Preparation of biaryl ring-linked aromatic heterocyclic derivatives and their application as immunomodulators

Abstract

The present invention provides a method for preparing and applying an aromatic heterocyclic derivative as an immunomodulator, specifically a compound as shown in the following formula I, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof, wherein the definitions of each group are as described in the specification, and the compound of formula I can be used to treat diseases associated with the PD-1 / PD-L1 signaling pathway. [Formula 1] JPEG2023525116000109.jpg66163

Description

[Technical Field]

[0001] The present invention relates to the field of small molecule drugs, and in particular, the present invention provides small molecule compounds that can be used to treat diseases associated with the PD-1 / PD-L1 signaling pathway. [Background technology]

[0002] The immune system plays a crucial role in the control and treatment of many diseases, including various cancers and diseases caused by viruses. However, cancer cells can rapidly proliferate by evading or inhibiting the immune system through several pathways. One of these pathways is by altering the activating and inhibitory molecules expressed on immune cells. Blockade of inhibitory immune checkpoints, such as PD-1, has proven to be a highly effective method for inhibiting cancer cells.

[0003] PD-1, also known as CD279, is a programmed cell death protein-1 (PD-1) expressed primarily on activated T and B cells. Its function is to inhibit cell activation, a normal self-stabilizing mechanism of the immune system. PD-1 acts as a defense against autoimmune diseases, as excessive T and B cell activation can lead to autoimmune diseases. PD-1 is a type I transmembrane glycoprotein consisting of 268 amino acids. Its structure primarily consists of an outer immunoglobulin variable region, a hydrophobic transmembrane region, and an intracellular region. The intracellular region contains two phosphorylation sites located in the immunoreceptor tyrosine-based inhibitory motif and immunoreceptor tyrosine-based transducing motif, respectively, demonstrating that PD-1 can reversely regulate T cell receptor-mediated signals. PD-1 has two ligands, PD-L1 and PD-L2, which have different expression patterns. PD-L1 is upregulated on a variety of tumor cells, and it binds to PD-1 on T cells, inhibiting T cell proliferation and activation, rendering T cells in an inactivated state and ultimately inducing immune escape.

[0004] PD-1 / PD-L1 plays a role in inverse immune regulation. After PD-1 binds to PD-L1, it can induce tyrosine polyphosphorylation at the T cell immunoreceptor tyrosine transduction motif domain, and the phosphorylated tyrosine can bind to the phosphatases protein tyrosinase 2 and protein tyrosinase 1. This inhibits the activation of extracellular signal-regulated kinase (EPK) and phosphoinositide 3-kinase (PI3K) and serine-threonine protein kinase (Akt), thereby inhibiting T lymphocyte proliferation and related cytokine secretion. PD-1 / PD-L1 signaling inhibits T cell activation and proliferation while also inducing the secretion of cytokines such as interleukin 2, interferon-γ, and IL-10. Furthermore, PD-1 / PD-L1 signaling has a similar immune function on B cells. After PD-1 binds to the B cell antigen receptor, the PD-1 cytoplasmic region interacts with tyrosinase, which contains a protein tyrosinase 2-binding site, thereby inhibiting B cell activation.

[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 inhibitors have potent antitumor activity against various tumors. Currently available PD-1 / PD-L1 antibody inhibitors include BMS's Ninolumab, Merck's Lambrolizumab, and Roche's Atezolizumab. In addition, there are more PD-1 / PD-L1 antibody inhibitors under investigation, including CureTech's Pidilizumab, GSK's AMP-224, and AstraZeneca's MEDI-4736.

[0006] Tumor immunotherapy is considered to be a new generation revolution in cancer treatment following targeted therapy, but the PD-1 monoclonal antibody drugs currently on the market and under investigation have their own drawbacks, including the fact that they can only be administered by injection, cannot be administered orally, are unstable in the body, are easily degraded by proteases, are prone to immune cross-reactivity, are relatively difficult to purify, and have high production costs. Therefore, small molecule inhibitors of the PD-1 / PD-L1 interaction are a better option for tumor immunotherapy.

[0007] In summary, there is an urgent need in the art to develop novel small molecule inhibitors of the PD-1 / PD-L1 interaction. Summary of the Invention [Problem to be solved by the invention]

[0008] It is an object of the present invention to provide novel small molecule inhibitors of the PD-1 / PD-L1 interaction. [Means for solving the problem]

