CN112457329B - Preparation and application of aromatic heterocyclic derivative as immunomodulator - Google Patents

Abstract

The invention provides a nitrogen-containing heterocyclic compound, a preparation method, a pharmaceutical composition and application thereof, in particular to a compound 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 compounds of formula I are useful in the treatment of diseases associated with the PD-1/PD-L1 signaling pathway.

Description

Preparation and application of aromatic heterocyclic derivatives as immunomodulators

Technical Field

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

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 suppress 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 inhibitory immune checkpoints, such as PD-1, has proven to be a very effective way to inhibit cancer cells.

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 inhibit 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 immunoreceptor tyrosine inhibition motif and the immunoreceptor 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, inhibits T cell proliferation and activation, and makes T cells inactive, ultimately inducing immune escape.

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 inhibiting 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.

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 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 inhibitors under development, including Cure Tech's Pidilizumab, GSK's AMP-224, and AstraZeneca's MEDI-4736.

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 inhibitors of PD-1/PD-L1 interaction are a better choice for tumor immunotherapy.

In summary, there is an urgent need to develop novel small molecule inhibitors of PD-1/PD-L1 interaction in this field.

Summary of the invention

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

In the first aspect of the present invention, there is provided a compound as shown in the following formula I, or its optical isomers, cis-trans isomers, hydrates, solvates, or pharmaceutically acceptable salts thereof:

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

_L1 is selected from the group consisting of 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-, 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

_M1 and _M2 are each independently selected from the following group: C( _R6 ) ₂ , _NR6 , O, S, SO, _SO2 ; wherein _R6 is selected from the following group: H, chlorine, bromine, fluorine, iodine, cyano, hydroxyl, nitro, NRf, substituted or unsubstituted _C1 - _C6 alkyl, substituted or unsubstituted _C3 - _C8 cycloalkyl, substituted or unsubstituted _C6 - _C10 aryl, substituted or unsubstituted _C6 - _C10 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 - _C10 cycloalkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 -C _-C (═O)-substituted or unsubstituted C ₂ -C ₆ alkynyl, -C(═O)-substituted or unsubstituted C 2 -C 6 alkenyl, -C(═O)-substituted or unsubstituted C ₂ -C ₆ alkynyl;

M ₃ , M ₄ , and M ₅ are each independently selected from the group consisting of a chemical bond, CR ₆ , N, NR ₆ , O, S, SO, and SO ₂ ;

M ₆ are each independently selected from the following group: CR ₆ , N;

And the It is an aromatic ring;

A group having the following structure:

in,

^Z1 is selected from the following group: O, S, NRf, NO-Rf; wherein, the Rf is selected from the following group: H, substituted or unsubstituted _C1 - _C6 alkyl, substituted or unsubstituted _C3 - _C8 cycloalkyl, substituted or unsubstituted _C6 - _C10 aryl, substituted or unsubstituted _C6 - _C10 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 _{-C10 cycloalkyl} , -C(＝O)-substituted or unsubstituted C2 _{- C6} alkenyl, -C(＝O)-substituted or unsubstituted _C2 - _C6 alkynyl;

Z ² , Z ³ , and Z ⁴ are each independently selected from the group consisting of N, CH ₂ , NO, SO, SO ₂ , C(═O), NRa, and CRa; wherein Ra is selected from the group consisting of H, chlorine, bromine, fluorine, iodine, cyano, hydroxyl, nitro, NRf, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, substituted or unsubstituted C ₆ -C ₁₀ heteroaryl, -C(═O)-NRdRe, -C(═O)-substituted or unsubstituted C ₁ -C ₆ alkoxy, -C(═O)-substituted or unsubstituted C ₁ -C ₆ alkyl, -C(═O)-substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C _-C (═O)-substituted or unsubstituted C ₂ -C ₆ alkynyl, -C(═O)-substituted or unsubstituted C 2 -C 6 alkenyl, -C(═O)-substituted or unsubstituted C ₂ -C ₆ alkynyl;

Y ¹ , Y ² , and Y ³ are each independently selected from the group consisting of CH, CH ₂ , NH, N, NO, CF, CRa, O, S, SO, or SO ₂ ;

is a single bond or a double bond;

and is an aromatic or non-aromatic fragment;

R is H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, cyano, -C(＝O)-NRdRe, -C(＝O)-substituted or unsubstituted C ₁ -C ₆ alkoxy, -C(＝O)-substituted or unsubstituted C ₁ -C ₆ alkyl, -C(＝O)-substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, -C(＝O)-substituted or unsubstituted C ₂ -C ₆ alkenyl, -C(＝O)-substituted or unsubstituted C ₂ -C ₆ alkynyl, or -(L _1a ) _r -(L _2a ) _s -(L _3a ) _s -, wherein,

Each L _1a is independently a group selected from the group consisting of a chemical bond, a substituted or unsubstituted C ₁ -C ₇ alkylene group, a substituted or unsubstituted C 2 -C 4 alkenylene group, a substituted or unsubstituted C 2 -C 4 alkynylene group, -S-, -O-, a substituted or unsubstituted -NH-, -S(O)-, -S(O) ₂ -,

L _2a is selected from the group consisting of a substituted or unsubstituted C6-C12 arylene group, a substituted or unsubstituted 5-12-membered heteroarylene group having 1-3 heteroatoms, a substituted or unsubstituted C3-C8 cycloalkylene group, and a substituted or unsubstituted 5-10-membered heterocyclylene group having 1-3 heteroatoms;

L _3a is selected from the group consisting of H, substituted or unsubstituted C1-C10 alkyl, C1-C10 aryl, -CN, hydroxy, amino, carboxyl, -CO-NH-SO ₂ -R _g , -NH-SO ₂ -R _g , -SO ₂ -NH-CO-R _g ;

r is 1, 2, 3, 4, 5, 6;

s is 0, 1, and 2 respectively;

Rd, Re and Rg are each independently selected from the group consisting of H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl;

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 5-12 membered C5-C12 cyclyl, wherein the 5-12 membered heteroaryl and 5-12 membered heterocyclyl have 1-4 heteroatoms selected from B, P, N, O, S, wherein P, N, O as ring atoms may be oxo-substituted and one or more ring carbon atoms may be replaced by a carbonyl group;

A divalent group formed by a ring selected from the group consisting of: Wherein, the bonding position of the ring can be N or C;

R ₁ , R ₂ , R ₂ 'and R ₄ are each independently selected from the group consisting of 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 or -(L _1a ) _r -(L _2a ) _s -(L _3a ) _s -;

