CN111039942B

CN111039942B - Nitrogen-containing heterocyclic compound, and preparation method, pharmaceutical composition and application thereof

Info

Publication number: CN111039942B
Application number: CN201811191323.8A
Authority: CN
Inventors: 王喆; 张基勇; 曾志宏
Original assignee: Shanghai Longwood Biopharmaceuticals Co Ltd
Current assignee: Shanghai Longwood Biopharmaceuticals Co Ltd
Priority date: 2018-10-12
Filing date: 2018-10-12
Publication date: 2023-04-14
Anticipated expiration: 2038-10-12
Also published as: CN111039942A

Abstract

The invention provides a nitrogen heterocyclic compound, a preparation method, a pharmaceutical composition and application thereof,specifically, the invention provides a compound shown as the following formula I, or an optical isomer, a hydrate, a solvate or a pharmaceutically acceptable salt thereof; wherein, the definition of each group is described in the specification. The compounds of formula I are useful for treating diseases associated with the PD-1/PD-L1 signaling pathway.

Description

Nitrogen-containing heterocyclic compound, and preparation method, pharmaceutical composition and application thereof

Technical Field

The invention relates to the field of small molecule medicines, and particularly provides a small molecule compound which can be used for treating diseases related to a PD-1/PD-L1 signal pathway.

Background

The immune system plays a crucial role in controlling and curing many diseases, such as various cancers, diseases caused by viruses, and the like. Cancer cells often multiply rapidly by evading or suppressing the immune system in some way. One such way is to alter the expression of activating and inhibitory molecules on immune cells. Blocking the suppressive immune checkpoint, like PD-1, has proven to be a very effective method of suppressing cancer cells.

PD-1 is programmed cell death protein-1, also known as CD279. It is expressed mainly in activated T cells and B cells and functions to inhibit the activation of cells, which is a normal homeostatic mechanism of the immune system, and PD-1 is a protective wall of our human body because excessive T/B cell activation causes autoimmune diseases. PD-1 is a type I transmembrane glycoprotein composed of 268 amino acids, and its structure mainly includes 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 inhibitory motif and immunoreceptor tyrosine switch motif, which also demonstrates that PD-1 is capable of back-regulating T cell receptor-mediated signaling. PD-1 has two ligands, PD-L1 and PD-L2, which differ in their expression pattern. PD-L1 is up-regulated in various tumor cells, and is combined with PD-1 on T cells to inhibit the proliferation and activation of the T cells, so that the T cells are in an inactivated state, and finally immune escape is induced.

PD-1/PD-L1 exerts a reverse immunomodulatory effect. When PD-1 is combined with PD-L1, tyrosine of an immune receptor tyrosine conversion motif structure domain of the T cell can be phosphorylated, and the phosphorylated tyrosine can be combined with phosphatase protein tyrosinase 2 and protein tyrosinase 1. This can block activation of extracellular signal-regulated kinases and also block activation of phosphoinositide 3-kinase (PI 3K) and serine-threonine protein kinase (Akt), thereby inhibiting T-lymphocyte proliferation and secretion of relevant cytokines. PD-1/PD-L1 signal can inhibit T cell activation and proliferation, and can make cytokine interleukin 2, interferon gamma and IL-10 secrete. In addition, PD-1/PD-L1 signal has similar immune function to B cell too, after PD-1 binds with B cell antigen receptor, PD-1 cytoplasmic area reacts with tyrosinase containing protein tyrosinase 2 binding site, thus hinder the activation of B cell.

PD-1/PD-L1 based immunotherapy is a new generation of immunotherapy of great interest. In recent years, a series of surprising results of research have demonstrated that PD-1/PD-L1 inhibitors have potent antitumor activity against a variety of tumors. PD-1/PD-L1 antibody inhibitors that are currently on the market are Ninolumab from BMS, lambolizumab from Merck, and Atezolizumab from Roche. In addition to this, there are many PD-1/PD-L1 antibody inhibitors in the research including Cure Tech's Pidilizumab, GSK's AMP-224, and AstraZeneca's MEDI-4736.

Although tumor immunotherapy is considered as a new generation of revolution for targeted post-treatment cancer therapy. However, the currently marketed and researched PD-1 single-resistant drugs have their own defects, including injection administration, oral administration failure, instability in vivo, easy protease decomposition, easy immune cross reaction, difficult purification, high production cost and the like. Therefore, small molecule inhibitors of the PD-1/PD-L1 interaction are a better choice for tumor immunotherapy.

In view of the foregoing, there is a pressing need in the art to develop novel small molecule inhibitors of the PD-1/PD-L1 interaction.

Disclosure of Invention

The invention aims to provide a novel small-molecule inhibitor of PD-1/PD-L1 interaction.

In a first aspect of the present invention, there is provided a compound represented by the following formula I, or an optical isomer, hydrate, solvate, or a pharmaceutically acceptable salt thereof:

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

L ₁ 、L ₂ and L ₃ Each independently selected from the group consisting of: a bond, substituted or unsubstituted C1-C4 alkylene, substituted or unsubstituted C2-C4 alkenylene substituted or unsubstituted C2-C4 alkynylene, -S-, -O-, substituted or unsubstituted-NH-, -S (O) ₂ -, substituted or unsubstituted-NHC (O) NH-, or,

Substituted or unsubstituted>

Substituted or unsubstituted>

Substituted or unsubstituted->

Or substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

Z ¹ selected from the group consisting of: o, S, NRf, N-O-Rf; wherein, said Rf is selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₆ -C ₁₀ Aryl, substituted or unsubstituted C ₆ -C ₁₀ Heteroaryl, 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;

Z ² 、Z ³ 、Z ⁴ each independently selected from the group consisting of: NH, N, CH ₂ N-O, SO or SO ₂ ；

Y ¹ 、Y ² 、Y ³ Each independently selected from the group consisting of: CH. CH (CH) ₂ NH, N-O, CH, O, S, SO or SO ₂ ；

Is a single bond or a double bond; />

And is provided with

Is an aromatic or non-aromatic moiety;

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 -, in which,

each L _1a Each independently is a group selected from the group consisting of; chemical bond, substituted or unsubstituted C ₁ -C ₇ Alkylene, substituted or unsubstituted C2-C4 alkenylene, substituted or unsubstituted C2-C4 alkynylene-S-, -O-, substituted or unsubstituted-NH-, -S (O) ₂ -、

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

L _3a selected from the group consisting of: substituted or unsubstituted C1-C10 alkyl, C1-C10 aryl, -CN, hydroxy, amino, carboxy, -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, 1,2 respectively;

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;

selected from the group consisting of: 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 said heterocyclyl has 1-3 heteroatoms; or said->

Is none;

or the above-mentioned

Is equal to +>

Each independently selected from the group consisting of: substituted or unsubstituted C6-C10 arylene, or substituted or unsubstituted 5-12 membered (preferably 5-7 membered) heteroarylene having 1 to 3 heteroatoms, substituted or unsubstituted 5-12 membered heterocyclylene, substituted or unsubstituted 5-12 membered C5-C12 cycloalkylene; or said->

Is none;

and said

Is not absent at the same time;

R ₁ 、R ₂ 、R ₃ and R ₄ 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, hydroxy, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted- (C1-C6 alkylene) -NH- (C1-C6 alkylene), carboxy, 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 substituted or unsubstitutedIs/are>

Or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s -；

Unless otherwise specified, "substituted" means substituted with one or more (e.g., 2,3, 4, etc.) substituents selected from the group consisting of: 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, hydroxy, amino, C1-C6 alkylamino, carboxy, -NHAc, a group selected from the group consisting of unsubstituted or substituted with one or more substituents 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 substituent is selected from the following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

in the formulae above, any of the heteroatoms is selected from the group consisting of: n, S and O.

In another preferred embodiment, the above

A bicyclic structure of 5-7 membered ring and 5-6 membered ring.

In another preferred embodiment, the

Is a bicyclic structure selected from the group consisting of: a 6-membered and 6-membered ring, a 6-membered and 5-membered ring, a 5-membered and 6-membered ring.

In another preferred embodiment, the

Is a structure shown in the following formula (II):

wherein:

X ¹ 、X ² 、X ³ 、X ⁴ 、X ⁵ 、X ⁶ each independently selected from the group consisting of: NH, N, CH ₂ Or CH = CH;

is a single bond or a double bond.

