Biphenyl-substituted five-membered heterocyclic compounds, methods of making, pharmaceutical compositions, and uses thereof

Abstract

The application discloses a kind of biphenyl-substituted five-membered heterocyclic compounds and preparation method, pharmaceutical composition and application thereof.The structure of the biphenyl-substituted five-membered heterocyclic compounds is as shown in formula (I), also includes stereoisomer, mesomer, racemate, prodrug, crystal, pharmaceutically acceptable salt or their mixture thereof.The PD-L1 inhibitor activity is had, can significantly inhibit PD-1 / PD-L1 protein-protein interaction, blocks PD-1 / PD-L1 signal pathway, so it can be widely applied in the preparation of the immunomodulator drug for preventing and / or treating tumor, infectious disease, inflammatory disease, autoimmune disease and organ transplant rejection, and the compound preparation method is conducive to structure expansion.

Description

Technical Field

[0001] This invention relates to a class of biphenyl-substituted five-membered heterocyclic compounds, their preparation methods, pharmaceutical compositions, and applications, specifically to a class of biphenyl-substituted five-membered heterocyclic compounds with PD-1 / PD-L1 protein-protein interaction inhibitory activity, their preparation methods, pharmaceutical compositions, and applications. Background Technology

[0002] Cancer is a group of genetically modified diseases that seriously threaten human life and health. Programmed death receptor (PD-1) and its ligands PD-L1 / 2 are important co-inhibitory molecules in immune checkpoints, playing a negative regulatory role in the host's innate and adaptive immune responses, and thus playing a crucial regulatory role in the body's immune homeostasis. Studies have found that various tumors upregulate PD-L1 expression in themselves and the tumor microenvironment, continuously activating the PD-1 / PD-L1 signaling pathway, inhibiting T cell function, and leading to tumor immune escape. Tumor immunotherapy utilizes the body's own immune system to kill tumor cells. The continuous success of tumor immunotherapy in clinical practice has made it a highly sought-after treatment for cancer.

[0003] Immune escape is a fundamental characteristic of malignant tumors. Under normal physiological conditions, the human immune system can recognize and eliminate foreign molecules in a timely manner. However, in cancer patients, due to the decline in the body's immune capacity and the special biological characteristics of tumor cells, tumor cells can evade the recognition and killing of the immune system through various mechanisms, ultimately surviving and developing within the body—this is the phenomenon of immune escape. Tumor immune escape is a complex pathological process involving the patient's immune system and the biological characteristics of tumor cells, such as tumor heterogeneity, energy metabolism, and tumor stem cells. Although our understanding of the mechanisms of tumor immune escape is not yet fully complete, it is recognized that immune checkpoints are closely related to it. Therefore, the development of drugs based on immune checkpoints has become the fastest-progressing category in tumor immunotherapy, with the development of PD-1 / PD-L1 monoclonal antibodies and small molecule inhibitors becoming a hot topic in the global new drug development field.

[0004] Cytotoxic T-lymphocyte-associated molecule-4 (CTLA-4), programmed cell death receptor-1 (PD-1), and programmed cell death ligand-1 (PD-L1) are the most widely studied and recognized inhibitory checkpoint pathways. Co-stimulatory molecules of immune checkpoints mainly include CD27, CD40, OX40, GITR, CD137, OX40, and ICOS, while co-inhibitory molecules mainly include CTLA-4, PD-1, PD-L1, PD-L2, TIM-3, VISTA, and IDO. Co-stimulatory molecules can enhance the immune response, thus facilitating the clearance of "foreign molecules" by immune cells; while co-inhibitory molecules play a negative regulatory role in immunity, thereby maintaining immune homeostasis and preventing excessive immunization that could cause tissue damage. Currently, PD-1 / PD-L1 has been well-established as a target for tumor immunotherapy. Besides being expressed on mature T cells, PD-1 is also expressed at low levels on CD4-CD8-T cells, B cells, dendritic cells (DCs), and natural killer (NK) cells in the thymus. PD-1 has two ligands: PD-L1 is mainly expressed on mature T cells, B cells, and some non-hematopoietic cell types, but it can be expressed on various cell types under the induction of inflammatory factors (such as IFN-γ, TNF-α, and VEGF). PD-L2 has a relatively narrow expression range, primarily expressed on macrophages and DCs. When PD-1 binds to its ligand, it induces phosphorylation of tyrosine residues in the ITSM domain of the cytoplasm, thereby recruiting SHP-2 phosphatase near the TCR, inhibiting the activation of proximal TCR kinase, leading to reduced phosphorylation levels of TCR-CD3 molecules and Lck-mediated ZAP-70, and subsequently activating downstream signaling pathways. The negative regulation of immunity by PD-1 / PD-L is mainly achieved by inhibiting the PI3K-AKT and RAS signaling pathways, blocking the activation of transcription factors that play an important role in T cell activation, proliferation, function and survival, such as activator protein-1 (AP-1), nuclear factor activating T cells (NFAT) and NF-κB.

[0005] Furthermore, numerous studies have shown that abnormalities in the PD-1 / PD-L signaling pathway are closely related to the occurrence, development, and poor prognosis of various human tumors. In the tumor microenvironment, when the PD-1 / PD-L1 signaling pathway is overactivated, tumor cells can survive by using anti-apoptotic signals and inhibiting the activity of antigen-specific T lymphocytes. Additionally, blocking the PD-1 / PD-L1 signaling pathway with PD-1 or PD-L1 antibodies can inhibit tumor cell growth. By reversing the effects on T lymphocyte signal transduction, reactivating T lymphocytes, promoting the generation of effector T lymphocytes and memory T lymphocytes, and inhibiting the differentiation of regulatory T lymphocytes, the immune killing capacity of T lymphocytes in the tumor microenvironment is ultimately enhanced, thereby achieving the goal of tumor treatment. Currently, there are already [number missing] [products / technology missing] globally. and Several PD-1 / PD-L1 monoclonal antibody drugs have been launched and are widely used in clinical treatment of various solid tumors and hematological cancers, including malignant melanoma, non-small cell lung cancer, gastric cancer, liver cancer, kidney cancer, and bladder cancer, with significant clinical efficacy. Furthermore, the indications for these drugs are continuously expanding. At the same time, clinical trials of PD-1 / PD-L1 monoclonal antibody drugs in combination with other drugs for the treatment of various malignant tumors are also actively underway.

[0006] However, PD-1 / PD-L1 monoclonal antibody drugs also have significant drawbacks, such as easily causing excessive T cell activation and triggering immune-related side effects. Furthermore, they cannot be administered orally, leading to poor patient compliance, and are difficult to prepare and purify, resulting in high costs. Therefore, the development of small molecule inhibitors of PD-1 / PD-L1 is of great value. Summary of the Invention

[0007] Objectives of the Invention: The first objective of this invention is to provide a biphenyl-substituted five-membered heterocyclic compound; the second objective is to provide a method for preparing the compound; the third objective is to provide a pharmaceutical composition containing the compound; and the fourth objective is to provide a pharmaceutical application of the compound and the pharmaceutical composition thereof.