[0009] A first aspect of the present invention provides a compound as shown in formula I below, or an optical isomer, cis-trans isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof: [ka] wherein n, m, p, and q are each independently selected from 0, 1, 2, 3, or 4; L1 and L2 each independently represent a chemical bond, a substituted or unsubstituted C1-C4 alkylene group, a substituted or unsubstituted C2-C4 alkenylene group, a substituted or unsubstituted C2-C4 alkynylene group, -S-, -O-, substituted or unsubstituted -NH-, -S(O)-, -S(O)2-, substituted or unsubstituted -NHC(O)NH-, [ka] , substituted or unsubstituted [ka] , substituted or unsubstituted [ka] , substituted or unsubstituted [ka] is selected from the group consisting of M1 and M2 are each independently selected from the group consisting of C(R6)2, NR6, O, S, SO, and SO2, where R6 is H, chlorine, bromine, fluorine, iodine, a cyano group, a hydroxyl group, a nitro group, or NR f , a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted C6-C 10 Aryl groups, substituted or unsubstituted C6-C 10 Heteroaryl group, -C(=O)-NR d R e , -C(=O)-substituted or unsubstituted C1-C6 alkoxy group, -C(=O)-substituted or unsubstituted C1-C6 alkyl group, -C(=O)-substituted or unsubstituted C3-C 10 selected from the group consisting of a cycloalkyl group, a substituted or unsubstituted C2-C6 alkenyl group, a substituted or unsubstituted C2-C6 alkynyl group, a -C(=O)-substituted or unsubstituted C2-C6 alkenyl group, and a -C(=O)-substituted or unsubstituted C2-C6 alkynyl group; M3, M4, and M5 are each independently selected from the group consisting of a chemical bond, CR6, N, NR6, O, S, SO, and SO2; each M6 is independently selected from the group consisting of CR6 and N; And the above [ka] is an aromatic ring, [ka] teeth, [ka] and having a group as shown in the structure where: X6, X7, X8, X9, X 10 , X 11 , Z 2 , Z 3 , Z 4 are independently N, NO, and CR a wherein R a is H, chlorine, bromine, fluorine, iodine, cyano group, hydroxyl group, nitro group, NR f , a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted C6-C 10 Aryl groups, substituted or unsubstituted C6-C 10 Heteroaryl group, -C(=O)-NR d R e , -C(=O)-substituted or unsubstituted C1-C6 alkoxy group, -C(=O)-substituted or unsubstituted C1-C6 alkyl group, -C(=O)-substituted or unsubstituted C3-C 10 selected from the group consisting of a cycloalkyl group, a substituted or unsubstituted C2-C6 alkenyl group, a substituted or unsubstituted C2-C6 alkynyl group, a -C(=O)-substituted or unsubstituted C2-C6 alkenyl group, and a -C(=O)-substituted or unsubstituted C2-C6 alkynyl group; Y 1 , Y 2 are each independently CH, CH2, NH, N, NO, CF, or CR a , O, S, SO, or SO2; [ka] is a single or double bond, and [ka] is an aromatic or non-aromatic fragment, [ka] is a substituted or unsubstituted 5- to 12-membered heteroaryl group, a substituted or unsubstituted C6-C 10 Aryl groups, substituted or unsubstituted 5- to 12-membered heterocyclic groups, substituted or unsubstituted 5- to 12-membered C5-C 12 wherein the 5- to 12-membered heteroaryl group and the 5- to 12-membered heterocyclic group have 1 to 4 heteroatoms selected from B, P, N, O, and S, and wherein P, N, and O as ring atoms are substituted by oxo, and one or more ring carbon atoms can be substituted by a carbonyl group; [ka] teeth, [ka] wherein the attachment point of the ring can be N or C; R1, R2, R2', and R4 are each independently H, halogen, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C2-C6 alkenyl group, a substituted or unsubstituted C2-C6 alkynyl group, a substituted or unsubstituted C1-C6 alkoxy group, a substituted or unsubstituted C3-C8 cycloalkyl group, oxo (i.e., =O), =NR f , -CN, a hydroxyl group, NRdRe (e.g., an amino group), a substituted or unsubstituted C1-C6 amine group, a substituted or unsubstituted -(C1-C6 alkylene)-NH-(C1-C6 alkylene), a carboxyl group, a substituted or unsubstituted C6-C 10 aryl groups, substituted or unsubstituted 5- to 12-membered heteroaryl groups having 1 to 3 heteroatoms, substituted or unsubstituted 5- to 12-membered heterocyclic groups having 1 to 4 heteroatoms, substituted or unsubstituted [ka] , substituted or unsubstituted [ka] or -(L 1a ) r-(L 2a ) s -(L 3a ) s - selected from the group consisting of R3 and R3' are each independently [ka] wherein Rb, Rc, and Rd are each independently selected from the group consisting of H and a substituted or unsubstituted C1-C8 alkyl group, or Rb and Rc together with the adjacent N atom form a substituted or unsubstituted 5- to 10-membered heterocyclic group having 1 to 3 heteroatoms selected from N, S, and O; Each L 1a are each 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-, substituted or unsubstituted -NH-, -S(O)-, and -S(O)2-; L 2a is a substituted or unsubstituted C6-C 12 an arylene group, a substituted or unsubstituted 5- to 12-membered heteroarylene group having 1 to 3 heteroatoms, a substituted or unsubstituted C3-C8 cycloalkylene group, and a substituted or unsubstituted 5- to 10-membered heterocyclylene group having 1 to 3 heteroatoms; L 3a is H, substituted or unsubstituted C1-C 10 Alkyl groups, C1-C 10 Aryl group, -CN, hydroxyl group, amino group, carboxyl group, -CO-NH-SO2-R g , -NH-SO2-R g , -SO2-NH-CO-R g is selected from the group consisting of R d , R e and R g are each independently H, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C 10 Cycloalkyl groups, substituted or unsubstituted C6-C 10aryl groups; R f is H, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C8 cycloalkyl group, a substituted or unsubstituted C6-C 10 Aryl groups, substituted or unsubstituted C6-C 10 Heteroaryl group, cyano group, -C(=O)-NR d R e , -C(=O)-substituted or unsubstituted C1-C6 alkoxy group, -C(=O)-substituted or unsubstituted C1-C6 alkyl group, -C(=O)-substituted or unsubstituted C3-C 10 and a cycloalkyl group, a -C(=O)-substituted or unsubstituted C2-C6 alkenyl group, and a -C(=O)-substituted or unsubstituted C2-C6 alkynyl group, and unless otherwise specified, the term "substituted" refers to, for example, halogen including, but not limited to, -F, Cl, Br, -CH2Cl, -CHCl2, -CCL3, -CH2F, -CHF2, -CF3, oxo, -CN, a hydroxyl group, an amino group, a C1-C6 alkylamino group, a carboxyl group, -NHAc, an unsubstituted or C1-C6 alkyl group, a C1-C6 alkoxy group, a C6-C 10 Aryl groups, C3-C8 cycloalkyl groups, halogenated C6-C 10 aryl groups, 5-10 membered heteroaryl groups having 1-3 heteroatoms selected from N, S, and O, and 5-10 membered heterocyclic groups having 1-3 heteroatoms selected from N, S, and O, wherein the substituents are selected from the group consisting of halogen, hydroxyl groups, carboxyl groups, cyano groups, C1-C6 alkoxy groups, and C1-C6 alkylamino groups; In each of the above formulas, any of the heteroatoms is selected from the group consisting of B, P, N, S and O.