In another preferred embodiment, the above

Is equal to->

In another preferred embodiment, the

Is a ring-forming group selected from the group consisting of: benzene, pyridine, pyrimidine, pyridazine, tetrazine, triazine, pyrrole, thiophene, furan, tetrazole, triazole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, oxadiazole, thiadiazole, naphthalene, indole, indazole, quinoline, isoquinoline, morpholine, dihydropiperidine, thiomorpholine, piperidine, piperazine, tetrahydropyran, dihydropyran, pyrroline, tetrahydrothiophene, tetrahydrofuran, oxetane, thietane, azetidine, benzofuran, benzothiophene, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, benzisoxazole, benzotriazole.

In another preferred embodiment, the above

Is a ring-forming group selected from the group consisting of:

In another preferred embodiment, the above

Each independently selected from the group consisting of: substituted or unsubstituted phenylene, substituted or unsubstituted 5-7 membered heteroarylene having 1-3 heteroatoms, substituted or unsubstituted 5-7 membered heterocyclylene, substituted or unsubstituted C5-C7 cycloalkylene.

In another preferred embodiment, the

Each independently a ring-forming group selected from the group consisting of: benzene, pyridine, pyrimidine, pyridazine, tetrazine, triazine, pyrrole, thiophene, furan, tetrazole, triazole, imidazole, thiazole, oxazole, pyrazole, isothiazole, isoxazole, oxadiazole, thiadiazole, naphthalene, indole, indazole, quinoline, isoquinoline, morpholine, dihydropiperidine, thiomorpholine, piperidine, piperazine, tetrahydropyran, dihydropyran, pyrroline, tetrahydrothiophene, tetrahydrofuran, oxetane, thietane, azetidine, benzofuran, benzothiophene, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, benzisoxazole, benzotriazole.

In another preferred embodiment, the above

Each independently a ring-forming group selected from the group consisting of: />

In another preferred embodiment, the

Each independently is a structure represented by the following formula (III):

wherein:

Z ^1a 、Z ^2a 、Z ^3a 、Z ^4a each independently selected from the group consisting of: n and CH.

In another preferred embodiment, the structure of formula II or formula III may be substituted or unsubstituted.

In another preferred embodiment, the structure of formula III is

In another preferred embodiment, R is ₂ Selected from the group consisting of: C1-C6 alkyl, CN.

In another preferred embodiment, the ring is

Having a substituent represented by formula IV below:

wherein, L is ₄ Selected from the group consisting of: substituted or unsubstituted C1-C4 alkylene, -S-, -O-, -NRa-, -S (O) ₂ -; preferably a substituted or unsubstituted C1-C4 alkylene group;

selected from the group consisting of: a substituted or unsubstituted 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; preferably, said->

Is a 5-7 membered nitrogen containing heterocyclyl;

R ₅ 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.

In another preferred embodiment, R is ₄ Selected from the group consisting of: -O (substituted or unsubstituted C1-C6 alkyl), -O (CH) ₂ ) _n - (substituted or unsubstituted 5-to 7-membered heteroaryl having 1 to 3 heteroatoms selected from N, S and O).

In another preferred embodiment, the compound of formula I is selected from a compound of formula Ia, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof:

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

L ₁ selected from the group consisting of: a bond, substituted or unsubstituted C1-C4 alkylene, substituted or unsubstituted C2-C4 alkenylene, substituted or unsubstituted C2-C4 alkynylene, -S-, -O-, substituted or unsubstituted-NH-, -S (O) ₂ -, substituted or unsubstituted-NHC (O) NH-, (II),

Substituted or unsubstituted->

Substituted or unsubstituted>

Substituted or unsubstituted>

Or substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

Z ² 、Z ³ 、Z ⁴ each independently selected from the group consisting of: NH, N, CH ₂ N-O, SO or SO2;

Y ¹ 、Y ² 、Y ³ each independently selected from the group consisting of: CH. CH (CH) ₂ NH, N-O, CH, O, S, SO, or SO2;

is a single bondOr a double bond;

and is

Is an aromatic or non-aromatic moiety;

each L is _1a Each independently is a group selected from the group consisting of; chemical bond, substituted or unsubstituted C ₁ -C ₇ Alkylene, substituted or unsubstituted C2-C4 alkenylene, substituted or unsubstituted C2-C4 alkynylene-S-, -O-, substituted or unsubstituted-NH-, -S (O) ₂ -、

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

s is 0, 1,2 respectively;

rd, re are each independently selected from the group consisting of: H. substituted or notGeneration C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₃ -C ₁₀ Cycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ An aryl group;

Is absent; or said +>

Is equal to +>

R ₁ 、R ₂ 、R ₃ And R ₄ 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, hydroxy, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted- (C1-C6 alkylene) -NH- (C1-C6 alkylene), carboxy, 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 substituted or unsubstituted>

Or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s -；

Unless otherwise specified, "substituted" means substituted with one or more (e.g., 2,3, 4, etc.) substituents selected from the group consisting of: 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, hydroxy, amino, C1-C6 alkylamino, carboxy, -NHAc, a group selected from the group consisting of unsubstituted or substituted with one or more substituents 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 following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

Preferably, the compound is selected from compounds 1-125.

In another preferred embodiment, the compound of formula I is selected from compounds of formula Ib, or optical 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;

Substituted or unsubstituted->

Substituted or unsubstituted->

Substituted or unsubstituted->

Or substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

Z ² 、Z ³ 、Z ⁴ each independently selected from the group consisting of: NH (NH)、N、CH、CH ₂ N-O, SO or SO2;

is a single or double bond;

and is

Is an aromatic or non-aromatic moiety;

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

s is 0, 1,2 respectively;

Is none; or said +>

Is equal to->

R ₁ 、R ₂ 、R ₃ And R ₄ 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, hydroxy, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted- (C1-C6 alkylene) -NH- (C1-C6 alkylene), carboxy, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-to 12-membered heteroaryl having 1 to 3 heteroatoms, substituted or unsubstituted having 1 to 4 heteroatoms5-to 12-membered heterocyclic groups of the subgroups, substituted or unsubstituted

Substituted or unsubstituted->

Or substituted or unsubstituted>

Or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s -；

Preferably, the compound is selected from compounds 126-202.

In another preferred embodiment, the compound of formula I is selected from compounds of formula Ic, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof:

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

L ₁ selected from the group consisting of: a bond, substituted or unsubstituted C1-C4 alkylene, substituted or unsubstituted C2-C4 alkenylene substituted or unsubstituted C2-C4 alkynylene, -S-, -O-, substituted or unsubstituted-NH-, -S (O) ₂ -, substituted or unsubstituted-NHC (O) NH-, (II),

Substituted or unsubstituted->

Substituted or unsubstituted->

Substituted or unsubstituted->

Or substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

Z ¹ selected from the group consisting of: o, S, NRf, N-O-Rf; wherein, said Rf is selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₆ -C ₁₀ Aryl, substituted or unsubstituted C ₆ -C ₁₀ Heteroaryl, 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 An alkynyl group;

is a single bond or a double bond;

and is

Is an aromatic or non-aromatic moiety;

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

s is 0, 1,2 respectively;

selected from the group consisting of: 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 said heterocyclyl has 1-3 heteroatoms; or said>

Is absent; or said +>

Is equal to->

R ₁ 、R ₂ 、R ₃ And R ₄ Each independently selected from the group consisting of: H. halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstitutedC2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, oxo (i.e = O), = NRf, -CN, hydroxy, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted- (C1-C6 alkylene) -NH- (C1-C6 alkylene), carboxy, 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 substituted or unsubstituted>

Or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s -；

Preferably, the compound is selected from the group consisting of compounds 203-313.

In another preferred embodiment, the compound of formula I is selected from the group consisting of formulae Id-1 and Id-2, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof:

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

L ₁ and L ₃ Each independently selected from the group consisting of: a bond, substituted or unsubstituted C1-C4 alkylene, substituted or unsubstituted C2-C4 alkenylene substituted or unsubstituted C2-C4 alkynylene, -S-, -O-, substituted or unsubstituted-NH-, -S (O) ₂ -, substituted or unsubstituted-NHC (O) NH-, or,

Substituted or unsubstituted->

Substituted or unsubstituted->

Substituted or unsubstituted->

Or substituted or unsubstituted>

Rf is selected from the group consisting of: H. substituted or unsubstituted C ₁ -C ₆ Alkyl, substituted or unsubstituted C ₆ -C ₁₀ Aryl, substituted or unsubstituted C ₆ -C ₁₀ Heteroaryl, 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) -substitutionOr unsubstituted C ₂ -C ₆ An alkynyl group;

Z ² and Z ³ Each independently selected from the group consisting of: NH, N, CH ₂ N-O, SO or SO2;

Y ¹ and Y ² Each independently selected from the group consisting of: CH. CH (CH) ₂ NH, N-O, CH, O, S, SO, or SO2;

is a single bond or a double bond;

and is

Is an aromatic or non-aromatic moiety;

L _3a selected from the group consisting of: substituted orUnsubstituted C1-C10 alkyl, C1-C10 aryl, -CN, hydroxy, amino, carboxy, -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, 1,2 respectively;

Is absent; or said +>

Is equal to->

R ₁ 、R ₂ And R ₃ 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, hydroxy, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted- (C1-C6 alkylene) -NH- (C1-C6 alkylene), carboxy, substituted or unsubstituted C6-C610 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 substituted or unsubstituted>

Or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s -；

Preferably, the compound is selected from the group consisting of compounds 314-355.

