Biphenyl Compounds, Pharmaceutical Compositions, and Methods for Their Preparation and Use

Abstract

The present application provides a biphenyl compound, a pharmaceutical composition, and a method for preparing and using the same. The biphenyl compound has a structure represented by formula (I). The biphenyl compound according to the present application can bind well to multiple target point proteins, has good bioavailability, and can be used for the preparation of a drug for preventing or treating tumors, autoimmune diseases, inflammatory diseases, neurodegenerative diseases, or an anti-aging agent.

Description

Technical Field

[0001] The present application relates to the field of pharmaceutical technology, and specifically, to biphenyl compounds, pharmaceutical compositions, and their preparation methods and uses.

Background Art

[0002] According to medical data, it has been clarified that inflammation is a risk factor for causing tumors. For example, chronic hepatitis may also be a direct cause of inducing liver cancer, Helicobacter pylori infection in the stomach tends to increase the risk of gastric cancer, and in cervical cancer, there are many patients infected with papillomavirus. Other factors, for example, autoimmune intestinal diseases are closely related to colon cancer, and PM2.5 in the air is also a cause of inducing lung cancer. Tumors are defined as non-communicable diseases similar to heart disease, chronic respiratory diseases or diabetes. They are often chronic diseases over a long period of time and have a slow progression. The relationship between inflammation and tumors was first proposed by Galenus more than 1,800 years ago, and many studies have demonstrated that persistent inflammation can progress lesions from infection or autoimmune inflammation to tumors.

[0003] Biphenyl compounds widely exist in active natural products, have various biological activities such as antiviral, antioxidant, anti-inflammatory, antitumor, anti-spasmodic, metabolic antagonism, etc., and have a very broad research perspective. We have been engaged in the research on the structural optimization of such natural products, antitumor activity, treatment of autoimmune diseases, and structure-activity relationship with anti-inflammatory and metabolic antagonism for a long time. The structural modification of natural products contributes to the discovery of precursors with stronger tumor suppression activity, activity in treating autoimmune diseases and inflammatory diseases, and lower toxicity compared with the original natural products, and further becomes antitumor, anti-inflammatory immune agents and metabolic antagonist drugs.

[0004] The potential target points of the biphenyl compounds designed according to the content of this application in antitumor and anti-inflammatory immunity are summarized as follows.

[0005] Epidermal Growth Factor Receptor (EGFR): The mutation activation of EGFR is an important factor that causes abnormal biological activities of tumor cells. Here, the T790M mutation in EGFR is a change from cytosine (C) to thymine (T) in one base pair, that is, the 790th threonine in the function of EGFR tyrosine kinase is substituted by methionine. Such a mutation may cause EGFR to be reactivated, resulting in drug resistance to tyrosine kinase inhibitors (TKIs). On the other hand, after docking such compounds and derivatives with the protein after this mutation by means of computer-aided design, it is found that the scores of such compounds are all high, suggesting that biphenyl compounds may be candidates for this target point.

[0006] Vascular Endothelial Growth Factor (VEGF) is a signaling protein that stimulates angiogenesis within cells and has the function of promoting angiogenesis and regeneration. VEGF binds to the vascular endothelial growth factor receptor (VEGFR, also called tyrosine kinase receptor) on the cell membrane surface, and biological effects occur through a series of signal transduction pathways, ultimately causing angiogenesis.

[0007] Histone-lysine N-methyltransferase EZH2: EZH2 is an enzyme encoded by one human EZH2 gene. It has already been identified that two transcript variants transcribed by this gene encode different subtypes, and the changes in gene sequences are essentially different from abnormal epigenetic modifications. The reason is that when mutations occur in the DNA sequence, the gene is difficult to repair, and it is also difficult to remove the mutated gene product. However, epi Genetic modifications can potentially be reversed by inhibitors of the associated chromatin modification enzyme system. Therefore, it is very important to clarify the mechanism of action of the epigenetic modification enzyme system in tumor cells, and furthermore, to provide corresponding therapeutic means to prevent epigenetic modifications. In 2020, Epizyme's EZH2 inhibitor Tazemetestat, approved by the FDA, was launched and used for the treatment of metastatic or locally advanced epithelioid sarcoma. Currently, a total of five drugs are undergoing clinical phase I / II studies.

[0008] Histone deacetylase (HDAC): A type of protease that plays an important role in the structural modification of chromosomes and the regulation of gene expression. Generally, histone acetylation contributes to the dissociation of DNA and the histone octamer, and by relaxing the structure of the nucleosome, each transcription factor and synergistic transcription factor can specifically bind to the DNA binding site, activating gene transcription. In the cell nucleus, the histone acetylation and histone deacetylation processes are in a dynamic balance and are jointly regulated by histone acetyltransferase (HAT) and histone deacetylase (HDAC).

[0009] Src kinase: A type of non-receptor protein kinase that is widely present in cancer cells and plays an important role in various processes of cell growth and proliferation, such as gene transcription, cell differentiation, migration, angiogenesis, and prevention of apoptosis. The research on Src inhibitors has already become a hot spot in anti-tumor drug research. Currently, a series of Src inhibitors are in the clinical research stage.

[0010] Cancer immunotherapy is an increasingly effective treatment strategy. T cells play an important role in immunotherapy, and many immune checkpoints are treasures to be explored. Since the therapeutic effects of CTLA-4 and PD-1 / PD-L1 monoclonal antibodies have been affirmed, competition in the orbit of monoclonal antibodies here is fierce. Therefore, more attention should be paid to the target points of other immune checkpoints, and their functions determine the therapeutic effect of immunotherapy.

[0011] The survival and development of T cells are affected by TCR signals, and the TCR signaling pathway depends on Src family kinases (SFK). Lck is an important member of SFK and is expressed throughout most of the T cell life cycle. In addition, Lck plays an important role in activating the TCR signaling pathway to activate T cells. CSK is an important regulator of SFK, which inactivates Lck by phosphorylating Lck (Tyr505), and the latter suppresses the activation of T cells by TCR. Therefore, CSK and p-Lck (Tyr505) may be effective target points for future immunomodulatory therapy.

[0012] CD73 is a 5'-nucleotide hydrolase that can hydrolyze extracellular adenosine monophosphate (AMP) to adenosine. Adenosine is a strong immunosuppressive molecule that + suppresses the activation of CD8 T cells and further contributes to allowing cancer cells to escape from "killing" by T cells. NK cells infiltrating tumors increase the expression of CD73, and the occurrence frequency of these CD73+ NK cells is related to the size of tumors in breast cancer patients. The research results support the fact that tumors can hijack NK cells for immune evasion, and the expression of CD73 defines an inducible NK cell population and has immunomodulatory properties in the tumor microenvironment.

[0013] KIR: Killer cell immunoglobulin-like receptor is a receptor expressed on the surface of NK cells and some T cells. It specifically recognizes cell surface MHC-I molecules and exerts an immunomodulatory function, and can play a role in the occurrence and development of tumors and immune inflammatory diseases.

[0014] LAG-3: Lymphocyte activation gene 3 protein is an immune negative regulatory molecule receptor that can bind to MHC-II molecules and FGL1 distributed in activated T cells, NK cells, and dendritic cells. It has a function of maintaining the stability of the internal environment and participating in immune regulation, and is closely related to the occurrence and development of tumors, immune inflammatory diseases, and metabolic diseases. Currently, several LAG3 monoclonal antibodies are in the clinical trial stage.

[0015] 4-1BB, also known as CD137, is a member of the TNF family expressed on the surface of activated T cells and an inducible T cell surface receptor. 4-1BB and its ligand are another important co-stimulatory molecule outside the CD28 / B7 co-stimulatory signaling pathway.

[0016] PI3K-AKT-mTOR pathway: Phosphatidylinositol 3-kinase (PI3Ks) belongs to the lipid kinase family, participates in metabolic processes such as cell glycolysis and lipogenesis, and can regulate tumor angiogenesis. The PI3K / Akt / mTOR signaling pathway can induce the occurrence of tumors and can be involved in cell autophagy through multiple mechanisms, such as suppressing the occurrence of autophagy by 3-methyladenine (3-MA), a specific inhibitor of PI3K, suppressing the structural change of Bax, phosphorylating other apoptosis-related components such as Bad and caspase 9 at the mitochondrial membrane potential level to suppress cell apoptosis, and down-regulating the expression of the tumor suppressor protein p53 in the cell nucleus.

[0017] ACAT1 belongs to the specific thiol enzyme superfamily, named acetoacetyl-CoA thiolase, also known as acetyl-CoA acetyltransferase (ACAT). According to many reports, the expression of ACAT1 in tumor cells is usually abnormal, and it has been shown to play an extremely important role in the occurrence and development of tumors. The mechanism of the increased activity of ACAT1 in different human cancer cells is worthy of research. The high expression of ACAT1 reduces its overall survival rate, and the recycling of ketone bodies by the overexpression of ACAT1 in MDA-MB-231 human breast cancer cells drives tumor progression and metastasis. Therefore, research on biphenyl compounds targeting ACAT1 may lead to the discovery of new antitumor drugs.

[0018] Biphenyl compounds have been confirmed to have the above-mentioned broad active effects, but their exact mechanism of action and structure-activity relationship need further study. It is necessary to synthesize their structural derivatives and study the mode of action of the drugs and the diseases that can be treated, especially inflammation-related, tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases, and aging.

Summary of the Invention

Problems to be Solved by the Invention

[0019] This application provides biphenyl compounds, pharmaceutical compositions, and their preparation methods and uses. The biphenyl compounds, their isomers, pharmaceutically acceptable salts, prodrugs, polymorphs or co-crystals according to this application can be used in the preparation of drugs or anti-aging drugs for preventing and / or treating tumors, autoimmune diseases, inflammatory diseases, neurodegenerative diseases, and metabolic resistance diseases.

[0020] In a first aspect, this application provides a biphenyl compound having a structure represented by the following formula I.

Chemical Formula

[0021] (However, each of R1 and R2 is independently hydrogen, halogen, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkoxycarbonyl group, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkyl group, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkoxy group, trihalo C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkyl group, trihalo C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkoxy group, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkoxymethyleneoxy group, methoxy C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, and is any one selected therefrom, n is selected from 0, 1, 2, 3 or 4, X is selected from CH2, O or NH, Y is CH2, O, OH, NH, NH2, NH(CH2) n’ NH2, NH(CH2) n’ NHCOCF3, OBn, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkyl group, C1-C8 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, C8, etc.) alkoxy group, trihalomethylamine group, dimethylamine group, methylamine group or trihaloalkoxy group, and is any one selected therefrom, provided that n' is selected from 1, 2, 3, 4 or 5, M is any one selected from CH2, O, OH, NH, NH2, trihalomethylamine group, trihalomethylacyl group, dimethylamino group, methylamine group, carboxyl group, trihaloalkyl group or C1-C8 alkoxymethyleneoxy group, and either M and Y are linked by a bond, or M and Y do not form a bond.)

[0022] In the present application, the "trihalo" in the trihalo C1-C8 alkyl group and the trihalo C1-C8 alkoxy group means containing three halogens. Among them, the trihalo C1-C8 alkyl group includes F3C1-C8 alkyl group, Br3C1-C8 alkyl group, Cl3C1-C8 alkyl group, I3C1-C8 alkyl group, F2Br C1-C8 alkyl group, F2Cl C1-C8 alkyl group, F2I C1-C8 alkyl group, FBr2C1-C8 alkyl group, FCl2C1-C8 alkyl group, FI2C1-C8 alkyl group, ClBr2C1-C8 alkyl group, ICl2C1-C8 alkyl group and ClI2C1-C8 alkyl group. The trihalo C1-C8 alkoxy group includes F3C1-C8 alkoxy group, Br3C1-C8 alkoxy group, Cl3C1-C8 alkoxy group, I3C1-C8 alkoxy group, F2Br C1-C8 alkoxy group, F2Cl C1-C8 alkoxy group, F2I C1-C8 alkoxy group, FBr2C1-C8 a lkoxy group, FCl2C1-C8 alkoxy group, FI2C1-C8 alkyl group, ClBr2C1-C8 alkoxy group, ICl2C1-C8 alkoxy group and ClI2C1-C8 alkoxy group. Preferably, the trihalo C1-C8 alkyl group includes an F3C1-C4 alkyl group, a Br3C1-C4 alkyl group, a Cl3C1-C4 alkyl group, an I3C1-C4 alkyl group, an F2Br C1-C4 alkyl group, an F2Cl C1-C4 alkyl group, an F2I C1-C4 alkyl group, an FBr2C1-C4 alkyl group, an FCl2C1-C4 alkyl group, an FI2C1-C4 alkyl group, a ClBr2C1-C4 alkyl group, an ICl2C1-C4 alkyl group, and a ClI2C1-C4 alkyl group, and the trihalo C1-C8 alkoxy group includes an F3C1-C4 alkoxy group, a Br3C1-C4 alkoxy group, a Cl3C1-C4 alkoxy group, an I3C1-C4 alkoxy group, an F2Br C1-C4 alkoxy group, an F2Cl C1-C4 alkoxy group, an F2I C1-C4 alkoxy group, an FBr2C1-C4 alkoxy group, an FCl2C1-C4 alkoxy group, an FI2C1-C4 alkyl group, a ClBr2C1-C4 alkoxy group, an ICl2C1-C4 alkoxy group, and a ClI2C1-C4 alkoxy group.