[0010] In another preferred example, R1, R2, R2', and R4 are each independently H, halogen, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C2-C6 alkenyl group, a substituted or unsubstituted C2-C6 alkynyl group, a substituted or unsubstituted C1-C6 alkoxy group, a substituted or unsubstituted C3-C8 cycloalkyl group, oxo (i.e., =O), =NR f , -CN, a hydroxyl group, NRdRe (e.g., an amino group), a substituted or unsubstituted C1-C6 amine group, a substituted or unsubstituted -(C1-C6 alkylene)-NH-(C1-C6 alkylene), a carboxyl group, a substituted or unsubstituted C6-C 10 aryl groups, substituted or unsubstituted 5- to 12-membered heteroaryl groups having 1 to 3 heteroatoms, substituted or unsubstituted 5- to 12-membered heterocyclic groups having 1 to 4 heteroatoms, substituted or unsubstituted [ka] , substituted or unsubstituted [ka] , or -(L 1a ) r -(L 2a ) s -(L 3a ) s - is selected from the group consisting of

[0011] In another preferred embodiment, the ring [ka] and / or [ka] has the substituents as shown in formula IV below: [ka] wherein each L4 is independently selected from the group consisting of a substituted or unsubstituted C1-C4 alkylene group, -S-, -O-, -NRa-, -S(O)-, and -S(O)2-, and is preferably a substituted or unsubstituted C1-C4 alkylene group, provided that the structure formed by each L4 together is chemically stable, [ka] is a substituted or unsubstituted C5-C 10 a cycloalkyl group, and a substituted or unsubstituted 3- to 10-membered heterocyclic group having 1 to 3 heteroatoms selected from B, P, N, S, and O, and preferably [ka] is a 3- to 8-membered nitrogen-containing heterocyclic group, Each R5 is independently selected from the group consisting of a substituted or unsubstituted C1-C6 alkyl group, -CN, a hydroxyl group, an amino group, and a carboxyl group, wherein the substituents are selected from the group consisting of a halogen, a hydroxyl group, a carboxyl group, a cyano group, and a C1-C6 alkoxy group.

[0012] In another preferred embodiment, the compound of formula I has the structure as shown in the following formula: [ka]

[0013] In another preferred embodiment, [ka] is a structure as shown in the formula: [ka] wherein X1, X2, and Y4 are each independently selected from CH (wherein CH can be substituted by R1 or R3), N, O, S, and NH (wherein NH can be substituted by R1 or R3).

[0014] In another preferred example, M1 and M2 are each independently CH2.

[0015] In another preferred embodiment, the compound has a structure as shown in formula Ia or Ib: [ka] where: R3 and R3' are each independently [ka] wherein Rb and Rc together with the adjacent N atom form a substituted or unsubstituted 5- to 10-membered heterocyclic group (preferably a 5- to 7-membered heterocyclic group) having 1 to 3 heteroatoms selected from N, S and O; The remaining groups are defined as above.

[0016] In another preferred example, in the compound, R3 and R3' are each independently [ka] is.

[0017] In another preferred embodiment, the compound is selected from the group consisting of: [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0018] A second aspect of the present invention provides a process for preparing a compound of formula I according to the first aspect of the present invention, said process comprising the steps of: [ka] (a) Suzuki coupling reaction of intermediates 1 and 2 catalyzed by a suitable palladium catalyst to obtain target product I; Here, the definitions of each group are as described in the first aspect of the present invention.

[0019] In another preferred embodiment, the method for preparing the intermediate 1 is as follows: [ka] Starting from compound 1-1, chiral alcohol 1-2 is formed under the action of a chiral auxiliary (e.g., R / S-CBS) and a reducing agent (e.g., borane), (b) 1-2 (or 1-5) and 1-3 are subjected to a coupling reaction (e.g., Suzuki, Buchwald, etc.) under suitable palladium catalyst and ligand conditions to give intermediate 1-4 (or 1-6), (c) Compound 1-1 and R / St-butylsulfonimide are used as starting materials to form a protected chiral amino compound under the action of a Lewis acid (e.g., ethyl tetratitanate) and a reducing agent (e.g., lithium aluminum tetrahydrogen, sodium borohydride, etc.), and then the protecting group is removed under acidic conditions to obtain chiral amino compound 1-5; (d) Starting from 1-4 (or 1-6) and bis-pinacolatodiboron, Suzuki coupling reaction can be carried out under suitable palladium catalyst and ligand conditions to give intermediate 1.

[0020] In another preferred embodiment, the method for preparing the intermediate 2 is as follows: Method 1: [ka] Starting from compounds 2-1 and 2-2, intermediate 2 is formed under the action of a dehydrating agent (eg, concentrated sulfuric acid, PPA, etc.).

[0021] Method 2: [ka] Starting from compounds 2-3 and 2-4, intermediate 2 is formed by affinity substitution reaction under acidic or basic conditions.

[0022] A third aspect of the present invention provides a pharmaceutical composition comprising: (1) a compound according to 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. In another preferred embodiment, the pharmaceutical composition is used to treat a disease selected from the group consisting of cancer, infectious diseases, and autoimmune diseases.

[0023] In another preferred example, the cancer is selected from the group consisting of 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, urological tumors, blood tumors, glioma, digestive system tumors, reproductive system tumors, lymphoma, nervous system tumors, brain tumors, and head and neck cancer.

[0024] In another preferred example, the cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), myelodysplastic syndrome (MDS), myeloproliferative disorder (MPD), chronic myelogenous leukemia (CML), multiple myeloma (MM), non-Hodgkin's lymphoma (NHL), mantle cell lymphoma (MCL), follicular lymphoma, Waldestromm's macroglobulinemia (WM), T-cell lymphoma, B-cell lymphoma, or diffuse large B-cell lymphoma (DLBCL).

[0025] In another preferred embodiment, the infectious disease is selected from bacterial infections and viral infections. In another preferred embodiment, the autoimmune disease is selected from an organ-specific autoimmune disease and a systemic autoimmune disease.

[0026] In another preferred embodiment, the organ-specific autoimmune disease includes chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, pulmonary hemorrhagic nephritic syndrome, primary biliary cirrhosis, multiple encephalomyelitis, and acute idiopathic polyneuropathy. In another preferred embodiment, the systemic autoimmune disease includes rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, and autoimmune hemolytic anemia.