In another preferred embodiment, the compound of formula I is selected from a compound of formula Ie, or an optical isomer, hydrate, solvate, or pharmaceutically acceptable salt thereof:

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

Substituted or unsubstituted>

Substituted or unsubstituted->

Substituted or unsubstituted>

Or substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

is a single bond or a double bond; />

And is

Is an aromatic or non-aromatic moiety;

each L _1a Each independently is a group selected from the group consisting of; chemical bond, substituted or unsubstituted C ₁ -C ₇ Alkylene, substituted or unsubstituted C2-C4 alkenylene, substituted or unsubstituted C2-C4 alkynylene, -S-, -O-, substituted or unsubstituted-NH-、-S(O)-、-S(O) ₂ -、

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

s is 0, 1,2 respectively;

Is none; or said->

Is equal to +>

R ₁ 、R ₂ 、R ₃ And R ₄ Each independentlySelected 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, hydroxy, NRdRe (e.g., amino), substituted or unsubstituted C1-C6 amine, substituted or unsubstituted- (C1-C6 alkylene) -NH- (C1-C6 alkylene), carboxy, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-to 12-membered heteroaryl having 1 to 3 heteroatoms

Substituted or unsubstituted 5-12 membered heterocyclic group having 1-4 hetero atoms, substituted or unsubstituted

Substituted or unsubstituted>

Or substituted or unsubstituted>

Or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s -；

Preferably, the compound is selected from compounds 356-360.

In another preferred embodiment, the pharmaceutically acceptable salts include salts formed in combination with inorganic acids, organic acids, alkali metal ions, alkaline earth metal ions, or organic bases capable of providing physiologically acceptable cations, as well as 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, lycic acid, maleic tartaric acid, fumaric acid, citric acid or lactic acid; the alkali metal ions are selected from lithium ions, sodium ions and potassium ions; the alkaline earth metal ions are selected from calcium ions and magnesium ions; the organic base capable of providing a physiologically acceptable cation is selected from methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris (2-hydroxyethyl) amine.

In a second aspect of the invention, there is provided a process for the preparation of a compound of formula I according to the first aspect of the invention, said process comprising a step selected from those shown in schemes 1,2 or 3:

synthesis scheme 1

(e) Taking halide 1-1 and appropriate coupling reagent 1-2 (such as boric acid, boric acid ester, tin reagent or Grignard reagent) as basic raw materials, and obtaining an intermediate compound 1-3 through coupling reaction (such as Suzuki, stille or Kumada coupling) catalyzed by palladium or copper;

(f) Taking the intermediate 1-3 as a raw material, and reacting with carboxylic acid 1-4 under the action of a condensing agent (such as HATU, EDCI or HBTU) to obtain an amide intermediate 1-5;

(g) Taking the intermediate 1-5 as a raw material, and removing a protecting group (Boc) under an acidic condition to obtain an intermediate 1-6;

(h) Taking the intermediate 1-6 as a raw material, and carrying out nucleophilic substitution reaction with a halide under an alkaline condition, or carrying out reductive amination reaction with aldehyde or ketone under the action of a reducing agent to obtain a target compound I;

synthesis scheme 2:

(e) Taking carboxylic ester 2-1 as a raw material, and carrying out an aminolysis reaction with amine 2-2 under the catalysis of Lewis acid to obtain an intermediate compound 2-3;

(f) Taking the intermediate 2-3 and a proper coupling reagent 2-4 (such as boric acid, boric acid ester, a tin reagent or a Grignard reagent) as basic raw materials, and obtaining an intermediate compound 2-5 through coupling reaction (such as Suzuki, stille or Kumada coupling) catalyzed by palladium or copper;

(g) Taking the intermediate 2-5 as a raw material, and removing a protecting group (Boc) under an acidic condition to obtain an intermediate 2-6;

(h) Taking the intermediate 2-6 as a raw material, and carrying out nucleophilic substitution reaction with a halide under an alkaline condition, or carrying out reductive amination reaction with aldehyde or ketone under the action of a reducing agent to obtain a target compound I;

synthesis scheme 3:

(e) Using carboxylic ester 3-1 as a raw material, and carrying out Buchwald-Hartwig coupling or Ullmann coupling with amine 3-2 under the catalysis of palladium or copper to obtain an intermediate compound 3-3;

(f) Taking the intermediate 3-3 and a proper coupling reagent 3-4 (such as boric acid, boric acid ester, a tin reagent or a Grignard reagent) as basic raw materials, and obtaining an intermediate compound 3-5 through a coupling reaction (such as Suzuki, stille or Kumada coupling) catalyzed by palladium or copper;

(g) Taking the intermediate 3-5 as a raw material, and removing a protecting group (Boc) under an acidic condition to obtain an intermediate 3-6;

(h) Taking the intermediate 3-6 as a raw material, and carrying out nucleophilic substitution reaction with a halide under an alkaline condition, or carrying out reductive amination reaction with aldehyde or ketone under the action of a reducing agent to obtain a target compound I;

said Cy is

Is->

Y ¹ 、Y ² 、Z ¹ 、Z ² 、Z ³ And R is as defined above.

In a third aspect of the invention, there is provided a pharmaceutical composition comprising (1) 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; (2) a pharmaceutically acceptable carrier.

In a fourth aspect of the present invention, there is provided a use of the compound according to the first aspect of the present invention or a stereoisomer or a tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or the pharmaceutical composition according to the second aspect of the present invention, for the preparation of a pharmaceutical composition for the prevention and/or treatment of a disease associated with an activity or an expression amount of PD-1/PD-L1.

In a fifth aspect of the invention, there is provided 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.

In another preferred embodiment, the pharmaceutical composition is used for treating a disease selected from the group consisting of: cancer, infectious disease, autoimmune disease.

In another preferred embodiment, the cancer is selected from skin cancer, lung cancer, urinary tumor, hematological tumor, breast cancer, glioma, digestive tumor, reproductive tumor, lymphoma, nervous system tumor, brain tumor, head and neck cancer.

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, systemic autoimmune disease.

In another preferred embodiment, the organ-specific autoimmune disease comprises 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, acute idiopathic polyneuritis.

In another preferred embodiment, the systemic autoimmune disease comprises rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia.

In a sixth aspect of the invention, there is provided a method of inhibiting the PD-1/PD-L1 interaction in vitro, comprising the steps of: 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 a PD-L1 protein.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Detailed Description

The inventors of the present invention have conducted extensive and intensive studies and have found a class of PD-1/PD-L1 interaction inhibitors having an excellent inhibitory effect. On the basis of this, the inventors have completed the present invention.

Definition of

As used herein, the term "alkyl" includes straight or branched chain alkyl groups. Such as C ₁ -C ₈ Alkyl represents a straight or branched chain alkyl group having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, etc.

As used herein, the term "alkenyl" includes straight or branched chain alkenyl groups. Such as C ₂ -C ₆ Alkenyl means straight-chain or branched alkenyl having 2 to 6 carbon atoms, e.g. vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl2-butenyl, or the like.

As used herein, the term "alkynyl" includes straight or branched chain alkynyl groups. E.g. C ₂ -C ₆ Alkynyl means straight or branched chain alkynyl 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 cycloalkyl groups having 3 to 10 carbon atoms. It may be a single ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may also be in the form of a double ring, such as a bridged or spiro ring.

As used herein, the term "C ₁ -C ₈ Alkylamino "refers to a substituted amino group ₁ -C ₈ The amino group substituted by the alkyl can be mono-substituted or di-substituted; 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 chain 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 following groups N, S and O" refers to a saturated or partially saturated cyclic group having 3-10 atoms and wherein 1-3 atoms are heteroatoms selected from the following groups N, S and O. It may be monocyclic or may be in the form of a double ring, for example in the form of a bridged or spiro ring. Specific examples may be oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, pyrrolidinyl, and the like.