[0008] Technical solution: The biphenyl-substituted five-membered heterocyclic compound of the present invention has the structure of formula (I), and further comprises its stereoisomer, meso compound, racemic compound, prodrug, crystal, pharmaceutically acceptable salt, or mixture thereof.

[0009]

[0010] in:

[0011] A and B are selected from C, N, or O;

[0012] X is selected from N, O, or S;

[0013] R 1Selected from C1-C4 alkyl, cyano, hydroxyl, or halogen;

[0014] R 2 Selected from hydrogen, halogen, nitro, cyano, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl or -O(CH2)2O-;

[0015] R 3 R 4 Each is independently selected from hydrogen, C1-C8 alkyl, C1-C8 alkoxy, C1-C8 alkylamine, C3-C8 cycloalkyl, 5-7 membered heterocyclic group, or R 3 R 4 Together with the nitrogen atoms attached thereto, they form 5-7 membered heterocyclic groups; the 5-7 membered heterocyclic groups contain one or more heteroatoms selected from O, S or N; the C1-C8 alkyl, C1-C8 alkoxy, C1-C8 alkylamine, C3-C8 cycloalkyl or 5-7 membered heterocyclic groups are substituted by one or more Y groups;

[0016] Y is selected from hydrogen, halogen, hydroxyl, mercapto, methylthio, carbonyl, carboxyl, amino, guanidinyl, furanyl, tetrahydropyrrolyl, morpholinyl, N-methylpiperazinyl, C1-C4 alkyl, -CO2R 5 -NHCOR 5 -NR 6 R 7 or -CONR 6 R 7 The C1-C4 alkyl group is substituted with one or more hydrogens, hydroxyl groups, or halogens.

[0017] R 5 Selected from C1-C8 alkyl groups;

[0018] R 6 R 7 Each is independently selected from hydrogen, C1-C8 alkyl, C1-C8 alkoxy, C3-C8 cycloalkyl, or R 6 R 7 Together with the nitrogen atoms attached to them, they form 5-7 membered heterocyclic groups; the C1-C8 alkyl, C1-C8 alkoxy, C3-C8 cycloalkyl or 5-7 membered heterocyclic groups are substituted by one or more Z groups;

[0019] Z is selected from hydrogen, halogen, hydroxyl, mercapto, carboxyl, amino, or acetamide.

[0020] Preferably, in the structure:

[0021] A, B, and X are selected from any of the following scenarios:

[0022] (1) When A is N, B is N or O, and X is N, O or S;

[0023] (2) When A is C, B is N or C, and X is O or S;

[0024] R 1 Selected from methyl or halogen;

[0025] R 2 Selected from hydrogen, halogens, or -O(CH2)2O-;

[0026] R 3 R 4 Each is independently selected from hydrogen, C1-C5 alkyl, or R 3 R 4 Together with the nitrogen atoms attached to them, they form 5-6 membered N-containing heterocyclic groups; the C1-C5 alkyl or 5-6 membered heterocyclic groups are substituted by one or more Y groups;

[0027] Y is selected from hydrogen, hydroxyl, carbonyl, carboxyl, guanidinyl, C1-C4 alkyl, -CO2R 5 -NR 6 R 7 or -CONR 6 R 7 The C1-C4 alkyl group is substituted with one or more hydrogen or hydroxyl groups;

[0028] R 5 Selected from C1-C4 alkyl groups;

[0029] R 6 R 7 Each is independently selected from hydrogen or C1-C4 alkyl groups.

[0030] Preferably, in the structure:

[0031] Selected from

[0032] R 1 Selected from methyl or halogen;

[0033] R 2 Selected from hydrogen, halogens, or -O(CH2)2O-;

[0034] R 3 R 4 Each is independently selected from hydrogen, C1-C5 alkyl, or R 3 R 4 Together with the nitrogen atoms attached to them, they form 5-6 membered N-containing heterocyclic groups; the C1-C5 alkyl or 5-6 membered heterocyclic groups are substituted by one or more Y groups;

[0035] Y is selected from hydrogen, hydroxyl, carbonyl, carboxyl, guanidinyl, C1-C4 alkyl, -CO2CH3, amino, or -CONH2; wherein the C1-C4 alkyl is substituted by one or more hydrogen or hydroxyl groups.

[0036] Preferably, in the structure:

[0037] Selected from

[0038] Preferably, in the structure:

[0039] Selected from

[0040] Preferably, the biphenyl-substituted five-membered heterocyclic compound of the present invention is selected from any of the following compounds:

[0041]

[0042]

[0043] Preferably, the pharmaceutically acceptable salt is a salt formed by the compound with an acid or base, wherein the acid is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, and ferulic acid; and the base is selected from inorganic bases containing alkali metal cations, alkaline earth metal cations, or ammonium cations, or choline, piperazine, morpholine, triethylamine, diisopropylamine, and trimethylamine.

[0044] "Pharmaceutically acceptable salts" refer to salts of compounds prepared by reacting a compound with a relatively non-toxic acid or base, containing specific substituents. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming carbonates or bicarbonates), phosphoric acid (forming phosphates, monohydrogen phosphates, dihydrogen phosphates, sulfuric acid (forming sulfates or bisulfates), hydroiodic acid, phosphorous acid, etc.); and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid. Acids such as citric acid, tartaric acid, and methanesulfonic acid; organic acid salts also include salts of organic acids such as amino acids (e.g., arginine) and glucuronic acid. Certain compounds contain both basic and acidic functional groups, thus allowing them to be converted into either a base or acid addition salt. Preferably, the salt is contacted with a base or acid in a conventional manner, and then the parent compound is separated, thereby regenerating the free form of the compound. The free form of the compound differs from its various salt forms in certain physical properties, such as different solubilities in polar solvents.

[0045] Pharmaceutically acceptable salts can be synthesized from parent compounds containing an acid radical or a base using conventional chemical methods. Generally, such salts are prepared by reacting these compounds, in their free acid or base form, with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of both. Non-aqueous media such as ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile are generally preferred.

[0046] Preferably, the stereoisomer is R 3 R 4 Isomers introduced by neutral chiral C or N.

[0047] Preferably, the prodrug is R 3 R 4 The ester prodrug introduced by the carboxyl group is more preferably a C1-C4 alkyl ester.

[0048] The method for preparing the biphenyl-substituted five-membered heterocyclic compound of the present invention is selected from any of the following methods:

[0049] Method 1: Compound a-1 is prepared by Suzuki coupling, esterification, condensation, acylation, cyclization and condensation reaction to obtain compound (I), or by further alkaline hydrolysis to obtain compound (I).