[0027] In another preferred embodiment, the pharmaceutical composition is also used to improve T cell function in patients with chronic hepatitis B (CHB). In another preferred embodiment, the inhibitor further comprises at least one therapeutic agent selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, or ipilimumab.

[0028] A fourth aspect of the present invention provides the use of a compound according to the first aspect of the present invention or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to the third aspect of the present invention, for use in the preparation of a pharmaceutical composition for the prevention and / or treatment of a disease associated with the activity or expression level of PD-1 / PD-L1.

[0029] A fifth aspect of the present invention provides a PD-1 / PD-L1 inhibitor, which comprises a compound according to the first aspect of the invention, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0030] A sixth aspect of the present invention provides a method of inhibiting PD-1 / PD-L1 interaction in vitro, comprising contacting a compound according to the first aspect of the invention, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, with PD-L1 protein. [Effects of the Invention]

[0031] 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 (e.g., in the Examples) can be combined with each other to form new or preferred technical solutions, which will not be repeated here due to space limitations. DETAILED DESCRIPTION OF THE INVENTION

[0032] As a result of extensive and detailed research, the present inventors have discovered an inhibitor of PD-1 / PD-L1 interaction with excellent inhibitory effects, and have completed the present invention based on this discovery.

[0033] definition As used herein, the term "alkyl group" includes straight-chain or branched-chain alkyl groups. For example, a C1-C8 alkyl group refers to a straight-chain or branched-chain alkyl group having 1 to 8 carbon atoms, such as a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, t-butyl group, etc.

[0034] As used herein, the term "alkenyl group" includes straight-chain and branched-chain alkenyl groups. For example, a C2-C6 alkenyl group refers to a straight-chain or branched-chain alkenyl group having from 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.

[0035] As used herein, the term "alkynyl group" includes straight-chain and branched-chain alkynyl groups. For example, a C2-C6 alkynyl group refers to a straight-chain or branched-chain alkynyl group having from 2 to 6 carbon atoms, such as an ethynyl group, a propynyl group, a butynyl group, or the like.

[0036] As used herein, "C3-C 10 The term "cycloalkyl group" refers to a cycloalkyl group having from 3 to 10 carbon atoms. It may be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may be bicyclic, such as bridged or spirocyclic.

[0037] As used herein, the term "C1-C8 alkylamino group" refers to an amino group substituted with a C1-C8 alkyl group, which may be mono- or di-substituted, such as methylamino group, ethylamino group, propylamino group, isopropylamino group, butylamino group, isobutylamino group, t-butylamino group, dimethylamino group, diethylamino group, dipropylamino group, diisopropylamino group, dibutylamino group, diisobutylamino group, di-t-butylamino group, and the like.

[0038] As used herein, the term "C1-C8 alkoxy group" refers to a straight or branched chain alkoxy group having 1 to 8 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, t-butoxy, and the like.

[0039] As used herein, the term "3- to 10-membered heterocycloalkyl group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O" refers to a saturated or partially saturated cyclic group having 3 to 10 atoms, of which 1 to 3 heteroatoms are selected from the group consisting of N, S, and O. It may be a monocyclic or bicyclic group in the form of a bridged ring or a spirocyclic ring. Specific examples include oxetane, azetidine, tetrahydro-2H-pyranyl group, piperidinyl group, tetrahydrofuranyl group, morpholinyl group, and pyrrolidinyl group.

[0040] As used herein, "C6-C 10 The term "aryl group" refers to an aryl group having 6 to 10 carbon atoms, such as a phenyl group or a naphthyl group or similar group.

[0041] As used herein, the term "5- to 10-membered heteroaryl group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O" refers to a cyclic aromatic group having 5 to 10 atoms, 1 to 3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or fused ring form. Specific examples include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl, (1,2,4)-triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, etc.

[0042] Unless otherwise specified, the groups described in the present invention are "substituted or unsubstituted", and all groups in the present invention are substituted or unsubstituted, including halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-amino, 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 It may be substituted by a substituent selected from the group consisting of an aryl group, a C1-C6 alkoxy-C1-C6 alkyl group, a C1-C6 alkoxy-carbonyl group, a phenoxycarbonyl group, a C2-C6 alkynyl-carbonyl group, a C2-C6 alkenyl-carbonyl group, a C3-C6 cycloalkyl-carbonyl group, a C1-C6 alkyl-sulfonyl group, and the like.

[0043] 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" refers to substitution with an atom selected from F, Cl, Br, and I.

[0044] Unless otherwise specified, the structural formulae depicted in the present invention are intended to include all isomeric forms (e.g., enantiomers, diastereomers, and geometric (or conformational) isomers), including asymmetric centers, such as the R and S configurations, and (Z) and (E) isomers of double bonds. Thus, all individual stereochemical isomers of the compounds of the present invention or mixtures of the enantiomers, diastereomers, or geometric (or conformational) isomers thereof are within the scope of the present invention.

[0045] As used herein, the term "tautomer" refers to structural isomers of different energies that can be interconverted across a low energy barrier. For example, proton tautomers (i.e., protonation) include interconversions via migration of a proton, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversions via recombination of some bond electrons.

[0046] As used herein, the term "solvate" refers to a compound of the present invention that coordinates with solvent molecules to form a complex of a specific ratio. As used herein, the term "hydrate" refers to a complex formed when a compound of the present invention is coordinated with water. It should be understood that the definition of each group herein is intended to create a chemically stable structure.

[0047] Active ingredient As used herein, "compounds of the invention" refers to compounds shown in Formula I and further includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of Formula I. Preferred compounds of the present invention include compounds 1 to 360 (including various R-configuration and / or S-configuration stereoisomers of each compound, and / or E- / Z-cis-trans isomers). In another preferred example, the pharmaceutically acceptable salts include salts formed by combining with an inorganic acid, an organic acid, an alkali metal ion, an alkaline earth metal ion, or an organic base capable of providing a physiologically acceptable cation, and ammonium salts.