As used herein, the term "C ₆ -C ₁₀ Aryl "means 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 group having 1-3 heteroatoms selected from the following groups of N, S and O" refers to a cyclic aromatic group having 5-10 atoms, wherein 1-3 atoms are heteroatoms selected from the following groups of N, S and O. It may be a single ring or a condensed ring form. Specific examples thereof may be pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1, 2, 3) -triazolyl and (1, 2, 4) -triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl and the like.

Unless specifically stated to be "substituted or unsubstituted", the groups of the present invention may be substituted with a substituent selected from the group consisting of: halogen, nitrile group, nitro group, hydroxyl group, amino group, C ₁ -C ₆ Alkyl-amino, C ₁ -C ₆ Alkyl radical, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ Alkynyl, C ₁ -C ₆ Alkoxy, halo C ₁ -C ₆ Alkyl, halo C ₂ -C ₆ Alkenyl, halo C ₂ -C ₆ Alkynyl, halo C ₁ -C ₆ Alkoxy, allyl, benzyl, C ₆ -C ₁₂ Aryl radical, C ₁ -C ₆ alkoxy-C ₁ -C ₆ Alkyl radical, 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 Alkyl-sulfonyl, and the like.

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 formulae depicted herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, the asymmetric center-containing R and S configuration, and the double bond (Z) and (E) isomers. Thus, individual stereochemical isomers of the compounds of the present invention or mixtures of enantiomers, diastereomers or geometric isomers (or conformers) thereof are within the scope of the present invention.

As used herein, the term "tautomer" means that structural isomers having different energies may exceed the low energy barrier, thereby converting each other. For example, proton tautomers (i.e., proton transmutations) include interconversion by proton shift, such as 1H-indazoles and 2H-indazoles. Valence tautomers include interconversion by some recombination of bonding electrons.

As used herein, the term "solvate" refers to a complex of a compound of the present invention coordinated to solvent molecules in a specific ratio.

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

Active ingredient

As used herein, "compounds of the 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 classes of R configuration and/or S configuration stereoisomers of each compound, and/or E-/Z-cis-trans isomers).

All such salts within the scope of the present invention may be prepared by conventional methods. During the preparation of the compounds of formula I and solvates and salts thereof, different crystallization conditions may occur, either as polycrystals or as a co-crystal.

Preparation of Compounds of formula I

For the preparation of the compounds of the general formula I according to the invention, the preparation of the compounds of the general formula I according to the invention can be obtained by the following synthetic route, depending on the structure of the general formula I.

(a) Taking halide 1-1 and appropriate coupling reagent 1-2 (such as boric acid, boric acid ester, tin reagent or Grignard reagent) as basic raw materials, and obtaining an intermediate compound 1-3 through coupling reaction (such as Suzuki, stille or Kumada coupling) catalyzed by palladium or copper;

(b) Taking the intermediate 1-3 as a raw material, and reacting with carboxylic acid 1-4 under the action of a condensing agent (such as HATU, EDCI or HBTU) to obtain an amide intermediate 1-5;

(c) Taking the intermediate 1-5 as a raw material, and removing a protecting group (Boc) under an acidic condition to obtain an intermediate 1-6;

(d) Taking the intermediate 1-6 as a raw material, and carrying out nucleophilic substitution reaction with a halide under an alkaline condition, or carrying out reductive amination reaction with aldehyde or ketone under the action of a reducing agent to obtain a target compound I;

the second method comprises the following steps:

(a) Taking carboxylic ester 2-1 as a raw material, and carrying out an aminolysis reaction with amine 2-2 under the catalysis of Lewis acid to obtain an intermediate compound 2-3;

(b) Taking the intermediate 2-3 and a proper coupling reagent 2-4 (such as boric acid, boric acid ester, a tin reagent or a Grignard reagent) as basic raw materials, and obtaining an intermediate compound 2-5 through a coupling reaction (such as Suzuki, stille or Kumada coupling) catalyzed by palladium or copper;

(c) Taking the intermediate 2-5 as a raw material, and removing a protecting group (Boc) under an acidic condition to obtain an intermediate 2-6;

(d) Taking the intermediate 2-6 as a raw material, and carrying out nucleophilic substitution reaction with a halide under an alkaline condition, or carrying out reductive amination reaction with aldehyde or ketone under the action of a reducing agent to obtain a target compound I;

the method 3 comprises the following steps:

(a) Using carboxylic ester 3-1 as a raw material, and carrying out Buchwald-Hartwig coupling or Ullman coupling with amine 3-2 under the catalysis of palladium or copper to obtain an intermediate compound 3-3;

(b) Taking the intermediate 3-3 and a proper coupling reagent 3-4 (such as boric acid, boric acid ester, a tin reagent or a Grignard reagent) as basic raw materials, and obtaining an intermediate compound 3-5 through coupling reaction (such as Suzuki, stille or Kumada coupling) catalyzed by palladium or copper;

(c) Taking the intermediate 3-5 as a raw material, and removing a protecting group (Boc) under an acidic condition to obtain an intermediate 3-6;

(d) Taking the intermediate 3-6 as a raw material, and carrying out nucleophilic substitution reaction with a halide under an alkaline condition, or carrying out reductive amination reaction with aldehyde or ketone under the action of a reducing agent to obtain a target compound I;

said Cy is

Is->

Y ¹ 、Y ² 、Z ¹ 、Z ² 、Z ³ And R is as defined above.

In addition, the starting materials and intermediates in the above reactions are readily available, and the reactions in each step can be readily synthesized according to reported literature or by conventional methods in organic synthesis to those skilled in the art. The compounds of formula I may exist in the form of solvates or non-solvates and crystallization using different solvents may result in different solvates.

Pharmaceutical compositions and methods of administration

Since the compound of the present invention has excellent inhibitory activity of PD-1/PD-L1 interaction, the compound of the present invention and various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates thereof, and pharmaceutical compositions containing the compound of the present invention as a main active ingredient can be used for the prevention and/or treatment (stabilization, alleviation or cure) of diseases associated with PD-1/PD-L1 interaction (e.g., cancer, infectious diseases, autoimmune diseases).

The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention in combination with 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. Typically, the pharmaceutical composition contains 1-2000mg of a compound of the invention per dose, more preferably, 10-200mg of a compound of the invention per dose. Preferably, said "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 be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and between the compounds of the present invention without significantly diminishing the pharmaceutical effectiveness of the compounds. Examples of pharmaceutically acceptable carrier moieties are 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, glycerol, mannitol, sorbitol, etc.), emulsifiers (e.g. propylene glycol, glycerol, mannitol, sorbitol, etc.)

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular or subcutaneous).

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, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) Binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) Disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) Wetting agents, such as cetyl alcohol and glycerol 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, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such a composition may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in 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 compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, and oils, in particular, cottonseed, groundnut, corn germ, olive, castor, and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions can also contain 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 as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise 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 nonaqueous carriers, diluents, solvents or vehicles 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 (e.g., other anti-cancer agents).

When administered in combination, the pharmaceutical composition further comprises 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 may be used simultaneously, separately or sequentially with the compounds of the invention for the prevention and/or treatment of a disease associated with the PD-1/PD-L1 interaction.

When using pharmaceutical compositions, a safe and effective amount of a compound of the present invention is administered to a mammal (e.g., a human) in need of treatment at a dosage that is pharmaceutically considered to be effective, typically 1 to 2000mg, preferably 20 to 500mg per day for a human of 60kg body weight. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The main advantages of the present invention include:

(1) The compound has high inhibitory activity on PD-1/PD-L1 interaction, has strong binding capacity with PD-L1 protein, and has the capacity of relieving the inhibition of IFN gamma by PD-L1.

(2) The compound of the invention has better solubility; has very low toxicity to normal cells, and thus can be applied to a subject in a wide dose range.

(3) Compared with the prior art, the compound of the invention has better solubility and good drug forming property, and the compound of the invention has good bioavailability in vivo experiments, and in addition, compared with the prior art, the compound of the invention can be easily prepared into pharmaceutically acceptable salts, thereby being beneficial to further preparation.

(4) The in vivo efficacy research shows that the compound can obviously inhibit the growth of subcutaneous tumors no matter on the tumor volume or the tumor weight, and can obviously increase the number of lymphocytes in the blood and the spleen of a mouse.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.

The test materials and reagents used in the following examples are commercially available without specific reference.

General materials and test methods:

the instruments and materials involved in the examples are described below:

the NMR spectrum was obtained by analysis with a Bruker AV-400 (400 MHz) NMR spectrometer.

Chemical shifts are reported in ppm units (CDC 1) using tetramethylsilane as an internal standard ₃ Delta 7.26 ppm). The data reported are the chemical shifts and their split and coupling constants (s: singlet; d: doublet; t: triplet; q: quartet; br: broad)(ii) a m: multiple peaks).

