A fully symmetric biphenyl derivative, its preparation method and application

Abstract

This invention relates to a fully symmetrical biphenyl derivative, its preparation method, and its applications. In particular, this invention relates to a fully symmetrical biphenyl derivative having the structure of formula (I), its preparation method, and pharmaceutical compositions containing it. The series of compounds of this invention can be widely used in the preparation of drugs for the prevention and / or treatment of cancer or tumors, immune-related diseases and disorders, infectious diseases, or metabolic diseases mediated by the PD-1 / PD-L1 signaling pathway, and holds promise for development into a new generation of PD-1 / PD-L1 inhibitors.

Description

Technical Field

[0001] This invention belongs to the field of drug synthesis, specifically relating to a fully symmetrical biphenyl derivative, its preparation method, and its application. Prior Technology

[0002] The immune system plays a crucial role in controlling and eliminating diseases such as cancer. However, tumor cells often develop strategies to evade or suppress the surveillance of the immune system to promote their malignant growth. One important mechanism is the alteration of the expression of co-stimulatory and co-inhibitory immune checkpoint molecules on immune cells. Blocking the signaling pathway of immune checkpoint molecules, such as PD-1, has proven to be a promising and effective therapeutic approach.

[0003] Programmed cell death molecule 1 (PD-1), also known as CD279, is a receptor molecule expressed on the surface of activated T cells, natural killer T cells, B cells, and macrophages. Its structure includes an extracellular immunoglobulin variable region-like domain, a transmembrane region, and an intracellular region. The intracellular region contains two phosphorylation sites, located in an immunoreceptor tyrosine kinase-based inhibitory domain and an immunoreceptor tyrosine kinase-based switching domain, suggesting that PD-1 can negatively regulate T cell receptor-mediated signaling pathways.

[0004] PD-1 has two ligands, PD-L1 and PD-L2, which differ in their expression profiles. PD-L1 protein is upregulated in macrophages and dendritic cells after treatment with lipopolysaccharide (LPS) and granulocyte-macrophage colony-stimulating factor (GM-CSF), and is also upregulated in T cells and B cells after stimulation by T cell receptor and B cell receptor signaling pathways. It is also highly expressed in almost all tumor cells, and its expression is upregulated after interferon (IFN) gamma stimulation. In fact, the expression status of tumor PD-L1 is considered to have a prognostic correlation in many tumor types. PD-L2 expression, on the other hand, is more concentrated, mainly expressed on dendritic cells.

[0005] When PD-1-expressing T cells come into contact with cells expressing its ligands, the functional activities following antigen stimulation, such as cell proliferation, cytokine release, and cell lysis, are inhibited. Therefore, the interaction between PD-1 and its ligands functions as an intrinsic negative feedback regulatory mechanism to prevent excessive T cell activation during infection, immune tolerance, or tumorigenesis, thereby reducing autoimmune diseases and promoting autoimmune tolerance. Prolonged antigen stimulation, such as that occurring in tumors or chronic infections, leads to high levels of PD-1 expression in T cells, resulting in a lack of activity and function in responses to these long-term antigens—a condition known as T cell exhaustion. B cells also experience inhibitory effects and corresponding functional decline due to PD-1 and its ligands.

[0006] Some evidence from preclinical animal studies suggests that PD-1 and its ligands negatively regulate immune responses. PD-1-deficient mice develop lupus-like acute proliferative glomerulonephritis and dilated cardiomyopathy. Blocking the PD-1 / PD-L1 interaction with PD-L1 antibodies has been shown to restore and enhance T cell activation in many systems. Monoclonal antibodies against PD-L1 may also benefit patients with advanced cancer. Several preclinical animal tumor models have also shown that blocking the PD-1 / PD-L1 signaling pathway with monoclonal antibodies enhances immune responses and leads to immune responses to a range of histologically distinct tumors. Using a long-term infected LCMV model, the PD-1 / PD-L1 interaction has been found to inhibit the activation, expansion, and effector cell acquisition of virus-specific CD8 T cells. In addition to enhancing immune responses to long-term antigens, blocking the PD-1 / PD-L1 pathway has also been found to enhance vaccine responses, including responses to therapeutic vaccines in long-term infection settings.

[0007] In summary, besides existing monoclonal antibodies, developing compounds that block the protein-protein interactions of PD-1 / PD-L1 could serve as an effective therapeutic approach to enhance or restore T cell function by blocking the PD-1 / PD-L1-mediated inhibitory signaling pathway. Therefore, compounds targeting and blocking PD-1 / PD-L1 interactions are expected to show promising efficacy in the immunotherapy of various cancers and other immune-related diseases. Summary of the Invention

[0008] The purpose of this invention is to provide an overall fully symmetrical biphenyl derivative, its preparation method and application, thereby potentially leading to the development of a new generation of PD-1 / PD-L1 inhibitors.

[0009] The first aspect of the present invention provides a compound of formula (I), its stereoisomers, or a pharmaceutically acceptable salt thereof:

[0010] Wherein, the two R1s may be the same or different, and each is independently selected from hydrogen, deuterium, halogen, cyano, C1-4 alkyl, and C3-6 cycloalkyl, wherein the C1-4 alkyl and C3-6 cycloalkyl are optionally further substituted by one or more substituents selected from deuterium, halogen, cyano, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterated C1-4 alkyl, cyclopropyl, hydroxyl, and C1-4 alkoxy;

[0011] The two R2s may be the same or different, and each is independently selected from hydrogen, deuterium, C1-4 alkyl, C3-6 cycloalkyl, and hydroxyl, wherein the C1-4 alkyl and C3-6 cycloalkyl are optionally further substituted by one or more substituents selected from deuterium, halogen, cyano, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterated C1-4 alkyl, cyclopropyl, hydroxyl, and C1-4 alkoxy.

[0012] The two R3s may be identical or different, and each is independently selected from hydrogen, deuterium, C1-10 alkyl, C3-10 cycloalkyl, and 3-10 heterocyclic groups, wherein the C1-10 alkyl, C3-10 cycloalkyl, and 3-10 heterocyclic groups may optionally be further substituted by one or more substituents selected from deuterium, halogen, cyano, hydroxyl, carboxyl, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterated C1-4 alkyl, C3-6 cycloalkyl, 3-6 heterocyclic groups, C6-10 aryl, 5-8 heteroaryl, C3-6 cycloalkoxy, 3-6 heterocyclic, and C1-4 alkoxy.

[0013] The two m's are either the same or different, and each is independently 0, 1, 2, 3, or 4;

[0014] The two n values ​​are either the same or different, and each is independently 0, 1, 2, 3 or 4.

[0015] As a preferred embodiment, the two R1s in the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts are the same or different, and are each independently selected from hydrogen, deuterium, halogen, cyano, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterated C1-4 alkyl, cyclopropylsubstituted C1-4 alkyl and C3-6 cycloalkyl.

[0016] As a further preferred embodiment, the two R1s in the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts are the same or different, and each is independently selected from hydrogen, deuterium, fluorine, chlorine, cyano, methyl, ethyl, isopropyl, trifluoromethyl, trideuterylmethyl, cyclopropylmethyl and cyclopropyl.

[0017] As a preferred embodiment, the two R2s in the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts are the same or different and are each independently selected from hydrogen, deuterium, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterium-substituted C1-4 alkyl, cyclopropyl-substituted C1-4 alkyl, C3-6 cycloalkyl and hydroxyl.

[0018] As a further preferred embodiment, the two R2s in the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts are the same or different, and are each independently selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideuteryl, cyclopropylmethyl, cyclopropyl and hydroxyl.

[0019] As a preferred embodiment, the two R3s in the compound of formula (I), its stereoisomers, or its pharmaceutically acceptable salts are identical or different, and each is independently selected from hydrogen, deuterium, C1-4 alkyl, C3-6 cycloalkyl, and 3-6 membered heterocyclic groups, wherein the C1-4 alkyl, C3-6 cycloalkyl, and 3-6 membered heterocyclic groups are optionally further substituted by one or more substituents selected from deuterium, halogen, cyano, hydroxyl, carboxyl, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterated C1-4 alkyl, C3-6 cycloalkyl, 3-6 membered heterocyclic groups, C3-6 cycloalkoxy, and C1-4 alkoxy.

[0020] As a further preferred embodiment, the two R3s in the compound of formula (I), its stereoisomers, or its pharmaceutically acceptable salts are identical or different, and each is independently selected from hydrogen, deuterium, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, and azircyclobutyl, wherein the methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, oxecyclobutyl, and azircyclobutyl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, cyano, hydroxyl, carboxyl, methyl, ethyl, isopropyl, trifluoromethyl, trideuterylmethyl, cyclopropyl, cyclobutyl, oxecyclobutyl, azircyclobutyl, cyclopropyloxy, cyclobutoxy, methoxy, ethoxy, and isopropoxy.

[0021] As a further preferred embodiment, the two R3s in the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts are the same or different, and are each independently selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideuteryl, cyclopropylmethyl, cyclopropyl, cyclobutyl, oxacyclobutyl and aziridine.