[0048] In another preferred example, the inorganic acid is selected from hydrochloric acid, hydrobromic acid, phosphoric acid, and sulfuric acid; the organic acid is selected from methanesulfonic acid, p-toluenesulfonic acid, trifluoroacetic acid, lysinic acid, maleic acid, tartaric acid, fumaric acid, citric acid, and lactic acid; the alkali metal ion is selected from lithium ion, sodium ion, and potassium ion; the alkaline earth metal ion is selected from calcium ion and magnesium ion; and the organic base capable of providing a physiologically acceptable cation is selected from methylamine, dimethylamine, trimethylamine, piperidine, morpholine, and tris(2-hydroxyethyl)amine.

[0049] All such 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 and its solvates and salts, polycrystals or eutectics can occur under different crystallization conditions.

[0050] Preparation of Compounds of Formula I In order to prepare the compounds according to general formula I of the present invention, according to the structure of general formula I, the preparation of the compounds of general formula I in the present invention can be obtained by the following synthetic route: [ka]

[0051] (a) Starting from intermediates 1 and 2, Suzuki coupling reaction catalyzed by a suitable palladium catalyst gives the desired product I; Here, the preparation method of intermediate 1 (see WO2018195321, WO2018005374 and WO2019023575) is as follows: [ka]

[0052] Starting from compound 1-1, chiral alcohol 1-2 is formed under the action of a chiral auxiliary (e.g., R / S-CBS) and a reducing agent (e.g., borane), (b) 1-2 (or 1-5) and 1-3 are subjected to a coupling reaction (e.g., Suzuki, Buchwald, etc.) under suitable palladium catalyst and ligand conditions to give intermediate 1-4 (or 1-6), (c) Compound 1-1 and R / St-butylsulfonimide are used as starting materials to form a protected chiral amino compound under the action of a Lewis acid (e.g., ethyl tetratitanate) and a reducing agent (e.g., lithium aluminum tetrahydrogen, sodium borohydride, etc.), and then the protecting group is removed under acidic conditions to obtain chiral amino compound 1-5; (d) Starting from 1-4 (or 1-6) and bis-pinacolatodiboron, Suzuki coupling reaction can be carried out under suitable palladium catalyst and ligand conditions to give intermediate 1.

[0053] Intermediate 2 can be prepared as follows. Method 1: [ka] Starting from compounds 2-1 and 2-2, intermediate 2 is formed under the action of a dehydrating agent (eg, concentrated sulfuric acid, PPA, etc.). Method 2: [ka] Starting from compounds 2-3 and 2-4, intermediate 2 is formed by affinity substitution reaction under acidic or basic conditions.

[0054] Furthermore, the starting materials and intermediates in the above reactions are readily available, and the reactions at each step can be easily synthesized according to published literature or by conventional methods in organic synthesis known to those skilled in the art.The compounds described in general formula I can exist in solvated or non-solvated forms, and different solvates may be obtained by crystallization using different solvents.

[0055] Pharmaceutical Compositions and Methods of Administration The compounds of the present invention have excellent inhibitory activity against PD-1 / PD-L1 interaction. Therefore, the compounds of the present invention and their various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, as well as pharmaceutical compositions containing the compounds of the present invention as a main active ingredient, are used for preventing and / or treating (stabilizing, alleviating, or curing) diseases associated with PD-1 / PD-L1 interaction (e.g., cancer, infectious diseases, autoimmune diseases).

[0056] The pharmaceutical compositions of the present invention contain a compound of the present invention and a pharmaceutically acceptable excipient or carrier within a safe and effective amount. Here, "safe and effective amount" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1 to 2000 mg of the compound / agent of the present invention, more preferably 10 to 200 mg of the compound / agent of the present invention. Preferably, the "single agent" is one capsule or tablet.

[0057] "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that must be of sufficient purity and sufficiently low toxicity to be suitable for human use. "Compatibility" refers to the ability of the components of the composition to blend with each other without significantly reducing the efficacy of the compounds of the present invention and with each other. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween®), wetting agents (e.g., sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0058] The administration route of the compound or pharmaceutical composition of the present invention is not particularly limited, and typical administration routes include (but are not limited to) oral and parenteral (intravenous, intramuscular, or subcutaneous).

[0059] 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 (a) a filler or compatibilizer, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) a binder, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) a humectant, such as glycerin; or (d) agar, calcium carbonate, potato starch. It is mixed with ingredients such as disintegrating agents such as potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, (e) retarders such as paraffin, (f) absorption accelerators such as quaternary amine compounds, (g) wetting agents such as cetyl alcohol and glyceryl monostearate, (h) adsorbents such as kaolin, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0060] Solid dosage forms such as tablets, sugar pills, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials known in the art. They can contain opacifying agents, and the release of the active compound or compounds of such compositions can be delayed in a specific part of the digestive tract. Examples of embedding materials that can be used include polymeric substances and waxes. If necessary, the active compound can be formed into microcapsules with one or more of the above-mentioned excipients.

[0061] 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 can contain an inert diluent conventionally used in the art, such as water or other solvents, and solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0062] Besides these inert diluents, compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and perfuming agents. In addition to the active compound, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and dehydrated sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0063] Compositions for parenteral injection can 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. The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (eg, other anti-cancer agents).

[0064] When administered in combination, the pharmaceutical composition further comprises one or more (two, three, four, or more) other pharmaceutically acceptable compounds, which may be used simultaneously, separately, or sequentially with the compounds of the present invention to prevent and / or treat diseases associated with PD-1 / PD-L1 interaction.

[0065] When a pharmaceutical composition is used, a safe and prevalent amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, and the dosage at the time of administration is the considered effective dose, and for a person weighing 60 kg, the daily dose is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage must also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0066] The main advantages of the present invention are: (1) The compounds of the present invention have high inhibitory activity against PD-1 / PD-L1 interaction, strong binding ability to PD-L1 protein, and the ability to relieve IFNγ inhibition by PD-L1. (2) The compounds of the present invention have superior solubility and very low toxicity to normal cells, and therefore can be applied to therapeutic subjects within a wider dose range. (3) Compared with the compounds of the prior art, the compounds of the present invention have better solubility, and therefore have good drug discovery potential; compared with the existing compounds, the compounds of the present invention show good bioavailability in in vivo experiments; and further, compared with the existing compounds, the compounds of the present invention can be easily prepared into pharmaceutically acceptable salts, which facilitates the further formation of formulations. (4) In vivo pharmacodynamic studies show that the compounds of the present invention can significantly inhibit the growth of subcutaneous tumors, regardless of tumor volume or weight, and significantly increase the number of lymphocytes in the blood and spleen of mice.