Mass spectrometry data were analyzed using a liquid chromatograph-mass spectrometer from the Finnigan advanced LCQ company (Finnigan LCQ Advantage), all reactions operated under anhydrous and anaerobic conditions under dry argon protection, among other requirements. The solid organometallic compound was stored in an argon protected dry box.

The tetrahydrofuran and the diethyl ether are obtained by distillation, and metal sodium and benzophenone are added into the tetrahydrofuran and the diethyl ether during the distillation. Methylene chloride, pentane and hexane were treated with calcium hydride.

The special raw materials and intermediates involved in the invention are provided by custom-made processing of Tianjin Changsen pharmaceutical industry Co., ltd, and all other chemical reagents are purchased from reagent suppliers such as Shanghai chemical reagent company, aldrich company (Aldrich), acros company (Acros), and the like. If the intermediates or products needed in the reaction during the synthesis process are not enough for the next experiment, the synthesis is repeated for a plurality of times until the intermediates or products are enough.

The raw materials and reagents related to the invention can be obtained by commercial or customized processing and purchase except for special instructions.

The compounds of the invention may contain one or more asymmetric centres and so the series of compounds may be in racemic or single enantiomeric form. The compound prepared by the invention is a heterocyclic compound with the purity higher than 95 percent, and the structural representation of each final product is respectively represented by MS or/and hydrogen spectrum nuclear magnetic resonance ( ¹ H NMR) analysis. The synthesis of the various compounds and intermediates of the invention is illustrated by the examples below.

Example 1: synthesis of (R) -3' - ((7- ((3-hydroxypyrrol-1-yl) methyl) quinolin-4-yl) oxy) -2,2' -dimethyl- [1,1' -diphenyl ] -3-carboxylic acid

Step 1:

diethyl ethoxymethylenemalonate (150mL, 749mmol) and m-toluidine (80mL, 738mmol) were placed in a flask, and the mixture was heated at 110 ℃ for 1 hour. The reaction mixture was cooled to room temperature. The flask was placed in a refrigerator and the precipitated solid was collected and washed with hexane to give the product as a white solid (120 g, 58% yield).

¹ H NMR(400MHz,CDCl ₃ )δ10.99(d,J＝13.8Hz,1H),8.54(d,J＝13.8Hz,1H),7.30–7.25(m,1H),7.02–6.92(m,3H),4.37–4.22(m,4H),2.38(s,3H),1.37(dt,J＝19.5,7.1Hz,6H).ESI m/z 278.2(M+H).

Step 2:

diethyl 2- ((m-tolylamino) methylene) malonate (148g, 534mmol) was suspended in diphenyl ether (200 mL) and the mixture was refluxed for 45 minutes. The solution was allowed to cool to room temperature and the solid formed was collected and bubbled with hexane to afford the product as a white solid (26.7 g, 22% yield).

¹ H NMR(400MHz,DMSO-d ₆ )δ12.19(s,1H),8.49(d,J＝6.5Hz,1H),8.04(d,J＝8.2Hz,1H),7.38(s,1H),7.24(dd,J＝8.3,1.5Hz,1H),4.21(q,J＝7.1Hz,2H),2.80(s,1H),2.44(s,3H),1.28(t,J＝7.1Hz,3H).ESI m/z 232.0(M+H).

And step 3:

ethyl-4-hydroxy-7-methylquinoline-3-carboxylate (43g, 187mmol) was added to 5M sodium hydroxide (200 mL) and heated under reflux for 1 hour. The mixture was cooled to room temperature and acidified to pH 4 by addition of 2M HCl. The solid formed was collected, washed with water and dried to give 4-hydroxy-7-methylquinoline-3-carboxylic acid as a white solid (37.8 g,90% yield).

ESI m/z 204.0(M+H).

And 4, step 4:

4-hydroxy-7-methylquinoline-3-carboxylic acid (35g, 172mmol) was suspended in diphenyl ether (100 mL). The mixture was refluxed for 1.5 hours and then cooled to room temperature. The precipitated solid was collected and washed with hexane to give 7-methylquinolin-4-ol as a white solid (16.8g, 62% yield).

ESI m/z 160.0(M+H).

And 5:

to a solution of 7-methylquinolin-4-ol (26g, 162mmol) in chloroform (150 mL) at 0 deg.C was added phosphorus oxychloride (50 mL). The mixture was then refluxed for 3 hours, then cooled to room temperature and neutralized with 2M NaOH. The resulting solid was collected, washed with water and dried to give 4-chloro-7-methylquinoline as a white solid (17g, 59% yield).

¹ H NMR(400MHz,CDCl ₃ )δ8.76(d,J＝4.8Hz,1H),8.14(d,J＝8.5Hz,1H),7.97–7.91(m,1H),7.50(dd,J＝8.6,1.7Hz,1H),7.45(d,J＝4.8Hz,1H),2.61(d,J＝0.9Hz,3H).ESI m/z 178.0(M+H).

And 6:

to CCl of 4-chloro-7-methylquinoline (7g, 39mmol) ₄ NBS (8g, 45mmol) and benzoyl peroxide (0.8g, 3.3mmol) were added to the solution (60 mL). The solution was refluxed for 5 hours. After completion of the reaction, the solvent was evaporated, and the obtained residue was purified by flash chromatography to give the product as a white solid (2.48g, 25% yield).

¹ H NMR(400MHz,CDCl ₃ )δ8.82(d,J＝4.7Hz,1H),8.27(d,J＝8.7Hz,1H),8.15–8.11(m,1H),7.72(dd,J＝8.7,1.8Hz,1H),7.53(d,J＝4.7Hz,1H),4.71(s,2H).ESI m/z 258.0(M+H).

And 7:

(R) -pyrrolidin-3-ol (957mg, 11mmol) was added to 7- (bromomethyl) -4-chloroquinoline (2.56g, 10mmol) and Et 3N (2.16ml, 15mmol) in dichloromethane (20 ml). After stirring at room temperature for 4 hours, the solvent was removed and the resulting residue was purified by flash chromatography to give the product (2.35g, 90% yield). ESI M/z 263.0 (M + H).

Step 8

To a solution of 3' -hydroxy-2, 2' -dimethyl- [1,1' -biphenyl ] -3-carboxylic acid methyl ester (256mg, 1mmol) in DMF (3 mL) at 0 deg.C was added NaH (80mg, 2mmol). The mixture was stirred at this temperature for 30 minutes, then (R) -1- ((4-chloroquinolin-7-yl) methyl) pyrrolidin-3-ol (262mg, 1mmol) was added. The reaction was heated at 120 ℃ overnight and then cooled to room temperature. The mixture was purified by prep-HPLC to give the product (79mg, 17% yield).

ESI m/z 469.2(M+H).

Example 2:5- (((trans) -4- (methoxycarbonyl) cyclohexyl) methyl) -1-methyl-4, 5,6, 7-tetrahydro-1H-imidazo [4,5-c ] pyridine-2-carboxylic acid

The synthesis route is as follows:

step 1:

dissolving the compound 1 (474mg, 2mmol) in 10mL anhydrous THF under the protection of nitrogen, cooling to-78 deg.C, slowly adding 1mL 2.5M butyl lithium n-hexane solution dropwise, continuing to stir for 30min after the dropwise addition is finished, then adding benzyl chloroformate (511mg, 3mmol) dropwise, continuing to stir for 1h after the dropwise addition is finished, heating to room temperature, quenching with saturated sodium bicarbonate solution, post-treating and performing column chromatography to obtain the product.

And 2, step:

compound 2 (300 mg) was added to 10mL of 4N HCl/dioxane solution, stirred at room temperature for 1h, followed by disappearance of starting material by TLC, and concentrated under reduced pressure to give a solid which was used directly in the next reaction.

And step 3:

under the protection of argon, DMSO (3.12g, 40mmol) is added into DCM (50 ml), after stirring for 10min, the mixture is cooled to-78 ℃, oxalyl chloride (2.54g, 20mmol) is added dropwise, after stirring for 10min, a DCM solution of a compound 4 (3.44g, 20mmol) is added dropwise, after stirring for 10min, triethylamine (5.26g, 52mmol) is slowly added dropwise, after stirring for 10min, the mixture is moved to room temperature and is continuously stirred for about 30min, DCM and water are added for quenching, extraction, drying and concentration are carried out, and then oily liquid is obtained.

And 4, step 4:

under argon protection, hydrochloride (31mg, 0.1mmol) of compound 3, compound 5 (22.3mg, 0.13mmol) and potassium carbonate (13.9mg, 0.1mmol) were added to a solution of DCM (1.0 ml), placed in an ice bath, stirred for 30min and NaBH (OAc) was added portionwise ₃ (49.1mg, 0.23mmol) and then transferred to room temperature and stirred overnight. Conversion to product was detected by TLC and LCMS.