[0022] As a further preferred embodiment, the compound of formula (I), its stereoisomers, or its pharmaceutically acceptable salts contain two identical R1s, and are selected from hydrogen, deuterium, fluorine, chlorine, cyano, methyl, ethyl, isopropyl, trifluoromethyl, trideuterylmethyl, cyclopropylmethyl, and cyclopropyl;

[0023] The two R2s are identical and are selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideuterium, cyclopropylmethyl, cyclopropyl, and hydroxyl;

[0024] The two R 3s are identical and are selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideuteryl, cyclopropylmethyl, cyclopropyl, cyclobutyl, oxetyl, and aziridine;

[0025] The two m's are the same, and are 0, 1, or 2;

[0026] The two n's are the same, and are either 1 or 2.

[0027] As the most preferred embodiment, the compound of formula (I), its stereoisomers, or pharmaceutically acceptable salts thereof include, but are not limited to, the following compounds: or .

[0028] A second aspect of the present invention provides a pharmaceutical composition comprising the aforementioned compound of formula (I), its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0029] A third aspect of the present invention provides the use of the aforementioned compound of formula (I), its stereoisomer, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diseases mediated by the PD-1 / PD-L1 signaling pathway.

[0030] As a preferred embodiment, the diseases mediated by the PD-1 / PD-L1 signaling pathway are selected from cancer or tumors, immune-related diseases and disorders, infectious diseases, infectious diseases, or metabolic diseases.

[0031] As a further preferred embodiment, the cancer or tumor is selected from lymphomas (including but not limited to lymphocytic lymphoma, primary central nervous system lymphoma, T-cell lymphoma, diffuse large B-cell lymphoma, follicular center lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, or primary mediastinal large B-cell lymphoma), sarcomas (including but not limited to Kaposi's sarcoma, fibrosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, leiomyosarcoma, rhabdomyosarcoma, soft tissue sarcoma, angiosarcoma, or lymphangioma). Sarcoma, melanoma, glioblastoma, synovial tumor, meningioma, biliary tract tumor, thymic tumor, nerve tumor, seminoma, nephroblastoma, pleomorphic adenoma, hepatocellular papilloma, renal tubular adenoma, cystadenoma, papilloma, adenoma, leiomyoma, rhabdomyoma, hemangioma, lymphangioma, osteoma, chondroma, lipoma, fibroma, central nervous system tumors, spinal axonoma, brainstem glioma, pituitary adenoma, multiple myeloma, ovarian tumor, myelodysplastic syndrome or mesothelioma, anterior Prostate cancer, recurrent or drug-resistant prostate cancer, thyroid cancer, parathyroid cancer, anal cancer, testicular cancer, urethral cancer, penile cancer, bladder cancer, ureteral cancer, uterine cancer, ovarian cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, adrenal cancer, Merkel cell carcinoma, embryonal carcinoma, and chronic or acute leukemia (including but not limited to acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, and chronic lymphocytic leukemia). The following cancers are included: bronchial cancer, esophageal cancer, nasopharyngeal carcinoma, hepatocellular carcinoma, renal cell carcinoma, small cell lung cancer, basal cell carcinoma, lung cancer, breast cancer, adenocarcinoma, papillary carcinoma, cystadenocarcinoma, squamous non-small cell lung cancer, non-squamous non-small cell lung cancer, rectal cancer, colon cancer, colorectal cancer, gastric cancer, pancreatic cancer, head and neck squamous cell carcinoma, head and neck cancer, gastrointestinal cancer, bone cancer, skin cancer, small bowel cancer, endocrine system cancer, renal pelvis cancer, epidermoid carcinoma, abdominal wall cancer, renal cell carcinoma, transitional cell carcinoma or choriocarcinoma, and metastatic tumors;

[0032] The immune-related diseases and disorders mentioned are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia, fibromyalgia, Alzheimer's disease, congestive heart failure, stroke, aortic stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infection, Crohn's disease, ulcerative colitis, allergic contact dermatitis and eczema, systemic sclerosis, and multiple sclerosis;

[0033] The infectious disease is selected from bacterial infectious diseases, viral infectious diseases, or fungal infectious diseases;

[0034] The infectious disease or infectious disease is selected from sepsis, liver infection, HIV, hepatitis A, hepatitis B, hepatitis C, hepatitis D, herpes simplex virus, human papillomavirus, or influenza;

[0035] The metabolic diseases mentioned are selected from diabetes mellitus, diabetic ketoacidosis, hyperglycemic hyperosmolar syndrome, hypoglycemia, gout, malnutrition, vitamin A deficiency, scurvy, vitamin D deficiency, or osteoporosis.

[0036] The fourth aspect of the present invention provides a compound of formula (I), its stereoisomer or a pharmaceutically acceptable salt thereof, which is used as a medicament for treating cancer or tumors, immune-related diseases and disorders, infectious diseases, or metabolic diseases mediated by the PD-1 / PD-L1 signaling pathway.

[0037] The present invention also relates to a method for treating cancer or tumor, immune-related diseases and disorders, infectious diseases, infectious diseases or metabolic diseases mediated by the PD-1 / PD-L1 signaling pathway, comprising administering to a patient in need a clinically effective amount of a compound of formula (I), its stereoisomer or a pharmaceutically acceptable salt thereof. Implementation

[0038] Through extensive and in-depth research, the inventors of this application have, for the first time, developed a series of biphenyl compounds with the general formula (I). These compounds exhibit strong inhibitory effects on the PD-1 / PD-L1 interaction and can be widely used in the preparation of drugs for the prevention and / or treatment of cancers or tumors, immune-related diseases and disorders, infectious diseases, and metabolic diseases mediated by the PD-1 / PD-L1 signaling pathway. They hold promise for development into a new generation of PD-1 / PD-L1 inhibitors. Based on this, this invention was completed.

[0039] Detailed explanation: Unless otherwise stated or specifically stated, the following terms used in the specification and the claims have the following meanings.

[0040] "alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group, preferably including straight-chain alkyl groups with 1 to 10 or 1 to 4 carbon atoms, and branched alkyl groups, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tributyl, dibutyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3- Methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl or their various branched isomers, etc. "C 1-10 alkyl" refers to straight-chain alkyl and branched alkyl with 1 to 10 carbon atoms, and "C 1-4 alkyl" refers to straight-chain alkyl and branched alkyl with 1 to 4 carbon atoms.

[0041] The alkyl group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 ynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0042] "Cycloalkyl" or "carbocyclic" refers to a substituent in a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon. A partially unsaturated cyclic hydrocarbon is one that may contain one or more (preferably 1, 2, or 3) double bonds, but none of the rings has a fully conjugated π-electron system. Cycloalkyl groups are classified as monocyclic or polycyclic, preferably comprising 3 to 10 or 3 to 6 carbon atoms. For example, "C 3-10 cycloalkyl" refers to a cycloalkyl group comprising 3 to 10 carbon atoms, and "C 3-6 cycloalkyl" refers to a cycloalkyl group comprising 3 to 6 carbon atoms.

[0043] Monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptanetrienyl, and cyclooctyl.

[0044] Polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups. "Spirocycloalkyl" refers to a polycyclic group in which a single carbon atom (called a spiro atom) is shared between the rings. These groups may contain one or more (preferably 1, 2, or 3) double bonds, but none of the rings has a fully conjugated π-electron system. Based on the number of shared spiro atoms between the rings, spirocycloalkyl groups are classified into monospirocycloalkyl, bispirocycloalkyl, or polyspirocycloalkyl groups. Spirocycloalkyl groups include, but are not limited to: .

[0045] "Fused cyclic alkyl" refers to a polycyclic aromatic hydrocarbon group in which each ring shares a pair of adjacent carbon atoms with the other rings in the system. One or more rings may contain one or more (preferably 1, 2, or 3) double bonds, but no ring has a fully conjugated π-electron system. Based on the number of constituent rings, fused cyclic alkyl groups can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic. Fused cyclic alkyl groups include, but are not limited to: .

[0046] "Bridged cycloalkyl" refers to a polycyclic aromatic hydrocarbon group in which any two rings share two non-directly bonded carbon atoms. These groups may contain one or more (preferably 1, 2, or 3) double bonds, but none of the rings has a fully conjugated π-electron system. Based on the number of constituent rings, they can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl groups. Bridged cycloalkyl groups include, but are not limited to: .

[0047] The cycloalkyl ring can be fused to an aryl, heteroaryl, or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl ring, including but not limited to indanyl, tetrahydronaphthyl, benzocycloheptyl, etc.

[0048] "Cycloalkyl" or "carbocyclic" may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 ynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0049] "Heterocyclic group" or "heterocyclic" refers to a substituent for a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon. The partially unsaturated cyclic hydrocarbon refers to a cyclic hydrocarbon that may contain one or more (preferably 1, 2, or 3) double bonds, but none of the rings has a fully conjugated π-electron system. One or more (preferably 1, 2, 3, or 4) ring atoms in the heterocyclic group are selected from N, O, N·O, or S(O)r (where r is an integer 0, 1, or 2), but do not include the -OO-, -OS-, or -SS- ring portions. The remaining ring atoms are carbon. Heterocyclic groups preferably include 3 to 10 or 3 to 6 ring atoms; for example, "3-6 membered heterocyclic group" refers to a heterocyclic group containing 3 to 6 ring atoms, and "3-10 membered heterocyclic group" refers to a heterocyclic group containing 3 to 10 ring atoms.