_R3 is Wherein, Rb, Rc and _Rd are each independently selected from the following group: H, substituted or unsubstituted C ₁ -C ₈ 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;

Unless otherwise specified, the term "substituted" refers to substitution 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, -CH ₂ Cl, -CHCl ₂ , -CCl ₃ , -CH ₂ F, -CHF ₂ , -CF ₃ , oxo, -CN, hydroxyl, amino, C1-C6 alkylamino, carboxyl, -NHAc, a group selected from the group consisting of 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 substituents are selected from the group consisting of halogen, hydroxyl, carboxyl, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

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

In another preferred embodiment, the ring and/or Having a substituent as shown in the following formula IV:

Wherein, each L ₄ is independently selected from the following group: substituted or unsubstituted C1-C4 alkylene, -S-, -O-, -NRa-, -S(O)-, -S(O) ₂ -; preferably substituted or unsubstituted C1-C4 alkylene, provided that the structure formed by each L ₄ together is chemically stable;

is selected from the following group: a substituted or unsubstituted C5-C10 cycloalkyl group, a substituted or unsubstituted 3-10 membered heterocyclic group having 1-3 heteroatoms selected from B, P, N, S and O; preferably, the is a 3-8 membered nitrogen-containing heterocyclic group;

Each R ₅ is independently selected from the following group: substituted or unsubstituted C1-C6 alkyl, -CN, hydroxyl, amino, carboxyl; wherein the substituent is selected from the following group: halogen, hydroxyl, carboxyl, cyano, C1-C6 alkoxy.

In another preferred embodiment, R ₁ , R ₂ , R ₂ 'and R ₄ are each independently 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 Or -(L _1a ) _r -(L _2a ) _s -(L _3a ) _s -.

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

In another preferred embodiment, the The structure is as shown below:

Wherein, X ₂ and Y ₄ are each independently selected from: CH (the CH may be substituted by R ₁ or R ₃ ), N, O, S, NH (the NH may be substituted by R ₁ or R ₃ ).

In another preferred embodiment, M ₁ and M ₂ are each independently CH ₂ .

In another preferred embodiment, the compound has a structure shown in the following formula:

in,

R ₅ is selected from the group consisting of -(L _1a ) _r -(L _2a ) _s -(L _3a ) _s ; wherein,

Each L _1a is independently a group selected from the group consisting of a chemical bond, a substituted or unsubstituted C ₁ -C ₇ alkylene group, a substituted or unsubstituted C 2 -C 4 alkenylene group, a substituted or unsubstituted C 2 -C 4 alkynylene group, -S-, -O-, a substituted or unsubstituted -NH-, -S(O)-, -S(O) ₂ -,

L _2a is selected from the following group: substituted or unsubstituted C3-C8 cycloalkylene, substituted or unsubstituted 3-10 membered heterocyclylene having 1-3 heteroatoms; preferably, the heterocyclylene contains at least one N atom as a ring atom;

L _3a is selected from the group consisting of H, substituted or unsubstituted C1-C10 alkyl, C1-C10 aryl, -CN, hydroxy, amino, carboxyl, -CO-NH-SO ₂ -R _g , -NH-SO ₂ -R _g , -SO ₂ -NH-CO-R _g ;

r is 1, 2, 3, 4, 5, 6;

s is 0, 1, and 2 respectively;

The remaining groups are as defined above.

In another preferred embodiment, the compound is selected from the following group:

The second aspect of the present invention provides a method for preparing the compound of formula I as described in the first aspect of the present invention, wherein the method comprises steps selected from those shown in synthesis schemes 1, 2 or 3:

(a) Intermediates 1 and 2 are used as raw materials to obtain the target product I through Suzuki coupling reaction catalyzed by a suitable palladium catalyst.

In another preferred embodiment, the intermediate 1 is prepared by the following method:

Using compound 1-1 as a raw material, under the action of a chiral auxiliary (such as R/S-CBS) and a reducing agent (such as borane), chiral alcohol 1-2 is formed;

(b) Using 1-2 (or 1-5) and 1-3 as raw materials, a coupling reaction (such as Suzuki, Buchwald, etc.) occurs under the conditions of a palladium catalyst and a ligand to obtain an intermediate 1-4 (or 1-6);

(c) Compound 1-1 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 1-5;

(d) Using 1-4 (or 1-6) and pinacol diboron as raw materials, a Suzuki coupling reaction occurs in the presence of a palladium catalyst and a ligand to obtain intermediate 1.

In another preferred embodiment, the intermediate 2 is prepared by the following method:

(a) Compounds 2-1 and 2-2 are used as raw materials, and in the presence of a dehydrating agent (such as concentrated sulfuric acid, PPA, etc.), intermediate 2-3 is formed;

(b) Using 2-3 (or 2-4) and 2-5 as raw materials, an affinity substitution reaction occurs under alkaline conditions (organic base or inorganic base) to form intermediate 2-8 (or 2-6);

(c) Using 2-6 and 2-7 as raw materials, a Suzuki coupling reaction occurs under the conditions of a palladium catalyst and a ligand to obtain an intermediate 2-8;

(d) Compound 2-8 is used as a raw material, and a substitution reaction occurs under the action of an iodination reagent (such as elemental iodine, NIS, etc.) to form an intermediate 2-9;

(e) Compound 2-9 is used as a raw material to form an active chlorinated product under the action of a chlorinating agent (such as phosphorus oxychloride, thionyl chloride, etc.), and then an affinity addition elimination reaction occurs under the action of an amino compound to form an intermediate 2-10;

(f) Compounds 2-10 and 2-11 are used as raw materials, and a coupling reaction occurs in the presence of a catalyst and a ligand to form intermediate 2.

The third aspect of the present invention provides a pharmaceutical composition comprising (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.

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

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

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

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.

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).

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

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

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.

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.

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.

In the sixth aspect of the present invention, there is provided an in vitro method for inhibiting PD-1/PD-L1 interaction, 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.

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

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

definition

As used herein, the term "alkyl" includes linear or branched alkyl groups. For example, _C1 - _C8 alkyl groups represent linear or branched alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.

As used herein, the term "alkenyl" includes straight or branched alkenyl groups. For example, _C2 - _C6 alkenyl refers to straight or branched alkenyl groups having 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or similar groups.

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 to 6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.

As used herein, the term "C ₃ -C ₁₀ 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.

As used herein, the term "C ₁ -C ₈ alkylamino" refers to an amine group substituted by a C ₁ -C ₈ 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.

As used herein, the term "C ₁ -C ₈ 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.

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.

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

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.