And 5:

dissolving compound 6 (100mg, 0.24mmol) in THF (5 ml), adding 10% Pd/C (100 mg), hydrogenating at normal pressure for 5h, filtering, concentrating the solvent to obtain a solid.

LCMS found 336[M+H] ⁺

Example 3: synthesis of 5- (2- ((tert-butyldiphenylsilyl) oxy) ethyl) -1-methyl-4-oxo-4, 5-dihydro-1H-pyrrolo [3,2-c ] pyridine-2-carboxylic acid

The synthetic route is as follows:

step 1:

adding raw material aldehyde 1 (25g, 1.0eq) into methanol (250 ml), protecting with argon, cooling (the internal temperature is-10 ℃ to-15 ℃) and adding azide raw material 2 (57g, 2.5eq), wherein the system is clear and does not heat up; dropwise adding 30% sodium methoxide methanol solution (81g, 2.5 eq) to control the temperature at minus 12 +/-2 ℃, and maintaining the temperature for 1h; standing overnight at 0 deg.C; the system was cloudy and the viscosity gradually increased. And (3) treatment: pouring the reaction solution into a mixed system of ice water and solid ammonium chloride, stirring for 30 minutes, filtering, rinsing the solid for 3 times by adding water, and pulling the solid to dry by an oil pump to obtain 58g of brown yellow solid (the yield is quantitative), and directly putting the brown yellow solid into the next step.

¹ H NMR(400MHz,CDCl ₃ )δ8.49(dd,J＝7.6,1.9Hz,1H),8.11(dd,J＝5.0,1.9Hz,1H),7.25(d,J＝11.9Hz,1H),6.93(dd,J＝7.7,4.9Hz,1H),3.99(s,3H),3.92(s,3H).

LCMS found 208[M+H-N ₂ ] ⁺

Step 2:

adding the raw material 3 (26 g) into o-dichlorobenzene (500 ml), and reacting for 1 hour at 145 ℃ by external bath; and (3) processing: cooling to-10 deg.C, stirring for 10min, filtering, rinsing the solid with n-heptane, and drying to obtain light yellow solid 7g (yield: 50%)

¹ H NMR(400MHz,DMSO-d ₆ )δ12.36(s,1H),7.83(d,J＝5.9Hz,1H),7.13(d,J＝2.0Hz,1H),7.02(d,J＝5.9Hz,1H),3.97(s,3H),3.87(s,3H).

LCMS:found 207[M+H] ⁺

And 3, step 3:

adding the raw material 4 (6.8g, 1.0eq) into DMAc (70 ml), dissolving, adding potassium carbonate (9.12g, 2.0eq), dropwise adding methyl iodide (7g, 1.5eq), and reacting at room temperature overnight; and (3) processing: water (50 ml), ammonia (20 ml), EA (100ml × 3) was added for extraction, and column: ethyl acetate/n-heptane =15/85 elution gave 6.05g of a yellow solid (yield: 83.4%).

¹ H NMR(400MHz,CDCl ₃ )δ7.94(d,J＝6.1Hz,1H),7.39(d,J＝0.9Hz,1H),7.26(s,1H),6.97–6.83(m,1H),4.11(s,3H),4.04(s,3H),3.91(s,3H).

LCMS:found 221[M+H] ⁺

And 4, step 4:

adding 5 (5g, 1.0eq) of raw material into acetic acid (82g, 30eq), adding 47% of hydrobromic acid aqueous solution (55g, 30eq), heating to 95 ℃ after dissolving, and reacting for 2 hours; and (3) treatment: concentration to give 9g (yield: quantitative) of pale yellow solid, which was directly fed to the next step.

LCMS:found 193[M+H] ⁺

And 5:

adding raw material 6 (4.36g, 1.0eq) into methanol (90 ml) for insolubilization, adding concentrated sulfuric acid (5g, 1.0eq), and carrying out reflux reaction at the temperature of 85 ℃ in an external bath overnight; and (3) processing: under ice bath, 70ml of saturated sodium bicarbonate is dropwise added, pH is kept at 7-8, solid is precipitated, the solid is filtered, rinsed by water and dried to obtain 4.43g of white solid (yield: 94.8%).

¹ H NMR(400MHz,DMSO-d ₆ )δ11.02(s,1H),7.24(s,1H),7.20(t,J＝6.5Hz,1H),6.58(d,J＝7.4Hz,1H),3.93(s,3H),3.81(s,3H).

LCMS found:207[M+H] ⁺

And 6:

adding 7 (4.2g, 1.0eq) raw material into DMAc (50 ml), protecting with argon, adding 13.3g,2.0eq cesium carbonate, adding 8 (9.62g, 1.3eq) silanol raw material, and reacting at room temperature overnight; and (3) treatment: adding water (100 ml), and 1M to obtain hydrochloric acid (80 ml) to adjust the pH to be 5-6; EA extraction 3 times, column pass: 20% EA/n-heptane elution, concentrated to give a white solid 4.5g, yield: 45.4 percent.

¹ H NMR(400MHz,DMSO-d ₆ )δ7.54(d,J＝7.5Hz,1H),7.45(d,J＝7.3Hz,4H),7.39(t,J＝7.5Hz,2H),7.32–7.23(m,5H),6.66(d,J＝7.5Hz,1H),4.13(t,J＝5.0Hz,2H),3.95(s,3H),3.86(t,J＝5.0Hz,2H),3.83(s,3H),0.93(s,9H).

LCMS found:489[M+H] ⁺ 。

Step 7

Taking raw material 9 (4.5g, 1.0eq) and adding THF (45 ml), adding water (45 ml) after dissolving, adding hydrated lithium hydroxide (0.97g, 2.5eq) and reacting for 2 hours; and (3) treatment: 1M hydrochloric acid (50 ml) and water (50 ml) were added and a white solid precipitated, which was stirred for 30 minutes, the solid was filtered and dried to give 4.05g of 5- (2- ((tert-butyldiphenylsilyl) oxy) ethyl) -1-methyl-4-oxo-4, 5-dihydro-1H-pyrrolo [3,2-c ] pyridine-2-carboxylic acid as a white solid, yield: 92.6 percent.

¹ H NMR(400MHz,DMSO-d ₆ )δ12.82(s,1H),7.52(d,J＝7.5Hz,1H),7.47–7.42(m,4H),7.43–7.36(m,2H),7.28(t,J＝7.4Hz,4H),7.21(s,1H),6.65(d,J＝7.5Hz,1H),4.13(t,J＝5.0Hz,2H),3.94(s,3H),3.86(t,J＝4.9Hz,2H),0.93(s,9H).

LCMS found:475[M+H] ⁺ 。

Example 4: synthesis of 5- (2- ((tert-butyldiphenylsilyl) oxy) ethyl) -4-oxo-4, 5-dihydrofuran [3,2-c ] pyridine-2-carboxylic acid

The synthesis route is as follows:

step 1:

sequentially adding compound 1 (8.7g, 56.7mmol, 1.0eq) to a reaction flask, 30% meona-MeOH solution (40 mL); heating to 75 ℃ in an external bath, and stirring; TLC monitoring reaction is completed; cooling, adding water, and extracting with ethyl acetate for 3 times; the organic phases were combined, washed 4 times with 15% saline and 1 time with saturated saline. Drying with anhydrous sodium sulfate; and (5) spin-drying. 7.6g of a yellow solid are obtained, yield 89.7%.

LCMS found:150[M+H] ⁺

And 2, step:

2 (7.6g, 51.0mmol, 1.0eq) is added into the reaction bottle in sequence, anhydrous THF is steamed, and the inert gas is used for protection; cooling the dry ice-acetone to-78 ℃; n-BuLi (30.5mL, 76.5mmol,1.5eq, 2.5M) was added dropwise; stirring for 30min at low temperature; introducing carbon dioxide dried by concentrated sulfuric acid and anhydrous calcium chloride into the reaction solution; reacting for 3 hours, and slowly heating to 0 ℃ below zero; adding saturated solution of ammonium chloride for quenching; adding water and ethyl acetate; separating out an organic phase; the aqueous phase was washed once with ethyl acetate; adjusting the pH value of the water phase to 2; a large amount of solid was precipitated, filtered and dried to obtain 9.4g of solid with a yield of 95.5%.

¹ H NMR(400MHz,DMSO-d ₆ )δ8.15(d,J＝6.0Hz,1H),7.60(s,1H),7.39(dd,J＝6.0,0.9Hz,1H),4.02(s,3H).