[0050]

[0051] Method 2: Compound a-2 is prepared by Suzuki coupling, addition, cyclization and condensation reactions to obtain compound (I), or by further alkaline hydrolysis to obtain compound (I);

[0052]

[0053] Method 3: Compound e-1 is prepared into compound (I) by cyclization and condensation reactions, or by further alkaline hydrolysis.

[0054]

[0055] Method 4: Compound a-4 is prepared by two-step coupling, bromination, amination, condensation, cyclization and condensation reaction to obtain compound (I), or by further alkaline hydrolysis to obtain compound (I).

[0056]

[0057] Method 5: Compound a-4 is prepared by a three-step coupling reaction followed by a reduction amination reaction to obtain compound (I), or by further alkaline hydrolysis to obtain compound (I);

[0058]

[0059] Method 6: Compound c-5 is prepared into compound (I) by coupling reaction and reductive amination reaction, or further prepared into compound (I) by alkaline hydrolysis;

[0060]

[0061] Among them, R 1 R 2 R 3 R 4 The definition of X is as described above;

[0062] The compound of formula (I) obtained by the above method is salted with the corresponding acid or base to obtain a pharmaceutically acceptable salt.

[0063] The pharmaceutical composition of the present invention comprises the biphenyl-substituted five-membered heterocyclic compound of the present invention and a pharmaceutically acceptable carrier.

[0064] Preferably, the dosage form is tablet, capsule, powder, pill, granule, injection, oral liquid, syrup, inhaler, ointment, patch or suppository.

[0065] "Pharmaceutically acceptable carriers" are excipients widely used in the pharmaceutical manufacturing industry. Excipients primarily serve to provide a safe, stable, and functional pharmaceutical composition, and may also provide methods to facilitate the dissolution of the active ingredient at a desired rate after administration to a subject, or to promote the effective absorption of the active ingredient after administration to a subject. The pharmaceutical excipients may be inert fillers or provide a function, such as stabilizing the overall pH of the composition or preventing the degradation of the active ingredient. The pharmaceutical excipients may include one or more of the following: binders, suspending agents, emulsifiers, diluents, fillers, granulators, adhesives, disintegrants, lubricants, anti-adhesion agents, flow aids, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

[0066] The pharmaceutical compositions described in this invention can be prepared using any method known to those skilled in the art, based on the disclosure. For example, conventional mixing, dissolving, granulation, emulsification, grinding, encapsulation, embedding, or lyophilization processes.

[0067] The pharmaceutical compositions of this invention can be administered in any form, including by injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical, or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra-arterial, intramuscular) administration. The pharmaceutical compositions of this invention can also be controlled-release or sustained-release dosage forms (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, tablets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of parenteral formulations include, but are not limited to, solutions for injection, dry powder formulations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories; and tablets.

[0068] The biphenyl-substituted five-membered heterocyclic compounds or their pharmaceutical compositions described in this invention are used in the preparation of PD-L1 inhibitor drugs.

[0069] The biphenyl-substituted five-membered heterocyclic compounds or their pharmaceutical compositions described in this invention are also used in the preparation of immunomodulatory drugs.

[0070] Preferably, the drug is for the prevention or treatment of tumors, infectious diseases, inflammatory diseases, organ transplant rejection, or autoimmune diseases.

[0071] More preferably, the tumor is one or more of the following: malignant melanoma, lung cancer, breast cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, lymphoma, leukemia, prostate cancer, testicular cancer, kidney cancer, brain cancer, head and neck cancer, ovarian cancer, cervical cancer, endometrial cancer, mesothelial carcinoma, thyroid tumor, liver cancer, and esophageal cancer.

[0072] More preferably, the infectious disease is an infection caused by one or more of the following: human immunodeficiency virus, hepatitis B virus, hepatitis C virus, influenza virus, poliovirus, cytomegalovirus, Coxsackievirus, human papillomavirus, Epstein-Barr virus, and varicella-zoster virus.

[0073] Further preferably, the autoimmune disease is one or more of the following: rheumatoid arthritis, systemic lupus erythematosus, dermatomyositis, scleroderma, nodular vasculitis, multiple sclerosis, myasthenia gravis, mixed connective tissue disease, psoriasis, and autoimmune reactions caused by infection.

[0074] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0075] The compounds designed in this invention possess PD-L1 inhibitory activity, thereby effectively inhibiting PD-1 / PD-L1 protein-protein interactions and suppressing the activity IC50. 50 The concentrations are at nanomolar levels, even below 100 nanomolar levels, which are superior to existing clinical drugs and show promising application prospects. Furthermore, the compound preparation method is highly versatile, facilitating the development of various structures. Detailed Implementation

[0076] The technical solution of the present invention will be further described below with reference to the embodiments.

[0077] Reagents and materials: Unless otherwise specified, all reagents required for the experiment are commercially available chemically pure or analytically pure products.

[0078] instrument: 1¹H NMR was determined using a Bruker AV-300MHz NMR spectrometer. Chemical shift values ​​(δ) are expressed in ppm, coupling constant values ​​(J) are expressed in Hz, and TMS was used as an internal standard. Mass spectrometry (MS) was performed using a Shimadzu LCMS-2020 mass spectrometer. Thin-layer chromatography (TLC) was performed using HG / T2354-92 GF254 silica gel produced by Qingdao Ocean Chemical Co., Ltd., with color development at 254nm using a ZF7 three-way UV analyzer. Column chromatography was performed using 300-400 mesh coarse-pore (ZCX-II) silica gel from Qingdao Ocean Chemical Plant.

[0079] Example 1: Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazol-2-yl)methyl)glycine (1) and its hydrochloride (1s)

[0080]

[0081] Synthesis of 2-methyl-[1,1'-biphenyl]-3-carboxylic acid (1A)

[0082] 3-Bromo-2-methylbenzoic acid (10.00 g, 46.50 mmol), phenylboronic acid (11.34 g, 93.00 mmol), potassium carbonate (19.28 g, 139.51 mmol), and Pd(PPh3)4 (0.54 g, 0.47 mmol) were added to 100 mL of 1,4-dioxane and 10 mL of water. The reaction was carried out under N2 atmosphere at 80 °C for 12 h. The mixture was concentrated under reduced pressure, the pH was adjusted to 2 with 4 M HCl, filtered, and dried to give 9.53 g of a white solid, with a yield of 96%. MS (EI) m / z 213 [M+H] + ; 1 H NMR(300MHz,Chloroform-d)δ(ppm)7.74-7.34(m,1H),7.50-7.38(m,3H),7.38-7.34(m,2H),7.33-7.28(m,2H),2.29(s,3H).