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, C ₁ -C ₆ alkyl-amino, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C _{2 -C 6 alkynyl, C 1 -C 6 alkoxy} , halogenated C ₁ -C ₆ alkyl, halogenated C ₂ -C ₆ alkenyl, halogenated C ₂ -C ₆ alkynyl, halogenated C ₁ -C ₆ alkoxy, allyl, benzyl, C ₆ -C ₁₂ aryl, C ₁ -C ₆ alkoxy-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy-carbonyl, phenoxycarbonyl, C ₂ -C ₆ alkynyl-carbonyl, C ₂ -C ₆ alkenyl-carbonyl, C ₃ -C ₆ cycloalkyl-carbonyl, C ₁ -C 6 _6- alkyl-sulfonyl, etc.

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.

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.

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.

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.

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

It is to be understood that herein, the definitions of the various groups are intended to create chemically stable structures.

Active ingredients

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.

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

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.

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.

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.

Preparation of compounds of formula I

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.

(a) intermediates 1 and 2 are used as raw materials to obtain the target product I through a Suzuki coupling reaction catalyzed by a suitable palladium catalyst;

Among them, the preparation method of intermediate 1 (refer to WO 2018195321, WO2018005374 and WO2019023575) is as follows:

Using compound 1-1 as a raw material, under the action of a chiral auxiliary (such as R/S-CBS) and a reducing agent (such as borane), chiral alcohol 1-2 is formed;

(b) Using 1-2 (or 1-5) and 1-3 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 1-4 (or 1-6);

(c) Compound 1-1 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 1-5;

(d) using 1-4 (or 1-6) and pinacol diboron as raw materials, under the conditions of suitable palladium catalyst and ligand, a Suzuki coupling reaction occurs to obtain intermediate 1;

The preparation method of intermediate 2 is as follows:

(a) Compounds 2-1 and 2-2 are used as raw materials, and in the presence of a dehydrating agent (such as concentrated sulfuric acid, PPA, etc.), intermediate 2-3 is formed;

(b) Using 2-3 (or 2-4) and 2-5 as raw materials, an affinity substitution reaction occurs under alkaline conditions (organic base or inorganic base) to form intermediate 2-8 (or 2-6);

(c) Using 2-6 and 2-7 as raw materials, a Suzuki coupling reaction occurs under the conditions of a suitable palladium catalyst and a ligand to obtain an intermediate 2-8;

(d) Compound 2-8 is used as a raw material, and a substitution reaction occurs under the action of an iodination reagent (such as elemental iodine, NIS, etc.) to form an intermediate 2-9;

(e) Compound 2-9 is used as a raw material to form an active chlorinated product under the action of a chlorinating agent (such as phosphorus oxychloride, thionyl chloride, etc.), and then an affinity addition elimination reaction occurs under the action of an amino compound to form an intermediate 2-10;

(f) Compounds 2-10 and 2-11 are used as raw materials, and a coupling reaction occurs under the conditions of a suitable catalyst and ligand to form intermediate 2;

In addition, the starting materials and intermediates in 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.

Pharmaceutical compositions and methods of administration

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).

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.

"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.

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).

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 capsules, tablets and pills, the dosage forms may also comprise buffering agents.

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.

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.

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

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.

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.

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

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 diseases related to PD-1/PD-L1 interaction.

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.

The main advantages of the present invention include:

(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.

(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.

(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.

(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.

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.

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

Common materials and test methods:

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

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

Chemical shifts were recorded with tetramethylsilane as internal standard and expressed in ppm (CDC1 ₃ : δ 7.26 ppm). The recorded data information is as follows: chemical shift and its splitting and coupling constants (s: singlet; d: doublet; t: triplet; q: quartet; br: broad peak; m: multiplet).

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.

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

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.

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

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%, and the structural characterization of each final product is determined by MS and/or hydrogen nuclear magnetic resonance ( ^1H NMR) analysis. The synthesis of various compounds and intermediates of the present invention is illustrated by examples below.

Example 1 Synthesis of Compound LW1005-001

Step 1-1:

Compound 1 (5.0 g, 23.6 mmol, WO2012158550.), 2 (4.4 g, 25.9 mmol), Xantphos-PdCl2 (892 mg, 1.18 mmol) and Cs ₂ CO ₃ (15.4 g, 47.2 mmol) were added to dioxane (100 mL), N2 protected, and reacted at 95 degrees for 2 hours. The reaction was complete on TLC plate. The reaction solution was filtered, the filtrate was dried by spin drying, and column chromatography (EA:HEP=1:30) was performed to obtain 4.8 g of light yellow solid. MS-APCI: 348 [M+H] ⁺ . Step 1-2:

Compound 3 (4.8 g, 13.8 mmol) and 4 (2.5 g, 20.7 mmol, 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, 20.7 mmol) was added. After 30 minutes, the reaction was complete on TLC. The reaction solution was washed with saturated NaHCO 3 , extracted with DCM, dried with DCM, spin-dried, and purified by column (MeOH:DCM=1:20) to obtain 5.1 g of product, a light yellow oily viscous substance. MS-APCI: 419[M+H] ⁺ .

Steps 1-3:

Compound 5 (5.1 g, 12.2 mmol), Bpin ₂ (3.4 g, 13.4 mmol), Pd(dppf)Cl ₂ .DCM (498 mg, 0.61 mmol), KOAc (2.4 g, 24.4 mmol) were added to dioxane (100 mL), N ₂ 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] ⁺ .

Steps 1-4:

Compound 7 (51 g, Journal of Medicinal Chemistry, 2019, 62, 276-287) and 2-methyl-3-bromobenzoic acid (95.56 g) were added to PPA (500 g) in sequence, and mechanically stirred at 140 ° C for 6 hours. After the reaction is complete, ice water is poured into the reaction bottle, and after dilution, it is poured out and slurried for 30 minutes and filtered. 500 ml of water is added to the filtered solid, and sodium hydroxide solid is added under mechanical stirring. The aqueous phase is adjusted to pH = 6-8 and filtered. The filter cake solid is dried at 55 ° C to obtain 80 g of off-white solid product. MS-APCI: 305 [M + H] ⁺ .

Steps 1-5:

Add raw material 8 (20 g) and cesium carbonate (42 g) to 300 ml DMF, add bromoethanol (16 g), heat and stir at 50°C for 16 hours. TLC detection shows that the reaction is complete. The reaction solution is filtered and concentrated, slurried with water, filtered, and dried to obtain 19 g of off-white solid. MS-APCI: 349 [M+H] ⁺ .