[0050] Monocyclic heterocyclic groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperidine, morpholinyl, thiomorpholinyl, homopiperidine, oxobutyl, tetrahydrofuranyl, etc.

[0051] Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups. A "spiroheterocyclic group" refers to a polycyclic heterocyclic group in which one or more (preferably 1, 2, 3, or 4) ring atoms are selected from N, O, N·O, or S(O)r (where r is an integer 0, 1, or 2), and the remaining ring atoms are carbon. These may contain one or more double bonds (preferably 1, 2, or 3), but none of the rings has a fully conjugated π-electron system. Spiroheterocyclic groups are classified into monospirocyclic, bispirocyclic, or polyspirocyclic groups based on the number of shared spiro atoms between rings. Spiroheterocyclic groups include, but are not limited to: .

[0052] "Fused heterocyclic group" refers to a polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system. One or more (preferably 1, 2, 3, or 4) rings may contain one or more (preferably 1, 2, or 3) double bonds, but no ring has a fully conjugated π-electron system. One or more (preferably 1, 2, 3, or 4) ring atoms are selected from heteroatoms of N, O, N·O, or S(O)r (where r is an integer 0, 1, or 2), and the remaining ring atoms are carbon. Based on the number of constituent rings, they can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic alkyl groups. Fused heterocyclic groups include, but are not limited to: .

[0053] "Bridged heterocyclic groups" refer to polycyclic heterocyclic groups in which any two rings share two non-directly bonded atoms. These groups may contain one or more (preferably 1, 2, or 3) double bonds, but none of the rings has a fully conjugated π-electron system. One or more (preferably 1, 2, 3, or 4) ring atoms are selected from heteroatoms of N, O, N·O, or S(O)r (where r is an integer of 0, 1, or 2), and the remaining ring atoms are carbon. Based on the number of rings, they can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic groups. Bridged heterocyclic groups include, but are not limited to: .

[0054] The heterocyclic ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, including but not limited to: .

[0055] The "heterocyclic group" or "heterocycle" may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 ynyl, C3-10 cycloalkyl, 3-10 membered heterocyclic group, C6-10 aryl, 5-10 membered heteroaryl, =O, =S and -SF5.

[0056] "Aryl" or "aromatic ring" refers to an all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group, a polycyclic group with a conjugated π-electron system (i.e., a ring with adjacent carbon atom pairs), preferably an all-carbon aryl group containing 6-10 carbons. For example, "C 6-10 aryl" refers to an all-carbon aryl group containing 6-10 carbons, including but not limited to phenyl and naphthyl. The aryl ring can be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, including but not limited to: .

[0057] The “aryl” or “aromatic ring” can be substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 ynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0058] "Heteroaryl" or "heteroary ring" refers to a heteroaryl system containing one or more (preferably 1, 2, 3, or 4) heteroatoms, including N, O, N·O, and S(O)r (where r is an integer 0, 1, or 2), preferably a heteroaryl system containing 5-10 or 5-8 ring atoms. For example, "5-8-membered heteroaryl" refers to a heteroaryl system containing 5-8 ring atoms, and "5-10-membered heteroaryl" refers to a heteroaryl system containing 5-10 ring atoms, including but not limited to furanyl, thiophene, pyridinyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyridine, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, including but not limited to: .

[0059] The "heteroaryl" or "heteroary ring" may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 ynyl, C3-10 cycloalkyl, 3-10 membered heterocyclic group, C6-10 aryl, 5-10 membered heteroaryl, =O, =S and -SF5.

[0060] "Alkenyl" refers to an alkyl group as defined above, consisting of at least two carbon atoms and at least one carbon-carbon double bond, preferably a straight-chain or branched alkenyl group containing 2-10 carbon atoms. For example, "C 2-10 alkenyl" refers to a straight-chain or branched alkenyl group containing 2-10 carbon atoms. This includes, but is not limited to, vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl groups.

[0061] "Alkenyl" can be substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups, independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0062] "Alkynyl" refers to an alkyl group as defined above, consisting of at least two carbon atoms and at least one carbon-carbon triple bond, preferably a straight-chain or branched alkynyl group containing 2-10 carbon atoms. For example, "C2-10 alkynyl" refers to a straight-chain or branched alkynyl group containing 2-10 carbon atoms. This includes, but is not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl.

[0063] "Alkynyl" can be substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups, independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0064] "Alkoxy" refers to -O-alkyl, where alkyl is defined as described above. For example, "C 1-4 alkoxy" refers to alkyloxy groups containing 1-4 carbons, including but not limited to methoxy, ethoxy, propoxy, butoxy, etc.

[0065] "Alkoxy" can be optionally substituted or unsubstituted. When substituted, the substituent, preferably one or more (preferably 1, 2, 3 or 4) of the following groups, are independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0066] "Cycloalkoxy" or "cycloalkyloxy" refers to -O-cycloalkyl, where cycloalkyl is defined as described above. For example, "C 3-6 cycloalkoxy" refers to cycloalkyloxy containing 3-6 carbons, including but not limited to cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexoxy, etc.

[0067] "Cycloalkoxy" or "cycloalkyloxy" may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 cycloalkyl, 3-10 heterocyclic, C6-10 aryl, 5-10 heteroaryl, =O, =S and -SF5.

[0068] "Heterocyclic oxy group" or "heterocyclic oxy group" refers to -O-heterocyclic oxy group, wherein the definition of heterocyclic oxy group is as described above, including but not limited to azirrocyclobutyoxy group, oxocyclobutyoxy group, azirrocyclopentyloxy group, nitrogen, oxocyclohexyloxy group, etc.

[0069] The "heterocyclic oxy group" or "heterocyclic oxy group" may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more (preferably 1, 2, 3 or 4) groups independently selected from deuterium, halogen, cyano, nitro, azide, C1-10 alkyl, halosubstituted C1-10 alkyl, deuterated C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 cycloalkyl, 3-10 membered heterocyclic group, C6-10 aryl, 5-10 membered heteroaryl, =O, =S and -SF5.

[0070] "Halogenated C1-4 alkyl" refers to 1-4 carbon alkyl groups in which the hydrogen atoms on the alkyl group are optionally replaced by fluorine, chlorine, bromine, or iodine atoms, including but not limited to difluoromethyl, dichloromethyl, dibromomethyl, trifluoromethyl, trichloromethyl, tribromomethyl, etc.

[0071] "Deuterated C1-4 alkyl" refers to alkyl groups on an alkyl group in which hydrogen is optionally replaced by a deuterium atom, consisting of 1-4 carbons. This includes, but is not limited to, monodeuteryl, dideuteryl, trideuteryl, etc.

[0072] "Halogen" refers to fluorine, chlorine, bromine, or iodine; "NaOH" refers to sodium hydroxide; "BH 3·Me 2S" refers to dimethyl sulfide borane; "MeOH" refers to methanol; "NaBH 3CN" refers to sodium cyanoboronhydride; "HOAc" refers to acetic acid; "DMF" refers to N,N-dimethylformamide; "THF" refers to tetrahydrofuran; and "Dess-Martin reagent" refers to Dess-Martin reagent.

[0073] The terms "optional" or "optionally located" mean that the event or environment described thereafter may but does not have to occur. This description includes situations where the event or environment occurs or does not occur, that is, it includes both substituted and unsubstituted cases. For example, "optionally alkyl-substituted heterocyclic groups" means that the alkyl group may but does not have to be present. This description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0074] "Substituted" refers to one or more hydrogen atoms in a group being independently replaced by a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, conforming to the valence bond theory in chemistry, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated bond (such as an alkene).

[0075] Stereoisomers, also known as cis-trans isomers, are isomers formed by different spatial arrangements of atoms in a molecule. They can be classified into two main categories: cis-trans isomers and enantiomers, or enantiomers and diastereomers. Stereoisomers resulting from the rotation of single bonds are called conformational stereo-isomers, sometimes also called rotamers. Stereoisomers resulting from bond length, bond angle, the presence of double bonds, or rings within the molecule are called configurational stereo-isomers. Configurational stereo-isomers are further divided into two categories. Isomers resulting from the inability of single bonds in double bonds or cyclic carbon atoms to rotate freely are called geometrical isomers, also known as cis-trans isomers, and have two configurations: Z and E. For example, cis-2-butene and trans-2-butene are a pair of geometric isomers. Stereoisomers with different optical properties due to the lack of anti-axial symmetry in their molecules are called optical isomers, which are classified into R and S configurations. In this invention, unless otherwise specified, "stereoisomer" can be understood to include one or more of the enantiomers, configuration isomers, and conformational isomers mentioned above.

[0076] In this invention, "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable acid addition salt, including inorganic acid salts and organic acid salts, which can be prepared by methods known in the art.

[0077] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0078] The present invention will be further described in detail and completely below with reference to the embodiments, but this is by no means a limitation of the present invention, nor is the present invention limited to the contents of the embodiments.

[0079] The structures of the compounds of this invention were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 / 500 NMR spectrometer, with deuterated dimethyl monoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as solvents, and tetramethylsilane (TMS) as the internal standard.