[0083] Synthesis of methyl 2-methyl-[1,1'-biphenyl]-3-carboxylic acid ester (1B)

[0084] 1A (9.53 g, 44.90 mmol) was added to 100 mL of methanol, and 5 mL of concentrated sulfuric acid was added dropwise with stirring. The mixture was refluxed for 4 h. After cooling, the solution was concentrated under reduced pressure, 300 mL of water was added, and the solution was extracted with ethyl acetate. The extract was washed with saturated brine, and the organic phases were combined to give 9.96 g of a yellow oily liquid, with a yield of 98%. MS (EI) m / z 227 [M+H] + ; 1H NMR (300MHz, Chloroform-d) δ (ppm) 7.45-7.32 (m, 4H), 7.30-7.15 (m, 3H), 7.07 (d, J = 7.5Hz, 1H), 3.98 (s, 3H), 2.32 (s, 3H).

[0085] Synthesis of 2-methyl-[1,1'-biphenyl]-3-carbazide (1C)

[0086] 1B (9.96 g, 44.02 mmol) was added to 100 mL of ethanol, followed by 5 mL of hydrazine hydrate. The mixture was refluxed for 5 h. After cooling, the mixture was concentrated under reduced pressure, and 100 mL of ice water was added. A solid precipitated, which was filtered and dried to give 9.24 g of a white solid, with a yield of 93%. MS (EI) m / z 227 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 8.02 (s, 1H), 7.45 (d, J = 7.5Hz, 3H), 7.37 (s, 1H), 7.33 (d, J = 1.8Hz, 3H), 2.52 (s, 3H).

[0087] Synthesis of N'-(2-chloroacetyl)-2-methyl-[1,1'-biphenyl]-3-carbamoylhydrazide (1D)

[0088] Chloroacetyl chloride (0.50 g, 4.46 mmol) was dissolved in 5 mL of anhydrous dichloromethane and added dropwise to a dichloromethane solution of 1C (1.00 g, 4.42 mmol) and triethylamine (1.34 g, 13.26 mmol) cooled in an ice bath. The reaction was carried out for 3 h, filtered, and dried to give 1.21 g of a white solid, with a yield of 90%. MS (EI) m / z 303 [M+H] + MS(EI)m / z 301[M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.64-7.52 (m, 3H), 7.51-7.31 (m, 5H), 5.06 (s, 2H), 2.33 (s, 3H).

[0089] Synthesis of 2-chloromethyl-5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazole (1E)

[0090] 1D (0.70 g, 2.31 mmol) and 5 mL of phosphorus oxychloride were refluxed for 12 h. The mixture was concentrated under reduced pressure, dissolved in ethyl acetate, and extracted dropwise into ice water. The extract was dried over anhydrous sodium sulfate and purified by column chromatography to give 0.47 g of a white solid, yield 71%. MS (EI) m / z 285 [M+H]+ ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.62-7.56 (m, 4H), 7.50-7.32 (m, 4H), 5.04 (s, 2H), 2.39 (s, 3H).

[0091] Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazol-2-yl)methyl)glycine methyl ester (1m)

[0092] IE (0.30 g, 1.05 mmol), K₂CO₃ (0.51 g, 3.69 mmol), and glycine methyl ester hydrochloride (0.33 g, 2.63 mmol) were added to 5 mL of acetonitrile, and the mixture was refluxed for 8 h. After cooling, the mixture was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 0.14 g of a white solid (41% yield). MS (EI) m / z 338 [M+H] + ; 1 H NMR(300MHz,Chloroform-d)δ(ppm)7.84(dt,J=7.2,1.8Hz,1H),7.56-7.47(m,4H),7.4 5-7.40(m,1H),7.39-7.33(m,2H),4.68(s,2H),4.12(s,2H),3.67(s,3H),2.33(s,3H).

[0093] Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazol-2-yl)methyl)glycine ethyl ester (1e)

[0094] Following the preparation method of 1m, a white solid was obtained from glycine ethyl ester hydrochloride in 39% yield. MS(EI)m / z 352[M+H] + ; 1 H NMR(300MHz,Chloroform-d)δ(ppm)7.86(dt,J=7.2,1.8Hz,1H),7.55-7.48(m,4H),7.47-7.43(m,1H), 7.41-7.36(m,2H),4.65(s,2H),4.19(q,J=7.2Hz,2H),4.10(s,2H),2.34(s,3H),1.26(t,J=6.9Hz,3H).

[0095] Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazol-2-yl)methyl)glycine isopropyl ester (1i)

[0096] Following the preparation method of 1m, a white solid was obtained from glycine isopropyl hydrochloride in 35% yield. MS(EI)m / z 366[M+H] + ; 1 H NMR(300MHz,Chloroform-d)δ(ppm)7.82(dt,J=7.2,1.5Hz,1H),7.55-7.49(m,3H),7.48-7.42(m,2H),7.41-7.34(m,2H),5.10(p,J=6.3Hz 1H), 4.62 (s, 2H), 4.10 (s, 2H), 2.34 (s, 3H), 1.20 (d, J = 6.9Hz, 6H).

[0097] Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazol-2-yl)methyl)glycine (1) and its hydrochloride (1s)

[0098] 1 M (0.35 g, 1.04 mmol) and LiOH (0.75 mg, 3.11 mmol) were added to 3 mL of methanol and reacted at room temperature for 5 h. The mixture was concentrated under reduced pressure, 2 mL of water was added, and the pH was adjusted to 3 with 4 M hydrochloric acid. The mixture was filtered and dried to give 0.31 g of a white solid (1), with a yield of 92%. 1 M (0.31 mg, 0.94 mmol) was added to 1 mL of 1,4-dioxane (4 M) hydrochloric acid solution and reacted overnight at room temperature. The mixture was concentrated under reduced pressure, washed with anhydrous diethyl ether, filtered, and dried to give 0.30 g of a white solid. MS (ESI) m / z 324 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.21 (s, 1H), 7.92 (dt, J = 7.2, 1.8Hz, 1H), 7.54-7.47 ( m,4H),7.46-7.43(m,1H),7.40-7.36(m,2H),4.67(s,2H),4.09(s,2H),2.44(s,3H).

[0099] The following compounds were prepared using a procedure similar to that in Example 1:

[0100]

[0101]

[0102] Example 13: Synthesis of (3-(2-methyl-[1,1'-biphenyl]-3-yl)-1,2,4-oxadiazol-5-yl)methyl)glycine (13) and its hydrochloride (13s)

[0103]

[0104] Synthesis of 3-bromo-2-methylbenzonitrile (2A)

[0105] 3-Bromo-2-methylbenzonitrile (10.00 g, 51.01 mmol), phenylboronic acid (12.44 g, 102.02 mmol), potassium carbonate (7.76 g, 56.11 mmol), and Pd(PPh3)4 (0.59 g, 0.51 mmol) were added to 100 mL of 1,4-dioxane and 10 mL of water. The reaction was carried out at 90 °C for 12 h under N2 atmosphere. After cooling, the mixture was concentrated under reduced pressure, and extracted with 300 mL of water and ethyl acetate. The extract was washed with saturated brine, and the organic phases were combined to give 8.87 g of a white solid, 90% yield. MS (EI) m / z 194 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.62 (d, J = 7.5Hz, 1H), 7.47 (d, J = 8.4Hz, 2H), 7.45-7.27 (m, 5H), 2.46 (s, 3H).