[0067] Hereinafter, the present invention will be further described in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and do not limit the scope of the present invention. In the following examples, experimental methods without specific conditions are usually in accordance with conventional conditions or conditions suggested by manufacturers. Unless otherwise specified, percentages and parts are calculated by weight percentages and parts by weight. Experimental materials and reagents used in the following examples are available from commercial channels unless otherwise specified.

[0068] Common materials and test methods: A description of the equipment and materials referred to in the examples follows. 1 H NMR spectra are analyzed by a Bruker AV-400 (400 MHz) NMR instrument. Chemical shifts are recorded using tetramethylsilane as an internal standard and are expressed in ppm (CDC13: δ 7.26 ppm). The recorded data information is as follows: chemical shifts and their splitting and coupling constants (s: singlet, d: doublet, t: triplet, q: quartet, br: broad peak, m: multiplet).

[0069] Mass spectrometry data were analyzed using a Finnigan LCQ Advantage liquid mass spectrometer, unless otherwise specified. All reactions were performed under anhydrous, oxygen-free conditions under the protection of dry argon gas. Solid metal-organic compounds were stored in a dry oven under the protection of argon gas. Tetrahydrofuran and ethyl ether are obtained by distillation, adding sodium metal pre-benzophenone during the distillation. Dichloromethane, pentane and hexane are treated with calcium hydride.

[0070] The special raw materials and intermediates mentioned in this invention are processed and provided by Tianjin Changsen Pharmaceutical Co., Ltd., etc., and all other chemical reagents are purchased from reagent suppliers such as Shanghai Chemical Reagent Company, Aldrich Company, Acros Company, etc. If the intermediates or products required for the reaction during synthesis are not sufficient for the next step, the synthesis will be repeated several times until a sufficient amount is reached. Materials and reagents referred to in this invention may be purchased commercially or by order unless otherwise specified.

[0071] The compounds of the present invention may contain one or more asymmetric centers, and therefore the series of compounds may be in the form of racemates or single enantiomers. The compounds prepared according to the present invention are heterocyclic compounds with purity greater than 95%, and the structure of each final product is characterized by MS and / or hydrogen nuclear magnetic resonance (NMR). 1 The synthesis of various compounds and intermediates of the present invention is described below by way of examples.

[0072] Example 1. Synthesis of Compound 1 [ka] Phase 1-1: [ka] Compound 1-1 (5.0 g, 23.6 mmol, WO2012158550), 1-2 (4.4 g, 25.9 mmol), Xantphos-PdCl2 (892 mg, 1.18 mmol), and Cs2CO3 (15.4 g, 47.2 mmol) were added to dioxane (100 mL), protected with N2, and reacted at 95 °C for 2 hours. The reaction completion was detected by TLC point plate. The reaction solution was filtered, and the filtrate was spin-dried. 4.8 g of a pale yellow solid was obtained by column chromatography (EA:HEP = 1:30). MS-APCI: 348 [M+H] + .

[0073] Phase 1-2: [ka] Compounds 1-3 (4.8 g, 13.8 mmol) and 1-4 (2.5 g, 20.7 mmol) were added to DCM, followed by 2.8 g of TEA. After stirring at room temperature for 1 hour, NaBH(OAc)3 (4.4 g, 20.7 mmol) was added. After 30 minutes, the reaction was detected as complete by TLC point plate. The reaction mixture was washed with saturated NaHCO3, extracted with DCM, the DCM was dried, spin-dried, and purified by column (MeOH:DCM = 1:20) to obtain 5.1 g of a pale yellow oily viscous product. MS-APCI: 419 [M+H] + .

[0074] Stages 1-3: [ka] Compound 1-6 (200 mg, 0.435 mmol, WO2018119266), 1-5 (219 mg, 0.522 mmol), Pd(dppf)2Cl2 (36 mg, 0.044 mmol), and Na2CO3 (138 mg, 1.3 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and reacted at 90 °C for 2 hours. The reaction completion was detected by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and column-dried to obtain 42.6 mg of a white solid. MS-APCI: 672 [M+H] + .

[0075] Example 2. Synthesis of Compound 2 [ka] Stage 2-1: [ka] Compound 1-7 (200 mg, 0.43 mmol, WO2018119286), 1-5 (218 mg, 0.52 mmol), Pd(dppf)2Cl2 (35 mg, 0.043 mmol), and Na2CO3 (138 mg, 1.3 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and reacted at 90 °C for 2 hours. The reaction completion was detected by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and subjected to column chromatography to obtain 12.6 mg of a pale yellow solid. MS-APCI: 673 [M+H] + .

[0076] Example 3. Synthesis of Compound 3 [ka] Phase 3-1: [ka] Compound 1-3 (1 g, 2.87 mmol) and 3-1 (661 mg, 5.74 mmol, dissolved in 1.5 mL of AcOH) were added to DCM, followed by the addition of 0.5 g of TEA. After stirring at room temperature for 1 hour, NaBH(OAc)3 (1.82 g, 8.61 mmol) was added. After 12 hours, the reaction was detected as complete by TLC point plate. The reaction mixture was washed with saturated NaHCO3, extracted with DCM, the DCM was dried, spin-dried, and purified by column chromatography (MeOH:DCM = 1:20) to obtain 0.62 g of a pale yellow oily viscous product. MS-APCI: 447 [M+H] + .

[0077] Phase 3-2: [ka] Compound 3-2 (200 mg, 0.447 mmol), 1-6 (218 mg, 0.52 mmol, WO2018119266), Pd(dppf)2Cl2 (37 mg, 0.0447 mmol), and Na2CO3 (138 mg, 1.3 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and reacted at 90 °C for 2 hours. The reaction completion was detected by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and column purified to obtain 18.5 mg of a pale yellow solid. MS-APCI: 700 [M+H] + .