LCMS found:194[M+H] ⁺

And step 3:

compound 3 (9.0g, 46.6mmol, 1.0eq), ethanol (270 mL) and concentrated sulfuric acid (11.5 mL) were added to a reaction flask, and heated to reflux; stirring overnight, evaporating to remove ethanol, cooling the residue with ice water bath, and adding water dropwise to precipitate solid; filtration and drying gave 7.4g of solid, yield 76.7%.

¹ H NMR(400MHz,DMSO-d ₆ )δ11.68(s,1H),7.61(d,J＝2.3Hz,1H),7.50(d,J＝7.5Hz,1H),6.73(d,J＝7.4Hz,1H),4.33(q,J＝7.1Hz,2H),1.31(td,J＝7.1,2.9Hz,3H).

LCMS found:208[M+H] ⁺

And 4, step 4:

4 (7.0g, 33.8mmol, 1.0eq), 70mL of DMAc, cesium carbonate (22.0g, 67.5mmol, 2.0eq) and compound 5 (14.7g, 40.5mmol, 1.2eq) are added into a reaction bottle, stirred at room temperature for 2 hours, water and ethyl acetate are added, ethyl acetate is separated, the water phase is extracted once by ethyl acetate, the organic phases are combined, and the mixture is washed by saturated sodium chloride and dried by screwing to obtain a compound 6 which is directly fed into the next step without purification.

LCMS found 490[M+H] ⁺

And 5:

crude compound 6 from the previous step, ethanol (70 mL) and water (18 mL) were added to the reaction flask and LiOH solid (1.22g, 20.9mmol, 1.5eq) was added. Stirring at room temperature for 0.5h, concentrating ethanol, and adding water and ethyl acetate to the residue; separating out ethyl acetate; water phase ice water bath, adjusting the pH value to 5, and separating out a large amount of solid; filtration and drying gave 10.3g of solid 7 in 66.1% yield (two steps).

¹ H NMR(400MHz,DMSO-d ₆ )δ7.82(d,J＝7.5Hz,1H),7.54(d,J＝0.9Hz,1H),7.48–7.34(m,7H),7.29(t,J＝7.3Hz,4H),6.80(dd,J＝7.5,0.9Hz,1H),4.19(t,J＝4.9Hz,2H),3.88(t,J＝4.9Hz,2H),0.92(s,9H).

LCMS found:462[M+H] ⁺

Step 6: 2-amino-5- (2- ((tert-butyldiphenylsilyl) oxy) ethyl) furan [3,2-c ] pyrido-4 (5H) -one

Adding the compound 7 (462mg, 1mmol) into anhydrous THF (5 mL), then adding triethylamine (233mg, 2.3mmol) and azido diphenyl phosphate (316mg, 1.15mmol), stirring at room temperature for 2h, then heating to reflux reaction for 2h, then adding water (1.0 mL), continuing to reflux reaction overnight, cooling, adding EA for extraction, concentrating, and separating by column chromatography to obtain the product. LCMS found:462[ solution of [ M ] +H ]] ⁺

Example 5: synthesis of 5- (2- ((tert-butyldiphenylsilyl) oxy) ethyl) -7-cyano-1-methyl-4-oxo-4, 5-dihydro-1H-pyrrolo [3,2-c ] pyridine-2-carboxylic acid

The synthetic route is as follows:

step 1:

adding 25g of the compound 1 into 75ml of methanol, cooling to 0 ℃, dropwise adding 62.89g of thionyl chloride, and heating to room temperature for reacting for 2d after dropwise adding; and (3) sampling LCMS, completely reacting the raw materials, evaporating most of methanol, pouring ice water into the residue, extracting by EA, combining organic phases, washing with salt, drying, and spin-drying to obtain 22.7g of a crude product. The crude product is directly put into the next step.

And 2, step:

adding 10.7g of compound 2 into 54ml of methanol, adding 14.6g of sodium methoxide/methanol (30 percent by weight), and after the addition is finished, heating and refluxing overnight (the system turns turbid from clear); sample LCMS and starting material was essentially reacted to completion and worked up. The methanol was evaporated, the residual DCM diluted, washed with ice water, brine, dried and spin dried to give 8.9g of crude yellow solid. The crude product is directly put into the next step. Yield: 84.7 percent

Step 3

Cooling 180ml of anhydrous THF to below-10 ℃ under the protection of argon; adding 2.8g of LiAiH4 in batches; dissolving 8.9g of compound 3 in 50ml of THF, and dropwise adding at a temperature below-10 ℃; keeping the temperature for reaction for 30min after dripping, sampling LCMS, basically completely reacting the raw materials, controlling the temperature to be less than-10 ℃, stirring 3ml of water and anhydrous sodium sulfate into sand, and adding the sand in batches; after the addition, filtering, and washing a filter cake EA; the combined organic phases were salted, dried overnight, spun dry, and column-mixed to give 4.7g white solids (15% EA in HEP out product). Yield: 59.5 percent

Step 4

Dissolving 3.5g of compound 4 in 70ml of DCM, adding 10.2g of dess-Martin in batches, and reacting for 2h at room temperature after the addition is finished (the system is white and turbid); sample LCMS showed the starting material was essentially reacted to completion, poured into ice sodium bicarbonate + sodium thiosulfate (1) saturated aqueous solution, and aqueous phase was extracted 1 time with DCM; the combined organic phases were washed with saturated brine, dried, spun-dried and chromatographed to give 3.1g of a white solid. Yield: 90.6 percent

Step 5

Dissolving 4g of compound 5 in 40ml of methanol, and cooling to the temperature of minus 15 ℃ to minus 10 ℃ under the protection of argon; 4.8g of ethyl azidoacetate are added; controlling the temperature to be-15 to-10 ℃, and dropwise adding 10g of sodium methoxide/methanol (accounting for 30 wt%); after dripping, the reaction is kept for 2h (the system is changed from clear to white and turbid); carrying out external bath at-10 ℃, and keeping the temperature to react overnight; sampling LCMS, pouring the raw materials into ice saturated ammonium chloride aqueous solution after the raw materials are basically completely reacted, filtering the solid, extracting the aqueous phase for 2 times by EA, combining the organic phase soluble solid, washing with salt, drying and spin-drying to obtain 6g of light yellow solid. The crude product is directly put into the next step. Yield: 97.8 percent

Step 6

Dissolving 10.1g of compound 6 in 300ml of THF (tetrahydrofuran), and cooling to-40 to-30 ℃ under the protection of argon; 7.0g of methanesulfonyl chloride was added; controlling the temperature to be between 40 ℃ below zero and 30 ℃ below zero, dripping 21.6g of triethylamine, and keeping the temperature to react for 1.5h (the system turns turbid from clear); LCMS was sampled and the material remained-7%, poured into ice water, EA extracted, combined organic phases, brine washed, dried, spun dry, and run through column to give 4.8g pale yellow solids (10% EA in HEP out product). Yield: 50.2 percent

Step 7

Dissolving 4.8g of compound 7 in 200ml of o-dichlorobenzene, heating to 140 ℃ under the protection of argon, and reacting for 1h (the system is clear); LCMS is sampled, the raw materials react completely, and the oil pump is decompressed and concentrated to dry o-dichlorobenzene. The crude product is directly put into the next step.

Step 8

Dissolving 4.37g of compound 8 in 50ml of DMAc, adding 4.24g of potassium carbonate under the protection of argon, and dropwise adding 2.61g of methyl iodide at room temperature; after dripping, reacting at room temperature for 1h; sample LCMS, the raw material reaction is complete, pour into ice ammonia water, EA extract, combine organic phase, salt wash, dry, spin dry, mix sample and get 4.7g light yellow solid. Yield: 100 percent

Step 9

Dissolving 2.8g of compound 9 in 33ml of acetic acid, adding 33ml of hydrobromic acid, and after the addition is finished, heating to 93 ℃ (outer oil bath) to react for 1h; sampling LCMS, cooling the residual 3 percent of the raw material and 20 percent of saponified acid to room temperature, and drying the raw material at 60 ℃ by an oil pump to obtain yellow solid; dissolving in 30ml methanol, dripping 3ml concentrated sulfuric acid, heating and refluxing, and monitoring by LCMS until the saponified acid content is less than 8 percent, namely treating: most of the methanol is removed by rotation, EA is diluted, and the yellow solid crude product is obtained by salt washing, drying and rotary drying. Yield: 89.8 percent

Step 10

Dissolving 1.36g of compound 10 in 70ml of DMAc, adding 3.11g of cesium carbonate, adding 2.08g of B, and reacting at room temperature for 3 hours after the addition is finished; sample LCMS and the material was essentially reacted completely, poured into ice water, extracted with 2-MeTHF, combined organic phases, washed with salt, dried, spun dry, and chromatographed on a column to give 2.4g of a pale yellow oil. Yield: 88.6 percent

Step 11

Dissolving 4.3g of compound 11 in 86ml of NMP, adding 2.0g of cuprous cyanide, and after the addition, heating to 170 ℃ (external oil bath) for reaction for 6h; sampling LCMS, cooling the residual 3 percent of the raw materials to room temperature, pouring into ice water, extracting by EA, combining organic phases, washing by salt, drying, spin-drying, mixing the sample and passing through a column to obtain 1.8g of light yellow solid. Yield: 46.2 percent

¹ H NMR(400MHz,CDCl ₃ )δ7.77(s,1H),7.52(s,1H),7.51–7.47(m,4H),7.37–7.30(m,2H),7.28(dd,J＝7.2,1.1Hz,4H),4.33(s,3H),4.13(dd,J＝5.4,3.9Hz,2H),3.97(dd,J＝5.4,3.9Hz,2H),3.91(s,3H),1.06(s,9H).