Steps 1-6:

The raw material 9 (15 g) was suspended in DCM (300 mL), and then Et ₃ N (13 g) and BzCl (12 g) were added in sequence, stirred at room temperature, reacted for one hour, TLC showed that the reaction was complete, 200 ml of water was added, extracted, separated, dried, concentrated, and slurried to obtain 18 g of white solid. MS-APCI: 453 [M+H] ⁺ .

Steps 1-7:

Add raw material 10 (18 g) and NIS (10.7 g) to MeCN (230 mL) in sequence, add 5 ml of TFA dropwise, and stir in an oil bath at 55°C for 1.5 hours. After the reaction is completed by TLC, cool, filter, and dry to obtain 13.8 g of yellow solid. MS-APCI: 579 [M+H] ⁺ .

Steps 1-8:

Add raw material 11 (1.0 g) to POCl ₃ (10 mL), react at 100°C for 1 hour, concentrate to dryness, add methylamine aqueous solution to the residual liquid, stir under ice bath, green solid appears, filter, dry to obtain 960 mg solid. MS-APCI: 592 [M+H] ⁺ .

Steps 1-9:

Add raw material 12 (960 mg) and K ₂ CO ₃ (770 mg) to MeOH (20 mL) in sequence, stir at 55° C., monitor by TLC, until the reaction is complete, filter, and dry to obtain 900 mg of a green solid. MS-APCI: 488 [M+H] ⁺ . Steps 1-10:

The raw material 13 (48 mg), ZnCN ₂ (22 mg), Pd ₂ (dba) ₃ (11 mg), and dppf (15 mg) were mixed in a 50 mL single-mouth bottle, and N ₂ was used for protection. 5 mL of DMF was added and stirred at 95°C for 16 hours. The reaction was completed by TLC. The reaction solution was filtered, the filtrate was washed with water, and extracted with EA. After column chromatography, 28 mg of green solid was obtained. MS-APCI: 387 [M + H] ⁺ .

Steps 1-11:

Compound 14 (100 mg), 6 (139 mg), Pd(dppf)Cl ₂ (15 mg), and Na ₂ CO ₃ (53 mg) were placed in a reaction bottle, degassed with N ₂ , and 3 mL of dioxane/0.6 mL of water were injected into the reaction bottle. The reaction was allowed to react at 90 degrees for 2 hours. The reaction was monitored by TLC. After the reaction was complete, the reaction solution was washed with water, extracted with EA, dried, spin-dried, and filtered to obtain 150 mg of a white solid. MS-APCI: 647 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d6) δ10.04(s,1H),9.75-9.61(m,2H),9.02(s,1H),8.17(d,J＝7.7Hz,1H),7.83(d,J＝8.1Hz,1H),7.67–7.45(m,3H),7.39(t,J＝7.5Hz,1H) ),7.25(d,J＝7.4Hz,1H),6.58(d,J＝6.5Hz,1H),6.52(t,J＝7.0Hz,1H),5.54-5.40(m,1H),5.32- 5.15(m,1H),4.53-4.38(m,3H),4.36-4.31(m,1H),4.28-4.22(m,1H),4.01(s,3H),3.81(s,2H),3.61(s,3H),3.24(s,3H),3.10-3.02(m,1H),2. 90-2.80(m,1H),2.74-2.58(m,2H),2.46(s,3H),2.15-2.06(m,1H),2.02-1.92(m,1H),1.87-1.78(m,1H).

Example 2 Synthesis of Compound LW1005-002

Step 2-1:

Compound 11 (1.0 g, 1.73 mmol) was added to POCl ₃ (15 ml), reacted at 90°C for 1 hour, concentrated to dryness and set aside. NH ₂ OMe/HCl (10 g) was dissolved in 6 mL of water, NaOH (4.79 g) was added and stirred for 10 minutes, filtered, the filtrate was cooled in an ice bath and poured into the residue after reaction 1, stirred in an ice bath, solid appeared, filtered and dried to obtain 1 g of crude solid. MS-APCI: 608 [M+H] ⁺ .

Step 2-2:

Compound 15 (3 g, 4.9 mmol) was dissolved in MeOH/THF (30 mL/30 mL), and 5N sodium methoxide solution (2.5 mL, 12.3 mmol) was added, stirred at room temperature, and monitored by TLC. After the reaction was completed, water was added to quench the reaction, and the solvent was dried by spin drying. A solid appeared, which was filtered and dried to obtain 2.7 g of a light yellow solid. MS-APCI: 504 [M+H] ⁺ .

Step 2-3:

The raw material 16 (7.8 g, 15.5 mmol), Zn(CN) ₂ (1.09 g, 9.28 mmol), Xantphos-PdCl2 (1.17 g, 1.55 mmol), and Cs ₂ CO ₃ (7.56 g, 23.2 mmol) were mixed in a 250 mL three-necked flask, replaced with N ₂ for 3 times, injected with 150 mL DMF, stirred at 90°C for 2 h, and the reaction was complete after LCMS detection. Filter, concentrate the filtrate to dryness, and slurry the solid with (DCM/MeOH=100 mL/2 mL) to obtain 4.5 g of light yellow solid. MS-APCI: 403 [M+H] ⁺ .

Steps 2-4:

4 (100 mg, 0.25 mmol), 5 (139 mg, 0.3 mmol), Pd (dppf) ₂ Cl ₂ (20 mg, 0.025 mmol), Na ₂ CO ₃ (53 mg, 0.5 mmol) were placed in a reaction flask, N ₂ was degassed, 3 mL dioxane/0.6 mL water was injected into the reaction flask, and the reaction was carried out at 90 degrees for 2 hours. The TLC plate showed that the reaction was complete. The reaction solution was washed with water, extracted with EA, dried, spin-dried, and passed through a column (DCM: MeOH = 10: 1) to obtain 180 mg of a crude product, and prepared for separation to obtain 29.5 mg of a light yellow solid. MS-APCI: 663 [M + H] ⁺ . ¹ H NMR (400MHz, DMSO-d6) δ9.95(m,1H),8.25(s,1H),7.95(dd,J＝7.8,1.4Hz,1H),7.83(d,J＝8.1Hz,1H),7.53–7.45(m,2H),7.39(dt,J＝14.8,7.4Hz,2H),7.23( d,J＝7.4Hz,1H),6.57(d,J＝7.3Hz,1H),6.52(t,J＝8.8Hz,1H),5.49(d,J＝33.1Hz,1H),4.9 8(s,1H),4.44(d,J＝25.2Hz,1H),4.30(d,J＝31.6Hz,1H),4.06-3.94(m,5H),3.80-3.69(m,5H),3.63-3.50(m,1H),3.24(s,1H),2.87-2.80(m,1H), 2.76–2.54(m,3H),2.38(d,J＝14.2Hz,3H),2.15-2.06(m,1H),2.03-1.92(m,1H),1.90–1.77(m,1H).

