according to claim 9, wherein the cancer is selected from skin cancer, lung cancer, hematological tumors, breast cancer, glioma, digestive system tumors, reproductive system tumors, lymphoma, nervous system tumors, head and neck cancer; the immune diseases are selected from diabetes, myasthenia gravis, rheumatoid arthritis, systemic lupus erythematosus and dermatitis.

11. A method of treating cancer or an immune disease, comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition of claim 6 or 7.