Aromatic heterocyclic compound containing tricyclic structure, preparation method and application thereof

Abstract

The present invention provides an aromatic heterocyclic compound containing a tricyclic structure, and a preparation method and application thereof. Specifically, the present invention provides a compound as shown in the following formula I, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof; wherein the definition of each group is as described in the specification. The compound of formula I can be used to treat diseases related to the PD-1 / PD-L1 signaling pathway.

Description

Technical Field

[0001] The present invention relates to the field of small molecule drugs. Specifically, the present invention provides a small molecule compound that can be used to treat diseases related to the PD-1 / PD-L1 signaling pathway. Background of the Invention

[0002] The immune system plays a vital role in controlling and curing many diseases, such as various cancers and diseases caused by viruses. However, cancer cells can often evade or modulate the immune system in some ways, thereby multiplying rapidly. One of the ways is to change the activation and inhibition molecules expressed on immune cells. Blocking the regulatory immune checkpoints, such as PD-1, has proven to be a very effective way to inhibit cancer cells.

[0003] PD-1 is programmed cell death protein-1, also known as CD279. It is mainly expressed in activated T cells and B cells, and its function is to regulate cell activation, which is a normal self-stabilizing mechanism of the immune system. Because excessive T / B cell activation can cause autoimmune diseases, PD-1 is a protective wall for our body. PD-1 is a type I transmembrane glycoprotein composed of 268 amino acids. Its structure mainly includes the outer immunoglobulin variable region, the hydrophobic transmembrane region, and the intracellular region. The intracellular region contains two phosphorylation sites, which are located in the immune receptor tyrosine regulatory motif and the immune receptor tyrosine conversion motif, which also proves that PD-1 can reversely regulate T cell receptor-mediated signals. PD-1 has two ligands, PD-L1 and PD-L2, which are expressed differently. PD-L1 is upregulated in a variety of tumor cells. It binds to PD-1 on T cells, regulates T cell proliferation and activation, and makes T cells inactive, ultimately inducing immune escape.

[0004] PD-1 / PD-L1 plays a reverse immunomodulatory role. When PD-1 binds to PD-L1, it can cause tyrosine polyphosphorylation in the tyrosine conversion motif domain of the T cell immune receptor, and the phosphorylated tyrosine can bind to the phosphatase protein tyrosinase 2 and protein tyrosinase 1. This can hinder the activation of extracellular signal-regulated kinases, and can also block the activation of phosphatidylinositol 3-kinase (PI3K) and serine-threonine protein kinase (Akt), thereby regulating T lymphocyte proliferation and the secretion of related cytokines. While PD-1 / PD-L1 signals inhibit T cell activation and proliferation, they can also cause the secretion of cytokines interleukin 2, interferon γ, and IL-10. In addition, PD-1 / PD-L1 signals also have similar immune functions on B cells. When PD-1 binds to the B cell antigen receptor, the PD-1 cytoplasmic region interacts with tyrosinase containing the protein tyrosinase 2 binding site, thereby hindering the activation of B cells.

[0005] PD-1 / PD-L1-based immunotherapy is a new generation of immunotherapy that has attracted much attention. In recent years, a series of surprising research results have confirmed that PD-1 / PD-L1 modulators have strong anti-tumor activity against a variety of tumors. Currently available PD-1 / PD-L1 antibody inhibitors include BMS's Ninolumab, Merck's Lambrolizumab, and Roche's Atezolizumab. In addition, there are many PD-1 / PD-L1 antibody modulators under development, including Cure Tech's Pidilizumab, GSK's AMP-224, and AstraZeneca's MEDI-4736.

[0006] Although tumor immunotherapy is considered to be a new generation of revolution in cancer treatment after targeted therapy, the PD-1 monoclonal antibodies currently on the market and under development have their own defects, including only being injected and not orally administered, being unstable in the body, being easily decomposed by proteases, being prone to immune cross-reactions, being difficult to purify and having high production costs, etc. Therefore, small molecule regulators of the PD-1 / PD-L1 interaction are a better choice for tumor immunotherapy or viral infection treatment.

[0007] In summary, there is an urgent need in this field to develop novel small molecule regulators of PD-1 / PD-L1 interaction. Summary of the invention

[0008] The purpose of the present invention is to provide a novel small molecule regulator of PD-1 / PD-L1 interaction.

[0009] The first aspect of the present invention provides a compound as shown in the following formula I, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof:

[0010]

[0011] wherein n, q, m, t and p are each independently selected from 0, 1, 2, 3 or 4;

[0012] X1, X2, X3, X4, X5 and X6 are each independently selected from the group consisting of N, O, S, SO, SO2, C(R)2, NR;

[0013] Y1, Y2, Y3, Y4, Y5 and Y6 are each independently selected from the group consisting of N, CH, C;

[0014] X7 is selected from the following group: N, CR;

[0015] X8 and X9 are each independently selected from the following groups: O, S, SO, SO2, C(R)2, NR; wherein R is selected from the following groups: H, C1-C6 alkyl, chlorine, bromine, fluorine, iodine, cyano, hydroxyl, nitro, NRf, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl, substituted or unsubstituted C6-C 10 heteroaryl, -C(=O)-NRdRe, -C(=O)-substituted or unsubstituted C1-C6 alkoxy, -C(=O)-substituted or unsubstituted C1-C6 alkyl, -C(=O)-substituted or unsubstituted C3-C 10 Cycloalkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, -C(=O)-substituted or unsubstituted C2-C6 alkenyl, -C(=O)-substituted or unsubstituted C2-C6 alkynyl;

[0016] M3, M4, and M5 are each independently selected from the group consisting of a chemical bond, CR, N, NR, O, S, SO, and SO2;

[0017] M6 is each independently selected from the following group: CR, N, C;

[0018] and represents a single or double bond;

[0019] Each is independently selected from the following group: a substituted or unsubstituted C5-C12 arylene group, or a substituted or unsubstituted 5-12-membered (preferably 5-7-membered) heteroarylene group having 1-3 heteroatoms, a substituted or unsubstituted 5-12-membered heterocyclylene group, or a substituted or unsubstituted C5-C12 cycloalkylene group;