Example 3 Synthesis of Compound LW1005-003

Step 3-1:

Mix raw material 8 (5.0 g, 16 mmol) and cesium carbonate (10.0 g, 32 mmol) in DMF (100 mL), add methyl 3-bromopropionate (8.0 g, 48 mmol), stir at 55°C for 6 hours, wash with water, filter, and purify by column chromatography to obtain 5.2 g of a yellow-brown solid. MS-APCI: 391 [M+H] ⁺ .

Step 3-2:

Add raw material 18 (5.0 g, 54.6 mmol) and NIS (4.3 g, 19.2 mmol) to MeCN (100 mL) in sequence, then drop 2 mL of TFA into the reaction solution and stir in an oil bath at 55°C for 2 hours. After the reaction is complete, cool, filter, rinse with n-heptane (15 mL x 3), and dry to obtain 5 g of white solid. MS-APCI: 517 [M+H] ⁺ .

Step 3-3:

Using compound 19 (1.0 g, 1.94 mmol), O-methylhydroxylamine hydrochloride (5 g, 60 mmol) and NaOH (2.4 g, 60 mmol) as reactants, according to the synthesis method of step 2-1, 1.1 g of a yellow solid was obtained.

MS-APCI: 546[M+H] ⁺ .

Step 3-4:

The raw material 20 (600 mg, 1.13 mmol) was dissolved in THF/H ₂ O (1:1, 50 mL), and lithium hydroxide (142 mg) was added. The reaction was allowed to react at room temperature for 0.5 hours and the reaction was monitored by TLC. After the reaction was complete, 20 ml of water was added, TFA was used to adjust the acidity, and DCM (20 ml x 3) was used for extraction, drying, and concentration to obtain 450 mg of a yellow solid. MS-APCI: 532 [M+H] ⁺ .

Steps 3-5:

Using compound 21 (120 mg, 0.23 mmol), Zn(CN) ₂ (26 mg, 0.23 mmol), Xantphos-PdCl ₂ (17 mg, 0.023 mmol), and Cs ₂ CO ₃ (150 mg, 0.46 mmol) as reactants, according to the synthesis method of step 2-3, 80 mg of yellow solid was obtained. MS-APCI: 431 [M+H] ⁺ .

Steps 3-6:

Compound 22 (120 mg, 0.28 mmol), 6 (156 mg, 0.33 mmol), Pd(dppf) ₂ Cl ₂ (23 mg, 0.028 mmol), and Na ₂ CO 3 (59 mg, 0.56 mmol) were used as reactants and the synthetic method of steps 2-4 was followed to obtain 5 mg of a light yellow solid. MS-APCI: 691 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d6) δ8.32(s,1H),7.90(d,J＝7.9Hz,1H),7.57(d,J＝7.9Hz,1H),7.45(d,J＝7.5Hz,2H),7.35(dt,J＝19.6,7.7Hz,2H),7.17(d,J＝7.5Hz,1H), 6.48(d,J＝7.1Hz,1H),6.36(t,J＝8.6Hz,1H),4.66(s,1H),4.16(s,1H),4. 07(t,J＝6.8Hz,2H),3.90(s,3H),3.74(s,3H),3.50-3.45(m,2H),2.73(t,J＝6.9Hz,2H),2.65-2.50(m,3H),2.42-2.37(m,1H),2.36-2.32(m,3H),2 .23-2.27(m,1H),2.08-2.00(m,1H),1.96-1.91(m,1H),1.50(dd,J＝9.9,5.7Hz,1H).

Example 4 Synthesis of Compound LW1005-004

Step 4-1:

Compound 8 (5.0 g, 16 mmol), methyl 4-bromobutyrate (3.56 g, 21.4 mmol) and cesium carbonate (10.7 g, 32 mmol) were used as reactants and the synthesis method of step 3-1 was followed to obtain 5.7 g of a gray solid. MS-APCI: 405 [M+H] ⁺ .

Step 4-2:

Compound 23 (5.7 g, 14.1 mmol) and NIS (6.35 g, 28.2 mmol) were used as reactants and according to the synthesis method of 3-2, 4.9 g of light pink solid was obtained. MS-APCI: 531 [M+H] ⁺ .

Step 4-3:

Using compound 24 (1.0 g, 1.88 mmol), O-methylhydroxylamine hydrochloride (5 g, 60 mmol) and NaOH (2.4 g, 60 mmol) as reactants, according to the synthesis method of step 2-1, 1.1 g of a yellow solid was obtained.

MS-APCI: 560[M+H] ⁺ .

Step 4-4:

The raw material 25 (830 mg, 1.49 mmol) was dissolved in 20 mL THF/H2O (1:1), and lithium hydroxide (125 mg) was added. The reaction was allowed to react at room temperature for 0.5 hours. The reaction was complete on TLC. THF was dried and TFA was added to adjust the acidity. Solids precipitated and filtered. The solids were dried to give 740 mg of yellow solids. MS-APCI: 546 [M+H] ⁺ .

Step 4-5:

26 (400 mg, 0.73 mmol), Zn(CN)2 (52 mg, 0.44 mmol), Xantphos-PdCl2 (56 mg, 0.074 mmol), Cs2CO3 (358 mg, 1.1 mmol) were mixed in a 50 mL two-necked bottle, 10 mL NMP was added, N2 was replaced 3 times, and the mixture was stirred at 85 °C for 1 hour. The reaction was completed by LCMS. Water was added, acid was adjusted with TFA, EA was extracted, dried, concentrated, and column chromatography was performed to obtain 120 mg of a yellow solid. MS-APCI: 445 [M+H] ⁺ .

Steps 4-6:

Compound 27 (120 mg, 0.27 mmol), 6 (139 mg, 0.3 mmol), Pd(dppf) ₂ Cl ₂ (23 mg, 0.028 mmol), and Na ₂ CO 3 (86 mg, 0.81 mmol) were used as reactants and the synthetic method of steps 2-4 was followed to obtain 40 mg of a light yellow solid. MS-APCI: 704 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d6) δ8.32(s,1H),7.91(d,J＝7.8Hz,1H),7.57(d,J＝7.9Hz,1H),7.45(t,J＝6.6Hz,2H),7.35(dt,J＝18.8,7.6Hz,2H),7.17(d,J＝7.4Hz,1H), 6.48(d,J＝7.4Hz,1H),6.36(t,J＝8.6Hz,1H),4.65(s,1H),4.16(s,1H),3.95- 3.91(m,1H),3.90(s,3H),3.74(s,3H),3.50-3.40(m,4H),2.83–2.71(m,1H),2.68-2.63(m,2H),2.60-2.53(m,2H),2.44-2.38(m,1H),2.35(d,J ＝14.9Hz,3H),2.30-2.23(m,2H),2.08-2.02(m,1H),2.00–1.91(m,3H),1.54-1.43(m,1H).