[0023] In the present application, "trihalo" in the trihalomethylamine group, trihalomethylacyl group, and trihaloalkyl group means containing three halogens.

[0024] In the present application, "C1-C8" in the methoxy C1-C8 methyleneoxy group means that the repeating unit of the methylene group is 1-8 (for example, it may be 1, 2, 3, 4, 5, 6, 7, 8, etc.).

[0025] In the present application, R1 and R2 are each independently hydrogen, fluorine, chlorine, bromine, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkoxycarbonyl group, C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkyl group, C1-C7 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, etc.) alkoxy group, trihalo C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkyl group, trihalo C1-C7 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, etc.) alkoxy group, C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkoxymethyleneoxy group, methoxy C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, and is any one selected therefrom, n is selected from 1, 2, 3 or 4, X is selected from CH2, O or NH, Y is CH2, O, OH, NH, NH2, NH(CH2) n’ NH2, NH(CH2) n’ NHCOCF3, OBn, C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkyl group, C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkoxy group, trihalomethylamine group, dimethylamine group, methylamine group or trihaloalkoxy group, and is any one selected therefrom, provided that n' is selected from 2, 3, 4 or 5, M is any one selected from CH2, O, OH, NH, NH2, trihalomethylamine group, trihalomethylacyl group, dimethylamine group, methylamine group, carboxyl group, trihaloalkyl group or C1-C6 (which may be, for example, C1, C2, C3, C4, C5, C6, etc.) alkoxymethyleneoxy group, and either M and Y are linked by a bond, or M and Y do not form a bond.

[0026] In the present application, R1 and R2 are each independently hydrogen, fluorine, chlorine, bromine, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C4 (which may be, for example, C1, C2, C3, C4, etc.) alkoxycarbonyl group, C1-C4 (which may be, for example, C1, C2, C3, C4, etc.) alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C4 (which may be, for example, C1, C2, C3, C4, etc.) alkyl group, C1-C7 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, etc.) alkoxy group, trihalo C1-C4 (which may be, for example, C1, C2, C3, C4, etc.) alkyl group, trihalo C1-C7 (which may be, for example, C1, C2, C3, C4, C5, C6, C7, etc.) alkoxy group, C1-C4 (which may be, for example, C1, C2, C3, C4, etc.) alkoxymethyleneoxy group, methoxy C1-C4 (which may be, for example, C1, C2, C3, C4, etc.) methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, and are each any one selected therefrom. n is selected from 1, 2, 3 or 4, X is selected from CH2, O or NH, Y is CH2, O, OH, NH, NH2, NH(CH2) n’ NH2, NH(CH2) n’NHCOCF3, Obn, a C1-C4 (which may be, for example, C1, C2, C3, or C4) alkyl group, a C1-C4 (which may be, for example, C1, C2, C3, or C4) alkoxy group, a trihalomethylamine group, a dimethylamine group, a methylamine group, or a trihaloalkoxy group, provided that n’ is selected from 2, 3, 4, or 5, M is one selected from CH2, O, OH, NH, NH2, a trihalomethylamine group, a trihalomethylacyl group, a dimethylamine group, a methylamine group, a carboxyl group, a trihaloalkyl group, or a C1-C4 alkoxymethyleneoxy group, and M and Y are linked by a bond or M and Y do not form a bond.

[0027] In the present application, R1 and R2 are each independently hydrogen, fluorine, chlorine, bromine, a hydroxy group, a dimethylamine group, a cyano group, a nitro group, a methoxycarbonyl group, an ethoxycarbonyl group, a methylcarbonyloxy group, an ethoxycarbonyl group, a methylamine group, a methylsulfonyl group, a dimethylsulfamoyl group, an amino group, a carboxyl group, a methyl group, an ethyl group, a methoxy group, an ethoxy group, a propoxy group, a 1-butanoxy group, a 1-pentanoxy group, a 1-hexanoxy group, a 1-heptanoxy group, a 2-pentanoxy group, a 2-hexanoxy group, a trifluoromethyl group, a trifluoroethyl group, a trifluoromethoxy group, a methoxymethyleneoxy group, a methoxyethyloxy group, a methoxypropyloxy group, a methoxyn-butoxy group, a cyclopentyloxy group, or a cyclohexyloxy group, n is selected from 1, 2, 3, or 4, X is selected from CH2, O, or NH, Y is CH2, O, OH, NH, NH2, NH(CH2) n’ NH2, NH(CH2) n’It is any one selected from NHCOCF3, Obn, methyl group, ethyl group, methoxy group, ethoxy group, trifluoromethylamine group, trichloromethylamine group, dimethylamine group, methylamine group or trifluoromethyloxy group, provided that n’ is selected from 2, 3, 4 or 5, M is any one selected from CH2, O, OH, NH, NH2, trifluoromethylamine group, trichloromethylamine group, trifluoromethylacyl group, trichloromethylacyl group, dimethylamine group, methylamine group, carboxyl group, trifluoromethyl group or methoxymethyleneoxy group, and M and Y are linked by a bond or M and Y do not form a bond.

[0028] In the present application, the compound is any one selected from the structures represented by the following formulas 1 to 20.

Chemical formula

[0029] As a second aspect, the present application provides an isomer of the biphenyl-based compound described in the first aspect or a pharmaceutically acceptable salt thereof.

[0030] Preferably, the pharmaceutically acceptable salt is hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, p-toluenesulfonate, acetate, trifluoroacetate, salicylate, amino acid salt, 2-O-β-D-glucopyranosyl-L-ascorbate, maleate, tartrate, fumarate, citrate, lactate, sodium salt, potassium salt, calcium salt, magnesium salt, lithium salt, ammonium salt, or a salt of an organic base capable of providing a physiologically acceptable cation, including any one or a combination of at least two of them.

[0031] Preferably, the salt of the organic base capable of providing the physiologically acceptable cation includes any one or at least a combination of two of methylamine salt, dimethylamine salt, trimethylamine salt, piperidine salt, morpholine salt, or tris(2-hydroxyethyl)amine salt.

[0032] All salts according to the present application can be prepared by common methods. In addition, during the preparation process of the compound solvate represented by formula (I) and its salts, polycrystals or co-crystals may appear under different crystal conditions.

[0033] In the present application, the term "halogen" means fluorine, chlorine, bromine, or iodine.

[0034] The present application includes all possible stereoisomers and mixtures of two or more isomers.

[0035] As a third aspect, the present application relates to a method for preparing a biphenyl-based compound according to the first aspect, wherein the preparation method includes the following steps: Synthesis method 1: (A) A nucleophilic addition reaction is carried out on the compound represented by formula II, the compound represented by formula III, and the compound represented by formula IV to produce a compound represented by formula V, and the reaction formula is as follows:

[0036]

Chemical formula

[0037] (The nucleophilic addition reaction is carried out in an organic solvent, the organic solvent includes any one or at least a combination of two of tetrahydrofuran, diethyl ether, toluene, or benzene, the temperature of the nucleophilic addition reaction is 0 to 30 °C (for example, it may be 0 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, etc.), and the time is 1 to 10 h (for example, it may be 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, etc.).) (B) React the compound represented by formula V with iodine in an iodination reaction to produce a compound represented by formula VI, and the reaction formula is as follows:

[0038] [Chemical formula]

[0039] (The iodination reaction is carried out under the participation of a fluorine reagent, and the temperature of the iodination reaction is 0 to 3 0 °C (for example, it may also be 0 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, etc.), and the time is 1 to 100 h (for example, it may also be 1 h, 10 h, 20 h, 30 h, 40 h, 50 h, 60 h, 70 h, 80 h, 90 h, 100 h, etc.).) (C) React the compound represented by formula VI by a coupling reaction under alkaline conditions and in the presence of zero-valent palladium to produce a compound represented by formula I, and the reaction formula is as follows:

[0040] [Chemical formula]

[0041] (The coupling reaction is carried out in the presence of an organic alkali and / or an inorganic alkali. The organic alkali includes any one or at least two combinations of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine or N,N - diisopropylethylamine. The inorganic alkali includes any one or at least two combinations of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate or potassium carbonate. The coupling reaction is carried out in a solvent. The solvent includes any one or at least two combinations of dimethyl sulfoxide, N,N - dimethylformamide or dioxane. The temperature of the coupling reaction is 30 - 120 °C (for example, it may be 30 °C, 50 °C, 70 °C, 90 °C, 100 °C, 110 °C, 120 °C, etc.), and the time is 3 - 20 h (for example, it may be 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, 17 h, 20 h, etc.).) (However, R1 and R2 are each independently selected from hydrogen, halogen, dimethylamine group, cyano group, nitro group, C1 - C8 alkoxycarbonyl group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1 - C8 alkyl group or trihalo C1 - C8 alkyl group. n is selected from 0, 1, 2, 3 or 4. X, Y, M are each independently selected from CH2.) Alternatively, synthesis method 2: ((D) Nucleophilic addition reaction is carried out between the compound represented by formula II’ and the compound represented by formula III’ to produce the compound represented by formula IV’, or nucleophilic addition reaction is carried out between the compound represented by formula II’ and the compound represented by formula II to produce the compound represented by formula IV’’, or nucleophilic addition reaction is carried out between the compound represented by formula II’ and the compound represented by formula III to produce the compound represented by formula IV’’’. The reaction formula is as follows,

[0042]

Chemical formula

[0043] (The nucleophilic addition reaction is carried out in an organic solvent, and the organic solvent includes any one or at least a combination of two of tetrahydrofuran, diethyl ether, toluene or benzene. The temperature of the nucleophilic addition reaction is 0 - 30 °C (for example, it may be 0 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, etc.), and the time is 1 - 10 h (for example, it may be 1 h, 3 h, 5 h, 7 h, 9 h, 10 h, etc.).) (E) Generate a compound represented by formula V' from the compound represented by formula IV' by an iodination reaction, or generate a compound represented by formula V'' from the compound represented by formula IV'' by an iodination reaction, or generate a compound represented by formula V''' from the compound represented by formula IV''' by an iodination reaction. The reaction formula is as follows,

[0044]

Chemical formula

[0045] (The iodination reaction is carried out under the participation of a fluorine reagent. The temperature of the iodination reaction is 0 - 30 °C (for example, it may be 0 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, etc.), and the time is 1 - 100 h (for example, it may be 1 h, 10 h, 20 h, 30 h, 40 h, 50 h, 60 h, 70 h, 80 h, 90 h, 100 h, etc.).) (F) After subjecting the compound represented by formula V' to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium, hydrolyze it under Lewis acid conditions to generate a compound represented by formula VI', or subject the compound represented by formula V'' to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium, and then hydrolyze it under Lewis acid conditions to generate a compound represented by formula VI'', or subject the compound represented by formula V''' to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium, and then hydrolyze it under Lewis acid conditions to generate a compound represented by formula VI'''. The reaction formula is as follows,

[0046] [Chemistry]

[0047] (The coupling reaction is carried out in the presence of an organic base and / or an inorganic base. The organic base includes any one or at least two combinations of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine, or N,N-diisopropylethylamine. The inorganic base includes any one or at least two combinations of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate, or potassium carbonate. The coupling reaction is carried out in a solvent, and the solvent includes any one or at least two combinations of dimethyl sulfoxide, N,N-dimethylformamide, or dioxane.) (The coupling reaction is carried out at a temperature of 30 to 120 °C (for example, it may be 30 °C, 50 °C, 70 °C, 90 °C, 100 °C, 110 °C, 120 °C, etc.), and the time is 3 to 20 h (for example, it may be 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, 17 h, 20 h, etc.).) (The Lewis acid includes boron tribromide and / or aluminum trichloride. The temperature of the hydrolysis is -70 to -40 °C (for example, it may be -70 °C, -60 °C, -50 °C, -40 °C, etc.), and the time is 1 to 10 h (for example, it may be 1 h, 3 h, 5 h, 7 h, 9 h, 10 h, etc.).) ((G) Reacting the compound represented by formula VI’ with an alkyl halide compound to produce the compound represented by formula I, or reacting the compound represented by formula VI’’ with an alkyl halide compound to obtain the compound represented by formula I, or reacting the compound represented by formula VI’’’ with an alkyl halide compound to obtain the compound represented by formula I. The alkyl halide compound includes alkyl bromide or alkyl chloride, and the reaction formula is as follows, including

[0048] [Chemistry]

[0049] (The alkylation reaction is carried out in the presence of an organic base and / or an inorganic base. The organic base includes any one or at least two combinations of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine, or N,N-diisopropylethylamine. The inorganic base includes any one or at least two combinations of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate, or potassium carbonate. The temperature of the alkylation reaction is 0 to 30 °C (for example, it may be 0 °C, 5 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, etc.), and the time is 1 to 10 h (for example, it may be 1 h, 3 h, 5 h, 7 h, 9 h, 10 h, etc.).) (However, R1 and R2 are each independently selected from hydrogen, halogen, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C8 alkoxycarbonyl group, C1-C8 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 alkyl group, C1-C8 alkoxy group, trihalo C1-C8 alkyl group, trihalo C1-C8 alkoxy group, C1-C8 alkoxymethyleneoxy group, methoxy C1-C8 methyleneoxy group, cyclopentyloxy group, or cyclohexyloxy group, and n is selected from 0, 1, 2, 3, or 4, and X, Y, M are each independently selected from CH2.) Alternatively, Synthetic Method 3: (H) The compound represented by formula VII is subjected to a hydrolysis reaction with an alkali to generate the compound represented by formula VIII. The alkali includes an organic base and / or an inorganic base, and the reaction formula is as follows:

[0050]

Chemical formula

[0051] (The organic alkali includes any one or at least a combination of two or more of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine or N,N - diisopropylethylamine; the inorganic alkali includes any one or at least a combination of two or more of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate or potassium carbonate; the temperature of the hydrolysis reaction is 10 - 100 °C (for example, it may be 10 °C, 30 °C, 50 °C, 70 °C, 90 °C, 100 °C, etc.), the time is 1 - 40 h (for example, it may be 1 h, 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, 17 h, 20 h, 25 h, 30 h, 35 h, 40 h, etc.), the hydrolysis reaction is carried out in the presence of a solvent, and the solvent includes any one or at least a combination of two or more of tetrahydrofuran, methanol, ethanol or water.) (I) The compound represented by formula VIII and the compound represented by formula IX are subjected to a condensation reaction under alkaline conditions to produce a compound represented by formula X, and the reaction formula is as follows:

[0052]

Chemical formula

[0053] (The condensation reaction is carried out in the presence of an organic base and / or an inorganic base, the organic base includes any one or at least a combination of two of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine or N,N-diisopropylethylamine, the inorganic base includes any one or at least a combination of two of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate or potassium carbonate, the temperature of the condensation reaction is 10-30 °C (for example, it may be 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, etc.), the time is 1-40 h (for example, it may be 1 h, 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, 17 h, 20 h, 25 h, 30 h, 35 h, 40 h, etc.), the condensation reaction is carried out in the presence of a solvent, the solvent includes any one or at least a combination of two of dichloromethane, tetrahydrofuran or N,N-dimethylformamide, the condensation reaction is carried out in the presence of a condensing agent, and the condensing agent includes any one or at least a combination of two selected from carbodiimide hydrochloride (EDCI), dicyclohexylcarbodiimide (DCC), 2-(7-azabenzotriazole)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (HATU) or 1-hydroxybenzotriazole (HOBt).) (J) The compound represented by formula X is subjected to a coupling reaction to produce the compound represented by formula XI, and the reaction formula is as follows,

[0054]

Chemical formula

[0055] (The coupling reaction is carried out in the presence of an organic alkali and / or an inorganic alkali, wherein the organic alkali includes any one or at least a combination of two of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine or N,N-diisopropylethylamine, and the inorganic alkali includes any one or at least a combination of two of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate or potassium carbonate. The temperature of the coupling reaction is 30-120 °C (for example, it may be 30 °C, 50 °C, 70 °C, 90 °C, 100 °C, 110 °C, 120 °C, etc.), and the time is 3-20 h (for example, it may be 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, 17 h, 20 h, etc.). The coupling reaction is carried out in the presence of a solvent, and the solvent includes any one or at least a combination of two of dimethyl sulfoxide, N,N-dimethylformamide, dioxane or ethylene glycol dimethyl ether.) The compound represented by the formula X and bis(pinacolato)diboron are subjected to a coupling reaction.) (K) The compound represented by the formula XI and the compound represented by the formula XII are subjected to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium to produce the compound represented by the formula I, or the compound represented by the formula XI and the compound represented by the formula XII are subjected to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium. After the reaction, it is hydrolyzed with an organic alkali or an inorganic alkali, and then stirred with hydrochloric acid or trifluoroacetic acid to obtain an amino hydrochloride. After separation, it is reacted with an acyl reagent to produce the compound represented by the formula I, and the reaction formula is as follows,

[0056]

Chemical formula

[0057] (The coupling reaction is carried out in the presence of an organic alkali and / or an inorganic alkali, the organic alkali includes any one or at least a combination of two of pyridine, piperidine, diisopropylamine, ethylenediamine, triethylamine or N,N-diisopropylethylamine, the inorganic alkali includes any one or at least a combination of two of sodium hydroxide, potassium hydroxide, lithium hydroxide, cesium carbonate or potassium carbonate, the temperature of the coupling reaction is 30~120 °C (for example, it may be 30 °C, 50 °C, 70 °C, 90 °C, 100 °C, 110 °C, 120 °C, etc.), the time is 3~20 h (for example, it may be 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, 17 h, 20 h, etc.), and the coupling reaction is carried out in the presence of a solvent and the solvent includes any one or at least a combination of two of dimethyl sulfoxide, N,N-dimethylformamide, dioxane or ethylene glycol dimethyl ether. (The concentration of the hydrochloric acid or trifluoroacetic acid is 0.8~1.2 N (for example, it may be 0.8 N, 0.9 N, 1 N, 1.1 N, 1.2 N, etc.).) (However, R1 and R2 are each independently selected from hydrogen, halogen, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C8 alkoxycarbonyl group, C1-C8 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 alkyl group, C1-C8 alkoxy group, trihalo C1-C8 alkyl group, trihalo C1-C8 alkoxy group, C1-C8 alkoxymethyleneoxy group, methoxy C1-C8 methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, n is selected from 0, 1, 2, 3 or 4, X is selected from CH2, O or NH, X' is selected from OH or NH2, and Y is CH2, O, OH, NH, NH2, NH(CH2) n’ NH2, NH(CH2) n’ NHCOCF3, Obn, C 1~8 alkyl group, C 1~8It is any one selected from an alkoxy group, a trihalomethylamine group, a dimethylamine group, a methylamine group or a trihaloalkoxy group, provided that n' is selected from 1, 2, 3, 4 or 5, M is any one selected from CH2, O, OH, NH, NH2, a trihalomethylamine group, a trihalomethylacyl group, a dimethylamino group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C8 alkoxymethyleneoxy group, and M' is any one selected from OH, NH2, a trihalomethylamine group, a trihalomethylacyl group, a dimethylamino group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C8 alkoxymethyleneoxy group.) A preparation method is provided.

[0058] As a fourth aspect, the present application is a pharmaceutical composition containing an active ingredient and a pharmacologically acceptable vector, The active ingredient contains the biphenyl-based compound described in the first aspect and / or an isomer of the biphenyl-based compound described in the second aspect or a pharmaceutically acceptable salt thereof, A pharmaceutical composition is provided.

[0059] Preferably, the active ingredient in the pharmaceutical composition is 0.1 to 95% by mass.

[0060] The pharmaceutical composition according to the present application can be prepared based on methods known in the art, and by combining the active ingredient with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants, it can be made into any dosage form suitable for use in humans or animals.

[0061] The biphenyl-based compound according to the first aspect of the present application, an isomer of the biphenyl-based compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect can be administered in the form of a unit dose, and the administration route may be enteral or parenteral, for example, oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0062] The dosage form may be a liquid dosage form, a solid dosage form or a semi-solid dosage form. The liquid dosage form may be a solution (including true solution and colloidal solution), an emulsion (including o / w type, w / o type and complex emulsion), a suspending agent, an injection (including aqueous injection, powder injection and infusion), eye drops, nasal drops, lotion and liniment, etc. The solid dosage form may be a tablet (including ordinary tablet, enteric-coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, enteric-coated capsule), granule, powder, pellet, drop, suppository, film, patch, (powder) aerosol, spray, etc. The semi-solid dosage form may be an ointment, gel, paste, etc.

[0063] The biphenyl-based compound according to the first aspect of the present application, the isomer of the biphenyl-based compound according to the second aspect or its pharmaceutically acceptable salt, or the pharmaceutical composition according to the fourth aspect may be prepared into a general preparation, or a sustained-release preparation, a release control preparation, a target preparation and each microparticle administration system.

[0064] To prepare the biphenyl compound according to the first aspect of the present application, an isomer of the biphenyl compound according to the second aspect, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect into tablets, various excipients known in the art can be widely used, including diluents, adhesives, wetting agents, disintegrants, lubricants, and glidants. The diluent may be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc. The wetting agent may be water, ethanol, isopropyl alcohol, etc. The adhesive may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic slurry, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone, polyethylene glycol, etc. The disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium hydrogen carbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfonate, etc. The lubricant and glidant may be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0065] Furthermore, the tablets may be made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double tablets and multilayer tablets.

[0066] To prepare the biphenyl compound according to the first aspect of the present application, the isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect into a capsule preparation, these may be mixed with a diluent and a glidant, and the mixture may be directly placed into a hard capsule or a soft capsule. The compound of the present application as an active ingredient may be made into granules or pellets with a diluent, an adhesive, and a disintegrant, and then placed into a hard capsule or a soft capsule. The varieties of each diluent, adhesive, wetting agent, disintegrant, and glidant for preparing tablets can also be used for preparing capsule preparations.

[0067] To prepare the biphenyl compound according to the first aspect of the present application, the isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect into an injection preparation, water, ethanol, isopropyl alcohol, propylene glycol or a mixture thereof may be used as a solvent, and an appropriate amount of solubilizers, solubilizing aids, pH adjusters, and osmotic pressure adjusters commonly used in this field may be added. The solubilizer or solubilizing aid may be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc. The pH adjuster may be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc. The osmotic pressure adjuster may be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, when preparing a freeze-dried powder injection preparation, mannitol, glucose, etc. may be added as a support agent.

[0068] Also, if necessary, a coloring agent, a preservative, a fragrance, a flavoring agent or other additives may be added to the pharmaceutical preparation may be added.

[0069] To achieve the purpose of drug administration and enhance the therapeutic effect, the drug or pharmaceutical composition of the present application can be administered by any known administration method.

[0070] As a fifth aspect, the present application provides the use of the biphenyl compound according to the first aspect, an isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect, in the preparation of a drug or an anti-aging agent for preventing and / or treating tumors, autoimmune diseases, inflammatory diseases, neurodegenerative diseases, and metabolic resistance diseases.

[0071] The target of action of the biphenyl compound according to the first aspect, the isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect of the present application is an animal, preferably a mammal, and more preferably a human.

[0072] In the present application, the tumor is any one or at least a combination of two or more selected from glioma, melanoma, gastric cancer, lung cancer, breast cancer, renal cancer, liver cancer, oral epithelial cancer, head and neck tumors, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, rectal adenocarcinoma, leukemia, or lymphoma.

[0073] Preferably, the autoimmune disease includes any one or at least a combination of two or more of rheumatoid arthritis, systemic lupus erythematosus, ulcerative colitis, psoriasis, dermatitis, or amyotrophic lateral sclerosis.

[0074] Preferably, the inflammatory disease includes any one or at least a combination of two or more of polyarteritis, phlebitis, or reflux esophagitis.

[0075] Preferably, the neurodegenerative disease includes senile dementia and / or Parkinson's disease.

[0076] Preferably, the metabolic disease includes diabetes, hyperuricemia or gout.

[0077] The dosage of the biphenyl compound according to the first aspect of the present application, the isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect can vary significantly depending on the nature and severity of the disease to be prevented or treated, the individual situation of the patient or animal, the administration route, dosage form, etc. Generally, the appropriate daily dosage range is 0.001 to 150 mg / Kg body weight, preferably 0.1 to 100 mg / Kg body weight, more preferably 1 to 70 mg / Kg body weight, and most preferably 2 to 30 mg / Kg body weight. The above dosage can be administered in one dosage unit or divided into several dosage units, and depends on the administration plan including the clinical experience of the doctor and the use of other treatment means.

[0078] The biphenyl compound according to the first aspect of the present application, the isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect may be taken alone or in combination with other therapeutic agents or symptomatic agents. When the biphenyl compound according to the first aspect of the present application, the isomer of the biphenyl compound according to the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect has a synergistic effect with other therapeutic agents, its dosage should be adjusted according to the actual situation.

Advantages of the Invention

[0079] Compared with the prior art, the present application has the following beneficial effects.

[0080] The compound according to the present application is a novel biphenyl derivative, such a compound can bind well to a plurality of target point proteins, and has good bioavailability, and can be used for the preparation of drugs or anti-aging drugs for preventing or treating tumors, autoimmune diseases, inflammatory diseases, neurodegenerative diseases. Among them, the tumor diseases are glioma, melanoma, gastric cancer, lung cancer, breast cancer, renal cancer, liver cancer, oral squamous cell carcinoma, head and neck tumors, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, rectal adenocarcinoma, leukemia, or lymphoma, etc.

Brief Description of the Drawings

[0081]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Mode for Carrying Out the Invention

[0082] Hereinafter, the technical solution of the present application will be further described with specific embodiments. Those skilled in the art should understand that the above examples are only for understanding the present application and should not be regarded as specifically limiting the present application.

[0083] Preparation Example 1 This preparation example provides compound 21, and the preparation method of the compound 21 includes the following steps.