[0078] Example 4. Synthesis of Compound 6 [ka] Stage 4-1: [ka] Compound 1-5 (200 mg, 0.477 mmol), 4-1 (253 mg, 0.52 mmol, WO2018119266), Pd(dppf)2Cl2 (39 mg, 0.0477 mmol), and Na2CO3 (138 mg, 1.3 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and reacted at 90 °C for 2 hours. The reaction completion was detected by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and subjected to column chromatography to obtain 21.5 mg of a pale yellow solid. MS-APCI: 700 [M+H] + .

[0079] Example 5. Synthesis of Compound 19 [ka] Stage 5-1: Compound 1-5 (1.5 g, 4.34 mmol) was dissolved in acetonitrile / dichloromethane (20 mL / 10 mL). Palau' chlorine (1 g, 4.77 mmol) and TFA (0.35 mL, 4.77 mmol) were added to the solution, and the mixture was stirred overnight at room temperature. The solution turned from clear to cloudy. After detecting the completion of the reaction by LC-MS, the mixture was filtered, the solid was rinsed twice with DCM, the filtrate was washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 1.46 g of a white solid. MS-APCI: 382 [M+H] + .

[0080] Stage 5-2: [ka] Compound 5-1 (1 g, 2.61 mmol) and 3-1 (600 mg, 5.22 mmol, dissolved in 1.5 mL of AcOH) were added to DCM, followed by the addition of 0.5 g of TEA. After stirring at room temperature for 1 hour, NaBH(OAc)3 (1.66 g, 7.83 mmol) was added. After 12 hours, the reaction was detected as complete by TLC point plate. The reaction mixture was washed with saturated NaHCO3, extracted with DCM, the DCM was dried, spin-dried, and purified by column chromatography (MeOH:DCM = 1:20) to obtain 0.53 g of a pale yellow oily viscous product. MS-APCI: 447 [M+H] + .

[0081] Stage 5-3: [ka] Compound 5-2 (230 mg, 0.477 mmol), 5-3 (275 mg, 0.57 mmol, WO2018119286), Pd(dppf)2Cl2 (39 mg, 0.0477 mmol), and Na2CO3 (138 mg, 1.3 mmol) were added to a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and reacted at 90 °C for 2 hours. The reaction completion was monitored by TLC. After completion of the reaction, the solution was adjusted to neutral by adding TFA and purified by reverse phase column chromatography to obtain 25.4 mg of a pale yellow solid. MS-APCI: 700 [M+H] + .

[0082] Example 6. Synthesis of Compound 27 [ka] Stage 6-1: [ka] Compound 5-1 (1 g, 2.61 mmol) and 1-4 (645 mg, 5.22 mmol) were added to DCM, followed by the addition of 0.5 g of TEA. After stirring at room temperature for 1 hour, NaBH(OAc)3 (1.66 g, 7.83 mmol) was added. After 12 hours, the reaction was detected as complete by TLC point plate. The reaction mixture was washed with saturated NaHCO3, extracted with DCM, the DCM was dried, spin-dried, and purified by column chromatography (MeOH:DCM = 1:20) to obtain 0.6 g of a pale yellow oily viscous product. MS-APCI: 453 [M+H] + .

[0083] Stage 6-2: [ka] Compound 6-1 (200 mg, 0.44 mmol), B2pin2 (135 mg, 0.53 mmol), Pd(dppf)2Cl2 (36 mg, 0.044 mmol), and KOAc (129 mg, 1.3 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane was added to the reaction flask and the reaction was allowed to proceed at 90 °C for 2 hours. The completion of the reaction was detected by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and purified by column chromatography to obtain 180 mg of a pale yellow solid. MS-APCI: 501 [M+H] + .

[0084] Stage 6-3: [ka] Compound 6-3 (500 mg, 2.32 mmol), 6-4 (310 mg, 2.56 mmol), DMAP (567 mg, 4.64 mmol), and EDCI (889 mmol, 4.64 mmol) were added to DCM (15 mL) in this order and stirred at room temperature for 1 h. The reaction was confirmed to be complete by TLC. After completion, the reaction mixture was washed with 0.5 M HCl solution (5 mL x 3). The organic layer was then dried over anhydrous sodium sulfate and purified by column chromatography to obtain 548 mg of a white solid. MS-APCI: 317 [MH] - .

[0085] Stage 6-4: [ka] Compound 6-5 (548 mg, 1.72 mmol) was dissolved in HCl in isopropanol (12 M, 3 mL) and stirred at room temperature for 30 min. The solution was then suspended in water and evaporated to dryness, azeotroped with toluene (10 mL) three times, and dried in vacuo to give 375 mg of a white solid.

[0086] Stage 6-5: [ka] Compounds 6-6 (300 g, 1.37 mmol) and 6-7 (497 mg, 1.37 mmol, WO2018119266) were added to DCM, followed by 0.5 g of TEA. After stirring at room temperature for 1 hour, NaBH(OAc)3 (871 mg, 4.11 mmol) was added. After 12 hours, the reaction was detected as complete by TLC point plate. The reaction mixture was washed with saturated NaHCO3, extracted with DCM, the DCM was dried, spin-dried, and purified by column chromatography (MeOH:DCM = 1:20) to obtain 0.41 g of a pale yellow oily viscous product. MS-APCI: 564 [M+H] + .

[0087] Stage 6-6: [ka] Compound 6-2 (150 mg, 0.3 mmol), 5-3 (169 mg, 0.3 mmol), Pd(dppf)2Cl2 (24 mg, 0.03 mmol), and Na2CO3 (95 mg, 0.9 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and the reaction was allowed to proceed at 90 °C for 2 hours. The completion of the reaction was confirmed by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, and purified by reverse phase column chromatography to give 11.8 mg of a pale yellow solid. MS-APCI: 858 [M+H] + .