[0080] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 6120 mass spectrometer. High-performance liquid chromatography (HPLC) was performed using an Agilent 1200DAD high-performance liquid chromatograph (Sunfire C18 150 × 4.6 mm column) and a Waters 2695-2996 high-performance liquid chromatograph (Gimini C18 150 × 4.6 mm column).

[0081] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The standard size for TLC is 0.15 mm to 0.20 mm, while the standard size for TLC separation and purification is 0.4 mm to 0.5 mm. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0082] The starting materials used in the embodiments of the present invention are known and commercially available, or can be synthesized using or in accordance with methods known in the art.

[0083] Unless otherwise specified, all reactions in this invention are carried out under continuous magnetic stirring, in a dry nitrogen or argon atmosphere, using a dry solvent, and the reaction temperature is expressed in degrees Celsius (°C).

[0084] [Specific Examples of Compound Preparation]

[0085] [Example] [1] [:] [4-(((6-((3'-(5-(((4-] [Carboxyl bicyclic] [[2.2.2]] [Octane] [-1-] [base] [)] [Amino] [)] [methyl] [)-4-] [Cyclopropylpyridineamino] [)-2,2'-] [Dimethyl] [-[1,1'-] [Biphenyl] []-3-] [base] [)] [Carbamoyl] [)-4-] [Cyclopropylpyridine] [-3-] [base] [)] [methyl] [)] [Amino] [)] [Second Ring Road] [[2.2.2]] [Octane] [-1-] [Preparation of Carboxylic Acids]

[0086] [first step:] [4-(((4-] [Cyclopropyl] [-6-((3'-(4-] [Cyclopropyl] [-5-(((4-(] [Methoxycarbonyl] [)] [Second Ring Road] [[2.2.2]] [Octane] [-1-] [base] [)] [Amino] [)] [methyl] [)] [Pyridylamino] [)-2,2'-] [Dimethyl] [-[1,1'-] [Biphenyl] []-3-] [base] [)] [Carbamoyl] [)] [Pyridine] [-3-] [base] [)] [methyl] [)] [Amino] [)] [Second Ring Road] [[2.2.2]] [Octane] [-1-] Synthesis of methyl carboxylate

[0087] N,N'-(2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)bis(4-cyclopropyl-5-methoxypyridinemethylamine) [prepared according to the synthesis method in WO2021008491A1] (200 mg, 0.358 mmol) was placed in DMF (5 mL), followed by the addition of methyl 4-aminobicyclo[2.2.2]octane-1-carboxylate (656.1 mg, 3.58 mmol) and HOAc (1.5 mL), and stirred at room temperature for 1 hour. Sodium cyanoborohydride (225.0 mg, 3.58 mmol) was then added, and the mixture was stirred overnight at room temperature. After the reaction was complete, reverse-phase column chromatography was used to separate methyl 4-(((4-cyclopropyl-6-((3'-(4-cyclopropyl-5-(((4-(methoxycarbonyl)bicyclo[2.2.2]octane-1-yl)amino)methyl)pyridinylamino)-2,2'-dimethyl-[1,1'-biphenyl]-3-yl)aminomethyl)pyridin-3-yl)methyl)amino)bicyclo[2.2.2]octane-1-carboxylic acid (180 mg, 0.20 mmol, yield: 56.3%). MS m / z (ESI): 893.6 [M+H]+.

[0088] [Step Two:] [4-(((6-((3'-(5-(((4-] [Carboxyl bicyclic] [[2.2.2]] [Octane] [-1-] [base] [)] [Amino] [)] [methyl] [)-4-] [Cyclopropylpyridineamino] [)-2,2'-] [Dimethyl] [-[1,1'-] [Biphenyl] []-3-] [base] [)] [Carbamoyl] [)-4-] [Cyclopropylpyridine] [-3-] [base] [)] [methyl] [)] [Amino] [)] [Second Ring Road] [[2.2.2]] [Octane] [-1-] [Synthesis of carboxylic acids]

[0089] Methyl 4-(((4-cyclopropyl-6-((3'-(4-cyclopropyl-5-(((4-(methoxycarbonyl)bicyclo[2.2.2]octane-1-yl)amino)methyl)pyridinylamino)-2,2'-dimethyl-[1,1'-biphenyl]-3-yl)aminomethyl)pyridinyl-3-yl)methyl)amino)bicyclo[2.2.2]octane-1-carboxylic acid (350 mg, 0.39 mmol) was placed in a mixture of MeOH (5 mL), THF (5 mL), and water (5 mL), and then NaOH (314 mg, 7.84 mmol) was added. Under nitrogen protection, the mixture was heated to 50°C and stirred overnight. After the reaction was complete, the pH of the reaction solution was adjusted to ~6 with formic acid, and the reaction solution was concentrated. The crude product was separated by reverse-phase column chromatography to obtain 4-(((6-((3'-(5-((((4-carboxybicyclo[2.2.2]octane-1-yl)amino)methyl)-4-cyclopropylpyridinylamino)-2,2'-dimethyl-[1,1'-biphenyl]-3-yl)aminomethyl)-4-cyclopropylpyridinyl-3-yl)methyl)amino)bicyclo[2.2.2]octane-1-carboxylic acid (180 mg, 0.187 mmol, yield: 47.8%). MS m / z (ESI): 865.6 [M+H]+.

[0090] 1H NMR (400 MHz, DMSO- d 6) δ 10.31 (s, 2H), 8.55 (s, 2H), 7.89 (d, J= 8.0 Hz, 2H), 7.57 (s, 2H), 7.31 (t, J= 7.8 Hz, 2H), 6.97 (d, J= 7.5 Hz, 2H), 3.87 (s, 4H), 2.28 (td, J= 8.3, 4.2 Hz, 2H), 2.00 (s, 6H), 1.77 (dd, J= 10.5, 5.3 Hz, 12H), 1.60 (dd, J= 10.3, 5.5 Hz, 12H), 1.12 (dt, J= 7.7, 4.9 Hz, 4H), 0.84 (q, J= 5.2 Hz, 4H).

[0091] [Example] [2] [:] [4,4'-((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [carboxylic acid] [)] [Preparation]

[0092] [first step:] [4-(] [Methoxycarbonyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [Synthesis of carboxylic acids]

[0093] Dimethylbicyclo[2.2.1]heptane-1,4-dicarboxylic acid ester (2.0 g, 9.42 mmol) was dissolved in THF (40 mL), and a methanol solution of NaOH (377 mg, 9.42 mmol) (8 mL) was added dropwise. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was carefully concentrated until the solvent was completely removed. The solid was washed with petroleum ether and filtered. The filter cake was dissolved in water (50 mL), and the aqueous solution was acidified to pH ~4 with 2M hydrochloric acid and then extracted with ethyl acetate. The organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated to give 4-(methoxycarbonyl)bicyclo[2.2.1]heptane-1-carboxylic acid (1.49 g, yield: 80.0%).

[0094] [Step Two:] [4-((()] [Benzyloxy] [)] [Carbonyl] [)] [Amino] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0095] 4-(methoxycarbonyl)bicyclo[2.2.1]heptane-1-carboxylic acid (700 mg, 3.53 mmol) was dissolved in toluene (30 mL), followed by the addition of diphenyl azidophosphate (0.91 mL, 4.24 mmol) and triethylamine (0.98 mL, 7.06 mmol). The reaction mixture was heated to 120°C for 2 hours under nitrogen protection. The reaction solution was cooled to room temperature, and then benzyl alcohol (0.73 mL, 7.06 mmol) was added. The reaction mixture was then heated to 100°C for 18 hours under nitrogen protection. After the reaction was completed, the reaction solution was concentrated, and the crude product was separated by column chromatography to obtain methyl 4-(((benzyloxy)carbonyl)amino)bicyclo[2.2.1]heptane-1-carboxylic acid (1000 mg, 3.30 mmol, yield: 93.4%). MS m / z (ESI): 304.0 [M+H]+.

[0096] [Step 3:] [4-] [Aminobicyclic] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0097] Methyl 4-(((benzyloxy)carbonyl)amino)bicyclo[2.2.1]heptane-1-carboxylate (1000 mg, 3.30 mmol) was dissolved in methanol (30 mL), and then 10% Pd / C (100 mg) was added. The mixture was stirred overnight at room temperature under a hydrogen atmosphere at atmospheric pressure. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated to give methyl 4-aminobicyclo[2.2.1]heptane-1-carboxylate (500 mg, 2.955 mmol, yield: 89.6%). MS m / z (ESI): 170.0 [M+H]+.

[0098] [Step 4: Dimethyl] [4,4'-((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [Carboxylic esters] [)] [The synthesis of]

[0099] N,N'-(2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)bis(4-cyclopropyl-5-methoxypyridinemethoxyamine) (200 mg, 0.358 mmol) was dissolved in DMF (5 mL), then methyl 4-aminobicyclo[2.2.1]heptane-1-carboxylate (484.7 mg, 2.864 mmol) and HOAc (1.5 mL) were added, and the mixture was stirred at room temperature for 60 minutes. Then NaBH 3CN (225.0 mg, 3.58 mmol) was added, and the mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was complete, the reaction solution was separated by column chromatography (water / acetonitrile / formic acid) to obtain dimethyl 4,4'-((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(azaalkyldiyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid ester) (230 mg, 0.266 mmol, yield: 74.3%). MS m / z (ESI): 865.4 [M+H]+.