Example 5 Synthesis of Compound LW1005-005

Step 5-1:

Compound 3 (4.8 g, 13.8 mmol), (R)-3-pyrrolidinecarboxylic acid (2.4 g, 20.7 mmol, dissolved in 1.5 mL AcOH) and NaBH(OAc) ₃ (4.4 g, 20.7 mmol) were used as reactants. According to the synthesis method of step 1-2, 5 g of light yellow oily viscous material was obtained. MS-APCI: 447 [M+H] ⁺ .

Step 5-2:

Compound 17 (650 mg, 1.62 mmol), Bpin2 (452 mg, 1.78 mmol), Pd(dppf)Cl2.DCM (132 mg, 0.162 mmol), KOAc (317.5 mg, 3.24 mmol) were used as reactants and the synthesis method of 1-3 was followed to obtain 500 mg of a white solid. MS-APCI: 451 [M+H] ⁺ .

Step 5-3:

Compound 28 (89 mg, 0.2 mmol), 29 (99 mg, 0.22 mmol), Pd(dppf) ₂ Cl ₂ (17 mg, 0.02 mmol) and Na ₂ CO ₃ (53 mg, 0.5 mmol) were used as reactants and according to the synthesis method of 2-4, 13 mg of yellow solid was obtained. MS-APCI: 691 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d6) δ8.21(s,1H),7.91(d,J＝9.2Hz,1H),7.57(d,J＝8.0Hz,1H),7.45(t,J＝7.0Hz,2H),7.35(dt,J＝19.7,7.5Hz,2H),7.17(d,J＝7.3Hz,1H), 6.48(s,1H),6.36(t,J＝8.7Hz,1H),4.94(s,1H),4.32(s,1H),3. 95(d,J＝5.4Hz,1H),3.90(s,3H),3.73(s,1H),3.70(s,3H),3.51-3.45(m,4H),2.91-2.85(m,1H),2.70-2.64(m,2H),2.62–2.55(m,2H),2.35(dd,J ＝15.0,5.9Hz,3H),2.05–2.00(m,1H),1.97-1.88(m,3H),1.43-1.35(m,1H).

Example 6 Synthesis of Compound LW1005-006

Step 6-1:

Compound 17 (2.6 g, 6.45 mmol) was dissolved in DCM (120 mL), and then Dess-Martin Reagent (8.2 g, 19.34 mmol) was added to the above solution at one time, stirred at room temperature for 30 minutes, and the reaction was monitored by TLC. After the reaction was complete, water was added to quench the reaction, and then the reaction solution was washed with saturated sodium bicarbonate, 10% sodium thiosulfate solution and saturated brine in turn, the organic layer was dried over anhydrous sodium sulfate, filtered, and the organic layer was poured into a 250 mL single-mouth bottle. 4-Carboxylic acid piperidine methyl ester (1.85 g, 12.9 mmol), acetic acid (1 drop) and sodium acetate borohydride (4.1 g, 19.35 mmol) were added to the above solution in turn, stirred at room temperature for 2 hours, and the reaction was monitored by TLC. After the reaction was complete, water was added to quench the reaction, and then the reaction solution was washed with saturated sodium bicarbonate and saturated brine in turn, the organic layer was dried over anhydrous sodium sulfate, and column chromatography was performed to obtain 1.02 g of light yellow solid. MS-APCI: 528[M+H] ⁺ .

Step 6-2:

The raw material 30 (200 mg, 0.378 mmol) was dissolved in 10 mL THF/H2O (2:1), sodium hydroxide (45 mg, 1.135 mmol) was added, and the reaction was allowed to react overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, TFA (0.08 mL, 1.135 mmol) was added to adjust the solution to neutrality, THF was dried, and water was added to stir and wash, solid precipitated, filtered, and the solid was dried to obtain 195 mg of light yellow solid. MS-APCI: 514 [M+H] ⁺ .

Step 6-3:

Compound 31 (80 mg, 0.155 mmol), 6 (90 mg, 0.2 mmol), Pd(dppf) ₂ Cl ₂ (17 mg, 0.02 mmol) and Na ₂ CO ₃ (53 mg, 0.5 mmol) were used as reactants and according to the synthesis method of 2-4, 23 mg of yellow solid was obtained. MS-APCI: 774 [M+H] ⁺ . ¹ H NMR(400MHz, DMSO-d6)δ8.23(d,J＝2.7Hz,1H),7.91(d,J＝7.1Hz,1H),7.62–7.52(m,1H),7.44(d,J＝6.7Hz,2H),7.40–7.27(m,2H),7.17(dd,J＝7.4,2.7Hz,1H) ,6.48(d,J＝7.2Hz,1H),6.40–6.27(m,1H),4.64(s,1H),4.16(s,1H),3.96(d,J＝7.2 Hz,2H),3.90(s,3H),3.73(s,3H),3.45(dd,J＝10.1,2.5Hz,2H),2.82(d,J＝10.7Hz,2H),2.68-2.53(m,6H),2.42–2.31(m,3H),2.30-2.27(m,1H),2.2 0-2.13(m,2H),2.08-2.02(m,3H),1.98-1.90(m,2H),1.80–1.70(m,2H),1.54–1.40(m,3H).

Example 7 Synthesis of Compound LW1005-007

Step 7-1:

32 (9.1 g, 1.0 eq; EP2848622) and Cs2CO3 (41.56 g, 3.0 eq) were added to the reaction flask, DMF was added to dissolve, 2-bromoethanol (6.03 mL, 2.0 eq) was added dropwise, and stirred at 50 °C for 18 h. The reaction was completed by LC-MS monitoring. DMF was spin-dried, and 100 mL of water was added to slurry for 1 h. Filtered, the filter cake was washed with 50 mL of water and 100 mL of n-heptane. 9.0 g of brown solid was obtained. MS-APCI: 258 [M+H] ⁺

Step 7-2:

33 (9.0 g, 1.0 eq), 34 (13.46 g, 1.3 eq; Chem, 2019, 5, 929-939), Cs2CO3 (22.73 g, 2.0 eq), Pd(PPh3)4 (2.02 g, 0.05 eq) were added to the reaction flask, N2 was protected, 1,4-dioxane/water was added to dissolve, and stirred at 100°C for 3 h. LC-MS monitoring showed that the reaction was complete. The reaction solution was washed with water, brine, dried, concentrated, and column chromatographed to obtain 10.1 g of solid. MS-APCI: 348 [M+H] ⁺