[0020] Selected from the following group: substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12 membered heterocyclyl, substituted or unsubstituted C5-C12 cycloalkyl, wherein the heterocyclyl has 1-3 heteroatoms;

[0021] L1 is selected from the following group: a chemical bond, a substituted or unsubstituted C1-C4 alkylene, a substituted or unsubstituted C2-C4 alkenylene, a substituted or unsubstituted C2-C4 alkynylene, -S-, -O-, a substituted or unsubstituted -NH-, -S(O)-, -S(O)2-, a substituted or unsubstituted -NHC(O)NH-, Substituted or unsubstituted Substituted or unsubstituted Substituted or unsubstituted

[0022] G is Wherein, Rb and Rc are each independently selected from the following group: H, substituted or unsubstituted C1-C8 alkyl; or said Rb and Rc together with the adjacent N atom constitute a substituted or unsubstituted 5-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O;

[0023] R1, R2, R3, R4, R5, G and R are each independently selected from the group consisting of H, -CN, trifluoromethyl, sulfonylamino, nitro, hydroxy, halogen, -S-R8, -S(O)-R8, -S(O)2-R8, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, oxo substituted or unsubstituted C1-C6 amine, substituted or unsubstituted -(C1-C6 alkylene)-NH-(C1-C6 alkylene), carboxyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12-membered heteroaryl having 1-3 heteroatoms, substituted or unsubstituted 5-12-membered heterocyclyl having 1-4 heteroatoms, substituted or unsubstituted Substituted or unsubstituted or -(L 1a ) r -(L 2a ) s -(L 3a ) s -;-C 0-8 -O-R8,-C 0-8 -C(O)OR8,-C 0-8 -OC(O)OR8,-C 0-8 -NR8R9,-C 0-8 -N(R8)C(O)R9,-C 0-8 -C(O)NR8R9;

[0024] R8 and R9 are each independently selected from the following group: H, hydroxyl, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, oxo (i.e., =O), =NRf, -CN, hydroxyl, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted -(C1-C6 alkylene)-NH-(C1-C6 alkylene), carboxyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12-membered heteroaryl having 1-3 heteroatoms, substituted or unsubstituted 5-12-membered heterocyclyl having 1-4 heteroatoms, substituted or unsubstituted Substituted or unsubstituted or -(L 1a ) r -(L 2a ) s -(L 3a ) s -;

[0025] Each L 1a Each is independently a group selected from the group consisting of: a chemical bond, a substituted or unsubstituted C1-C7 alkylene group, a substituted or unsubstituted C2-C4 alkenylene group, a substituted or unsubstituted C2-C4 alkynylene group, -S-, -O-, a substituted or unsubstituted -NH-, -S(O)-, or -S(O)2-;

[0026] L 2a Selected from the following group: substituted or unsubstituted C6-C12 arylene, substituted or unsubstituted 5-12-membered heteroarylene having 1-3 heteroatoms, substituted or unsubstituted C3-C8 cycloalkylene, substituted or unsubstituted 5-10-membered heterocyclylene having 1-3 heteroatoms;

[0027] L 3a Selected from the following group: substituted or unsubstituted C1-C10 alkyl, C1-C10 aryl, -CN, hydroxyl, amino, carboxyl, -CO-NH-SO2-R g 、-NH-SO2-R g 、-SO2-NH-CO-R g ;

[0028] r is 1, 2, 3, 4, 5, 6;

[0029] s is 0, 1, and 2 respectively;

[0030] Rd, Re and Rg are each independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C 10 Cycloalkyl, -C 0-8 -O-R8,-C 0-8 -C(O)OR8,-C 0-8 -OC(O)OR8,-C 0-8 -NR8R9,-C 0-8 -N(R8)C(O)R9,-C 0-8 -C(O)NR8R9, substituted or unsubstituted C6-C 10 Aryl; or Rd and Re together form a substituted or unsubstituted 5-10 membered cycloalkyl, or a 5-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O;

[0031] The Rf is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C 10 Aryl, substituted or unsubstituted C6-C 10 heteroaryl, cyano, -C(=O)-NRdRe, -C(=O)-substituted or unsubstituted C1-C6 alkoxy, -C(=O)-substituted or unsubstituted C1-C6 alkyl, -C(=O)-substituted or unsubstituted C3-C 10 Cycloalkyl, -C(=O)-substituted or unsubstituted C2-C6 alkenyl, -C(=O)-substituted or unsubstituted C2-C6 alkynyl;

[0032] Unless otherwise specified, the "substituted" refers to being substituted by one or more (e.g., 2, 3, 4, etc.) substituents selected from the following group: halogen: including but not limited to -F, Cl, Br, -CH2Cl, -CHCl2, -CCl3, -CH2F, -CHF2, -CF3, oxo, -CN, hydroxyl, amino, C1-C6 alkylamino, carboxyl, -NHAc, a group selected from the following group which is unsubstituted or substituted by one or more substituents selected from the following group: C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, C3-C8 cycloalkyl, halogenated C6-C10 aryl, 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; the substituent is selected from the following group: halogen, hydroxyl, carboxyl, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

[0033] In the above formulae, any of the heteroatoms is selected from the group consisting of B, P, N, S and O.

[0034] In another preferred embodiment, the It is a benzene ring. Preferably, R3 is methyl, C1-C6 alkyl or alkoxy; C3-C6 cycloalkyl; C2-C6 alkene; C2-C6 alkynyl; halogen: including -F, Cl, Br, -CH2Cl, -CH2Br, -CHCl2, -CCl3, -CH2F, -CHF2, -CF3; -CN, hydroxyl, amino or alkyl substituted amino.