[0100] [Step 5:] [4,4'-((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [carboxylic acid] [)] [The synthesis of]

[0101] Dimethyl 4,4'-((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(azaalkyldiyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid ester) (230 mg, 0.266 mmol) was added to a mixture of MeOH (5 mL), THF (5 mL), and water (5 mL), followed by the addition of NaOH (212.7 mg, 5.317 mmol). The mixture was heated to 50°C and stirred for 1 hour under nitrogen protection. After the reaction was complete, the pH of the reaction solution was adjusted to ~6 with formic acid, and the reaction solution was concentrated. The crude product was separated by reversed-phase column chromatography to give 4,4'-((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(azaalkyldiyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid) (160 mg, 0.181 mmol, yield: 67.9%). MS m / z (ESI): 837.4 [M+H]+.

[0102] 1H NMR (400 MHz, DMSO- d 6) δ 10.33 (s, 2H), 8.69 (s, 2H), 7.86 (d, J= 8.2 Hz, 2H), 7.63 (s, 2H), 7.32 (t, J= 7.8 Hz, 2H), 6.99 (dd, J= 7.6, 1.3 Hz, 2H), 4.38 – 4.02 (m, 4H), 2.31 – 2.24 (m, 2H), 2.12 – 1.46 (m, 26H), 1.15 (d, J= 7.9 Hz, 4H), 0.95 – 0.78 (m, 4H).

[0103] [Example] [3] [:] [4,4'-((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Methylazaalkyldiyl] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [carboxylic acid] [)] [Preparation]

[0104] 4,4'-((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(azaalkyldiyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid) (50 mg, 0.060 mmol) was dissolved in DMF (3 mL), then HOAc (1 mL) and 35% formaldehyde aqueous solution (37.5 mg, 0.597 mmol) were added. After stirring at room temperature for 0.5 hours, NaBH3CN (37.5 mg, 0.597 mmol) was added. The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was complete, the reaction solution was separated by reversed-phase column chromatography to obtain 4,4'-((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(methylazaalkyldiyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid) (12 mg, 0.013 mmol, yield: 21.6%). MS m / z (ESI): 865.6 [M+H]+.

[0105] 1H NMR (400 MHz, DMSO- d 6) δ 10.30 (s, 2H), 8.52 (s, 2H), 7.90 (d, J= 8.0 Hz, 2H), 7.59 (s, 2H), 7.31 (t, J= 7.8 Hz, 2H), 6.97 (d, J= 7.5 Hz, 2H), 3.81 (s, 4H), 2.40 (td, J= 8.2, 4.2 Hz, 2H), 2.11(s, 6H), 2.05 – 1.95 (m, 10H), 1.87 – 1.57 (m, 16H), 1.12 (d, J= 8.1 Hz, 4H), 0.84 (d, J= 5.2 Hz, 4H).

[0106] [Example] [4] [:] [4,4'-(((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Methylene] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [carboxylic acid] [)] [Preparation]

[0107] [first step:] [4-(] [Hydroxymethyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0108] 4-(methoxycarbonyl)bicyclo[2.2.1]heptane-1-carboxylic acid (800 mg, 4.04 mmol) was dissolved in dry THF (40 mL), and BH3·Me2S / THF solution (5.25 mL, 5.25 mmol) was slowly added dropwise under ice-water bath cooling. After the addition was complete, the mixture was stirred overnight at room temperature. MeOH (10 mL) was added dropwise to the reaction solution, and the mixture was quenched by refluxing for 4 hours. The mixture was extracted with ethyl acetate and water, and the organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate. The mixture was filtered, the filtrate was concentrated, and the crude product was separated by column chromatography to obtain methyl 4-(hydroxymethyl)bicyclo[2.2.1]heptane-1-carboxylic acid (600 mg, yield: 80.7%).

[0109] 1H NMR (400 MHz, CDCl 3) δ 3.70 (s, 2H), 3.67 (s, 3H), 2.06 – 1.93 (m, 2H), 1.73 – 1.62 (m, 4H), 1.58 – 1.54 (m, 2H), 1.43 – 1.34 (m, 2H).

[0110] [Step Two:] [4-] [Cetyldicyclic] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0111] 4-(hydroxymethyl)bicyclo[2.2.1]heptane-1-carboxylic acid methyl ester (600 mg, 3.27 mmol) was dissolved in CH₂Cl₂ (20 mL), and then Dess-Martin reagent (1.66 g, 3.91 mmol) was added. The reaction was stirred at room temperature, and the reaction was monitored by TLC until the starting material disappeared. After filtration through diatomaceous earth, the filtrate was concentrated and separated by column chromatography to obtain 4-methoxybicyclo[2.2.1]heptane-1-carboxylic acid methyl ester (500 mg, yield: 84.3%).

[0112] 1H NMR (400 MHz, CDCl 3) δ 9.81 (s, 1H), 3.70 (s, 3H), 2.13 – 2.00 (m, 4H), 1.86 – 1.82 (m, 2H), 1.79 – 1.68 (m, 2H), 1.60 – 1.52 (m, 2H).

[0113] [Step 3:] [4-((] [Benzylamino] [)] [methyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0114] 4-Methyl 4-methylbicyclo[2.2.1]heptane-1-carboxylate (200 mg, 1.10 mmol) and benzylamine (235 mg, 2.20 mmol) were added to a mixture of MeOH (20 mL) and HOAc (0.2 mL). The mixture was stirred at room temperature for 30 minutes under nitrogen protection, followed by the addition of NaBH(OAc)3 (465 mg, 2.20 mmol), and stirring continued overnight at room temperature. After the reaction was complete, a saturated aqueous solution of NaHCO3 was added, and the mixture was extracted with CH2Cl2. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The mixture was filtered, and the crude product was concentrated and separated by column chromatography to obtain 4-((phenylmethylamino)methyl)bicyclo[2.2.1]heptane-1-carboxylate (300 mg, yield: 100%). MS m / z (ESI): 274.2 [M+H]+.

[0115] [Step 4:] [4-(] [Aminomethyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0116] Methyl 4-((benzylamino)methyl)bicyclo[2.2.1]heptane-1-carboxylate (300 mg, 1.10 mmol) and 10% Pd / C (50 mg) were placed in methanol (30 mL) and stirred overnight at room temperature under a hydrogen atmosphere. The reaction solution was filtered, and the filtrate was concentrated to give methyl 4-(aminomethyl)bicyclo[2.2.1]heptane-1-carboxylate (194 mg, yield: 96.5%). MS m / z (ESI): 184.1 [M+H]+.

[0117] [Step 5: Methyl] [4-({[(4-] [Cyclopropyl] [-6-{[3-(3-{4-] [Cyclopropyl] [-5-[({[4-(] [Methoxycarbonyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [base] []] [methyl] [}] [Amino] [)] [methyl] []] [Pyridine] [-2-] [Acetamino] [}-2-] [Methylphenyl] [)-2-] [Methylphenyl] []] [Carbamoyl] [}] [Pyridine] [-3-] [base] [)] [methyl] []] [Amino] [}] [methyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [Synthesis of Carboxylic Esters]

[0118] N,N'-(2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)bis(4-cyclopropyl-5-methoxypyridinemethylamine) (115 mg, 0.206 mmol) and methyl 4-(aminomethyl)bicyclo[2.2.1]heptane-1-carboxylate (189 mg, 1.03 mmol) were placed in a mixture of DMF (3 mL) and HOAc (0.75 mL) and stirred at room temperature for 1 hour under nitrogen protection. Then, NaBH3CN (65 mg, 1.03 mmol) was added, and stirring was continued at room temperature overnight. The reaction was completed, and the reaction mixture was separated by reverse-phase column chromatography to give methyl 4-({[(4-cyclopropyl-6-{[3-(3-{4-cyclopropyl-5-[({[4-(methoxycarbonyl)bicyclo[2.2.1]heptane-1-yl]methyl}amino)methyl]pyridin-2-nitroamino}-2-methylphenyl)-2-methylphenyl]aminomethyl}pyridin-3-yl)methyl]amino}methyl)bicyclo[2.2.1]heptane-1-carboxylic acid ester (124 mg, yield: 67.5%). MS m / z (ESI): 893.5 [M+H]+.

[0119] [Step 6:] [4,4'-(((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Methylene] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [carboxylic acid] [)] [The synthesis of]

[0120] Methyl 4-({[(4-cyclopropyl-6-{[3-(3-{4-cyclopropyl-5-[({[4-(methoxycarbonyl)bicyclo[2.2.1]heptane-1-yl]methyl}amino)methyl]pyridine-2-amino}-2-methylphenyl]aminomethyl}pyridine-3-yl)methyl]amino}methyl)bicyclo[2.2.1]heptane-1-carboxylic acid ester (124 mg, 0.139 mmol) was placed in a mixture of MeOH (3 mL), THF (3 mL) and water (1.5 mL), and NaOH (83 mg, 2.08 mmol) was added. The mixture was heated to 50°C and stirred for 1 hour under nitrogen protection. The reaction was completed, and the reaction solution was adjusted to pH ~6 with formic acid. The reaction solution was concentrated, and the crude product was separated by reverse-phase column chromatography to obtain 4,4'-(((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(azaalkyldiyl))di(methylene))di(bicyclo[2.2.1]heptane-1-carboxylic acid) carboxylate (45.5 mg, yield: 34.1%). MS m / z (ESI): 865.5 [M+H]+.