[0106] Synthesis of N-hydroxy-2-methyl-[1,1'-biphenyl]-3-carboximide (2B)

[0107] 2A (8.86 g, 45.85 mmol), ammonium hydroxide (11.15 g, 0.16 mol), and sodium bicarbonate (15.41 g, 183.39 mmol) were added to 100 mL of methanol and reacted at 50 °C for 4 h. After cooling, the mixture was concentrated under reduced pressure, and 300 mL of water was added. The mixture was extracted with ethyl acetate, washed with saturated brine, and the organic phases were combined to give 9.5 g of a white crystalline solid, with a yield of 91%. MS (EI) m / z 227 [M+H] + ; 1 ¹H NMR (300 MHz, Chloroform-d) δ (ppm) 7.65 (d, J = 7.5 Hz, 1H), 7.49–7.25 (m, 7H), 2.29 (s, 3H). Synthesis of N-((chlorocarbonyl)oxy)-2-methyl-[1,1'-biphenyl]-3-carboxamide (2C)

[0108] Add 2B (9.50 g, 41.98 mmol) to 100 mL of dichloromethane, add triethylamine (8.50 g, 83.97 mmol) dropwise, cool in an ice bath for 5 min, add chloroacetyl chloride (5.69 g, 50.38 mmol) dropwise, and react at room temperature for 4 h. Then proceed directly to the next reaction step.

[0109] Synthesis of 5-(chloromethyl)-3-(2-methyl-[1,1'-biphenyl]-3-yl)-1,2,4-oxadiazole (2D)

[0110] Dissolve 2C in 50 mL of toluene and reflux at 120 °C for 12 h. Concentrate under reduced pressure, extract with ethyl acetate, dry to anhydrous sodium sulfate, and purify by column chromatography to give 8.8 g of a white solid, yield 71%. MS (EI) m / z 285 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.90 (dd, J = 6.9, 2.4Hz, 1H), 7.50-7.32 (m, 7H), 4.50 (dd, J = 8.7, 7.2Hz, 2H), 2.45 (s, 3H).

[0111] Synthesis of (3-(2-methyl-[1,1'-biphenyl]-3-yl)-1,2,4-oxadiazol-5-yl)methyl)glycine methyl ester (13m)

[0112] Following the method in Example 1, 2D and glycine methyl ester hydrochloride were subjected to a condensation reaction to obtain a white solid 13m, with a yield of 58%. MS(EI) m / z 338 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.84-7.81 (m, 1H), 7.50 (d, J = 1.5Hz, 1H), 7.47 (d, J = 4.2Hz, 1H), 7.45-7. 41(m,2H),7.39(q,J=2.1Hz,2H),7.35(t,J=1.5Hz,1H),4.19(s,2H),3.88(s,3H),3.42(s,2H),2.34(s,3H).

[0113] Synthesis of compound 13 and its hydrochloride (13S)

[0114] Following the method in Example 1, 13m was hydrolyzed to obtain 13, with a yield of 84%. Then, 13 was reacted with hydrochloric acid to form a salt, yielding 13s, with a yield of 78%. MS(EI) m / z 324 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.51 (s, 1H), 7.86-7.81 (m, 1H), 7.48 (d, J = 1.6Hz, 1H), 7.46 (d, J = 4.2Hz, 1H ),7.44-7.40(m,2H),7.38(q,J=2.1Hz,2H),7.36(t,J=1.5Hz,1H),4.15(s,2H),3.42(s,2H),2.36(s,3H).

[0115] The following compounds were prepared using a similar procedure to that in Example 13:

[0116]

[0117] Example 17: Synthesis of (5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-thiadiazol-2-yl)methyl)glycine (17) and its hydrochloride (17s)

[0118]

[0119] Synthesis of 2-chloromethyl-5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-thiadiazole (3A)

[0120] 1 D (1.00 g, 3.30 mmol) and Lawson's reagent (1.14 g, 3.34 mmol) were added to 10 mL of toluene and refluxed for 2 h. The mixture was concentrated under reduced pressure, dissolved in ethyl acetate, and extracted dropwise into ice water. The extract was dried over anhydrous sodium sulfate and purified by column chromatography to give 0.70 g of a white solid, yield 71%. MS (EI) m / z 301 [M+H] + ; 1 HNMR(300MHz,Chloroform-d)δ(ppm)7.62(s,1H),7.50-7.31(m,7H),5.04(s,2H),2.39(s,3H).

[0121] Synthesis of (5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-thiadiazol-2-yl)methyl)glycine methyl ester (17m)

[0122] Following the method in Example 1, 3A and glycine methyl ester hydrochloride were subjected to a condensation reaction to obtain a white solid 17m, with a yield of 58%. MS(EI) m / z 354 [M+H] + ; 1 H NMR(300MHz,DMSO-d6)δ(ppm)7.59(d,J=1.8Hz,1H),7.47(d,J=7.5Hz,2H),7.43( s,1H),7.38(d,J=8.1Hz,4H),4.23(s,2H),3.65(s,3H),3.51(s,2H),2.39(s,3H).

[0123] Synthesis of compound 17 and its hydrochloride (17S)

[0124] Following the method in Example 1, 17m was hydrolyzed to obtain 17, with a yield of 84%. Then, 17 was reacted with hydrochloric acid to form a salt, yielding 17s, with a yield of 78%. MS(EI) m / z 340 [M+H]+ ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.28 (s, 1H), 7.60 (d, J = 1.8Hz, 1H), 7.49 (d, J = 7.5H z,2H),7.45(s,1H),7.39(d,J=8.1Hz,4H),4.20(s,2H),3.56(s,2H),2.33(s,3H).

[0125] The following compounds were prepared using a similar procedure to that in Example 17:

[0126]

[0127] Example 19: Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,4-oxadiazol-2-yl)methyl)glycine (19) and its hydrochloride (19s)

[0128]

[0129] Synthesis of 1-(3-bromo-2-methylphenyl)ethane-1-one (4A)

[0130] 1,3-Dibromo-2-methylbenzene (10.00 g, 40.01 mmol) was dissolved in 85 mL of LDM. Tri-n-butyl(1-ethoxyethylene)tin (14.41 g, 40.01 mmol) and Pd(PPh3)4 (0.46 g, 0.40 mmol) were added with stirring. The reaction was carried out at 85 °C for 12 h under a nitrogen atmosphere. After cooling, the mixture was extracted with ethyl acetate and concentrated under reduced pressure. 20 mL of 4 M HCl was added, and the reaction was carried out at room temperature for 3 h. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 8.02 g of an oily product (94% yield). MS (EI) m / z 213 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.59 (d, J = 7.8Hz, 1H), 6.93 (s, 2H), 2.60 (s, 3H), 2.30 (s, 3H).