[0035] In another preferred embodiment, the It has the structure shown below:

[0036]

[0037] Wherein, Y7 and Y8 are each independently selected from: CH (the CH may be substituted by R1 or G), N, O, S, NH (the NH may be substituted by R1 or G);

[0038] R1 is selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, oxo (i.e., =O), =NRf, -CN, hydroxyl, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted -(C1-C6 alkylene)-NH-(C1-C6 alkylene), carboxyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-12-membered heteroaryl having 1-3 heteroatoms, substituted or unsubstituted 5-12-membered heterocyclyl having 1-4 heteroatoms, substituted or unsubstituted Substituted or unsubstituted

[0039] In another preferred embodiment, the compound of formula I has a structure as shown below:

[0040]

[0041] In another preferred embodiment, the compound of formula I has a structure as shown below:

[0042]

[0043] In another preferred embodiment, the Select from the following group:

[0044]

[0045] In another preferred embodiment, R4 is selected from the following group: NRdRe; said Rd and Re are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C 10 Cycloalkyl, -C 0-8 -O-R8, -C 0-8 -C(O)OR8, -C 0-8 -OC(O)OR8, -C 0-8 -NR8R9, -C 0-8 -N(R8)C(O)R9, -C 0-8 -C(O)NR8R9, substituted or unsubstituted C6-C 10 Aryl; or Rd and Re together form a substituted or unsubstituted 5-10 membered cycloalkyl, or a 5-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O; preferably, the substituent is a carboxyl group.

[0046] In another preferred embodiment, the compound is selected from the following table;

[0047]

[0048]

[0049]

[0050]

[0051]

[0052] The second aspect of the present invention provides a method for preparing the compound of formula I according to the first aspect of the present invention, wherein the method comprises steps selected from those shown in Synthesis Scheme 1 or 2:

[0053] Synthesis Scheme 1:

[0054]

[0055] (a) Using intermediates 1 and 2 as raw materials, a Sonogashira coupling reaction catalyzed by a palladium catalyst was performed to obtain the target product 3;

[0056] (b) Using 3 and 2-1 as raw materials, a coupling reaction occurs under the conditions of a palladium catalyst and a ligand (such as Suzuki, Buchwald, etc.) to obtain intermediate I-1;

[0057] Synthesis Scheme 2:

[0058]

[0059] (a) Compound 3-1 is used as a raw material and reacted with a nitrating agent (such as concentrated sulfuric acid / NaNO3, concentrated sulfuric acid / fuming nitric acid, etc.) to form intermediate 3-2;

[0060] (b) Using 3-2 as a raw material, a reduction reaction occurs under reducing conditions (Pd-C / H2; zinc powder / ammonium chloride; iron powder / acetic acid, etc.) to form intermediate 3-3;

[0061] (c) Using 3-3 and 3-4 as raw materials, a condensation reaction occurs under the condition of a shrinking agent, and an oxidant (such as DDQ, etc.) is added to undergo an oxidation reaction to obtain an intermediate 3-5;

[0062] (d) Using 3-5 and 2-1 as raw materials, a coupling reaction occurs in the presence of a catalyst and a ligand to form intermediate I-2.

[0063] The third aspect of the present invention provides a pharmaceutical composition, which comprises (1) a compound as described in the first aspect of the present invention or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof; and (2) a pharmaceutically acceptable carrier.

[0064] The fourth aspect of the present invention provides the use of the compound or its stereoisomer or tautomer as described in the first aspect of the present invention, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition as described in the third aspect of the present invention, characterized in that it is used to prepare a pharmaceutical composition for preventing and / or treating diseases related to the activity or expression of PD-1 / PD-L1.

[0065] In the fifth aspect of the present invention, a PD-1 / PD-L1 regulator is provided, which comprises the compound described in the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate.

[0066] In another preferred embodiment, the pharmaceutical composition is used to treat a disease selected from the following group: cancer, infectious diseases, and autoimmune diseases.

[0067] In another preferred embodiment, the cancer is selected from the following group: pancreatic cancer, bladder cancer, colorectal cancer, breast cancer, prostate cancer, kidney cancer, hepatocellular carcinoma, lung cancer, ovarian cancer, cervical cancer, gastric cancer, esophageal cancer, melanoma, neuroendocrine cancer, central nervous system cancer, brain cancer, bone cancer, soft tissue sarcoma, non-small cell lung cancer, small cell lung cancer or colon cancer, skin cancer, lung cancer, urinary system tumors, blood tumors, gliomas, digestive system tumors, reproductive system tumors, lymphomas, nervous system tumors, brain tumors, head and neck cancer.

[0068] In another preferred embodiment, the cancer is selected from the following group: acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), myelodysplastic syndrome (MDS), myeloproliferative disease (MPD), chronic myeloid leukemia (CML), multiple myeloma (MM), non-Hodgkin's lymphoma (NHL), mantle cell lymphoma (MCL), follicular lymphoma, Waldestrom macroglobulinemia (WM), T cell lymphoma, B cell lymphoma or diffuse large B cell lymphoma (DLBCL).

[0069] In another preferred embodiment, the infectious disease is selected from bacterial infection and viral infection.

[0070] In another preferred embodiment, the autoimmune disease is selected from organ-specific autoimmune disease and systemic autoimmune disease.

[0071] In another preferred embodiment, the organ-specific autoimmune diseases include chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, Goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis, and acute idiopathic polyneuritis.

[0072] In another preferred embodiment, the systemic autoimmune diseases include rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, and autoimmune hemolytic anemia.

[0073] In another preferred embodiment, the pharmaceutical composition is also used to improve T cell function in patients with chronic hepatitis B (CHB).

[0074] In another preferred embodiment, the inhibitor further comprises at least one therapeutic agent selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, or ipilimumab.

[0075] In the sixth aspect of the present invention, a method for regulating PD-1 / PD-L1 interaction in vitro is provided, comprising the steps of contacting the compound described in the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate with PD-L1 protein.

[0076] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0077] After extensive and in-depth research, the inventors have discovered a class of PD-1 / PD-L1 interaction regulators with excellent regulatory effects. On this basis, the inventors have completed the present invention.

[0078] definition

[0079] As used herein, the term "alkyl" includes straight or branched alkyl groups. For example, C1-C8 alkyl groups represent straight or branched alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, etc.