Step 7-3:

35 (5.0 g, 1.0 eq) and Et3N (6 mL, 3.0 eq) were added to dichloromethane, and BzCl was added dropwise at RT. The mixture was stirred for 2 h at RT. The reaction was monitored by TLC to be complete. The reaction solution was washed with saturated sodium bicarbonate solution and saturated brine, dried, concentrated, and column chromatographed. 6.4 g of a light yellow to off-white solid was obtained. MS-APCI: 452 [M+H] ⁺

Step 7-4:

36 (11.0 g, 1.0 eq) was dissolved in acetonitrile, NIS was added, TFA was added dropwise at RT, and after addition, the mixture was stirred at 55°C for 3 h, and a large amount of solid precipitated. TLC monitored the reaction completion, and the mixture was filtered, the filter cake was washed with n-heptane, the filtrate was dried by rotary evaporation, extracted with EA/water, washed with 10% Na2S2O3 solution, dried, concentrated, and mixed with the filter cake for column chromatography. 13 g of off-white solid was obtained. MS-APCI: 578 [M+H] ⁺

Step 7-5:

37 (1.0 g) was added to the reaction flask, and 5 mL of POCl3 was added. The mixture was stirred at 90 °C for 1.5 h. The mixture was spin-dried and pumped to a foamy solid. 30% aqueous methylamine solution (20 mL, 100 eq) was added, dissolved in DCM, and stirred at rt for 15 min. The raw material was completely converted by TLC monitoring, and the organic phase was separated, washed with brine, dried, concentrated, and column chromatographed to obtain 1.0 g of a brown solid. MS-APCI: 591 [M+H] ⁺

Step 7-6:

Disperse 38 (1.0 g 1.0 eq) in methanol/THF, add K2CO3 (700 mg 3.0 eq) solid, and stir at room temperature for 3 h. TLC shows that a large amount of raw materials remain, add K2CO3 (700 mg 3.0 eq), continue stirring at room temperature overnight, TLC shows that the raw materials are basically completely reacted. Spin dry, extract with DCM/water, wash the organic phase with brine, dry, concentrate, and column chromatography. Obtain 700 mg of brown solid. MS-APCI: 487 [M+H] ⁺ Step 7-7:

39 (2.7 g 1.0 eq), Zn(CN)2 (299.4 mg 0.46 eq), Cs2CO3 (2.17 g 1.2 eq), Xantphos-PdCl2 (418 mg 0.1 eq) were added to the reaction flask, N2 was used for protection, DMF was added to dissolve, and the mixture was stirred at 90 °C for 2 h. LC-MS monitoring showed that the reaction was complete, DMF was dried, and the organic phase was extracted with DCM/water. The organic phase was washed with brine, dried, concentrated, and column chromatographed. 2.2 g of brown solid was obtained. MS-APCI: 386 [M+H] ⁺

Step 7-8:

40 (300 mg 1.0 eq), 5 (352.5 mg 1.2 eq), Na2CO3 (164.6 mg 2.0 eq), Pd(dppf)2Cl2 (41 mg, 0.1 eq) were added to the reaction flask, N2 was used for protection, 1,4-dioxane/water was added to dissolve, and stirred at 90°C for 2 h. LC-MS monitoring showed that the reaction was complete. The reaction solution was extracted with DCM/water, and the organic phase was washed with brine, dried, concentrated, and column chromatographed to obtain 30 mg of a light yellow solid. MS-APCI: 646[M+H] ⁺

Example 8 Synthesis of Compound LW1005-008

Step 8-1:

Compound 37 (1.5 g, 2.59 mmol) was added to POCl ₃ (30 ml), reacted at 90°C for 2 hours, concentrated to dryness and set aside. NH ₂ OMe/HCl (22 g) was dissolved in 10 mL of water, NaOH (10.36 g) was added and stirred for 10 minutes, filtered, the filtrate was cooled in an ice bath and poured into the residue after reaction 1, stirred in an ice bath, solid appeared, filtered and dried to obtain 1.5 g of crude solid. MS-APCI: 607 [M+H] ⁺ .

Step 24-2:

Compound 41 (300 mg, 0.49 mmol) was dissolved in MeOH/THF (30 mL/30 mL), K2CO3 (28 mg, 0.98 mmol) was added, stirred at room temperature, and monitored by TLC. After the reaction was completed, the solvent was dried, and a solid appeared. The solid was filtered and dried to obtain 200 mg of a light yellow solid. MS-APCI: 503 [M+H] ⁺ . Step 8-3:

The raw material 42 (135 mg, 0.268 mmol), Zn(CN) ₂ (16 mg, 0.137 mmol), Xantphos-PdCl2 (20 mg, 0.0268 mmol), and Cs ₂ CO ₃ (131 mg, 0.402 mmol) were mixed in a 50 mL three-necked flask, replaced with N ₂ for 3 times, injected with 10 mL DMF, stirred at 90°C for 2 h, and the reaction was complete after LC-MS detection. Filter, concentrate the filtrate to dryness, mix the sample, and column chromatography to obtain 4.5 g of light yellow solid. MS-APCI: 402 [M+H] ⁺ .

Step 8-4:

Compound 43 (50 mg, 0.124 mmol) was dissolved in DCM (120 mL), and then Dess-Martin Reagent (158 mg, 0.373 mmol) was added to the above solution at one time, stirred at room temperature for 30 minutes, and the reaction was monitored by TLC. After the reaction was complete, water was added to quench the reaction, and then the reaction solution was washed with saturated sodium bicarbonate, 10% sodium thiosulfate solution and saturated brine in turn, the organic layer was dried over anhydrous sodium sulfate, filtered, and the organic layer was poured into a 250 mL single-mouth bottle. 4-Carboxylic acid piperidine methyl ester (0.05 mL, 0.373 mmol), acetic acid (1 drop) and sodium acetate borohydride (79 mg, 0.373 mmol) were added to the above solution in turn, stirred at room temperature for 2 hours, and the reaction was monitored by TLC. After the reaction was complete, water was added to quench the reaction, and then the reaction solution was washed with saturated sodium bicarbonate and saturated brine in turn, the organic layer was dried over anhydrous sodium sulfate, and column chromatography was performed to obtain 20 mg of light yellow solid. MS-APCI: 527[M+H] ⁺ .