[0121] 1H NMR (400 MHz, DMSO- d 6) δ 10.30 (s, 2H), 8.57 (s, 2H), 7.88 (d, J= 8.0 Hz, 2H), 7.60 (s, 2H), 7.31 (t, J= 7.8 Hz, 2H), 6.98 (m, 2H), 3.98 (s, 4H), 2.67 (s, 4H), 2.28 (m, 2H), 2.01 (m, 6H), 1.90 – 1.82 (m, 4H), 1.65 – 1.58 (m, 4H), 1.54 (t, J= 10.3 Hz, 4H), 1.45 (m, 4H), 1.32 (m, 4H), 1.15 – 1.09 (m, 4H), 0.88 – 0.82 (m, 4H).

[0122] [Example] [5] [:] [4-((((6-((3'-(5-(((4-)] [Carboxyl bicyclic] [[2.2.1]] [Gengji] [-1-] [base] [)] [Ethyl] [)] [Aminomethyl] [)-4-] [Cyclopropylmethylpyridinamide] [)-2,2'-] [Dimethyl] [-[1,1'-] [Biphenyl] []-3-] [base] [)] [Carbamoyl] [)-4-] [Cyclopropylpyridine] [-3-] [base] [)] [methyl] [)] [Amino] [)] [Ethyl] [)] [Second Ring Road] [[2.2.1]] [Gengji] [-1-] [Preparation of Carboxylic Acids]

[0123] [first step:] [4-(2-(] [Benzylamino] [)] [Ethyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0124] Methyl 4-(2-carbonylethyl)bicyclo[2.2.1]heptane-1-carboxylate (350 mg, 1.78 mmol) and benzylamine (382 mg, 3.57 mmol) were placed in a mixture of MeOH (20 mL) and HOAc (0.2 mL) and stirred at room temperature for 30 minutes under nitrogen protection. Then, NaBH(OAc)3 (756 mg, 3.57 mmol) was added, and stirring was continued overnight at room temperature. After the reaction was complete, a saturated aqueous solution of NaHCO3 was added, and the mixture was extracted with CH2Cl2. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated and separated by column chromatography to obtain methyl 4-(2-(phenylmethylamino)ethyl)bicyclo[2.2.1]heptane-1-carboxylate (425 mg, yield: 82.9%). MS m / z (ESI): 288.2 [M+H]+.

[0125] [Step Two:] [4-(2-)] [Aminoethyl] [)] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] Synthesis of methyl carboxylate

[0126] Methyl 4-(2-(benzylamino)ethyl)bicyclo[2.2.1]heptane-1-carboxylate (425 mg, 1.48 mmol) and 10% Pd / C (50 mg) were placed in methanol (30 mL) and stirred overnight at room temperature under a hydrogen atmosphere at atmospheric pressure. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated to give methyl 4-(2-aminoethyl)bicyclo[2.2.1]heptane-1-carboxylate (246 mg, yield: 84.3%). MS m / z (ESI): 198.1 [M+H]+.

[0127] [Step 3: Dimethyl] [4,4'-(((((((2,2'-)] [Dimethyl] [-[1,1'-] [Biphenyl] []-3,3'-] [Two Basics] [)] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Carbonyl] [))] [two] [(4-)] [Cyclopropylpyridine] [-6,3-] [Two Basics] [))] [two] [(] [Methylene] [))] [two] [(] [Zazaalkyldiyl] [))] [two] [(] [Ethyl] [-2,1-] [Two Basics] [))] [two] [(] [Second Ring Road] [[2.2.1]] [Heptane] [-1-] [Carboxylic esters] [)] [The synthesis of]

[0128] N,N'-(2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)bis(4-cyclopropyl-5-methoxypyridinemethylamine) (110 mg, 0.197 mmol) and methyl 4-(2-aminoethyl)bicyclo[2.2.1]heptane-1-carboxylate (233 mg, 1.18 mmol) were placed in a mixture of DMF (3 mL) and HOAc (0.75 mL) and stirred at room temperature for 1 hour under nitrogen protection. Then, NaBH3CN (74 mg, 1.18 mmol) was added, and stirring was continued at room temperature overnight. The reaction was completed, and the reaction mixture was separated by reverse-phase column chromatography to give dimethyl 4,4'-(((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridin-6,3-diyl))di(methylene))di(azaalkyldiyl))di(ethyl-2,1-diyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid ester) (100 mg, yield: 55.1%). MS m / z (ESI): 921.5 [M+H]+.

[0129] [Step 4:] [4-((((6-((3'-(5-(((4-)] [Carboxyl bicyclic] [[2.2.1]] [Gengji] [-1-] [base] [)] [Ethyl] [)] [Aminomethyl] [)-4-] [Cyclopropylmethylpyridinamide] [)-2,2'-] [Dimethyl] [-[1,1'-] [Biphenyl] []-3-] [base] [)] [Carbamoyl] [)-4-] [Cyclopropylpyridine] [-3-] [base] [)] [methyl] [)] [Amino] [)] [Ethyl] [)] [Second Ring Road] [[2.2.1]] [Gengji] [-1-] [Synthesis of carboxylic acids]

[0130] Dimethyl 4,4'-(((((((2,2'-dimethyl-[1,1'-biphenyl]-3,3'-diyl)di(azaalkyldiyl))di(carbonyl))di(4-cyclopropylpyridine-6,3-diyl))di(methylene))di(azaalkyldiyl))di(ethyl-2,1-diyl))di(bicyclo[2.2.1]heptane-1-carboxylic acid ester) (100 mg, 0.108 mmol) was placed in a mixture of methanol (3 mL), THF (3 mL) and water (1.5 mL), and then NaOH (90 mg, 2.25 mmol) was added. The mixture was heated to 50 degrees Celsius and stirred for 1 hour. The reaction was completed, and the reaction solution was adjusted to pH ~6 with formic acid. After concentration, the reaction solution was separated by reverse-phase column chromatography to obtain 4-((((6-((3'-(5-((((4-carboxybicyclo[2.2.1]heptyl-1-yl)ethyl)aminomethyl)-4-cyclopropylmethylpyridinylamino)-2,2'-dimethyl-[1,1'-biphenyl]-3-yl)aminomethyl)-4-cyclopropylpyridinyl-3-yl)methyl)amino)ethyl)bicyclo[2.2.1]heptyl-1-carboxylic acid (45.5 mg, yield: 34.1%). MS m / z (ESI): 893.5 [M+H]+.

[0131] 1H NMR (400 MHz, DMSO- d 6) δ 10.30 (s, 2H), 8.55 (s, 2H), 7.88 (m, 2H), 7.58 (s, 2H), 7.31 (t, J= 7.8 Hz, 2H), 6.98 (m, 2H), 3.96 (s, 4H), 2.63 (t, J= 7.8 Hz, 4H), 2.29 (m, 2H), 2.00 (s, 6H), 1.86 (m, 4H), 1.69 (t, J= 7.8 Hz, 4H), 1.56 – 1.30 (m, 16H), 1.16 – 1.10 (m, 4H), 0.87 – 0.82 (m, 4H).

[0132] [, Biological testing evaluation , ] [, , ]

[0133] [one,] [PD-1 / PD-L1 HTRF] [Binding activity assay]

[0134] The effect of the compounds in the embodiments of the present invention on the PD-1 / PD-L1 protein interaction was determined using the Cisbio PD-1 / PD-L1 binding assay kit (#64ICP01PEG or 64ICP01PEH). The specific experimental method is as follows: 1) Add the diluted compound, 4 μL of Tag1-PD-L1 protein, and 4 μL of Tag2-PD-1 protein to a 384-well plate; 2) Incubate at room temperature for 15 minutes, then add 5 μL of anti-Tag1-Eu3+ antibody and 5 μL of anti-Tag2-XL665 antibody; 3) After incubation at room temperature for 2 hours or at 4 degrees Celsius overnight, take readings on a Pelkin Elmer EnVision. Read the values ​​at 665 nm and 620 nm respectively, and use the ratio of these two values ​​as the reading for each well. 4) Compare the readings of each well after compound treatment with the readings of the wells treated with DMSO to obtain the percentage of compound inhibition; 5) The IC50 values ​​of the compounds in the embodiments of the present invention were determined by nonlinear regression analysis at different compound concentrations. Specific experimental results are shown in Table 1.

[0135] [two,] [Jurkat] [Reporter Gene Cell Activity Assay]

[0136] The effects of the compounds in this invention on the interaction between PD-1 / PD-L1 proteins expressed on the cell surface and the resulting impact on T cell function were determined using the Jurkat reporter gene cell activity assay. In short, the NF-κB-luc reporter gene plasmid and the human PD-1 plasmid were transfected into Jurkat cells to establish a stable cell line simultaneously expressing both PD-1 and the NF-κB-luc reporter gene. Flow cytometry was used to identify the surface expression level of PD-1, and the reporter gene expression level was identified by the reporter gene response to stimulation with OKT-3 and Raiji cells.