[0080] As used herein, the term "alkenyl" includes straight or branched alkenyls. For example, C2-C6 alkenyl refers to a straight or branched alkenyl having 2-6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.

[0081] As used herein, the term "alkynyl" includes straight or branched alkynyl groups. For example, C2-C6 alkynyl refers to a straight or branched alkynyl group having 2-6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.

[0082] As used herein, the term "C3-C 10 The term "cycloalkyl" refers to a cycloalkyl group having 3 to 10 carbon atoms. It may be a monocyclic ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may also be a bicyclic ring, such as a bridged ring or a spiro ring.

[0083] As used herein, the term "C1-C8 alkylamino" refers to an amine group substituted by a C1-C8 alkyl group, which may be monosubstituted or disubstituted; for example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di-tert-butylamino and the like.

[0084] As used herein, the term "C1-C8 alkoxy" refers to a straight or branched alkoxy group having 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy and the like.

[0085] As used herein, the term "3-10 membered heterocycloalkyl having 1-3 heteroatoms selected from the group consisting of N, S and O" refers to a saturated or partially saturated cyclic group having 3-10 atoms, wherein 1-3 atoms are heteroatoms selected from the group consisting of N, S and O. It may be a monocyclic or bicyclic form, such as a bridged ring or a spirocyclic form. Specific examples may include oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl and pyrrolidinyl, etc.

[0086] As used herein, the term "C6-C 10 The term "aryl" refers to an aromatic group having 6 to 10 carbon atoms, for example, phenyl or naphthyl and the like.

[0087] As used herein, the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S and O" refers to a cyclic aromatic group having 5-10 atoms, of which 1-3 atoms are heteroatoms selected from the group consisting of N, S and O. It may be a monocyclic ring or a condensed ring. Specific examples may include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2,4)-triazolyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl and the like.

[0088] Unless otherwise specified as "substituted or unsubstituted", the groups described in the present invention may be substituted by substituents selected from the following groups: halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-amine, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, etc.

[0089] As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl and Br. "Halogenated" means substituted with an atom selected from F, Cl, Br, and I.

[0090] Unless otherwise specified, the structural formulas described in the present invention are intended to include all isomeric forms (such as enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers or geometric isomers (or conformational isomers) are all within the scope of the present invention.

[0091] As used herein, the term "tautomer" means that structural isomers with different energies can interconvert across a low energy barrier. For example, proton tautomers (i.e., prototropic) include interconversion via proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion via reorganization of some bonding electrons.

[0092] As used herein, the term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

[0093] As used herein, the term "hydrate" refers to a complex formed by coordination of a compound of the present invention with water.

[0094] Active ingredients

[0095] As used herein, "compounds of the present invention" refers to compounds of Formula I, and also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of Formula I.

[0096] Preferred compounds of the present invention include compounds 1-360 (including various R-configuration and / or S-configuration stereoisomers of each compound, and / or E- / Z-cis-trans isomers).

[0097] In another preferred embodiment, the pharmaceutically acceptable salts include salts formed by combining with inorganic acids, organic acids, alkali metal ions, alkaline earth metal ions or organic bases that can provide physiologically acceptable cations, and ammonium salts.

[0098] In another preferred embodiment, the inorganic acid is selected from hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid; the organic acid is selected from methanesulfonic acid, p-toluenesulfonic acid, trifluoroacetic acid, citric acid, maleic tartaric acid, fumaric acid, citric acid or lactic acid; the alkali metal ion is selected from lithium ion, sodium ion, potassium ion; the alkaline earth metal ion is selected from calcium ion and magnesium ion; the organic base capable of providing physiologically acceptable cations is selected from methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris(2-hydroxyethyl)amine.

[0099] All of these salts within the scope of the present invention can be prepared by conventional methods. In the preparation process of the compound of general formula I, its solvate and its salt, different crystallization conditions may result in polymorphism or co-crystal.

[0100] Preparation of compounds of formula I

[0101] In order to prepare the compound of general formula I of the present invention, according to the structure of general formula I, the compound of general formula I of the present invention can be obtained by the following synthetic route.

[0102] Reaction 1:

[0103]

[0104] (a) Using intermediates 1 and 2 as raw materials, a Sonogashira coupling reaction catalyzed by a suitable palladium catalyst is performed to obtain the target product 3;

[0105] (b) Using 3 and 2-1 as raw materials, a coupling reaction occurs under the conditions of a suitable palladium catalyst and ligand (such as Suzuki, Buchwald, etc.) to obtain intermediate I-1

[0106] The preparation method of intermediate 1 is as follows:

[0107]

[0108] (a) Compound 1-1 is used as a raw material, and a coupling reaction occurs under the conditions of a suitable palladium catalyst and a ligand to form an intermediate 1-2;

[0109] (b) using 1-2 as a raw material, reacting with primary and secondary amines under the catalysis of a suitable Lewis acid, adding a suitable reducing agent, further reducing to form intermediate 1-3; or directly reducing amination reaction in the presence of a suitable reducing agent to form intermediate 1-3;

[0110] (c) Using 1-3 as the starting material, a halogenation reaction occurs under the catalysis of a suitable Lewis acid to obtain intermediate 1.

[0111] Among them, the preparation method of intermediate 2-1 (refer to WO2018195321, WO2018005374 and WO2019023575) is as follows:

[0112]

[0113] (a) Compound 2-4 is used as a raw material to form chiral alcohol 2-5 in the presence of a chiral auxiliary (such as R / S-CBS) and a reducing agent (such as borane);

[0114] (b) Using 2-5 (or 2-8) and 2-6 as raw materials, a coupling reaction (such as Suzuki, Buchwald, etc.) occurs under the conditions of a suitable palladium catalyst and a ligand to obtain an intermediate 2-7 (or 2-9);

[0115] (c) Compound 2-4 and R / S-tert-butylsulfonimide are used as raw materials, and a protected chiral amino compound is formed under the action of Lwesis acid (such as ethyl tetratitanate) and a reducing agent (such as lithium aluminum tetrahydride, sodium borohydride, etc.), and then the protecting group is removed under acidic conditions to obtain a chiral amino compound 2-8;

[0116] (d) Intermediate 2-7 (or 2-9) and pinacol diboronate are used as raw materials, and a Suzuki coupling reaction is carried out in the presence of a suitable palladium catalyst and a ligand to obtain intermediate 2-1.