[0084] Step A or D:

Chemical Formula

[0085] At 0 °C, under the protection of argon gas, pentane-1,5-di(magnesium bromide) (0.5 M) (100 mL) was added dropwise to a solution of N,4-dimethoxy-N-methylbenzamide (19.5 g, 100 mmol) in tetrahydrofuran (100 mL). The reaction mixture was stirred at room temperature for 3 hours, and then quenched by adding saturated ammonium chloride solution (100 mL) dropwise thereto. The mixture was extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the filtered solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:ethyl acetate = 10:1) to give a white solid 21 (8.4 g) (yield 50%).

[0086] 1 1H NMR (400 MHz, CDCl3) δ 7.97 - 7.90 (m, 4H), 6.96 - 6.90 (m, 4H), 3.87 (d, J = 1.9 Hz, 6H), 2.97 - 2.87 (m, 4H), 1.79 (dt, J = 20.7, 7.5 Hz, 4H), 1.53 - 1.41 (m, 2H).

[0087] Preparation Example 2 This preparation example provides compound 22, and the preparation method of the compound 22 includes the following steps as follows.

[0088] Step B or E:

Chemical Structure

[0089] At room temperature, 1,7-bis(4-methoxyphenyl)heptane-1,7-dione (23.84 g, 70 mmol) and a fluorine reagent (24.83 g, 70 mmol) were dissolved in acetonitrile (500 mL), and iodine (17.8 g, 70 mmol) was added to the solution. The mixture was stirred at room temperature for 3 days. The filtered solid was washed with water and ethanol respectively to obtain a white solid (30 g, yield 70%).

[0090] 11H NMR (400 MHz, CDCl3) δ 8.39 (d, J = 2.1 Hz, 2H), 7.95 (dd, J = 8.6, 2.2 Hz, 2H), 6.84 (d, J = 8.6 Hz, 2H), 3.95 (s, 6H), 2.92 (t, J = 7.3 Hz, 4H), 1.78 (dt, J = 15.0, 7.4 Hz, 4H), 1.47 (tt, J = 9.6, 6.4 Hz, 2H).

[0091] Preparation Example 3 This preparation example provides compound 24, and the preparation method of the compound 24 includes the following steps. Step H: [Chemical formula]

[0092] Methyl 3-bromo-4-hexyloxybenzoate 23 (2 g, 6.35 mmol) and sodium hydroxide (0.51 g, 12.7 mmol) were respectively added to a mixed solution of tetrahydrofuran / methanol / water (8 mL / 4 mL / 6 mL), and stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure, water (30 mL) was added, the pH value was adjusted to 5 with hydrochloric acid, and filtered to obtain a white solid (1.55 g) (yield 81%).

[0093] 1 1H NMR (400 MHz, MeOD-d4) δ 8.15 (d, J = 2.1 Hz, 1H), 7.96 (dd, J = 8.6, 2.1 Hz, 1H), 7.07 (d, J = 8.7 Hz, 1H), 4.12 (t, J = 6.3 Hz, 2H), 1.83 (dq, J = 12.5, 6.3 Hz, 2H), 1.62 - 1.47 (m, 2H), 1.44 - 1.29 (m, 4H), 1.01 - 0.85 (m, 3H).

[0094] Preparation Example 4 This preparation example provides compound 25, and the preparation method of the compound 25 includes the following steps.

[0095] Step I: [Chemical formula]

[0096] Respectively, 3-bromo-4-hexyloxybenzoic acid 24 (0.77 g, 2.57 mmol), N-boc-propanediamine (0.58 g, 3.34 mmol), triethylamine (1.43 mL, 10.27 mmol) and propylphosphonic anhydride (30%) (4.9 g, 7.7 mmol) were added to anhydrous dichloromethane (15 mL), and the mixture was stirred at room temperature for 16 hours. Water (30 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (30 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The obtained crude product was purified by column chromatography (dichloromethane:methanol = 100:1 → 50:1) and separated to obtain a yellow oily substance 25 (990 mg) (yield 84%).

[0097] Preparation Example 5 This preparation example provides compound 26, and the preparation method of the compound 26 includes the following steps.

Chemical formula

[0098] Respectively, 3-bromo-4-hexyloxybenzoic acid 24 (0.77 g, 2.57 mmol), N-boc-butanediamine (0.63 g, 3.34 mmol), triethylamine (1.43 mL, 10.27 mmol) and propylphosphonic anhydride (30%) (4.9 g, 7.7 mmol) were added to anhydrous dichloromethane (15 mL) and stirred at room temperature for 16 hours. Water (30 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (30 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The obtained crude product was purified by column chromatography (dichloromethane:methanol = 100:1 → 50:1) and separated to obtain a brown solid (1.1 g) (yield 91%).

[0099] Preparation Example 6 This preparation example provides compound 27, and the preparation method of the compound 27 includes the following steps. [Chemical formula]

[0100] Respectively, 3-bromo-4-hexyloxybenzoic acid 24 (0.9 g, 3 mmol), N-boc-pentanediamine (0.8 g, 4 mmol), triethylamine (1.39 mL, 10 mmol) and propylphosphonic anhydride (30%) (5.7 g, 9 mmol) were added to anhydrous dichloromethane (30 mL), and the mixture was stirred at room temperature for 16 hours. To the reaction solution water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The obtained crude product was purified by column chromatography (dichloromethane:methanol = 100:1 → 50:1) and separated to obtain a white solid (1.22 g) (yield 84%).

[0101] Preparation Example 7 This preparation example provides Compound 28, and the preparation method of the Compound 28 includes the following steps. [Chemical formula]

[0102] Respectively, 3-bromo-4-hexyloxybenzoic acid (0.9 g, 3 mmol), N-boc-pentanediamine (0.64 g, 4 mmol), triethylamine (1.39 mL, 10 mmol) and propylphosphonic anhydride (30%) (5.7 g, 9 mmol) were added to anhydrous dichloromethane (30 mL), and the mixture was stirred at room temperature for 16 hours. To the reaction solution, water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The obtained crude product was purified by column chromatography (dichloromethane:methanol = 100:1 → 50:1) and separated to obtain a white solid (1.07 g) (yield 81%).

[0103] Preparation Example 8 This preparation example provides compound 29, and the preparation method of the compound 29 includes the following steps. Step J:

Chemical formula

[0104] Methyl 3-bromo-4-hexyloxybenzoate 23 (2.5 g, 7.94 mmol), pinacol boronate (2.42 g, 9.52 mmol), and potassium acetate (2.34 g, 23.8 mmol) were each added to 1,4-dioxane (50 mL). After degassing with argon gas, dichloropalladium (II) 1,1'-bis(diphenylphosphino)ferrocene (0.58 g, 0.79 mmol) was added, and the reaction mixture was stirred at 90 °C for 16 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The resulting crude product was purified by column chromatography (ethyl acetate: petroleum ether = 1:80 → 1:40) to obtain a white solid (1.37 g) (yield 48%).

[0105] 1 H NMR (400 MHz, MeOD-d4) δ 8.23 (d, J = 2.2 Hz, 1H), 8.05 (dd, J = 8.7, 2.3 Hz, 1H), 6.97 (d, J = 8.8 Hz, 1H), 4.14 - 4.06 (m, 2H), 3.87 (s, 3H), 1.85 - 1.74 (m, 2H), 1.64 - 1.51 (m, 2H), 1.44 - 1.32 (m, 16H ), 1.28 - 1.20 (m, 3H).

[0106] Preparation Example 9 This preparation example provides compound 30, and the preparation method of the compound 30 includes the following steps. Step K: Coupling reaction

Chemical formula

[0107] Respectively, t-butyl (3-(3-bromo-4-hexyloxy)benzamide)propylamine carbonate (0.9 g, 1.97 mmol), methyl 4-hexyloxy-3-(pinacolboronate)benzoate (0.86 g, 2.37 mmol) and potassium carbonate (0.82 g, 5.92 mmol) were added to 1,4-dioxane (30 mL), degassed with argon gas, then 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium (0.14 g, 0.2 mmol) was added, and the reaction solution was stirred at 90 °C for 16 hours. Water (50 mL) was added to the reaction solution, extracted with ethyl acetate (50 mL × 3), backwashed with saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate and then concentrated, and the obtained crude product was purified by column chromatography (ethyl acetate:dichloromethane = 1:40 → 1:5) to separate and obtain 30 (1.15 g) of a white solid (yield 95%). [M+H] + :613.4

[0108] Preparation Example 10 This preparation example provides Compound 31, and the preparation method of the said Compound 31 includes the following steps. [Chemical formula]

[0109] Respectively, t-butyl (3-(3-bromo-4-hexyloxy)benzamide)butylamine carbonate 26 (0.78 g, 1.66 mmol), methyl 4-hexyloxy-3-(pinacolboronate)benzoate (0.5 g, 1.38 mmol) and Potassium carbonate (0.57 g, 4.14 mmol) was added to 1,4-dioxane (20 mL), and after degassing with argon gas, 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium (0.1 g, 0.14 mmol) was added. The reaction mixture was stirred at 90 °C for 16 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL × 3), backwashed with a saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The resulting crude product was purified by column chromatography (ethyl acetate:dichloromethane = 1:40 → 1:5) to give 31 (0.81 g) of a white solid (yield 94%). 1 1H NMR (400 MHz, MeOD-d4) δ 7.98 (dd, J = 8.7, 2.3 Hz, 1H), 7.82 (dt, J = 7.8, 1.7 Hz, 2H), 7.67 (d, J = 2.4 Hz, 1H), 7.04 (dd, J = 8.7, 4.5 Hz, 2H), 3.97 (dd, J = 11.1, 6.2 Hz, 4H), 3.84 (s, 3H), 3.34 (t, J = 6.8 Hz, 2H), 3.04 (t, J = 6.8 Hz, 2H), 1.63 - 1.52 (m, 4H), 1.39 (s, 6H), 1.29 - 1.11 (m, 10H), 0.80 (ddd, J = 9.4, 5.6, 2.3 Hz, 6H).

[0110] Preparation Example 11 This preparation example provides Compound 32, and the preparation method of the Compound 32 includes the following steps.

Chemical Structure

[0111] t-Butyl (3-(3-bromo-4-hexyloxy)benzamide) pentylamine carbonate 27 (1.22 g, 2.48 mmol), methyl 4-hexyloxy-3-(pinacol boronate)benzoate (1.08 g, 2.97 mmol), and potassium carbonate (1.03 g, 7.44 mmol) were each added to 1,4-dioxane (30 mL), and after degassing with argon gas, 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium (0.18 g, 0.25 mmol) was added, and the reaction solution was stirred at 90 °C for 16 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 3), backwashed with a saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate, and then concentrated. The obtained crude product was purified by column chromatography (ethyl acetate:dichloromethane = 1:40 → 1:5) to separate and obtain a yellow solid 32 (1.52 g) (yield 95%).

[0112] Preparation Example 12 This preparation example provides Compound 33, and the preparation method of the Compound 33 includes the following steps.

Chemical formula

[0113] Respectively, t-butyl (3-(3-bromo-4-hexyloxy)benzamide) ethylamine carbonate (1.07 g, 2.42 mmol), methyl 4-hexyloxy-3-(pinacol boronate)benzoate (1.05 g, 2.9 mmol) and potassium carbonate (1 g, 7.26 mmol) were added to 1,4-dioxane (30 mL). After degassing with argon gas, 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium (0.18 g, 0.25 mmol) was added, and the reaction solution was stirred at 90 °C for 16 hours. Water (50 mL) was added to the reaction solution, and it was extracted with ethyl acetate (50 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated. The obtained crude product was purified by column chromatography (ethyl acetate:dichloromethane = 1:40 → 1:5) to obtain 33 (1.18 g) of a yellow solid (yield 81%).

[0114] Preparation Example 13 This preparation example provides Compound 34, and the preparation method of the said Compound 34 includes the following steps. Step K: Hydrolysis reaction

Chemical formula

[0115] Respectively, methyl 5'-((3-((t-butoxycarbonyl)amino)butyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylate 31 (0.9 g, 1.44 mmol) and sodium hydroxide (0.12 g, 2.88 mmol) were added to a mixed solution of tetrahydrofuran / methanol / water (8 mL / 4 mL / 6 mL), and it was stirred at 50 °C for 16 hours. The reaction solution was concentrated under reduced pressure, water (30 mL) was added, the pH value was adjusted to 5 with hydrochloric acid, and it was extracted with dichloromethane (20 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated to obtain 34 (0.26 g) of a yellow oily crude product. A crude product 34 (0.26 g) in the form of an oil was obtained.

[0116] Preparation Example 14 This preparation example provides compound 35, and the preparation method of the compound 35 includes the following steps.

Chemical formula

[0117] 5'-((3-((tert-Butoxycarbonyl)amino)propyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid methyl 30 (0.5 g, 0.82 mmol) and sodium hydroxide (0.16 g, 4.09 mmol) were respectively added to a mixed solution of tetrahydrofuran / methanol / water (8 mL / 4 mL / 6 mL), and stirred at 50 °C for 16 hours. The reaction solution was concentrated under reduced pressure, water (30 mL) was added, the pH value was adjusted to 5 with hydrochloric acid, extracted with dichloromethane (20 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated to obtain a yellow oily crude product 35 (0.65 g).

[0118] Preparation Example 15 This preparation example provides compound 36, and the preparation method of the compound 36 includes the following steps.