[0137] In addition, a stable PD-L1 expression cell line was obtained by transfecting human PD-L1 expression plasmid into Raji cells. Then, Jurkat / NF-κB-luc / PD-1 cells and Raji-PD-L1 cells were co-cultured and stimulated with OKT-3. A compound was then added, and the inhibitory effect of the compound on the PD-1 / PD-L1 interaction on the T cell activation signaling pathway was reflected by the reporter gene response readings. The specific experimental methods are as follows: 1) Add 30 μL of compound or antibody at different dilutions to a white 96-well plate (corning, 3610), then add 10 μL of OKT3 (Biolegend, 317326) (final OKT3 concentration 1 μg / mL); 2) Add 20 μL of Raji-PD-L1 cell suspension to each well, 5 × 10⁴ cells per well, and incubate in an incubator for 20 minutes; 3) Add 20 μL of Jurkat / NF-κB-luc / PD-1 cell suspension to each well, 5 × 10⁴ cells per well, mix well, and detect Bright-glo (Promega, E2620) after 6 h; 4) Compare the readings of each well after compound treatment with the readings of the wells treated with DMSO to obtain the activation factor of the compound; 5) The EC50 values ​​of the compounds in the embodiments of this invention were determined by nonlinear regression analysis of the activation folds at different compound concentrations. Specific experimental results are shown in Table 1:

[0138] [surface] [1] [Biological test results] [Example Number] [PD-1 / PD-L1 HTRF] [Binding Activity] [IC, 50 , / nM ] [Cellular activity] [EC, 50 , / nM ] [1] 2.56 135 [2] 3.67 151 [3] 4.62 1383 [4] 1.46 247 [5] 4.19 163

[0139] Based on the bioactivity data of the compounds in the specific embodiments, the series of compounds of the present invention have a strong inhibitory effect on the protein interaction of PD-1 / PD-L1, and this inhibitory effect can enhance or restore T cell activation at the cellular level.

[0140] [III. Pharmacokinetic Evaluation] [(] [use] [Caco-2] [Cell line evaluation of candidate drug permeability and intestinal absorption potential] [)]

[0141] This experiment used Caco-2 cells from human colon cancer cells. Under appropriate culture conditions, Caco-2 cells can form differentiated monolayers with many characteristics, such as tight junctions, microvilli, and brush border enzyme expression, similar to normal intestinal epithelium. The permeability of compounds in the Caco-2 cell model is closely related to absorption in the human body. The Caco-2 cell model has been widely used to evaluate the permeability and active transport processes of compounds in the intestine. This experiment used the Caco-2 cell model to evaluate the permeability and efflux ratio of the compounds in the embodiments of this invention and comparative compounds. The specific methods are as follows: 1) The Caco-2 cell culture medium was a modified Eagle's medium (MEM) containing 10% inactivated fetal bovine serum and 1% non-essential amino acids. Cells were seeded on polycarbonate filters and cultured in a 37°C, 5% CO2 incubator. 2) Cells can be cultured for 21-28 days after seeding for transport experiments, and the density of cell monolayers can be characterized and verified by the apparent permeability coefficient (P app) of luciferin. 3) In the experiment, the compounds were dissolved in DMSO to prepare a 10 mM stock solution. Metoprolol, atenolol, and erythromycin were used as reference compounds to evaluate permeability, serving as positive compounds for high and low permeability, and as positive substrates for P-gp, respectively. The apparent permeability coefficients of the reference compounds obtained according to this experimental method are as follows: [Reference compound] [P, app , (10 , -6 , cm·s , -1 , ) ] [P, app , (BA) / P , app , (AB) ] Permeability evaluation AB BA [Metoprolol] 24.70 31.13 1.26 High permeability [Atenol] 0.42 0.57 1.37 Low permeability [Erythromycin] 0.20 21.65 107.3 High external displacement ratio

[0142] The working solution was prepared by dilution with Hanks' balanced salt solution (HBSS, Invitrogen) containing 25 mM HEPES (pH 7.4). The detection concentration of the test compound was 10 µM. This study was a bidirectional permeation experiment from the top membrane side to the basement side (AB) and from the basement side to the top membrane side (BA), requiring incubation at 37°C for 90 minutes. After incubation, the sample diluted with buffer was detected by LC-MS / MS. The concentration of the compound was quantified using a standard curve. 4) Apparent permeability coefficient Papp value (cm / s): P app= (VA / (Area × time)) × ([drug]accepter / (([drug]initial, donor) ×Dilution Factor)

[0143] Where “VA” refers to the volume in the receptor pore, “Area” refers to the surface area of ​​the membrane, and “time” is the total transport time in seconds. 5) The apparent permeability coefficient Papp and efflux ratio ER = Papp(BA) / Papp(AB) of the corresponding compounds were measured according to the above calculation formula. The results are shown in Table 2:

[0144] [surface] [2] [:] [Caco-2] [Results of membrane permeability test] [Example Number] [P, app , (10 , -6 , cm·s , -1, ) ] [P, app , (BA) / P , app , (AB) ] AB BA [1] 2.46 16.53 6.75 [4] 9.79 21.68 2.21 [5] 2.45 15.68 6.40 [Comparative Compounds] [1] <2.28 * <1.85 * / [Comparative Compounds] [2] 0.67 0.36 0.53 [Comparative Compounds] [3] N / A N / A / [Comparative Compounds] [4] <0.24 * 0.98 / [Comparative Compounds] [5] <1.79 <0.65 / [Comparative Compounds] [6] 0.34 5.51 16.26

[0145] *P app value is expressed as "<" when it indicates that the value is calculated using the minimum standard concentration at the receiver side, and depends on the actual concentration limit at the receiver side. "N / A" indicates not detected.

[0146] [surface] [3] [Compare compound structures] [Comparative Compounds] [Structure] [1] WO2019149183 Example 104 [2] WO2019149183 Example 105 [3] WO2019149183 Example 121 [4] WO2019149183 Example 122 [5] WO2019149183 Example 133 [6] WO2019149183 Example 134

[0147] Table 2 shows that the membrane permeability test data of the compounds in the embodiments of the present invention significantly improved the membrane permeability of the AB ends through modification with the terminal connecting groups on both sides of the compound. In contrast, compounds 2, 4, and 6, which also have symmetrical diacid structures, have very low AB values ​​and poor membrane permeability. In addition, the inventors further found through in vivo pharmacokinetic experiments in mice that these compounds are almost not absorbed by direct oral administration and can only be administered orally as prodrugs (comparative compounds 1, 3, and 5), which differs from the metabolic pathway of the compounds in the embodiments of the present invention.

[0148] [IV. Pharmacokinetic Evaluation in Mice and Rats]

[0149] [1.] [Research Objective]

[0150] The purpose of this experiment was to study the pharmacokinetic behavior of some compounds of the present invention. The administration methods were: single oral (PO) or intravenous (IV) administration to ICR mice or rats.

[0151] [2.] [Test Protocol]

[0152] [2.1] [Experimental Drug]

[0153] The compounds used in this experiment were derived from the compounds in the specific embodiments of this invention and the listed comparative compounds.

[0154] [2.2] [Experimental animals]

[0155] Male ICR mice (N=9) Original source: Shanghai Xipu-Bikai Experimental Animal Co., Ltd.

[0156] Male ICR rats (N=3) Original source: Shanghai Xipu-Bikai Experimental Animal Co., Ltd.

[0157] [2.3] [Drug Preparation and Administration]

[0158] The compounds were weighed and dissolved in their respective solvents, shaken, and sonicated to prepare colorless, clear solutions. Nine mice and three rats were given oral administration after fasting overnight. The dosage for mice was 10 mg / kg (or 2 mg / kg IV), and the dosage for rats was 5 mg / kg (or 1 mg / kg IV).

[0159] [2.4] [Sample Collection]

[0160] Approximately 90 μL of blood was collected via the submandibular vein at each time point. Heparin sodium was used for anticoagulation. After collection, the samples were placed on ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 8000 rpm, 6 minutes, 2-8 degrees Celsius). Blood collection time points were 0, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. Samples were stored at -20 degrees Celsius.

[0161] 40 μL of plasma sample was added to 160 μL of ice-cold acetonitrile containing an internal standard, vortexed for 3 minutes, and centrifuged at 11,000 rpm for 5 minutes. 100 μL of the supernatant was added to 100 μL of water, and 5 μL was injected into LC / MS / MS for analysis. The prodrug compounds and their corresponding prototype compounds 2, 4, and 6 were analyzed simultaneously with compounds 1, 3, and 5. Specific analytical results are shown in Table 4.