[0131] Synthesis of 1-(2-methyl-[1,1'-biphenyl]-3-yl)ethane-1-one (4B)

[0132] Following the method of Example 1, 4A and phenylboronic acid were coupled to obtain oily compound 4B in 87% yield. MS(EI) m / z 211[M+H] + ; 1H NMR (300MHz, DMSO-d6) δ (ppm) 7.70 (dd, J = 7.2, 2.4Hz, 1H), 7.50-7.26 (m, 7H), 2.58 (d, J = 2.7Hz, 3H), 2.22 (d, J = 3.0Hz, 3H).

[0133] Synthesis of 2-bromo-1-(2-methyl-1,1'-biphenyl)-3-yl)ethane-1-one (4C)

[0134] 4B (1.50 g, 7.13 mmol) and copper bromide (3.51 g, 14.27 mmol) were added to 20 mL of ethyl acetate, and the mixture was refluxed for 8 h. After cooling, the mixture was filtered and concentrated under reduced pressure. 10 mL of tetrahydrofuran, diethyl phosphite (0.26 g, 1.87 mmol), and triethylamine (0.21 g, 2.06 mmol) were added, and the mixture was reacted at room temperature for 2 h. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 1.34 g of a yellow oil (65% yield). MS (EI) m / z 289 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 7.80 (dd, J = 6.3, 2.7Hz, 1H), 7.50-7.36 (m, 5H), 7.35-7.26 (m, 2H), 4.91 (s, 2H), 2.21 (s, 3H).

[0135] Synthesis of 2-amino-1-(2-methyl-[1,1'-biphenyl]-3-yl)ethane-1-one (4D)

[0136] 4C (1.21 g, 4.15 mmol) and sodium dimethamide (0.39 g, 4.15 mmol) were added to 15 mL of acetonitrile and reacted at 75 °C for 12 h. The mixture was filtered while hot, concentrated under reduced pressure, and refluxed for 1 h after adding 5 mL of 4 M HCl. The mixture was then concentrated under reduced pressure and recrystallized from ethyl acetate to give 0.65 g of a white solid, in 70% yield. MS (EI) m / z 226 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 8.39 (s, 3H), 7.90-7.84 (m, 1H), 7.53-7.39 (m, 6H), 4.52 (d, J = 6.0Hz, 2H), 2.28 (s, 3H).

[0137] Synthesis of 2-chloro-N-(2-(2-methyl-[1,1'-biphenyl]-3-yl))-2-oxoethyl)acetamide (4E)

[0138] Chloroacetyl chloride (0.20 g, 1.79 mmol) was dissolved in 3 mL of anhydrous dichloromethane and added dropwise to a dichloromethane solution of 4D (0.40 g, 1.78 mmol) and triethylamine (0.74 g, 5.33 mmol) cooled in an ice bath. The reaction was carried out for 3 h, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 0.48 g of a yellow solid, with a yield of 90%. MS (EI) m / z 302 [M+H] + ; 1 ¹H NMR (300MHz, DMSO-d⁶) δ (ppm) 7.65 (dd, J = 7.8, 1.5Hz, 1H), 7.52–7.45 (m, 2H), 7.45–7.41 (m, 1H), 7.41 (d, J = 3.2Hz, 2H), 7.38–7.33 (m, 2H), 7.26 (dd, J = 7.8, 1.5Hz, 1H), 4.83 (s, 2H), 2.28 (s, 3H). Synthesis of 2-chloromethyl-5-(2-methyl-[1,1'-biphenyl]-3-yl)oxazole (4F)

[0139] 0.20 g (0.51 mmol) of 4E was added to 3 mL of acetic anhydride, followed by 4 drops of concentrated sulfuric acid. The mixture was reacted at 80 °C for 2 h. After cooling, the mixture was extracted with ethyl acetate and concentrated under reduced pressure to obtain 0.11 g of a yellow oily substance, with a yield of 62%. MS (EI) m / z 284 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 7.68 (s, 1H), 7.51-7.26 (m, 8H), 4.83 (s, 2H), 2.28 (s, 3H).

[0140] Synthesis of ((5-(2-methyl-[1,1'-biphenyl]-3-yl)oxazol-2-yl)methyl)glycine methyl ester (19m)

[0141] Following the method in Example 1, 4F and glycine methyl ester hydrochloride were subjected to a condensation reaction to obtain a white solid 19m, with a yield of 58%. MS(EI) m / z 337 [M+H] + ; 1 H NMR(300MHz,DMSO-d6)δ(ppm)7.65(dd,J=7.8,1.5Hz,1H),7.51-7.45(m,2H),7.44-7.41(m,1H),7.40(d,J=3.2Hz,2H),7 .37-7.33(m,2H),7.26(dd,J=7.8,1.5Hz,1H),5.12(p,J=6.3Hz,1H),4.46(s,2H),3.91(s,3H),3.86(s,2H),2.24(s,3H).

[0142] Synthesis of compound 19 and its hydrochloride (19S)

[0143] Following the method in Example 1, 19m was hydrolyzed to obtain 19, with a yield of 84%. Then, 19 was reacted with hydrochloric acid to form a salt, yielding 19s, with a yield of 78%. MS(ESI) m / z 323 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.51 (s, 1H), 7.69 (dd, J = 7.8, 1.5Hz, 1H), 7.50-7.47 (m, 2H), 7.46-7.41 (m, 1H), 7.39(d,J=3.2Hz,2H),7.36-7.30(m,2H),7.25(dd,J=7.8,1.5Hz,1H),3.96(s,2H),3.37(s,2H),2.25(s,3H).

[0144] The following compounds were prepared using a similar procedure to that in Example 19:

[0145]

[0146]

[0147] Example 25: Synthesis of ((2-(2-methyl-[1,1'-biphenyl]-3-yl)thiazolyl-5-yl)methyl)glycine (25) and its hydrochloride (25s)

[0148]

[0149] Synthesis of 3-bromo-2-methyl-1,1'-biphenyl (5A)

[0150] 1,3-Dibromo-2-methylbenzene (10.00 g, 40.01 mmol), phenylboronic acid (4.88 g, 40.01 mmol), potassium carbonate (11.06 g, 80.02 mmol), and Pd(PPh3)4 (0.46 g, 0.40 mmol) were added to 100 mL of 1,4-dioxane and 10 mL of water. The reaction was carried out under N2 atmosphere at 80 °C for 12 h. After cooling, the mixture was concentrated under reduced pressure, extracted with ethyl acetate, and dried over anhydrous sodium sulfate to give 8.90 g of a white oily substance, with a yield of 90%. MS (EI) m / z 247 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.73 (s, 1H), 7.51-7.32 (m, 7H), 2.39 (s, 3H).