[0117] Reaction 2:

[0118]

[0119] (a) Compound 3-1 is used as a raw material and reacted with a suitable nitrating agent (such as concentrated sulfuric acid / NaNO3, concentrated sulfuric acid / fuming nitric acid, etc.) to form intermediate 3-2;

[0120] (b) Using 3-2 as a raw material, under appropriate reducing conditions (Pd-C / H2; zinc powder / ammonium chloride; iron powder / acetic acid, etc.), a reduction reaction occurs to form intermediate 3-3;

[0121] (c) Using 3-3 and 3-4 as raw materials, a condensation reaction occurs under the condition of a suitable shrinking agent, and an appropriate oxidant (such as DDQ, etc.) is added to undergo an oxidation reaction to obtain an intermediate 3-5;

[0122] (d) Using 3-5 and 2-1 as raw materials, a coupling reaction occurs under the conditions of a suitable catalyst and ligand to form intermediate I-2.

[0123] In addition, the starting materials and intermediates in each of the above reactions are easily available, and each step of the reaction can be easily synthesized according to the reported literature or by conventional methods in organic synthesis for those skilled in the art. The compound described in general formula I may exist in the form of a solvate or an unsolvate, and different solvates may be obtained by crystallization using different solvents.

[0124] Pharmaceutical compositions and methods of administration

[0125] Since the compounds of the present invention have excellent inhibitory activity on PD-1 / PD-L1 interaction, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to prevent and / or treat (stabilize, alleviate or cure) PD-1 / PD-L1 interaction-related diseases (e.g., cancer, infectious diseases, autoimmune diseases).

[0126] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0127] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0128] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration or subcutaneous administration).

[0129] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0130] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0131] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0132] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0133] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0134] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0135] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds (eg, other anticancer agents).

[0136] When administered in combination, the pharmaceutical composition also includes one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds. One or more (2, 3, 4, or more) of the other pharmaceutically acceptable compounds can be used simultaneously, separately or sequentially with the compounds of the present invention to prevent and / or treat PD-1 / PD-L1 interaction-related diseases.

[0137] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0138] The main advantages of the present invention include:

[0139] (1) The compounds of the present invention have high inhibitory activity on PD-1 / PD-L1 interaction, have strong binding ability with PD-L1 protein, and have the ability to relieve PD-L1 from inhibiting IFNγ.

[0140] (2) The compounds of the present invention have better solubility and very low toxicity to normal cells, and thus can be applied to therapeutic subjects within a larger dosage range.

[0141] (3) Compared with the compounds of the prior art, the compounds of the present invention have better solubility and thus have good drugability. Compared with the existing compounds, the compounds of the present invention show good bioavailability in in vivo experiments. In addition, compared with the existing compounds, the compounds of the present invention are very easy to prepare into pharmaceutically acceptable salts, thus facilitating the further formation of preparations.

[0142] (4) In vivo efficacy studies have shown that the compounds of the present invention can significantly inhibit the growth of subcutaneous tumors, both in terms of tumor volume and weight, and can significantly increase the number of lymphocytes in the blood and spleen of mice.

[0143] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the unrecorded specific conditions in the following examples are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0144] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial channels.

[0145] General materials and test methods:

[0146] The instruments and raw materials involved in the embodiments are described as follows:

[0147] The hydrogen nuclear magnetic resonance spectrum was obtained by Bruker AV-400 (400 MHz) nuclear magnetic resonance spectra.

[0148] Chemical shifts were recorded with tetramethylsilane as internal standard and expressed in ppm (CDC13: δ7.26ppm). The data recorded are as follows: chemical shifts and their splitting and coupling constants (s: singlet; d: doublet; t: triplet; q: quartet; br: broad; m: multiplet).

[0149] Mass spectrometry data were analyzed by Finnigan LCQ Advantage LC / MS, and all reactions were performed under dry argon protection in anhydrous and oxygen-free conditions. Solid metal organic compounds were stored in an argon protection dry box.

[0150] Tetrahydrofuran and ether are obtained by distillation, in which sodium metal and benzophenone are added. Dichloromethane, pentane and hexane are treated with calcium hydride.

[0151] The special raw materials and intermediates involved in the present invention are provided by Tianjin Changsen Pharmaceutical Co., Ltd., and all other chemical reagents are purchased from reagent suppliers such as Shanghai Chemical Reagent Company, Aldrich, and Acros. If the intermediates or products required for the reaction during the synthesis are insufficient for the next test, the synthesis is repeated several times until a sufficient amount is obtained.

[0152] Unless otherwise specified, the raw materials and reagents involved in the present invention can be purchased commercially or customized.

[0153] The compounds of the present invention may contain one or more asymmetric centers, so the series of compounds may be in the form of racemates or single enantiomers. The compounds prepared by the present invention are heterocyclic compounds with a purity higher than 95%. The structure of each final product is characterized by MS and / or H NMR ( 1 The following examples illustrate the synthesis of various compounds and intermediates of the present invention.

[0154] Example 1 Synthesis of Compound 001

[0155]

[0156] Step 1-1:

[0157]

[0158] Compound 1 (5.0 g, WO2012158550), 2 (4.4 g), Pd(OAc)2 (265 mg) and Cs2CO3 (15.4 g) were added to dioxane (100 mL), N2 was protected, and the reaction was carried out at 95 degrees for 2 hours. The reaction was complete on TLC. The reaction solution was filtered, the filtrate was dried, and column chromatography (EA:HEP=1:30) was performed to obtain 4.8 g of light yellow solid. MS-APCI: 348 [M+H] + .