Chemical formula

[0119] 5'-((3-((tert-Butoxycarbonyl)amino)pentyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid methyl 32 (1.72 g, 2.69 mmol) and sodium hydroxide (0.22 g, 5.38 mmol) were respectively added to a mixed solution of tetrahydrofuran / methanol / water (10 mL / 8 mL / 5 mL), and stirred at 50 °C for 16 hours. The reaction solution was concentrated under reduced pressure, water (30 mL) was added, the pH value was adjusted to 5 with hydrochloric acid, extracted with dichloromethane (20 mL × 3), backwashed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated to obtain a yellow solid crude product 36 (0.96 g).

[0120] Preparation Example 16 This preparation example provides Compound 37, and the preparation method of the Compound 37 includes the following steps.

Chemical formula

[0121] Methyl 5'-((3-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylate 33 (1.18 g, 1.97 mmol) and sodium hydroxide (0.16 g, 3.95 mmol) were added to a mixed solution of tetrahydrofuran / methanol / water (10 mL / 8 mL / 5 mL), and the mixture was stirred at 50 °C for 16 hours. The reaction solution was concentrated under reduced pressure, water (30 mL) was added, the pH value was adjusted to 5 with hydrochloric acid, and the mixture was extracted with dichloromethane (20 mL × 3), backwashed with a saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and then concentrated to obtain a crude product 37 (1.07 g) of a yellow solid.

[0122] Preparation Example 17 This preparation example provides Compound 38, and the preparation method of the Compound 38 includes the following steps. Step K: Hydrolysis of Boc

Chemical formula

[0123] Trifluoroacetic acid (5 mL) was added to a dichloromethane solution (20 mL) of 5'-((3-((tert-butoxycarbonyl)amino)butyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid 34 (0.26 g, 0.42 mmol), and the reaction solution was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The obtained product 38 was directly used in the next reaction.

[0124] 11H NMR (500 MHz, MeOD-d4) δ 8.47 (brs, 1H), 7.99 (d, J = 8.0 Hz, 1H), 7.83 (d, J = 15.1 Hz, 2H), 7.68 (s, 1H), 7.03 (dd, J = 16.4, 8.5 Hz, 2H), 3.98 (s, 4H), 3.40 (s, 2H), 2.96 (s, 2H), 1.69 - 1.59 (m, 8H), 1.36 - 1.11 (m, 12H), 0.83 (s, 6H). [1 / 2M+H] + : 257.0

[0125] Preparation Example 18 This preparation example provides Compound 39, and the preparation method of the said Compound 39 includes the following steps.

Chemical Structure

[0126] To a dichloromethane solution (30 mL) of 5'-((3-((tert-butoxycarbonyl)amino)pentyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid 36 (0.96 g, 1.56 mmol) was added trifluoroacetic acid ( 5 mL), and the reaction solution was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The obtained product 39 was used directly in the next reaction.

[0127] Preparation Example 19 This preparation example provides Compound 40, and the preparation method of the said Compound 40 includes the following steps.

Chemical Structure

[0128] To a dichloromethane solution (30 mL) of 5'-((3-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid (1.07 g, 1.83 mmol) was added trifluoroacetic acid (5 mL), and the reaction solution was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The obtained product 40 was used directly in the next reaction.

[0129] Preparation Example 20 This preparation example provides Compound 41, and the preparation method of the Compound 41 includes the following steps. [Chemical formula]

[0130] Trifluoroacetic acid (5 mL) was added to a dichloromethane solution (20 mL) of 5'-((3-((tert-butoxycarbonyl)amino)propyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid 35 (650 mg, 1.1 mmol), and the reaction solution was stirred at room temperature for 6 hours and then concentrated under reduced pressure. The obtained product 41 was used directly in the next reaction.

[0131] Example 1 This example provides Compound 1, and the preparation method of the Compound 1 includes the following steps. Operation of Step C: [Chemical formula]

[0132] Under the protection of argon gas at room temperature, 1,7-di(3-iodo-4-methoxyphenyl)heptane-1,7-dione 22 (5.92 g, 10 mmol), pinacol borate (3.06 g, 12 mmol) and potassium acetate (9.8 g, 100 mmol) were dissolved in dimethyl sulfoxide (400 mL), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (0.732 g, 1 mmol) was added to the solution. The reaction solution was stirred at 100 °C for 7 hours. 500 mL of water was added, the filtered solid was washed with water, and the obtained solid was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1 → 2:1) to obtain white solid 1 (0.3 g) (yield 8.9%).

[0133] 11H NMR (400 MHz, MeOD-d4) δ 8.04 (d, J = 2.5 Hz, 2H), 7.92 (dd, J = 8.7, 2.5 Hz, 2H), 7.19 (d, J = 8.7 Hz, 2H), 3.97 (s, 6H), 2.47 - 2.32 (m, 2H), 2.07 - 1.80 (m, 6H), 1.68 - 1.51 (m, 2H).

[0134] Example 2 This example provides compound 2, and the preparation method of the compound 2 includes the following steps. Operation of step F:

Chemical formula

[0135] At room temperature, 1 6 ,2 6 -Dimethoxy-1,2(1,3)-dibenzena[6]annulene-3,9-dione 1 (670 mg, 2 mmol) was dissolved in anhydrous dichloroethane (30 mL), and aluminum trichloride (801 mg, 6 mmol) was added to the solution. The reaction solution was stirred at 80 °C for 2 hours. 30 mL of water was added, the filtered solid was washed with water, collected, the obtained liquid was extracted with dichloromethane (20 mL × 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the resulting solution was concentrated under reduced pressure and purified on a preparative silica gel plate (dichloromethane:methanol = 15:1), and the separated obtained solids were combined, filtered, and a pseudo-white solid 2 (370 mg) (yield 60%) was obtained.

[0136] 1 1H NMR (400 MHz, MeOD-d4) δ 8.21 (d, J = 2.4 Hz, 2H), 7.82 (dd, J = 8.5, 2.3 Hz, 2H), 7.01 (d, J = 8.5 Hz, 2H), 3.00 - 2.83 (m, 4H), 2.04 - 2.00 (m, 4H), 1.69 (m, 2H).

[0137] Example 3 This example provides compound 3, and the preparation method of the compound 3 includes the following steps. Operation of Step G:

Chemical formula

[0138] At room temperature, 1 6 , 2 6 -dihydroxy-1,2(1,3)-dibenzena[1,2-c]cyclononane-3,9-dione 2 (40 mg, 0.13 mmol) and potassium carbonate (107 mg, 0.78 mmol) were dissolved in acetonitrile (10 mL), and cyclopentyl bromide (33 mg, 0.22 mmol) was added to the solution. The reaction solution was stirred at 90 °C for 12 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative silica gel plate (dichloromethane:methanol = 30:1) to obtain 3 (20 mg) of a pseudo-white solid (yield 41%).

[0139] 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1H), 7.99 (dd, J = 19.7, 2.3 Hz, 2H), 7.79 (dd, J = 8.6, 2.4 Hz, 1H), 7.67 (dd, J = 8.5, 2.3 Hz, 1H), 7.18 (d, J = 8.8 Hz, 1H), 6.98 (d, J = 8.5 Hz, 1H), 5.03 (s, 1H), 3.17 (d, J = 5.1 Hz, 1H), 2.76 (d, J = 73.8 Hz, 2H), 2.02 - 1.68 (m, 10H), 1.65 - 1.43 (m, 4H).

[0140] Example 4 This example provides compound 4, and the preparation method of the compound 4 includes the following steps.

Chemical formula

[0141] At room temperature, 1 6 , 2 6 -dihydroxy-1,2(1,3)-dibenzena[1,2-c]cyclononane-3 ,9-Diketone 2 (2.9 g, 9.35 mmol) and potassium carbonate (7.75 g, 56.1 mmol) were dissolved in acetonitrile (100 mL), and hexyl bromide (9.26 g, 56.1 mmol) was added to the solution. The reaction mixture was stirred at 90 °C for 12 hours. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane:methanol = 100:1) to obtain white solid 4 (1.79 g) (yield 40%).

[0142] 1 H NMR (400 MHz, MeOD-d4) δ 8.02 (d, J = 2.5 Hz, 2H), 7.89 (dd, J = 8.7, 2.4 Hz, 2H), 7.15 (d, J = 8.7 Hz, 2H), 4.23 - 4.20 (m, 2H), 4.13 - 4.02 (m, 2H), 3.42 - 3.34 (m, 2H), 2.38 (dd, J = 20.5, 10.2 Hz, 2H), 1.96 - 1.76 (m, 8H), 1.70 - 1.59 (m, 2H), 1.48 - 1.41 (m, 4H), 1.36 - 1.22 (m, 8H), 0.93 - 0.84 (m, 6H).

[0143] Example 5 This example provides compound 5, and the preparation method of the compound 5 includes the following steps.

Chemical formula

[0144] At room temperature, 1 6 ,2 6 -Dihydroxy-1,2(1,3)-dibenzocyclononan-3,9-dione 2 (31 mg, 0.1 mmol) and potassium carbonate (83 mg, 0.6 mmol) were dissolved in acetonitrile (10 mL), and pentyl bromide (90 mg, 0.6 mmol) was added to the solution. The reaction mixture was stirred at 90 °C for 12 hours. The reaction solution was concentrated under reduced pressure and purified by preparative silica gel plate (dichloromethane:methanol = 70:1) to obtain white solid 5 (14 mg) (yield 31%).

[0145] 11H NMR (400 MHz, MeOD-d4) δ 7.98 (d, J = 2.4 Hz, 2H), 7.87 (dd, J = 8.7, 2.4 Hz, 2H), 7.12 (d, J = 8.7 Hz, 2H), 4.18 - 4.03 (m, 4H), 3.39 - 3.33 (m, 2H), 2.36 (dd, J = 20.4, 10.3 Hz, 2H), 2.00 - 1.74 (m, 8H), 1.63 - 1.60 (m, 2H), 1.46 - 1.32 (m, 8H), 0.89 (t, J = 7.2 Hz, 6H).

[0146] Example 6 This example provides compound 6, and the preparation method of the compound 6 includes the following steps. [Chemical formula]

[0147] At room temperature, 1 6 ,2 6 -dihydroxy-1,2(1,3)-dibenzocyclononan-3,9-dione 2 (31 mg, 0.1 mmol) and potassium carbonate (110 mg, 0.8 mmol) were dissolved in acetonitrile (20 mL), and heptyl bromide (144 mg, 0.8 mmol) was added to the solution. The reaction solution was stirred at 90 °C for 12 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative silica gel plate (dichloromethane:methanol = 70:1) to obtain separated white solid 6 (16 mg) (yield 32%).

[0148] 1 1H NMR (400 MHz, MeOD-d4) δ 7.99 (d, J = 2.4 Hz, 2H), 7.88 (dd, J = 8.7, 2.4 Hz, 2H), 7.13 (d, J = 8.7 Hz, 2H), 4.21 - 4.03 (m, 4H), 3.44 - 3.36 (m, 2H), 2.37 (dd, J = 20.5, 10.2 Hz, 2H), 2.04 - 1.74 (m, 8H), 1.69 - 1.56 (m, 2H), 1.46 - 1.40 (m, 4H), 1.38 - 1.24 (m, 12H), 0.86 (t, J = 6.9 Hz, 6H).

[0149] Example 7 This example provides compound 7, and the preparation method of the compound 7 includes the following steps.

Chemical formula

[0150] At room temperature, 1 6 ,2 6 -dihydroxy-1,2(1,3)-dibenzena[6.6]annulene-3,9-dione 2 (31 mg, 0.1 mmol) and potassium carbonate (110 mg, 0.8 mmol) were dissolved in acetonitrile (20 mL), and 1-bromo-4-methoxybutane (135 mg, 0.8 mmol) was added to the solution. The reaction solution was stirred at 90 °C for 12 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative silica gel plate (dichloromethane:methanol = 70:1) to separate and obtain white solid 7 (15 mg) (yield 31%).

[0151] 1 H NMR (400 MHz, MeOD-d4) δ 7.98 (d, J = 2.5 Hz, 2H), 7.87 (dd, J = 8.7, 2.4 Hz, 2H), 7.13 (d, J = 8.7 Hz, 2H), 4.30 - 4.03 (m, 4H), 3.41 - 3.30 (m, 6H), 3. 26 (s, 6H), 2.42 - 2.29 (m, 2H), 1.99 - 1.77 (m, 8H), 1.75 - 1.56 (m, 6H).

[0152] Example 8 This example provides compound 8, and the preparation method of the compound 8 includes the following steps.

Chemical formula

[0153] To a dichloromethane solution (10 mL) of 5'-((4-aminobutyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid trifluoroacetate 38, triethylamine (1 mL) and propylphosphonic anhydride (50%) (800 mg) were added. After stirring the reaction solution at room temperature for 3 days, it was concentrated under reduced pressure. The obtained product was subjected to preparative silica gel plate (ethyl acetate:dichloromethane:acetic acid = 1:2:0.1) to obtain a white pasty solid 8 (17 mg).

[0154] 1 H NMR(400MHz,MeOD-d4)δ12.64(s,1H),9.43(s,1H),8.34(s,1H),7.94-7.83(m,2H),7.73(dd,J=9.1,2.2Hz,2H),7.20-6.98(m,2H),3.99-3.94(m,4H),3.24-3.19(m,4H),1.52-1.51(m,8H),1.23-1.16(m,12H),0.81-0.77(m,6H).