[0151] Synthesis of 4,4,5,5-Tetramethyl-2-(2-methyl-[1,1'-biphenyl]-3-yl)-1,3,2-dioxoborane (5B)

[0152] 5A (9.00 g, 36.42 mmol), phenylboronic acid (18.50 g, 72.83 mmol), potassium acetate (10.76 g, 72.85 mmol), and Pd(dppf)Cl2 (0.53 g, 0.07 mmol) were added to 100 mL of 1,4-dioxane and reacted at 80 °C for 12 h under N2 atmosphere. After cooling, the mixture was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 8.00 g of a white oily substance, yield 74%. MS (EI) m / z 295 [M+H] + ; 1 H NMR (300MHz, Chloroform-d) δ (ppm) 7.66 (s, 1H), 7.56-7.32 (m, 7H), 2.33 (s, 3H), 1.21 (s, 12H).

[0153] Synthesis of 2-(2-methyl-[1,1'-biphenyl]-3-yl)thiazolyl-5-carboxaldehyde (5C)

[0154] 5B (3.05 g, 10.42 mmol), 2-bromothiazol-5-carboxaldehyde (1.00 g, 5.21 mmol), potassium carbonate (1.44 g, 10.42 mmol), and Pd(PPh3)4 (0.11 g, 0.10 mmol) were added to 10 mL of 1,4-dioxane and 1 mL of water. The mixture was reacted at 80 °C under a nitrogen atmosphere for 12 h. After cooling, the mixture was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 2.00 g of a white oily substance, in 80% yield. MS (EI) m / z 280 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.82 (s, 1H), 8.10 (s, 1H), 7.63 (dd, J = 7.6, 1.5Hz, 1H), 7.51-7.40 (m, 4H), 7.39-7.33 (m, 3H), 2.30 (s, 3H).

[0155] Synthesis of ((2-(2-methyl-[1,1'-biphenyl]-3-yl)thiazolyl-5-yl)methyl)glycine methyl ester (25m): 5C (0.15 g, 0.44 mmol) was added to 5 mL of DCM, followed by the sequential addition of glycine methyl ester hydrochloride (0.14 g, 0.89 mmol), TEA (0.13 g, 1.33 mmol), glacial acetic acid (0.13 g, 2.21 mmol), and sodium cyanoborohydride (0.14 g, 2.21 mmol). The reaction was carried out at room temperature for 4 h. 10 mL of water was added, and the mixture was extracted with ethyl acetate and purified by column chromatography to give 0.12 g of a white solid, with a yield of 60%. MS (ESI) m / z 353 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 8.10 (s, 1H), 7.63 (dd, J = 7.6, 1.5Hz, 1H), 7.51-7.40 (m, 4H), 7. 39-7.33(m,3H),4.38-4.22(m,2H),3.92(s,3H),3.88(s,1H),3.87-3.78(m,2H),2.30(s,3H).

[0156] Synthesis of compound 25 and its hydrochloride (25S)

[0157] Following the method in Example 1, 25m was hydrolyzed to obtain a white solid 25, with a yield of 60%. Then, 25 was reacted with hydrochloric acid to form a salt, yielding a white solid 25s, with a yield of 94%. MS (ESI) m / z 339 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 10.03 (s, 1H), 9.83 (s, 1H), 8.10 (s, 1H), 7.63 (dd, J = 7.6, 1.5Hz, 1H), 7 .51-7.40(m,4H),7.39-7.33(m,3H),4.38-4.22(m,2H),3.88(s,1H),3.87-3.78(m,2H),2.30(s,3H).

[0158] The following compounds were prepared using a similar procedure to that in Example 27:

[0159]

[0160] Example 27: Synthesis of (5-(2-methyl-[1,1'-biphenyl]-3-yl)thiophen-2-yl)methyl)glycine (27) and its hydrochloride (27s)

[0161]

[0162] Synthesis of 5-(2-methyl-[1,1'-biphenyl]-3-yl)thiophene-2-carboxaldehyde (6A)

[0163] 5B (3.07 g, 10.47 mmol), 5-bromothiophene-2-carboxaldehyde (1.00 g, 5.23 mmol), potassium carbonate (1.45 g, 10.47 mmol), and Pd(PPh3)4 (0.06 g, 0.05 mmol) were added to 10 mL of 1,4-dioxane and 1 mL of water. The reaction was carried out under N2 atmosphere at 80 °C for 12 h. After cooling, the mixture was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 2.00 g of a yellow oily substance, in 80% yield. MS (EI) m / z 279 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.85 (s, 1H), 7.47 (dd, J = 7.8, 6.3Hz, 2H), 7.43-7.39 (m, 1H), 7.39-7.38 (m, 1H) ),7.36(d,J=2.1Hz,2H),7.36-7.31(m,2H),7.25(dd,J=6.6,2.4Hz,1H),7.17(d,J=3.6Hz,1H),2.22(s,3H).

[0164] Synthesis of (5-(2-methyl-[1,1'-biphenyl]-3-yl)thiophene-2-yl)methyl)glycine methyl ester (27m)

[0165] Following the method of Example 27, 6A and glycine methyl ester hydrochloride were subjected to a condensation reaction to obtain a yellow solid 27m, with a yield of 58%. MS(ESI) m / z 352 [M+H] + ; 1 H NMR(300MHz, DMSO-d6)δ(ppm)7.57(dd,J=7.8,6.3Hz,2H),7.45-7.40(m,1H),7.39-7.37(m,1H),7.35(d,J=2.1Hz,2H),7 .34-7.31(m,2H),7.24(dd,J=6.6,2.4Hz,1H),7.19(d,J=3.6Hz,1H),4.45(s,2H),3.90(s,3H),3.88(s,2H),2.23(s,3H).

[0166] Synthesis of compound 27 and its hydrochloride (27S)

[0167] Following the method in Example 1, 27m was hydrolyzed to obtain a yellow solid 27, with a yield of 60%. Then, 27 was reacted with hydrochloric acid to form a salt, yielding a yellow solid 27s, with a yield of 94%. MS(ESI) m / z 338 [M+H] + ; 1 H NMR(300MHz, DMSO-d6)δ(ppm)7.67(dd,J=7.8,6.3Hz,2H),7.48-7.42(m,1H),7.40-7.38(m,1H),7.37(d,J=2.1Hz, 2H),7.35-7.32(m,2H),7.28(dd,J=6.6,2.4Hz,1H),7.20(d,J=3.6Hz,1H),4.42(s,2H),3.88(s,2H),2.24(s,3H).