[0159] Step 1-2:

[0160]

[0161] Compound 3 (4.8 g) and 4 (2.5 g, dissolved in 1.5 mL AcOH) were added to DCM, and then 2.8 g TEA was added. After stirring at room temperature for 1 h, NaBH(OAc)3 (4.4 g) was added. After 30 minutes, the reaction was complete on TLC. The reaction solution was washed with saturated NaHCO3, extracted with DCM, dried with DCM, and purified by column (MeOH:DCM=1:20) to obtain 5.1 g of product as a light yellow oily viscous substance. MS-APCI: 419[M+H] + .

[0162] Steps 1-3:

[0163]

[0164] Compound 5 (5.1 g), Bpin2 (3.4 g), Pd(dppf)Cl2.DCM (498 mg), KOAc (2.4 g) were added to dioxane (100 mL), N2 gas was protected, and the reaction was carried out at 90 degrees for 2.5 hours. LC-MS detected that the reaction was complete. The reaction solution was filtered, washed with water, extracted with DCM, dried, spin-dried, and passed through a column (DCM:MeOH=30:1) to obtain 5.2 g of the product as a gray solid. MS-APCI: 467 [M+H] + .

[0165] 1H NMR(400MHz,Chloroform-d)δ7.85(d,J=8.1Hz,1H),7.75(d,J=7.3Hz,1H),7.50 (d,J=7.5Hz,1H),7.21(t,J=7.4Hz,1H),6.45(dd,J=7.1,4.5Hz,1H),6.37(d,J= 8.0Hz,1H),4.58(s,1H),4.22(s,2H),4.01(s,3H),3.38-3.30(m,2H),3.15-3.0 7(m,2H),2.62-2.53(m,2H),2.45-2.35(m,2H),2.27-2.17(m,3H),1.33(s,12H).

[0166] Steps 1-4:

[0167]

[0168] 7 (30g) was dissolved in 400mL of acetonitrile-Water (1:1), nitrogen protection, the system was clear, ice bath, p-toluenesulfonic acid hydrate (901g) was added at an internal temperature of 0-5 degrees, stirred for 10 minutes, the system was pink and turbid, and NaNO2 aqueous solution (22g, 45mL) was added dropwise at an internal temperature of 0-5 degrees. The system was reacted for 30 minutes, and the system was clear; protected from light, potassium iodide aqueous solution (67g, 60mL) was added dropwise at 0-5 degrees, and the temperature was naturally raised to room temperature for reaction; TLC tracking; post-treatment: quenched by adding saturated sodium sulfite under ice bath, HEP extraction, washed once with saturated brine, mixed, column: HEP washed down the product, concentrated to obtain 46.9g of yellow-brown liquid, yield: 98.1%. MS-APCI: 296[M+H] + .

[0169] Steps 1-5:

[0170]

[0171] 8 (1g), CuI (0.03g), Pd (Ph3P) 2Cl2 (0.12g) were added to a 50mL reaction bottle, replaced with nitrogen 3 times, anhydrous DMF (20mL) was added, the system was reddish brown and clear; triethylamine (1.7g) was added, the system was brown and clear; nitrogen replaced 3 times, protected from light, 9 (0.52g) was added, and the reaction was carried out at room temperature; TLC tracking; post-treatment: quenched by adding saturated ammonium chloride in an ice bath, extracted with MTBE 2 times, washed with saturated brine 5 times, mixed, column: HEP washed down the product, concentrated to obtain 1.03g of yellow-brown liquid, yield: 95%. MS-APCI: 309 [M + H] + .

[0172] Steps 1-6:

[0173]

[0174] 10 (5 g) was dissolved in 50 mL THF, and 17 mL TBAF (1 M) was added at room temperature and reacted for 1 h. TLC tracking; post-treatment: spin-drying, mixing, column: HEP washed down the product, concentrated to obtain 3 g of light yellow liquid.

[0175] Steps 1-7:

[0176]

[0177] 12 (5 g) was dissolved in 50 mL DMF, and NIS (7.6 g, dissolved in 20 mL DMF) was added dropwise at 0 degrees for 2 hours, and the reaction was continued at 0 degrees for 1 hour. TLC showed that the raw material had reacted. The reaction solution was poured into water, extracted with EA, dried, and filtered to obtain 7.0 g of product as a light yellow solid. MS-APCI: 275.2 [M+H] + .

[0178] Steps 1-8:

[0179]

[0180] In a 100mL two-necked bottle, 13 (3.0g) was dissolved in DMF (30mL), and Zn(CN)2 (1.3g) and Pd(pph3)4 (2.5g) were added. The reaction was carried out at 105°C for 1h. The reaction was complete on TLC. After cooling, the reaction solution was diluted with EA / HEP and filtered. The filtrate was washed with saturated NaCl and extracted with EA. Purification by column gave 1.5g of pure product as a yellow solid. MS-APCI: 174.0 [M+H] + .

[0181] Steps 1-9:

[0182]

[0183] In a 25 mL two-necked bottle, 14 (400 mg) was suspended in DCM (5 mL), and NIS (520 mg) was added at room temperature. The reaction was continued for 10 minutes, and the raw material disappeared on the TLC plate. Post-treatment: water was added to the reaction solution, and DCM was extracted. The DCM layer was washed with Na2S2O3, dried, and spin-dried to obtain 600 mg of the product as a brown solid. MS-APCI: 300.2 [M+H] + .

[0184] Steps 1-10:

[0185]

[0186] In a 15 mL reaction tube, 15 (400 mg), 11 (337 mg), Pd(PPh3)2Cl2 (232 mg), CuI (13 mg), and DIPEA (838 mg) were dissolved in DMAc (3 mL), and reacted at 85°C for 2 h under N2 protection. TLC detection showed that 15 disappeared.

[0187] Post-treatment: The reaction solution was washed with water and extracted with EA. The EA was dried and purified by column to obtain 260 mg of the product as a light yellow solid. MS-APCI: 366.2 [M+H] + .

[0188] Steps 1-11:

[0189]

[0190] In a 15 mL reaction tube, 16 (150 mg), 17 (135 mg), NaBH3CN (39 mg), TEA (83 mg) were added to THF (5 mL) and reacted at 70 degrees overnight. TLC detection showed that 16 disappeared. Post-treatment: The reaction solution was spin-dried and extracted with EA. The EA was dried, spin-dried, and purified by column to obtain 120 mg of the product as a light yellow oil. MS-APCI: 479.2 [M+H] + .