[0155] Example 9 This example provides compound 9, and the preparation method of the compound 9 includes the following steps.

Chemical formula

[0156] To a dichloromethane solution (20 mL) of 5'-((4-aminopropyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid trifluoroacetate 41, triethylamine (1 mL) and propylphosphonic anhydride (50%) (2.2 g) were added. After stirring the reaction solution at room temperature for 3 days, it was concentrated under reduced pressure. The obtained product was subjected to preparative silica gel plate (ethyl acetate:dichloromethane:acetic acid = 1:2:0.1) to obtain a white pasty solid 9 (30 mg).

[0157] 11H NMR (400 MHz, MeOD-d4) δ 12.60 (s, 1H), 9.39 (s, 1H), 8.32 (s, 1H), 7.84 (ddd, J = 31.2, 8.6, 2.3 Hz, 2H), 7.69 (dd, J = 9.9, 9.0 Hz, 2H), 7.08 (dd, J = 9.9, 9.0 Hz, 2H), 3.93 (d, J = 6.3 Hz, 4H), 3.20 (dd, J = 9.9, 6.3 Hz, 4H), 1.72 - 1.62 (m, 2H), 1.52 - 1.47 (m, 4H), 1.19 - 1.13 (m, 12H), 0.76 - 0.73 (dd, J = 7.0, 6.1 Hz, 6H).

[0158] Example 10 This example provides compound 10, and the preparation method of the compound 10 includes the following steps.

Chemical formula

[0159] To a dichloromethane solution (20 mL) of 5'-((4-aminopentyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid trifluoroacetate 39, triethylamine (5 mL) and propylphosphonic anhydride (50%) (3 g) were added. After stirring the reaction solution at room temperature for 16 hours, it was concentrated under reduced pressure. The obtained product was separated on a silica gel plate (ethyl acetate:dichloromethane:acetic acid = 1:2:0.1) to obtain white solid 10 (20 mg).

[0160] 1 1H NMR (400 MHz, MeOD-d4) δ 8.34 (s, 1H), 8.01 (dd, J = 8.6, 2.2 Hz, 1H), 7.88 - 7.81 (m, 2H), 7.70 (d, J = 2.3 Hz, 1H), 7.06 (dd, J = 8.7, 1.4 Hz, 2H), 4.00 (td, J = 6.2, 3.6 Hz, 4H), 3.36 (dd, J = 9.5, 4.7 Hz, 2H), 3.27 (d, J = 7.2 Hz, 2H), 1.69 - 1.55 (m, 8H), 1.45 - 1.35 (m, 2H), 1.33 - 1.15 (m, 12H), 0.83 (t, J = 6.9 Hz, 6H).

[0161] Example 11 This example provides Compound 11, and the preparation method of the Compound 11 includes the following steps.

Chemical formula

[0162] To a dichloromethane solution (20 mL) of 5'-((4-aminoethyl)carbamoyl)-2',6-bis(hexyloxy)-[1,1'-biphenyl]-3-carboxylic acid trifluoroacetate 40, triethylamine (5 mL) and propylphosphonic anhydride (50%) (3 g) were added. After stirring the reaction solution at room temperature for 16 hours, it was concentrated under reduced pressure. The obtained product was separated on a silica gel plate (ethyl acetate:dichloromethane:acetic acid = 1:2:0.1) to obtain white solid 11 (18 mg). 1 H NMR (400 MHz, MeOD-d4) δ 8.01 (dd, J = 8.6, 2.1 Hz, 1H), 7.84 (dd, J = 8.9, 2.3 Hz, 2H), 7.70 (d, J = 2.4 Hz, 1H), 7.07 (dd, J = 8.7, 1.1 Hz, 2H), 4.00 (td, J = 6.2, 1.7 Hz, 4H), 3.51 (dd, J = 12.0, 4.8 Hz, 4H), 1.63 - 1.59 (m, 4H), 1.34 - 1.14 (m, 12H), 0.83 (td, J = 6.9, 1.4 Hz, 6H).

[0163] Example 12 This example provides Compound 12, and the preparation method of the Compound 12 includes the following steps as follows.

Chemical formula

[0164] Methyl 5’-((3-((tert-butoxycarbonyl)amino)butyl)carbamoyl)-2’,6-bis(hexyloxy)-[1,1’-biphenyl]-3-carboxylate 31 (1.1 g, 1.76 mmol) was dissolved in a hydrochloric acid solution of ethyl acetate (80 mL). After the reaction solution was stirred at room temperature for 4 hours, it was concentrated under reduced pressure to obtain a white solid 12 (900 mg) (yield: 91%). 1 H NMR (400 MHz, MeOD-d4) δ 8.48 (s, 1H), 8.06 (d, J = 8.1 Hz, 1H), 7.94 - 7.85 (m, 2H), 7.74 (brs, 1H), 7.12 (d, J = 7.7 Hz, 2H), 4.04 (d, J = 5.3 Hz, 4H), 3.91 (brs, 2H), 3.46 (brs, 2H), 1.75 (brs, 4H), 1.65 (brs, 4H), 1.28 (brs, 12H), 0.87 (d, J = 5.6 Hz, 6H).

[0165] Example 13 This example provides compound 13, and the preparation method of the compound 13 includes the following steps.

Chemical formula

[0166] Referring to the preparation method of compound 25 in Step I, compound 8 (60 mg, 0.1 mmol) was reacted with compound 45 (20 mg, 0.11 mmol) to produce compound 13 (white solid, 20 mg, yield 26%). 1 H NMR (400 MHz, DMSO-d6) δ 9.39 (s, 1H), 8.31 (s, 1H), 7.84 (d, J = 8.8 Hz, 1H), 7.69 (d, J = 15.3 Hz ,2H),7.07(d,J=8.7Hz,1H),4.21(t,J=6.5Hz,1H),3.94(t,J=6.3Hz,2H),3.30(s,2H),3.21(dd,J=16.1,6.5Hz,4H),1.63(t,J=7.5Hz,1H),1.50(d,J=16.7Hz,7H),1.36(q,J=7.4Hz,2H),1.20(d,J=22.6Hz,10H),0.90(t,J=7.4Hz,2H),0.83 - 0.72(m,4H).

[0167] Example 14 This example provides compound 14, and the preparation method of the compound 14 includes the following steps.

Chemical Structure

[0168] (60 mg, 0.1 mmol) of compound 8 was added with (63 mg, 0.2 mmol) of cesium carbonate in 5 mL of DMF solvent, benzyl bromide (0.1 mL) was added dropwise at room temperature, and the reaction was allowed to proceed sufficiently for 2 h at room temperature. Water and ethyl acetate (20 mL) were added, and liquid separation was carried out. The organic phase was taken, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and compound 14 (60 mg, pale yellow solid, yield 85.7%) was obtained by column chromatography. 1 H NMR(400MHz,Chloroform - d)δ8.05(dd,J = 8.6,2.3Hz,1H),7.93(d,J = 2.2Hz,1H),7.81(dd,J = 8.6,2.4Hz,1H),7.55(d,J = 2.3Hz,1H),7.42(d,J = 7.1Hz,2H),7.39 - 7.28(m,3H),7.09(s,1H),6.92(dd,J = 8.7,5.5Hz,2H),6.19(s,1H),5.32(s,2H),3.93(q,J = 6.8Hz,4H),3.44(dd,J = 14.1,6.3Hz,4H),1.80 - 1.49(m,10H),1.22(d,J = 17.5Hz,14H),0.81(h,J = 3.7Hz,6H).

[0169] Example 15 This example provides compound 15, and the preparation method of the compound 15 includes the following steps.

Chemical formula

[0170] Example 16 This example provides compound 16, and the preparation method of the compound 16 includes the following steps.

Chemical formula

[0171] Refer to the preparation method of compound 30 in the coupling reaction of step K. Compound 46 (40 mg, 0.1 mmol) was reacted with compound 48 (28 mg, 0.1 mmol) to obtain compound 16 (25 mg, white solid, yield 53.4%). ESI + , 468.2.

[0172] Example 17 This example provides compound 17, and the preparation method of the compound 17 includes the following steps.

Chemical formula

[0173] Refer to the preparation method of compound 30 in the coupling reaction of step K. Compound 49 (30 mg, 0.1 mmol) and compound 48 (28 mg, 0.1 mmol) were reacted to obtain compound 17 (26 mg, white solid, yield 72.2%). ESI + ,359.2.

[0174] Example 18 This example provides compound 18, and the preparation method of the compound 18 includes the following steps.

Chemical formula

[0175] Refer to the preparation method of compound 30 in the coupling reaction of step K. Compound 49 (30 mg, 0.1 mmol) and compound 48 (33 mg, 0.1 mmol) were reacted to obtain compound 18 (16 mg, white solid, yield 39%). ESI + ,419.2.

[0176] Example 19 This example provides compound 19, and the preparation method of the compound 19 includes the following steps.

Chemical formula

[0177] Refer to the preparation method of compound 30 in the coupling reaction of step K. Compound 51 (36 mg, 0.1 mmol) and compound 48 (33 mg, 0.1 mmol) were reacted to obtain compound 19 (18 mg, white solid, yield 36%). HR-ESI(M+H) C 29 H 42 O6N 500.3007, found 557.3030.

[0178] Example 20 This example provides compound 20, and the preparation method of compound 20 includes the following steps: [ka]

[0179] Refer to the preparation method of compound 30 in the coupling reaction of step K. Compound 51 (36 mg, 0.1 mmol) was reacted with compound 48 (40 mg, 0.1 mmol) to obtain compound 20 (20 mg, white solid, yield 35.7%). HR-ESI(M+H)C 32 H 49 O6N2557.3585,found 557.3605.

[0180] Pharmacology Experiments Experimental Example 1: Inhibition of binding between LAG3 and its ligand MHCII by biphenyl compounds (1) Experimental Method HTRF (Homogeneous Time-Resolved Fluorescence) Fluorescence (FRET) is a method for detecting fluorescence by fluorescence resonance energy transfer (FRET) and time-resolved fluorescence (TRF). The detection was based on two technologies: light (TRF, Time-Resolved Flourescence) and fluorescence (TRF). 2 μL of the test compound with the appropriate concentration was added to a 384-well plate, and 4 μL of Tag1-LAG3 protein (final concentration 10 nmol L -1 ) and 4 μL of Tag2-MHCII (final concentration 20 nmol L -1 ) and incubated at room temperature for 15 min. 10 μL of anti-Tag1-Tb working solution (25 μL of anti-Tag1-Tab stock solution added to 2475 μL of dilution buffer) was added, followed by 10 μL of anti-Tag2-XL665 working solution (25 μL of Tag2-XL665 stock solution added to 2475 μL of dilution buffer), sealed with a membrane, and incubated at room temperature for 1 h. Detection was performed with a Perkin Elma Envision instrument.

[0181] (2) Inhibitory IC of binding of LAG3 / MHCII by biphenyl compounds 50 was measured. The measurement results are shown in Table 1 below.

[0182]

Table 1

[0183] (3) Summary: In Table 1, Compounds 8, 9, 10, 11, and 13 have an inhibitory IC for the LAG3 / MHC 50 II interaction, all at the μmol·L -1 level. It was found that none of these compounds have a strong inhibitory effect on the binding of LAG3 / MHC II.

[0184] Experimental Example 2, Experiment on inhibition of binding between LAG3 and its ligand FGL1 by biphenyl compounds (1) Experimental method Preparation of reagents: (1) Tag1-LAG3 protein: A protein solution with a concentration of 50 nM was prepared with a diluent (the final concentration in a 20 μL system was 10 nM). (2) Tag2-FGL1 protein: A protein solution with a concentration of 100 nM was prepared with a diluent (the final concentration in a 20 μL system was 20 nM). (3) Anti-Tag1-Tb: The 100X stock solution was diluted to 1X with the detection buffer. (4) Anti-Tag2-XL665: The 100X stock solution was diluted to 1X with the detection buffer.

[0185] Measurement steps: (1) 2 μL of the compound solution (10X test concentration) was added to a white, non-light-transmissive 384-well plate, and then 4 μL of Tag1-LAG3 protein and 4 μL of Tag2-FGL1 protein were added respectively, and incubated at room temperature for 15 min. . (2) Anti-Tag1-Tb and anti-Tag2-XL665 were mixed at a ratio of 1:1 in advance, and 10 μL of the mixed solution was added to each well. (3) Gently and uniformly mixed, sealed, incubated at room temperature for 1 h, or detected after overnight incubation.

[0186] IC of the inhibition of the binding of LAG3 / FGL1 by biphenyl compounds 50 is shown in Table 2 below.

[0187]

Table 2

[0188] (III) Summary: In Table 2, compound 8 has an IC 50 of 10 μmol·L -1 for inhibiting the interaction of LAG3 / FGL1. It was found that compound 8 has a strong inhibitory effect on the binding of LAG3 / FGL1.