[0162] [2.5] [Test Results]

[0163] [surface] [4] [Testing mouse exposure and oral bioavailability of the compound] [Example Number] [Dosage] [C, max , (ng / mL) [AUC, last , (hr*ng / mL) [Oral bioavailability] [(F%)] [Example] [1] PO (10 mpk) 1912.1 8786.2 19.5% IV (2 mpk) 4003.3 8995.8 / [Example] [4] PO (10 mpk) 4546.7 17564.0 57.5% IV (2 mpk) 3523.3 6109.2 / [Example] [5] PO (10 mpk) 2983.3 14569.9 134% IV (2 mpk) 1040.3 2169.4 / [Comparative Compounds] [1] PO (10 mpk) 0 0 0% Detection and comparison of compound 2 1813.3 3968 33.7% [Comparative Compounds] [2] PO (10 mpk) 0 0 0% IV (2 mpk) 5640.0 2355 / [Comparative Compounds] [3] PO (10 mpk) 0 0 0% Detection and comparison of compound 4 412.3 592 / [Comparative Compounds] [5] PO (10 mpk) 30.4 / / Detection and comparison of compound 6 437.7 866 / [Comparative Compounds] [6] PO (10 mpk) 46.2 147 /

[0164] Experiments show that the compounds in the embodiments of the present invention have significantly improved oral exposure compared with the comparative compounds, and have good oral bioavailability, which can well support the development of oral drug delivery in the next step.

[0165] [surface] [5] [Testing the rat exposure and oral bioavailability of the compound] [Example Number] [Dosage] [C, max , (ng / mL) [AUC, last , (hr*ng / mL) [Oral bioavailability] [(F%)] [Example] [1] PO (5 mpk) 426.0 2222.6 18.9% IV (1 mpk) 3537 2351 / [Example] [4] PO (5 mpk) 1980 4836 44% IV (1 mpk) 1863 2197 / [Example] [5] PO (5 mpk) 492 1607 15.9% IV (1 mpk) 1883 2023 / [Comparative Compounds] [1] PO (5 mpk) 0 0 0% Detection and comparison of compound 2 266 629 1.6% [Comparative Compounds] [2] PO / / / IV (1 mpk) 13167 7642 /

[0166] Experiments show that the compounds in the embodiments of the present invention have significantly improved oral exposure compared with the comparative compounds, and have good oral bioavailability, which can well support the development of oral drug delivery in the next step.

[0167] All references to this invention are incorporated herein by reference as if each reference were individually incorporated herein by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of formula (I), its stereoisomer, or a pharmaceutically acceptable salt thereof: wherein, Both R1s are identical and selected from hydrogen, deuterium, halogen, cyano, C1-4 alkyl, halo-substituted C1-4 alkyl, deuterium-substituted C1-4 alkyl, cyclopropyl-substituted C1-4 alkyl, and C3-6 cycloalkyl; both R2s are identical and selected from hydrogen, deuterium, C1-4 alkyl, halo-substituted C1-4 alkyl, deuterium-substituted C1-4 alkyl, cyclopropyl-substituted C1-4 alkyl, C3-6 cycloalkyl, and hydroxyl. The two R3 groups are identical and selected from hydrogen, deuterium, C1-4 alkyl, C3-6 cycloalkyl, and 3-6 membered heterocyclic groups, wherein the C1-4 alkyl, C3-6 cycloalkyl, and 3-6 membered heterocyclic groups are optionally further substituted by one or more substituents selected from deuterium, halogen, cyano, hydroxyl, carboxyl, C1-4 alkyl, halosubstituted C1-4 alkyl, deuterated C1-4 alkyl, C3-6 cycloalkyl, 3-6 membered heterocyclic groups, C3-6 cycloalkoxy, and C1-4 alkoxy; the two m groups are identical and are 0, 1, or 2; the two n groups are identical and are 1 or 2.

2. The compound of formula (I) according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the two R1s are identical and selected from hydrogen, deuterium, fluorine, chlorine, cyano, methyl, ethyl, isopropyl, trifluoromethyl, trideuterylmethyl, cyclopropylmethyl and cyclopropyl.

3. The compound of formula (I) according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the two R2s are identical and selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideuterium, cyclopropylmethyl, cyclopropyl and hydroxyl.

4. The compound of formula (I) according to claim 1, its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that the two R3s are identical and selected from hydrogen, deuterium, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl and azircyclobutyl, wherein the methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, oxecyclobutyl and azircyclobutyl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, cyano, hydroxyl, carboxyl, methyl, ethyl, isopropyl, trifluoromethyl, trideuterylmethyl, cyclopropyl, cyclobutyl, oxecyclobutyl, azircyclobutyl, cyclopropyloxy, cyclobutoxy, methoxy, ethoxy and isopropoxy.

5. The compound of formula (I) according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the two R3s are identical and selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideuteryl, cyclopropylmethyl, cyclopropyl, cyclobutyl, oxacyclobutyl and aziridine.

6. A compound of formula (I) according to claim 1, its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that: two R1s are identical and selected from hydrogen, deuterium, fluorine, chlorine, cyano, methyl, ethyl, isopropyl, trifluoromethyl, trideutermethyl, cyclopropylmethyl and cyclopropyl; two R2s are identical and selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideutermethyl, cyclopropylmethyl, cyclopropyl and hydroxyl; two R3s are identical and selected from hydrogen, deuterium, methyl, ethyl, isopropyl, trifluoromethyl, trideutermethyl, cyclopropylmethyl, cyclopropyl, cyclobutyl, oxacyclobutyl and aziridine; two ms are identical and are 0, 1 or 2; two ns are identical and are 1 or 2.

7. The compound of formula (I) according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that it is selected from the following compounds: or.

8. A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1-7, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

9. The use of a compound of formula (I) as claimed in any one of claims 1-7, its stereoisomer, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diseases mediated by the PD-1 / PD-L1 signaling pathway.

10. The application according to claim 9, wherein the diseases mediated by the PD-1 / PD-L1 signaling pathway are selected from tumors, immune-related diseases and disorders, infectious diseases, or metabolic diseases.

11. The application according to claim 10, wherein the tumor is cancer.

12. The application according to claim 10, characterized in that the tumor is selected from lymphoma, sarcoma, melanoma, glioblastoma, synovoma, meningioma, biliary tract tumor, thymic tumor, neurotumor, seminoma, nephroblastoma, hepatocellular papilloma, papilloma, adenoma, leiomyoma, rhabdomyoma, hemangioma, lymphangioma, osteoma, chondroma, lipoma, fibroma, central nervous system tumor, spinal axonoma, brainstem glioma, multiple myeloma, ovarian tumor, myelodysplastic syndrome, mesothelioma, anal tumor, etc. Cancer of the uterus, testicles, urethra, penis, bladder, ureter, uterus, ovary, fallopian tube, endometrium, cervix, vagina, vulva, Merkel cell carcinoma, embryonal carcinoma, chronic or acute leukemia, bronchus, esophagus, nasopharyngeal carcinoma, hepatocellular carcinoma, renal cell carcinoma, basal cell carcinoma, lung cancer, adenocarcinoma, papillary carcinoma, rectal cancer, colon cancer, colorectal cancer, stomach cancer, head and neck cancer, bone cancer, skin cancer, small bowel cancer, endocrine system cancer, renal pelvis cancer, epidermoid carcinoma, abdominal wall cancer, transitional cell carcinoma, or choriocarcinoma; The immune-related diseases and disorders are selected from rheumatoid arthritis, renal failure, lupus erythematosus, asthma, psoriasis, ulcerative colitis, pancreatitis, allergies, fibrosis, anemia, fibromyalgia, Alzheimer's disease, congestive heart failure, stroke, aortic stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infections, Crohn's disease, allergic contact dermatitis and eczema, systemic sclerosis, and multiple sclerosis; the infectious diseases are selected from bacterial infectious diseases, viral infectious diseases, or fungal infectious diseases; the infectious diseases are selected from sepsis, liver infection, HIV, herpes simplex virus, human papillomavirus, or influenza; the metabolic diseases are selected from diabetes, diabetic ketoacidosis, hyperglycemic hyperosmolar syndrome, hypoglycemia, gout, malnutrition, vitamin A deficiency, scurvy, vitamin D deficiency, or osteoporosis.

13. The application according to claim 12, characterized in that: the lymphoma is selected from lymphocytic lymphoma, primary central nervous system lymphoma, T-cell lymphoma, diffuse large B-cell lymphoma, follicular center lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, and primary mediastinal large B-cell lymphoma; the sarcoma is selected from Kaposi's sarcoma, fibrosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, leiomyosarcoma, rhabdomyosarcoma, soft tissue sarcoma, angiosarcoma, and lymphangiosarcoma; the chronic or acute leukemia is selected from acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, and chronic lymphocytic leukemia; the adenoma is selected from pleomorphic adenoma, renal tubular adenoma, cystic adenoma, and pituitary adenoma; the adenocarcinoma is selected from prostate cancer, thyroid cancer, parathyroid cancer, adrenal cancer, breast cancer, cystic adenocarcinoma, and pancreatic cancer; The lung cancer is selected from small cell lung cancer, squamous non-small cell lung cancer, and non-squamous non-small cell lung cancer; the head and neck cancer is head and neck squamous cell carcinoma; the liver infection is selected from hepatitis A, hepatitis B, hepatitis C, and hepatitis D.

14. The application according to claim 13, characterized in that the prostate cancer is recurrent or has developed resistance to existing drugs.

15. The application according to claim 10, wherein the tumor is a metastatic tumor.

16. A compound of formula (I) according to any one of claims 1-7, its stereoisomer or a pharmaceutically acceptable salt thereof, used as a medicine for treating tumors, immune-related diseases and disorders, infectious diseases, or metabolic diseases mediated by the PD-1 / PD-L1 signaling pathway.