[0191] Steps 1-12:

[0192]

[0193] 18 (120 mg) was dissolved in THF / H2O (4 mL, 1:1), and LiOH (18 mg) was added. The mixture was reacted at room temperature for 1 h. TLC detected the disappearance of 18. Post-treatment: THF was removed from the reaction solution, and 2N HCl was added to adjust the pH to 4. Solids precipitated and were filtered. The obtained solids were dried by spin drying. The product was 100 mg, a white solid. MS-APCI: 465.2 [M+H] + .

[0194] Steps 1-13:

[0195]

[0196] Compound 19 (100 mg), 6 (110 mg), Pd(dppf)Cl2 (10 mg), and Na2CO3 (43 mg) were added to dioxane / H2O (2.5 mL, 4:1), and reacted at 90 degrees for 2 h under N2 protection. The reaction was complete when detected by TLC. The reaction solution was filtered, washed with water, extracted with EA, dried, spin-dried, and passed through a column (DCM:MeOH=10:1) to obtain 80 mg of the product. Pale yellow solid. MS-APCI: 725.3 [M+H] + .

[0197] The method for synthesizing compound 001 was used to synthesize the following compounds:

[0198]

[0199]

[0200]

[0201] Example 2 Synthesis of Compound 002

[0202] Step 2-1:

[0203]

[0204] Dissolve 4.5g of compound 1-3 in 200mL DCM (in a 500mL single-necked flask), add 6.3g NIS, and protect with argon. React at room temperature overnight. TLC spot plate 1-3 reacts completely. Add 100mL of 2M HCl to the reaction solution, separate the liquids, and dry. Spin dry and purify through a column (Hep-DCM 1:1, then DCM) to obtain 2g of product as a light yellow solid. MS-APCI: 334[M+H] + .

[0205] Step 2-2:

[0206]

[0207] Dissolve 2g of compound 2-1, 1.0g of zinc cyanide and 60mg of tetrakistriphenylphosphine palladium in 50mL of dry DMF, protect with argon, and react at 140 degrees. TLC spot plate 2-1 reacts completely. Pour the reaction solution into 150mL of water, extract twice with DCM (100mL), and dry. Spin dry and purify through a column (DCM-MeOH 100, 50:1, 30:1, 20:1, 10:1, 5:1) to obtain 800mg of product as a light yellow solid. MS-APCI: 233[M+H]+ .

[0208] Step 2-3:

[0209]

[0210] 500 mg of compound 2-2 was dissolved in 50 mL of THF, and 10 mg of Pd-C was added to react at room temperature under hydrogen. TLC showed that 2-2 reacted completely. Insoluble matter was removed. After drying, the product was purified by column (DCM-MeOH=100, 50:1, 30:1, 20:1, 10:1, 5:1) to obtain 50 mg of product. MS-APCI: 203 [M+H] + .

[0211] Steps 2-4:

[0212]

[0213] Dissolve 2.02g of compound 2-3 in 50mL DMAC, add 1.98g of 2-4, protect with argon, and react at room temperature for 3 hours. Add 3.0g DDQ and react at room temperature overnight. Pour the reaction solution into 150mL of water, extract twice with DCM (100mL), and dry. Spin dry and purify through a column (DCM-MeOH=100, 50:1, 30:1, 20:1, 10:1, 5:1) to obtain 200mg of product as a light yellow solid. MS-APCI: 381[M+H] + .

[0214] Steps 2-5:

[0215]

[0216] 125 mg of compound 2-5, 248 mg (methyl azetidine-3-carboxylate hydrochloride), 125 mg of Na(CN)BH3 were dissolved in 10 mL of THF, argon protection, 167 mg of triethylamine was added, and the reaction was carried out at 70°C for 24 hours. The reaction solution was added to 100 mL of ice water, extracted twice with EA (100 mL), and dried. Purification by column (DCM-MeOH = 100, 50:1, 10:1, 5:1) gave 53 mg of light yellow solid, MS-APCI: 480 [M+H] +

[0217] Steps 2-6:

[0218]

[0219] Take bromine raw material 2-6 (1.7g), borate raw material 6 (1g), Pd (dppf) Cl2 (0.12g) and sodium carbonate (0.62g) and add them to a 50mL reaction bottle, replace with nitrogen 3 times, add 1,4-dioxane (8mL) and water (2mL), react at 90 degrees; TLC tracking; post-treatment: add saturated ammonium chloride to quench in an ice bath, extract 2 times with MTBE, wash 5 times with saturated brine, mix the sample, pass through the column: (HEP-DCM = 100, 50:1, 10:1, 5:1) flush the product, and concentrate to obtain 0.6g of yellow solid. MS-APCI: 740 [M + H] +

[0220] Steps 2-7:

[0221]

[0222] 53 mg of raw material 2-7 was dissolved in 5 mL of MeOH, and 2 mL of NaOH (2N) was added. The reaction was allowed to react at 26 degrees for 5 hours. The reaction solution was dried at 40 degrees. 50 mL of water was added to the residual solid, and the pH value was adjusted to 7 with 2M HCl. The solid was filtered out, washed with water, washed with THF, dried, and further purified to obtain 5 mg of a yellow solid. MS-APCI: 726 [M+H] + .