[0189] Experimental Example 3, Interaction Experiments of Compounds 4, 8 and 9 with LAG3 Proteins of Different Species (I) Experimental Method 1. Experimental Principle Surface plasmons are electromagnetic waves on the metal surface, which are generated by the interaction between freely vibrating photons and electrons. Surface plasmon resonance (SPR) is an optical phenomenon that occurs at the surface of two media, and this phenomenon is induced by photons or electrons. When light is incident from an optically dense medium to an optically sparse medium and total reflection occurs, an evanescent wave is formed and enters the optically sparse medium. When the totally reflected evanescent wave meets the plasmon wave on the metal surface, resonance occurs and the reflected light energy decreases, and a resonance peak may appear in the reflected light energy spectrum. This resonance is called surface plasmon resonance. The incident angle that causes surface plasmon resonance is called the SPR angle. SPR biosensors provide a highly sensitive label-free detection technology for real-time monitoring of intermolecular interactions. What is detected by the sensor is the change in the SPR angle. Furthermore, SPR is related to the refractive index of the metal surface. When the analyte binds to the chip surface, the refractive index of the chip surface changes, causing a change in the SPR angle. This is the basic principle by which SPR biosensors detect intermolecular interactions in real time. During interaction analysis, the change in the SPR angle is recorded in real time in the sensorgram.

[0190] 2. Method According to the data, the theoretical isoelectric point of the LAG3 protein is about 9.82, and protein immobilization is planned to be performed using the amino coupling method. Since the main ligands of the LAG3 protein are FGL1 and MHCII proteins, after the LAG3 protein was coupled, the activity of the protein was verified here by FGL1 protein injection.

[0191] Protein coupling buffer solution: 1.05×PBS-P+; 1.02×PBS-P+

[0192] Interaction buffer solution: 1.0×PBS-P+, 5%DMSO; 1.0×PBS-P+, 2%DMSO

[0193] 3. Protein coupling This experiment adopted the CM5 chip amino coupling method to immobilize human LAG3 protein on the F c2 or Fc3 channel, and mouse LAG3 protein on the Fc3 or Fc4 channel.

[0194] When interacting with Compound 8: The coupling conditions of LAG3 protein were as follows: the concentration was about 25 μg / mL, the system was a sodium acetate solution with pH 5.5, the chip activation time was 420 s, and the blocking time was 420 s.

[0195] When interacting with Compound 4 and Compound 9: The coupling conditions of LAG3 protein were as follows: the concentration was about 50 μg / mL, the system was a sodium acetate solution with pH 4.5, the chip activation time was 420 s, and the blocking time was 420 s.

[0196] (2) Results 1. Interaction between human LAG3 protein and Compound 4 The coupling amount of human LAG3 was about 5000 RU, and the theoretical Rmax was about 50 RU. The measurement results indicated that Compound 4 interacted with human LAG3 protein, and the kinetic constants were shown in Figure 1.

[0197] 2. Interaction between mouse LAG3 protein and Compound 4 The coupling amount of mouse LAG3 was about 5000 RU, and the theoretical Rmax was about 50 RU. The measurement results indicated that Compound 4 interacted with mouse LAG3 protein, and the kinetic constants were shown in Figure 2.

[0198] 3. Interaction between human LAG3 protein and Compound 8 The coupling amount of human LAG3 was about 3000 RU, and the theoretical Rmax was about 30 RU. The measurement results indicated that Compound 8 interacted with human LAG3 protein, and the kinetic constants were shown in Figure 3.

[0199] 4. Interaction between Mouse LAG3 Protein and Compound 8 The coupling amount of mouse LAG3 was approximately 3600 RU, and the theoretical Rmax was approximately 36 RU. The measurement results indicated that Compound 8 interacted with mouse LAG3 protein, and the kinetic constants were shown in Figure 4.

[0200] 5. Interaction between Human LAG3 Protein and Compound 9 The coupling amount of human LAG3 was approximately 5000 RU, and the theoretical Rmax was approximately 50 RU. The measurement results showed that Compound 9 interacted with human LAG3 protein, as shown in Figure 5. For the "quick up and down" interaction characteristics, fitting in the steady state mode was necessary.

[0201] 6. Interaction between Mouse LAG3 Protein and Compound 9 The coupling amount of mouse LAG3 was approximately 5000 RU, and the theoretical Rmax was approximately 50 RU. The measurement results showed that Compound 9 did not interact with mouse LAG3 protein (KD = 0.68 M), as shown in Figure 6.

[0202] 7. Interaction between Human FGL1 Protein and Compound 4 The coupling amount of human FGL1 was approximately 5857 RU, and the theoretical Rmax was approximately 58 RU. The measurement results indicated that Compound 4 interacted with human FGL1 protein, and the kinetic constants were shown in Figure 7.

[0203] 8. Interaction between Human FGL1 Protein and Compound 8 The coupling amount of human FGL1 was approximately 5200 RU, and the theoretical Rmax was approximately 52 RU. The measurement results indicated that Compound 8 interacted with human FGL1 protein, and the kinetic constants were shown in Figure 8.

[0204] 9. Interaction between Human FGL1 Protein and Compound 9 The coupling amount of human FGL1 was about 5857 RU, and the theoretical Rmax was about 58 RU. The measurement results showed that compound 9 did not interact with human FGL1 protein, and the kinetic constants are shown in Figure 9.

[0205] (III) Summary This test aimed to measure the interaction affinity between LAG3 proteins of different species and compounds 4, 8, and 9, and was measured by capturing the proteins with a CM5 chip. From the positive results measured by manual injection of FGL1 protein (5 μg / mL), it was found that the activity of LAG3 protein was well maintained by this interaction measurement method. The measurement results of the binding affinity between the compounds and LAG3 protein are shown in Table 3, and the measurement results of the binding affinity between the compounds and FGL1 protein are shown in Table 4.

[0206]

Table 3

[0207]

Table 4

[0208] As can be seen from the data in Table 3 and Table 4, the affinity of compound 8 and compound 4 for human and mouse LAG3 was generally comparable (10 -5 ~10 -6 M). At the same time, the affinity of compound 8 and compound 4 for FGL1, a new ligand of LAG3, could also reach the level of 10 -5 M.

[0209] Experimental Example 4. Investigation of the in vivo antitumor effects of compound 4 and 8 1. In vivo antitumor effect of compound 4 on mouse liver cancer HEPA1-6 (1) Thirty-two 8-week-old female C57L mice were selected and divided into five cages. HEPA1-6 cells cultured in DMEM + 10% FBS for in vitro use were washed with PBS, digested with 0.25% trypsin for 3 min, terminated with serum-containing medium, centrifuged at 2500 rpm for 5 min, the supernatant was discarded, appropriate PBS was added and resuspended, and this was repeated twice. BioRad When counted with a counting device, the cell count was 1.14×10 7 cells / mL. The cell suspension was prepared in an Erlenmeyer flask in an ice bath. The tumor fluid was uniformly aspirated with a 1 mL sterile syringe and inoculated into mice at 0.2 mL / mouse. After the inoculation was completed, the mice were continued to be bred under the original breeding conditions.

[0210] (2) Grouping of animals: On the second day after the C57B / L mice were inoculated with the tumor fluid, 32 mice were randomly grouped. The control group had 8 mice, the 5 mg / kg group of compound 4 had 6 mice, the 10 mg / kg group of compound 4 had 6 mice, the 20 mg / kg group of compound 4 had 6 mice, and the 100 mg / kg group of the positive control drug cyclophosphamide CTX had 6 mice. The test drug groups were orally administered once a day, and the positive control drug CTX was intraperitoneally injected once. The mice were weighed daily for administration.

[0211] (3) Preparation of drugs: The corresponding amounts of the test drugs were weighed according to the final concentrations of each group in the experiment. For the 5 mg / kg group of compound 4, the 10 mg / kg group of compound 4, and the 20 mg / kg group of compound 4, they were prepared with 5% CMCNa + Tween80, and for CTX, it was prepared with physiological saline. For the control group, a 5% CMCNa solution was orally administered. Dosage: As the intragastric administration ratio, 0.4 mL was administered per 20 g of animal, and as the intraperitoneal injection ratio, 0.2 mL was administered per 20 g of animal.

[0212] (4) The animals were sacrificed, the tumors were removed, and the tumor weights were measured. One hour after the animals in each group were administered on the final day, they were immediately processed, the weights of the animals were measured, the tumor tissues were dissected, and the tumor weights were measured.

[0213] (5) Results Twenty-seven days after administration, the animals were sacrificed, and the tumors were dissected and weighed. The inhibitory effect of Compound 4 on the growth of mouse liver cancer HEPA1-6 in vivo is shown in Table 5 below.

[0214]

Table 5

[0215] As shown in Table 5, at the dosages of 5 mg / kg and 10 mg / kg of Compound 4, the inhibitory rates of tumor weight were 24.72% and 52%, respectively, indicating a certain dosage-dependence.

[0216] 2. In vivo antitumor effect of Compound 4 on mouse melanoma B16F10 (1) Forty-three 8-week-old female C57B / L mice were selected and divided into 5 cages. One preserved B16F10-bearing female C57L mouse was taken, sacrificed by cervical dislocation, the skin of the mouse was sprayed with 75% alcohol in a clean bench, the tumor tissue of the tumor-bearing mouse was separated with sterile surgical instruments, the tumor tissue was minced with ophthalmic scissors, and placed in a sterile 5 mL homogenizer. The tumor tissue was gradually ground into tumor fluid on ice. When counted with a BioRad counting device, the cell count was 6.85×10 7 cells / mL. The tumor solution was prepared in an Erlenmeyer flask in an ice bath. The tumor fluid was uniformly aspirated with a 1 mL sterile syringe and inoculated into C57 / L mice at 0.2 mL per mouse. After the inoculation was completed, the mice were continued to be bred under the original breeding conditions.

[0217] (2) Grouping and administration of animals: On the second day after the tumor fluid was inoculated into C57B / L mice, the 43 mice were randomly grouped. The control group had 10 mice, the 20 mg / kg group of Compound 4 had 8 mice, the 30 mg / kg group of Compound 4 had 8 mice, the 40 mg / kg group of Compound 4 had 8 mice, and the 100 mg / kg group of the positive control drug cyclophosphamide CTX had 9 mice. The animals were weighed and administered.

[0218] (3) Preparation of drugs: The corresponding amounts of drugs were weighed according to the final concentrations of each group in the experiment. For the 20 mg / kg group and 30 mg / kg group of Compound 4, they were prepared with 0.25% CMCNa + Tween 80 and orally administered once a day. For the 40 mg / kg of Compound 4, it was prepared with physiological saline + Tween 80 and intraperitoneally administered. For CTX, it was prepared with physiological saline and administered once by intraperitoneal injection. For the control group, a 0.25% CMCNa solution was orally administered.

[0219] (4) The animals were sacrificed, the tumors were removed, and the weights of the tumors were measured. One hour after the animals in each group were administered on the final day, they were immediately processed, and the body weights and tumor weights of the animals were measured.

[0220] (5) Results After 15 days of administration, after the mice were treated, the tumors were dissected and measured. The in vivo growth inhibitory effect of Compound 4 on mouse melanoma B16F10 is shown in Table 6 below.

[0221]

Table 6

[0222] As shown in Table 6, in the group orally administered with 20 mg / kg and the group intraperitoneally injected with 40 mg / kg of Compound 4, the tumor weights were significantly smaller than those of the blank (P < 0.01). It was found that the growth of mouse melanoma B16F10 could be inhibited at this dosage.

[0223] 3. In vivo antitumor effect of Compound 8 on mouse melanoma B16F10

[0224] By the same method as above, the in vivo antitumor effect of Compound 8 on mouse melanoma B16F10 was observed.

[0225] Animal grouping and administration: On the second day after tumor fluid was inoculated into C57BL mice, the cancer-bearing mice were randomly grouped. They were divided into a blank group, a 5 mg / kg group of Compound 8, a 10 mg / kg group of Compound 8, a 20 mg / kg group of Compound 8, and a 100 mg / kg group of the positive control drug cyclophosphamide CTX. Every day, the body weights of the animals were measured and the drugs were administered. The test drug was administered orally every day, and the positive control drug CTX was administered intraperitoneally. After the experiment was completed, the body weights and tumor weights of the animals were measured, and the tumor weight inhibition rate (%) was calculated.

[0226] Table 7 shows the results of the inhibitory effect of Compound 8 on the growth of mouse melanoma B16F10 in vivo.

[0227]

Table 7

[0228] As can be seen from the data in Table 7, when Compound 8 was orally administered at 20 mg / kg, it had an obvious inhibitory effect on the growth of mouse melanoma B16F10, and it was found that Compound 8 had a good antitumor effect.

[0229] In this application, the process method of this application has been described by the above examples. However, the applicant declares that this application is not limited to the above process steps, that is, this application does not mean that it must be implemented depending on the above process steps. Those skilled in the art should understand that any improvement to this application, equivalent substitution for the raw materials selected in this application, addition of auxiliary components, selection of specific forms, etc. are all included within the protection scope and disclosure scope of this application.