Step 8-5:

The raw material 104 (230 mg, 0.436 mmol) was dissolved in 10 mL THF/H2O (2:1), and LiOH (21 mg, 0.872 mmol) was added. The reaction was allowed to react at room temperature for 2 hours, and the reaction was monitored by TLC. After the reaction was complete, TFA (0.08 mL, 1.135 mmol) was added to adjust the solution to neutrality, THF was dried, and water was added to stir and wash. Solids precipitated, filtered, and the solids were dried to obtain 200 mg of light yellow solids. MS-APCI: 513 [M+H] ⁺ .

Step 8-6:

Compound 105 (200 mg), 5 (148 mg), Pd(dppf)2Cl2 (34 mg) and Na2CO3 (170 mg) were placed in a reaction bottle, degassed with N2, and 3 mL dioxane/0.6 mL water was injected into the reaction bottle. The reaction was carried out at 90 degrees for 2 hours. The reaction was complete on the TLC plate. The reaction solution was washed with water, extracted with EA, dried, spin-dried, and passed through a column (DCM:MeOH=10:1) to obtain 18 mg of a yellow solid. MS-APCI: 767 [M+H] ⁺

Table 3 According to the synthesis method of LW1005-001, the following compounds were prepared using the corresponding raw materials

Table 4 According to the synthesis method of LW1005-007, the following compounds were prepared using the corresponding raw materials

Table 4 According to the synthesis method of LW1005-006, the following compounds were prepared using the corresponding raw materials

Table 5 The following compounds were prepared using the corresponding raw materials according to the synthesis method of LW1005-008

Biological Testing

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

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 -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.

The IC50 values of the compounds illustrated in the examples are expressed in the following manner: IC50: + = ≤ 100 nM; ++ = 100 to 1000 nM; +++ = > 1000 nM

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.

Table 1.

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 (10)

1. A compound of formula I, or an optical isomer, a cis-trans isomer, or a pharmaceutically acceptable salt thereof:

Wherein the compound of formula I has a structure shown in the following formula:

n, m and p are each independently selected from 0, 1, 2, 3 or 4;

L ₁ is-O-;

Z ¹ selected from the group consisting of: o, NR _f 、N-O-R _f The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is as follows _f Selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₃ -C ₈ Cycloalkyl, -C (=o) -NRdRe, -C (=o) -substituted or unsubstituted C ₁ -C ₆ Alkoxy, -C (=o) -substituted or unsubstituted C ₁ -C ₆ Alkyl, -C (=o) -substituted or unsubstituted C ₃ -C ₁₀ Cycloalkyl, -C (=o) -substituted or unsubstituted C ₂ -C ₆ Alkenyl, -C (=o) -substituted or unsubstituted C ₂ -C ₆ Alkynyl; rd, re are each independently selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₃ -C ₁₀ Cycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ An aryl group;

R ₆ selected from the group consisting of: cyano, substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₃ -C ₈ Cycloalkyl;

Y ¹ is O;

Y ² is N;

R ₅ ' is substituted or unsubstituted C ₁ -C ₆ Alkyl, or

Wherein each L ₄ Independently selected from the group consisting of: substituted or unsubstituted C1-C4 alkylene, -NRa-; provided that each L ₄ The co-formed structure is chemically stable; the Ra is selected from the following group: H. substituted or unsubstituted C ₁ -C ₆ An alkyl group;selected from the group consisting of: substituted or unsubstituted C5-C10 cycloalkyl, substituted or unsubstituted 3-8 A membered nitrogen-containing heterocyclic group;

each R is ₅ Each independently selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, -CN, hydroxy, amino, carboxy; wherein the substituents are selected from the group consisting of: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy;

selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl;

R ₁ 、R ₂ and R is ₂ ' each independently selected from the group consisting of: 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;

R ₃ is thatWherein Rb, rc and R _d Each independently selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₈ An alkyl group; or said Rb and Rc together with the adjacent N atom form a substituted or unsubstituted 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O;

unless otherwise specified, the term "substituted" refers to substitution with one or more substituents selected from the group consisting of: halogen, -F, cl, br, -CH ₂ Cl、-CHCl ₂ 、-CCl ₃ 、-CH ₂ F、-CHF ₂ 、-CF ₃ Oxo, -CN, hydroxy, amino, C1-C6 alkylamino, carboxy, -NHAc, a group selected from the group consisting of: 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 substituents are selected from the group consisting of: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

Any of the above formulas is selected from the group consisting of: B. p, N, S and O.

2. The compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein R ₅ ' is a substituent having the formula IV:

wherein each L ₄ Independently selected from the group consisting of: substituted or unsubstituted C1-C4 alkylene, -NRa-; provided that each L ₄ The co-formed structure is chemically stable;

selected from the group consisting of: substituted or unsubstituted C5-C10 cycloalkyl, substituted or unsubstituted 3-8 membered nitrogen containing heterocyclyl;

each R is ₅ Each independently selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, -CN, hydroxy, amino, carboxy; wherein the substituents are selected from the group consisting of: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy.

3. The compound of claim 1, or an isomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein said compound isThe structure is as shown in the following formula:

wherein X is ₂ Is N.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound has a structure according to the formula:

Wherein Z is ¹ Selected from the group consisting of: NR (NR) _f 、N-O-R _f The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is as follows _f Selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₃ -C ₈ Cycloalkyl, -C (=o) -NRdRe, -C (=o) -substituted or unsubstituted C ₁ -C ₆ Alkoxy, -C (=o) -substituted or unsubstituted C ₁ -C ₆ Alkyl, -C (=o) -substituted or unsubstituted C ₃ -C ₁₀ Cycloalkyl;

the remaining groups are as defined above.

5. The compound of claim 1, or an optical isomer, a cis-trans isomer, or a pharmaceutically acceptable salt thereof, wherein said compound is selected from the group consisting of:

6. a process for the preparation of a compound of formula I according to claim 1, comprising the steps of:

(a) Intermediate 1 and intermediate 2 are used as raw materials, and a target product I is obtained through a Suzuki coupling reaction catalyzed by a proper palladium catalyst.

7. A pharmaceutical composition comprising (1) a compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof; (2) a pharmaceutically acceptable carrier.

8. Use of a compound according to claim 1 or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 7, for the preparation of a pharmaceutical composition for the prevention and/or treatment of a disease associated with the activity or expression of PD-1/PD-L1.

9. A PD-1/PD-L1 inhibitor, wherein the inhibitor comprises a compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof.

10. A method of inhibiting PD-1/PD-L1 interactions in vitro comprising the steps of: contacting a compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, with a PD-L1 protein.