[0088] Example 7. Synthesis of Compound 8 [ka] Stage 7-1: [ka] Compound 1-1 (1 g, 4.69 mmol) was dissolved in DMF (10 mL) and stirred in an ice-water bath. NaH (60%, 135 mg, 5.628 mmol) was then added and the mixture was stirred for 1 hour. 7-1 (816 mg, 4.69 mmol) was then added to the reaction mixture, which was then slowly warmed to room temperature and stirred overnight. The reaction was monitored for completion by TLC, and the mixture was quenched by adding water. The mixture was diluted to 60 mL with ethyl acetate, washed with saturated saline, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 1.2 g of a solid. MS-APCI: 350 [M+H] + .

[0089] Stage 7-2: [ka] Compound 7-2 (1 g, 2.85 mmol) was dissolved in CHOH (5 mL), and a methanol solution of CHONa (5 M, 0.7 mL, 0.34 mmol) was added. The mixture was stirred overnight at room temperature. The reaction was monitored for completion by TLC, and the mixture was quenched by adding water. The mixture was diluted to 60 mL with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 0.95 g of a solid. MS-APCI: 346 [M+H] + .

[0090] Stage 7-3: [ka] Compound 7-3 (0.9 g, 2.6 mmol) was dissolved in DCM (10 mL) and stirred in an ice-water bath. CpZrClH (1.17 g, 4.55 mmol) was then added and stirred for 1 hour while maintaining the temperature below 0 °C. Completion of the reaction was confirmed by TLC. After completion of the reaction, water (10 mL) was added and stirred for 20 minutes. The solid was then filtered and rinsed with DCM. The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, and subjected to column chromatography to obtain 0.5 g of a solid. MS-APCI: 349 [M+H] + .

[0091] Stage 7-4: [ka] Compound 7-4 (0.5 g, 1.43 mmol) and 1-4 (354 mg, 2.86 mmol) were added to DCM, followed by the addition of 0.5 g of TEA. After stirring at room temperature for 1 hour, NaBH(OAc)3 (909 mg, 4.29 mmol) was added. After 2 hours, the reaction was detected as complete by TLC point plate. The reaction mixture was washed with saturated NaHCO3, extracted with DCM, the DCM was dried, spin-dried, and purified by column (MeOH:DCM = 1:20) to obtain 0.6 g of the product as a pale yellow oily viscous substance. MS-APCI: 420 [M+H] + .

[0092] Stage 7-5: [ka] Compound 7-5 (200 mg, 0.476 mmol), 4-1 (253 mg, 0.52 mmol, WO2018119266), Pd(dppf)2Cl2 (39 mg, 0.0477 mmol), and Na2CO3 (138 mg, 1.3 mmol) were placed in a reaction flask and protected with N2 degassing. 3 mL of dioxane / 0.6 mL of water was added to the reaction flask and reacted at 90 °C for 2 hours. The reaction completion was detected by TLC. After completion of the reaction, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and column-dried to obtain 23 mg of a pale yellow solid. MS-APCI: 701 [M+H] + .

[0093] According to the above synthesis method, the following compounds were prepared using the corresponding raw materials, and the mass spectrum data are shown in Table 1. [Table 1]

[0094] Biological Testing Example A: PD-1 / PD-L1 Homogeneous Time-Resolved Fluorescence (HTRF) Binding Measurement The assay was performed in a standard black 384-well polystyrene plate with a final volume of 20 μL. The inhibitor was first serially diluted with DMSO and then added to the wells of the plate, followed by the addition of other reaction components. The final concentration of DMSO during the assay was 1%. The assay was performed at 25°C in PBS buffer (pH 7.4) containing 0.05% Tween-20 and 0.1% BSA. Recombinant human PD-L1 protein (19-238) with a C-terminal His tag was purchased from AcroBiosystems (PD1-H5229). Recombinant human PD-1 protein (25-167) with a C-terminal Fc tag was also purchased from AcroBiosystems (PD1-H5257). After diluting the PD-L1 and PD-1 proteins with assay buffer, 0.1 μL of the solution was extracted and added to the wells of the plate. The plate was centrifuged, and the proteins were pre-incubated with the inhibitors for 40 minutes. After incubation, 0.1 μl of HTRF detection buffer containing europium-blocked labeled anti-human IgG (PerkinElmer-AD0212) Fc-specific and anti-His SureLight®-allophycocyanin (APC, PerkinElmer-AD0059H) binding antibodies was added. After centrifugation, the well plate was incubated at 25°C for 60 minutes. Data (665 nm / 620 nm ratio) was read on a PHERAstar FS plate reader. The final concentrations during the assay were ~3 nM PD1, 10 nM PD-L1, 1 nM europium anti-human IgG, and 20 nM anti-His-allophycocyanin. Activity data was fitted using GraphPad Prism 5.0 software to obtain IC50 values for the inhibitors. The IC50 values of the compounds shown in the Examples are expressed as follows: IC50: + = ≦ 100 nM, ++ = 100-1000 nM, +++ = > 1000 nM. Data for the compounds of the Examples obtained using the PD-1 / PD-L1 homogeneous time-resolved fluorescence (HTRF) binding assay described in Example A are shown in Table 1. 2 will be provided to. [Table 2]

[0095] All documents mentioned in this application are incorporated by reference in the same manner as if each document were incorporated by reference individually. Furthermore, after reading the above teachings of the present invention, those skilled in the art will be able to make various changes or modifications to the present invention, and these equivalents will also be understood to be within the scope defined by the appended claims of this application.

Claims

1. Compounds 1 to 108: 【Chemical 1】 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 【Chemistry 7】 characterized in that the compound is selected from the group consisting of The compound, or an optical isomer, cis-trans isomer, hydrate, solvate or pharmaceutically acceptable salt thereof.

2. 1. A pharmaceutical composition comprising: (1) The pharmaceutical composition, comprising the compound of claim 1, or an optical isomer, cis-trans isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof, and (2) a pharmaceutically acceptable carrier.

3. 3. The pharmaceutical composition according to claim 2, for use in preventing and / or treating a disease associated with the activity or expression level of PD-1 / PD-L1, wherein the disease is selected from the group consisting of cancer, infectious diseases, and autoimmune diseases.

4. A PD-1 / PD-L1 inhibitor, The PD-1 / PD-L1 inhibitor, characterized in that the inhibitor comprises the compound of claim 1, or an optical isomer, cis-trans isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof.

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