[0223] The following compounds were synthesized using the method for synthesizing compound 002:

[0224]

[0225]

[0226]

[0227]

[0228] Biological Testing

[0229] Example A: PD-1 / PD-L1 Homogeneous Time-Resolved Fluorescence (HTRF) Binding Assay

[0230] The assay was performed in a standard black 384-well polystyrene plate in a final volume of 20 μL. The inhibitor was first serially diluted in DMSO and added to the wells before the other reaction components were added. The final concentration of DMSO in the assay was 1%. The assay was performed in PBS buffer (pH 7.4) containing 0.05% Tween-20 and 0.1% BSA at 25°C. Recombinant human PD-L1 protein (19-238) with a His tag at the C-terminus was purchased from AcroBiosystems (PD1-H5229). Recombinant human PD-1 protein (25-167) with an Fc tag at the C-terminus was also purchased from AcroBiosystems (PD1-H5257). PD-L1 and PD-1 proteins were diluted in assay buffer and 0.1 μl of the solution was extracted and added to the wells. The plate was centrifuged and the proteins were preincubated with the inhibitors for 40 minutes. After incubation, 0.1 μl of HTRF detection buffer containing europium-blocked anti-human IgG (PerkinElmer-AD0212) Fc-specific and anti-His Sure -Allophycocyanin (APC, PerkinElmer-AD0059H) conjugated antibodies. After centrifugation, the wells were incubated at 25 ° C for 60 minutes. The data were read in a PHERAstar FS plate reader (665nm / 620nm ratio). The final concentrations in the assay were ~3nM PD1, 10nM PD-L1, 1nM Europium anti-human IgG, and 20nM anti-His-allophycocyanin. The activity data were fitted using GraphPad Prism 5.0 software to obtain the IC50 values ​​of the inhibitors.

[0231] The IC50 values ​​of the compounds illustrated in the examples are expressed in the following manner: IC50: +: ≤100 nM; ++: 100 nM to 1000 nM; +++: >1000 nM.

[0232] Data for Example compounds obtained using the PD-1 / PD-L1 homogeneous time-resolved fluorescence (HTRF) binding assay described in Example A are provided in Table 1.

[0233] Table 1.

[0234] Compound No. PD-1 / PD-L1 HTRF IC50(nM) 001 + 002 + 003 + 004 + 010 + 011 + 012 + 014 + 015 + 016 + 017 + 018 + 021 + 022 + 025 + 027 + 028 + 032 + 033 + 034 + 035 + 039 + 042 +

[0235] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound as shown in the following formula I, or a pharmaceutically acceptable salt thereof: in, is a structure selected from the group consisting of: n, m and p are each independently selected from 0, 1, 2 or 3; X7 is selected from the group consisting of CH; X8 and X9 are each independently selected from the following group: C(R)2; wherein R is selected from the following group: H, C1-C6 alkyl, chlorine, bromine, fluorine, iodine; M3, M4, and M5 are each independently selected from the group consisting of: CR; M6 is C; and represents a single bond or a double bond; Each is independently selected from the group consisting of substituted or unsubstituted phenyl, or substituted or unsubstituted pyridyl; Selected from the following group: substituted or unsubstituted 6-membered heteroaryl, substituted or unsubstituted C6 aryl, wherein the heteroaryl is a nitrogen-containing heteroaryl; L1 is selected from the group consisting of -S-, -O-; G is Wherein, Rb and Rc are each independently selected from the following group: H, substituted or unsubstituted C1-C8 alkyl; or said Rb and Rc together with the adjacent N atom constitute a substituted or unsubstituted 5-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O; R4 is selected from the group consisting of NRdRe; R1, R2, R3 and R5 are each independently selected from the group consisting of H, -CN, trifluoromethyl, sulfonylamino, nitro, hydroxy, halogen, substituted or unsubstituted C1-C10 alkyl; Rd and Re are each independently selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C 10 Cycloalkyl; or Rd and Re together form a substituted or unsubstituted 5-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O; Unless otherwise specified, the term "substituted" refers to being substituted by one or more substituents selected from the group consisting of halogen, oxo, -CN, hydroxyl, amino, C1-C6 alkylamino, carboxyl, -NHAc, a group selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, and C3-C8 cycloalkyl which is unsubstituted or substituted by one or more substituents selected from the group consisting of halogen, hydroxyl, carboxyl, cyano, C1-C6 alkoxy, and C1-C6 alkylamino; In the above formulae, any of the heteroatoms is selected from the group consisting of N, S and O.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that The is a benzene ring; and R3 is a C1-C6 alkyl, halogen, -CN, or hydroxyl.

3. The compound according to claim 1, characterized in that The It has the structure shown below: Wherein, Y7 is independently selected from: CH, N; R1 is selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, -CN, hydroxyl.

4. The compound according to claim 1, characterized in that The Select from the following group:

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R4 is selected from the following group: NRdRe; said Rd and Re are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl; or Rd and Re together form a substituted or unsubstituted 5-10 membered heterocyclic group having 1-3 heteroatoms selected from N, S and O.

6. A compound selected from the group consisting of:

7. A method for preparing the compound of formula I as claimed in claim 1, characterized in that: The compound of formula I is a compound of formula I-1 or a compound of formula I-2, and the method comprises steps selected from those shown in synthesis scheme 1 or 2: Synthesis Scheme 1: (a) Using intermediates 1 and 2 as raw materials, a Sonogashira coupling reaction catalyzed by a palladium catalyst was performed to obtain the target product 3; (b) Using 3 and 2-1 as raw materials, a coupling reaction occurs under the conditions of a palladium catalyst and a ligand to obtain an intermediate I-1; Synthesis Scheme 2: (a) Compound 3-1 is used as a raw material and reacted with a nitrating agent to form an intermediate 3-2; (b) using 3-2 as a raw material, a reduction reaction occurs under reducing conditions to form an intermediate 3-3; (c) using 3-3 and 3-4 as raw materials, a condensation reaction occurs under the condition of a shrinking agent, and an oxidant is added to cause an oxidation reaction to obtain an intermediate 3-5; (d) Using 3-5 and 2-1 as raw materials, a coupling reaction occurs in the presence of a catalyst and a ligand to form intermediate I-2.

8. A pharmaceutical composition, characterized in that The method comprises: (1) the compound according to claim 1, or a pharmaceutically acceptable salt thereof; and (2) a pharmaceutically acceptable carrier.

9. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 8, characterized in that: Used for preparing a pharmaceutical composition for preventing and / or treating diseases related to the activity or expression of PD-1 / PD-L1.

10. A PD-1 / PD-L1 regulator, characterized in that The modulator comprises the compound according to claim 1, or a pharmaceutically acceptable salt thereof.

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