Claims

1. A biphenyl compound having a structure represented by the following formula I. 【Chemical 1】 (However, R １ and R ２ are each independently selected from hydrogen, halogen, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C8 alkoxycarbonyl group, C1-C8 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 alkyl group, C1-C8 alkoxy group, trihalo C1-C8 alkyl group, trihalo C1-C8 alkoxy group, C1-C8 alkoxymethyleneoxy group, methoxy C1-C8 methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, and n is selected from 0, 1, 2, 3, or 4, X is selected from CH ２ , O or NH, Y is CH ２ , O, OH, NH, NH ２ , NH(CH ２ ), ｎ’ NH ２ , NH(CH ２ ), ｎ’ NHCOCF ３ , OBn, a C1-C8 alkyl group, a C1-C8 alkoxy group, a trihalomethylamine group, a dimethylamine group, a methylamine group or a trihaloalkoxy group, provided that n' is selected from 1, 2, 3, 4 or 5, M is CH ２ , O, OH, NH, NH ２ , a trihalomethylamine group, a trihalomethylacyl group, a dimethylamino group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C8 alkoxymethyleneoxy group, and M and Y are either linked by a bond or do not form a bond.

2. R １ and R ２ each independently represents any one selected from hydrogen, fluorine, chlorine, bromine, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C6 alkoxycarbonyl group, C1-C6 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C6 alkyl group, C1-C7 alkoxy group, trihalo C1-C6 alkyl group, trihalo C1-C7 alkoxy group, C1-C6 alkoxymethyleneoxy group, methoxy C1-C6 methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, n is selected from 1, 2, 3, or 4, X is selected from CH ２ , O or NH, Y is CH ２ , O, OH, NH, NH ２ , NH(CH ２ ), ｎ’ NH ２ , NH(CH ２ ), ｎ’ NHCOCF ３ , OBn, a C1-C6 alkyl group, a C1-C6 alkoxy group, a trihalomethylamine group, a dimethylamine group, a methylamine group or a trihaloalkoxy group, provided that n' is selected from 2, 3, 4 or 5, M is CH ２ , O, OH, NH, NH ２ , a trihalomethylamine group, a trihalomethylacyl group, a dimethylamine group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C6 alkoxymethyleneoxy group, and M and Y are either linked by a bond or do not form a bond, The biphenyl compound according to Claim 1.

3. R １ and R ２ each independently represents any one selected from hydrogen, fluorine, chlorine, bromine, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C4 alkoxycarbonyl group, C1-C4 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C4 alkyl group, C1-C7 alkoxy group, trihalo C1-C4 alkyl group, trihalo C1-C7 alkoxy group, C1-C4 alkoxymethyleneoxy group, methoxy C1-C4 methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, n is selected from 1, 2, 3, or 4, X is selected from CH ２ , O or NH, Y is CH ２ , O, OH, NH, NH ２ , NH(CH ２ ), ｎ’ NH ２ , NH(CH ２ ), ｎ’ NHCOCF ３ , Obn, a C1-C4 alkyl group, a C1-C4 alkoxy group, a trihalomethylamine group, a dimethylamine group, a methylamine group or a trihaloalkoxy group, provided that n' is selected from 2, 3, 4 or 5, M is CH ２ , O, OH, NH, NH ２ , a trihalomethylamine group, a trihalomethylacyl group, a dimethylamine group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C4 alkoxymethyleneoxy group, and M and Y are either linked by a bond or do not form a bond, The biphenyl compound according to Claim 1 or 2.

4. R １ and R ２ are each independently one selected from hydrogen, fluorine, chlorine, bromine, hydroxy group, dimethylamine group, cyano group, nitro group, methoxycarbonyl group, ethoxycarbonyl group, methylcarbonyloxy group, ethoxycarbonyl group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, methyl group, ethyl group, methoxy group, ethoxy group, propoxy group, 1-butanoxy group, 1-pentanoxy group, 1-hexanoxy group, 1-heptanoxy group, 2-pentanoxy group, 2-hexanoxy group, trifluoromethyl group, trifluoroethyl group, trifluoromethoxy group, methoxymethyleneoxy group, methoxyethyloxy group, methoxypropyloxy group, methoxyn-butoxy group, cyclopentyloxy group or cyclohexyloxy group, n is selected from 1, 2, 3, or 4, X is selected from CH ２ , O or NH, Y is CH ２ , O, OH, NH, NH ２ , NH(CH ２ ), ｎ’ NH ２ , NH(CH ２ ), ｎ’ NHCOCF ３ , Obn, a methyl group, an ethyl group, a methoxy group, an ethoxy group, a trifluoromethylamine group, a trichloromethylamine group, a dimethylamine group, a methylamine group or a trifluoromethaneoxy group, provided that n' is selected from 2, 3, 4 or 5, M is CH ２ , O, OH, NH, NH ２ , a trifluoromethylamine group, a trichloromethylamine group, a trifluoromethylacyl group, a trichloromethylacyl group, a dimethylamine group, a methylamine group, a carboxyl group, a trifluoromethyl group or a methoxymethyleneoxy group, and M and Y are either linked by a bond or do not form a bond, The biphenyl compound according to any one of Claims 1 to 3.

5. The biphenyl compound according to any one of Claims 1 to 4, which is any one selected from the structures represented by the following 1 to 20. [Chemical 2] [Chemical Formula 3]

6. An isomer of the biphenyl compound according to any one of Claims 1 to 5 or a pharmaceutically acceptable salt thereof, Preferably, the pharmaceutically acceptable salt includes any one or at least a combination of two of hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, p-toluenesulfonate, acetate, trifluoroacetate, salicylate, amino acid salt, 2-O-β-D-glucopyranosyl-L-ascorbate, maleate, tartrate, fumarate, citrate, lactate, sodium salt, potassium salt, calcium salt, magnesium salt, lithium salt, ammonium salt, or a salt of an organic base capable of providing a physiologically acceptable cation, Preferably, the salt of the organic base capable of providing the physiologically acceptable cation includes any one or at least a combination of two of methylamine salt, dimethylamine salt, trimethylamine salt, piperidine salt, morpholine salt, or tris(2-hydroxyethyl)amine salt, An isomer of the biphenyl compound or a pharmaceutically acceptable salt thereof.

7. A method for preparing the biphenyl compound according to any one of Claims 1 to 5, including the following steps, Synthesis method 1: (A) Nucleophilic addition reaction of a compound represented by formula II, a compound represented by formula III, and a compound represented by formula IV to generate a compound represented by formula V, and the reaction formula is as follows, [Chemical Formula 4] React the compound represented by formula V with iodine in an iodination reaction to produce a compound represented by formula VI, and the reaction formula is as follows: 【Chemical Formula 5】 React the compound represented by formula VI in a coupling reaction under alkaline conditions and in the presence of zero-valent palladium to produce a compound represented by formula I, and the reaction formula is as follows: [Chemical Formula 6] (However, R １ and R ２ are each independently selected from hydrogen, halogen, dimethylamine group, cyano group, nitro group, C1-C8 alkoxycarbonyl group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 alkyl group or trihalo C1-C8 alkyl group, n is selected from 0, 1, 2, 3 or 4, and X, Y, M are each independently CH ２ selected from.) Or, Synthesis Method 2: Perform a nucleophilic addition reaction on the compound represented by formula II' and the compound represented by formula III' to produce a compound represented by formula IV', or perform a nucleophilic addition reaction on the compound represented by formula II' and the compound represented by formula II to produce a compound represented by formula IV'', or perform a nucleophilic addition reaction on the compound represented by formula II' and the compound represented by formula III to produce a compound represented by formula IV''', and the reaction formula is as follows: 【Chemical Formula 7】 React the compound represented by formula IV' in an iodination reaction to produce a compound represented by formula V', or react the compound represented by formula IV'' in an iodination reaction to produce a compound represented by formula V'', or react the compound represented by formula IV''' in an iodination reaction to produce a compound represented by formula V''', and the reaction formula is as follows: [Chemical 8] After reacting the compound represented by formula V' in a coupling reaction under alkaline conditions and in the presence of zero-valent palladium, hydrolyze it under Lewis acid conditions to produce a compound represented by formula VI', or after reacting the compound represented by formula V'' in a coupling reaction under alkaline conditions and in the presence of zero-valent palladium, hydrolyze it under Lewis acid conditions to produce a compound represented by formula VI'', or after reacting the compound represented by formula V''' in a coupling reaction under alkaline conditions and in the presence of zero-valent palladium, hydrolyze it under Lewis acid conditions to produce a compound represented by formula VI''', and the reaction formula is as follows: 【Chemical Formula 9】 Perform an alkylation reaction on the compound represented by formula VI' and an alkyl halide compound to produce a compound represented by formula I, or perform an alkylation reaction on the compound represented by formula VI'' and an alkyl halide compound to obtain a compound represented by formula I, or perform an alkylation reaction on the compound represented by formula VI''' and an alkyl halide compound to obtain a compound represented by formula I, and the alkyl halide compound includes alkyl bromide or alkyl chloride, and the reaction formula is as follows: 【Chemical Formula 10】 (However, R １ and R ２ are each independently hydrogen, halogen, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C8 alkoxycarbonyl group, C1-C8 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 alkyl group, C1-C8 alkoxy group, trihalo C1-C8 alkyl group, trihalo C1-C8 alkoxy group, C1-C8 alkoxymethyleneoxy group, methoxy C1-C8 methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, and n is selected from 0, 1, 2, 3 or 4, and X, Y, M are each independently CH ２ selected from.) Or, Synthesis Method 3: The compound represented by formula (H) VII is subjected to a hydrolysis reaction with an alkali to produce a compound represented by formula VIII. The alkali includes an organic alkali and / or an inorganic alkali, and the reaction formula is as follows: 【Chemical 11】 (I) The compound represented by formula VIII and the compound represented by formula IX are subjected to a condensation reaction under alkaline conditions to produce a compound represented by formula X, and the reaction formula is as follows: 【Chemical 12】 (J) The compound represented by formula X is subjected to a coupling reaction to produce a compound represented by formula XI, and the reaction formula is as follows: 【Chemical 13】 (K) The compound represented by formula XI and the compound represented by formula XII are subjected to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium to produce a compound represented by formula I, or the compound represented by formula XI and the compound represented by formula XII are subjected to a coupling reaction under alkaline conditions and in the presence of zero-valent palladium. After the reaction, it is hydrolyzed with an organic alkali or an inorganic alkali, and then stirred with hydrochloric acid or trifluoroacetic acid to obtain an amino hydrochloride salt. After separation, it is reacted with an acyl reagent to produce a compound represented by formula I, and the reaction formula is as follows: 【Chemical 14】 (However, R １ and R ２ are each independently selected from hydrogen, halogen, hydroxy group, dimethylamine group, cyano group, nitro group, C1-C8 alkoxycarbonyl group, C1-C8 alkylcarbonyloxy group, methylamine group, methylsulfonyl group, dimethylsulfamoyl group, amino group, carboxyl group, C1-C8 alkyl group, C1-C8 alkoxy group, trihalo C1-C8 alkyl group, trihalo C1-C8 alkoxy group, C1-C8 alkoxymethyleneoxy group, methoxy C1-C8 methyleneoxy group, cyclopentyloxy group or cyclohexyloxy group, n is selected from 0, 1, 2, 3 or 4, X is selected from CH ２ , O or NH, X' is selected from OH or NH ２ , Y is selected from CH ２ , O, OH, NH, NH ２ , NH(CH ２ ), ｎ’ NH ２ , N H(CH ２ ) ｎ’ NHCOCF ３ 、Obn、C １～８ alkyl group, C １～８ alkoxy group, trihalomethylamine group, dimethylamine group, methylamine group or trihaloalkoxy group, and n' is selected from 1, 2, 3, 4 or 5. M is CH ２ , O, OH, NH, NH ２ , a trihalomethylamine group, a trihalomethylacyl group, a dimethylamino group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C8 alkoxymethyleneoxy group, and M' is OH, NH ２ , a trihalomethylamine group, a trihalomethylacyl group, a dimethylamino group, a methylamine group, a carboxyl group, a trihaloalkyl group or a C1-C8 alkoxymethyleneoxy group. ) Preparation method.

8. A pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable vector, wherein the active ingredient comprises the biphenyl compound according to any one of claims 1 to 5 and / or an isomer of the biphenyl compound according to claim 6 or a pharmaceutically acceptable salt thereof, Preferably, the active ingredient in the pharmaceutical composition is 0.1 to 95% by mass. Pharmaceutical composition.

9. Use of the biphenyl compound according to any one of claims 1 to 5, an isomer of the biphenyl compound according to claim 6 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8, in the preparation of a medicament or an anti-aging agent for preventing and / or treating tumors, autoimmune diseases, inflammatory diseases, neurodegenerative diseases, and metabolic resistance diseases.

10. The tumor is any one selected from glioma, melanoma, gastric cancer, lung cancer, breast cancer, renal cancer, liver cancer, oral epithelial cancer, head and neck tumors, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, rectal adenocarcinoma, leukemia, or lymphoma, or a combination of at least two of them. Preferably, the autoimmune disease includes any one or at least a combination of two or more of rheumatoid arthritis, systemic lupus erythematosus, ulcerative colitis, psoriasis, dermatitis, or amyotrophic lateral sclerosis. Preferably, the inflammatory disease includes any one or at least a combination of two or more of polyarteritis, phlebitis, or reflux esophagitis. Preferably, the neurodegenerative disease includes senile dementia and / or Parkinson's disease. Preferably, the metabolic disease includes diabetes, hyperuricemia, or gout. The use according to claim 9.

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