[0168] The following compounds were prepared using a similar procedure to that in Example 27:

[0169]

[0170] Example 29: Synthesis of (5-(2-methyl-[1,1'-biphenyl]-3-yl)furan-2-yl)methyl)glycine (29) and its hydrochloride (29s)

[0171]

[0172] Synthesis of 5-(2-methyl-[1,1'-biphenyl]-3-yl)furan-2-carboxaldehyde (7A)

[0173] 5B (3.35 g, 11.43 mmol), 5-bromofuran-2-carboxaldehyde (1.00 g, 5.71 mmol), potassium carbonate (1.58 g, 11.43 mmol), and Pd(PPh3)4 (0.07 g, 0.06 mmol) were added to 10 mL of 1,4-dioxane and 1 mL of water. The reaction was carried out under N2 atmosphere at 80 °C for 12 h. After cooling, the mixture was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give 0.98 g of a yellow oily substance, in 65% yield. MS (EI) m / z 263 [M+H] + ; 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.85 (s, 1H), 7.49 (dd, J = 7.8, 6.3Hz, 2H), 7.43-7.42 (m, 1H), 7.39-7.37 (m, 1H) ),7.35(d,J=2.1Hz,2H),7.34-7.29(m,2H),7.23(dd,J=6.6,2.4Hz,1H),7.17(d,J=3.6Hz,1H),2.25(s,3H).

[0174] Synthesis of (5-(2-methyl-[1,1'-biphenyl]-3-yl)furan-2-yl)methyl)glycine methyl ester (29m)

[0175] Following the method described in Example 27, 7A and glycine methyl ester hydrochloride were subjected to a condensation reaction to obtain a yellow solid 29m, with a yield of 59%. MS (ESI) m / z 336 [M+H] + ; 1 H NMR(300MHz,DMSO-d6)δ(ppm)7.55(dd,J=7.8,6.3Hz,2H),7.46-7.43(m,1H),7.39-7.37(m,1H),7.36(d,J=2.1Hz,2H),7 .35-7.29(m,2H),7.26(dd,J=6.6,2.4Hz,1H),7.15(d,J=3.6Hz,1H),4.46(s,2H),3.91(s,3H),3.88(s,2H),2.25(s,3H).

[0176] Synthesis of compound 29 and its hydrochloride (29S)

[0177] Following the method in Example 1, 29m was hydrolyzed to obtain a yellow solid 29, with a yield of 60%. Then, 29 was reacted with hydrochloric acid to form a salt, yielding a yellow solid 29s, with a yield of 94%. MS(ESI) m / z 322[M+H] + ; 1 H NMR(300MHz, DMSO-d6)δ(ppm)7.56(dd,J=7.8,6.3Hz,2H),7.47-7.41(m,1H),7.39-7.38(m,1H),7.36(d,J=2.1Hz, 2H),7.35-7.31(m,2H),7.24(dd,J=6.6,2.4Hz,1H),7.18(d,J=3.6Hz,1H),4.42(s,2H),3.89(s,2H),2.23(s,3H).

[0178] Example 30: Inhibitory activity of the compounds of the present invention against PD-1 / PD-L1 protein-protein interactions

[0179] 1. Experimental Objective

[0180] The inhibitory activity of the compounds of this invention against PD-1 / PD-L1 protein-protein interactions was detected using the PD-1 / PD-L1 binding assay kit (BPS Bioscience).

[0181] 2. Main experimental materials

[0182] The PD-1 / PD-L1 binding assay kit was purchased from BPS Bioscience and contained reagents required for the experiment, including PD-1, PD-L1, Anti-tag1-Eu, Anti-tag2-XL665, Dilute Buffer, and Detection Buffer; 384-well microplates were purchased from Perkin Elmer; and the positive control (BMS-202) was purchased from Selleck.

[0183] 3. Instruments

[0184] Centrifuge (Eppendorf, model: 5430); Microplate reader (Perkin Elmer, model: EnVision)

[0185] 4. Experimental Methods

[0186] (1) Prepare 1×Assaybuffer.

[0187] (2) Compound loading: Using an Echo550 instrument, 200 nL of the compound was transferred to a 384 reaction plate according to different concentration gradients.

[0188] (3) Prepare PD-L1-Biotin working solution in 1×Assaybuffer.

[0189] (4) Add 5 μL of PD-L1-Biotin working solution to the compound well and the positive control well, respectively; add 5 μL of Assaybuffer to the negative control well.

[0190] (5) Centrifuge at 1000 rpm for 30 seconds and incubate at room temperature for 15 minutes.

[0191] (6) Prepare a mixture of PD-1-Eu and Dye labeled acceptor in 1×Assaybuffer.

[0192] (7) Add 15 μL of PD-1-Eu and Dye-labeled acceptor mixture.

[0193] (8) Centrifuge at 1000 rpm for 30 seconds and incubate at room temperature for 90 minutes.

[0194] (9) EnVision reads the 665nm / 615nm ratio. The inhibition rate of the compound on protein binding is calculated based on the fluorescence ratio.

[0195] 5. Calculation formula

[0196]

[0197] Where: Ratio sample It is the ratio of the sample wells; Ratio min : Mean ratio of negative control wells; Ratio max Mean ratio of positive control wells. Calculate the IC50 of the compound using Graphpad. 50 value.

[0198] 6. Experimental Results

[0199] The inhibitory activity of the compounds of this invention against PD-1 / PD-L protein-protein interactions is shown in Table 1. Experimental results indicate that the compounds of this invention exhibit significant inhibitory activity against PD-1 / PD-L1 protein-protein interactions. Wherein, A represents IC50. 50 =1~100nM; B represents IC 50 =100.01~500nM; C represents IC 50 >500nM.

[0200] Table 1. Inhibitory activity of the compounds of the present invention against PD-1 / PD-L1 interaction

[0201]

Claims

1. A biphenyl-substituted five-membered heterocyclic compound, characterized in that, The biphenyl-substituted five-membered heterocyclic compound is selected from any of the following compounds, or pharmaceutically acceptable salts thereof: 。 2. The biphenyl-substituted five-membered heterocyclic compound according to claim 1, characterized in that, The pharmaceutically acceptable salt is a salt formed by the compound with an acid or base, wherein the acid is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, and ferulic acid; and the base is selected from inorganic bases containing alkali metal cations, alkaline earth metal cations, or ammonium cations, or choline, piperazine, morpholine, triethylamine, diisopropylamine, and trimethylamine.

3. A pharmaceutical composition, characterized in that, It comprises the biphenyl-substituted five-membered heterocyclic compound of claim 1 and a pharmaceutically acceptable carrier.

4. A pharmaceutical composition, characterized in that, It comprises the biphenyl-substituted five-membered heterocyclic compound of claim 2 and a pharmaceutically acceptable carrier.

5. The use of the biphenyl-substituted five-membered heterocyclic compound of claim 1 in the preparation of PD-L1 inhibitor drugs.

6. The use of the biphenyl-substituted five-membered heterocyclic compound of claim 2 in the preparation of PD-L1 inhibitor drugs.

7. Use of the pharmaceutical composition of claim 3 in the preparation of a PD-L1 inhibitor drug.

8. Use of the pharmaceutical composition of claim 4 in the preparation of a PD-L1 inhibitor drug.

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