Step 12

Dissolving 1.7g of compound 12 in 17ml of THF, dissolving 0.4g of LiOH in 17ml of H2O, and dripping into the reaction solution to react for 1 hour at room temperature after dripping; sampling TLC, almost completely reacting the raw materials, removing THF at room temperature by spinning, adjusting pH to be =1 by 2M hydrochloric acid aqueous solution under ice bath, stirring for 10min, filtering, washing a filter cake by water, and carrying water in a solid by toluene to obtain 2g of light yellow solid crude product. Yield: 100 percent

Example 6: (R) -3- ((3- ((tert-butyldiphenylsilyl) oxy) pyrrol-1-yl) methyl) -8-chloro-1, 7-diazanaphthalene

The synthetic route is as follows:

step 1:

compound 1 (21.8g, 110.6mmol, 1eq) was dissolved in TFA-H ₂ SO ₄ (4. The yield thereof was found to be 97.5%.

¹ H NMR(400MHz,DMSO-d ₆ )δ8.54(d,J＝2.1Hz,1H),8.05(d,J＝2.1Hz,1H),7.96(s,1H),7.55(s,1H).，2.50(s,3H)

LCMS:found 215:217＝1:1[M+H] ⁺

The other method comprises the following steps:

compound 1-1 (18.4g, 85.2mmol, 1eq) and thionyl chloride (180mL, 10V) are mixed to form white turbidity, the mixture is stirred and heated to reflux, after about 1h, the system becomes a yellow solution, the mixture is stopped, heated and concentrated, THF is added into the residue to be dissolved, ammonia water is dropwise added into 1, 4-dioxane solution (0.4M, 178.9mL.2.1eq) under ice bath, after the dropwise addition is finished, the system is warmed to room temperature and stirred for 30min, the residue is added with water, IOAc is extracted for three times, organic phases are combined, the mixture is washed by saturated common salt water, anhydrous sodium sulfate is dried, the filtrate is filtered and concentrated, and the product is 20.3g of white foamy solid, and the yield is quantitative.

Step 2:

compound 2 (40.8g, 189.7mmol, 1eq) and DMF-DMA (33.9g, 284.6mmol, 1.5eq) were dissolved in toluene (612mL, 15V), heated to reflux overnight with stirring, LCMS indicated complete consumption of starting material, concentrated under reduced pressure, and the residue was used in the next reaction without isolation.

LCMS:found 270:272＝1:1[M+H] ⁺

And step 3:

dissolving compound 3 (49.4g, 182.9mmol, 1eq) in tetrahydrofuran (1.5L, KF 0.05%,30V 10mL/g), adding potassium tert-butoxide (30.8g, 274.4mmol, 1.5eq) after nitrogen replacement, stirring and heating to reflux after nitrogen replacement again, changing the system from reddish brown to brownish black, cooling to room temperature, adding water to quench the reaction, adjusting the pH to 7-8 with hydrochloric acid, concentrating, extracting the residue with 2-methyltetrahydrofuran three times, combining organic phases, washing with saturated brine, drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and carrying out column chromatography on the residue to obtain 12g of compound 4 which is yellow to reddish brown solid. The total yield of the two steps is 29 percent.

¹ H NMR(400MHz,DMSO-d ₆ )δ11.63(s,1H),8.80(d,J＝2.2Hz,1H),8.48(d,J＝2.2Hz,1H),7.32(d,J＝7.1Hz,1H),6.51(d,J＝7.1Hz,1H).

LCMS:found 225:227＝1:1[M+H]

And 4, step 4:

after compound 4 (13.5g, 60.0mmol, 1eq) and phosphorus oxychloride (135mL, 10V) were mixed, stirred, heated and refluxed for 2h, LCMS showed that the reaction was completed, the reaction was concentrated, the residue was diluted with water, EA was added for beating for 1h, the filtration was performed, the filter cake was washed with water, and the product was obtained after drying as a pale yellow solid, 9.6g, yield 65.7%.

¹ H NMR(400MHz,DMSO-d ₆ )δ9.21(d,J＝2.3Hz,1H),8.93(d,J＝2.3Hz,1H),8.46(d,J＝5.6Hz,1H),7.92(d,J＝5.6Hz,1H).

LCMS:found 243:245:247＝3:3:1[M+H] ⁺ .

And 5:

the starting compound 5 (9.6g, 39.4mmol, 1eq), vinyl boronic acid pinacol ester (6.68g, 43.4mmol, 1.1eq), and sodium carbonate (20.9g, 197.1mmol, 5eq) were added to a THF-water (4, 480mL, 50V) mixed solvent, after nitrogen substitution, palladium tetratriphenylphosphine (911.2mg, 0.79mmol, 0.02eq) was added, after nitrogen substitution again, the mixture was stirred to reflux, and after 1.5h TLC and LCMS analysis, a small amount of the starting material remained. After stopping heating, the system was concentrated, and the residue was extracted three times with EA, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to column chromatography (EA in Hep 15%) to obtain the product compound 6 as a pale yellow solid 5.9g with a yield of 76.1%.

¹ H NMR(400MHz,DMSO-d ₆ )δ9.31(d,J＝2.2Hz,1H),8.50(d,J＝2.2Hz,1H),8.38(d,J＝5.5Hz,1H),7.89(d,J＝5.5Hz,1H),7.00(dd,J＝17.7,11.1Hz,1H),6.31(d,J＝17.7Hz,1H),5.67(d,J＝11.1Hz,1H).

LCMS:found 191/193＝3:1[M+H] ⁺

Step 6:

compound 6 (4.0g, 21.1mmol, 1.0eq), NMO (4.9g, 41.8mmol, 2.0eq) and K were sequentially mixed ₂ OsO ₄ (0.38g, 1.03mmol, 0.049eq) THF H is added ₂ O =3 in solvent 1 (100 ml) overnight with stirring at room temperature, TLC monitoring the reaction (PE: EA = 1); extracted with ethyl acetate (4 times) and the combined organicsThe phases are washed with saturated brine, the solvent is dried by spinning, and THF H is added ₂ Dissolving in a solvent with O =3, and adding NaIO ₄ Solid (11.2g, 52.4mmol, 2.49eq), stirred, TLC monitored reaction was complete, water was added to dissolve the solid, extracted with ethyl acetate (4 times), the organic phases were combined, washed with saturated chloride, dried with anhydrous sodium sulfate, solvent was spin dried to give product 7 as a flocculent solid, 3.45g, 85.3% yield.

1H NMR(400MHz,DMSO-d6)δ10.32(s,1H),9.48(d,J＝2.0Hz,1H),9.10(d,J＝2.0Hz,1H),8.54(d,J＝5.5Hz,1H),8.18(d,J＝5.5Hz,1H).

LCMS found 193:195＝3:1[M+H]

Step 7

Under nitrogen protection, compound 7 (3.45g, 17.9mmol, 1.0eq), compound 8 (6.5g, 17.96mmol, 1.0eq) and Et were added ₃ N (7.35g, 72.6mmol, 4.1eq) was added to DCM (100 ml), the temperature was reduced in an ice-water bath, and after stirring for about 15min, naBH (OAc) was added ₃ (5.7g, 26.9mmol, 1.5eq) was added to the reaction flask, stirred in an ice water bath for-2 h and then warmed to room temperature and stirred overnight, the TLC monitoring reaction (DCM: meOH = 20); the yield thereof was found to be 62.3%.

¹ H NMR(400MHz,DMSO-d ₆ )δ9.04(d,J＝2.1Hz,1H),8.39(d,J＝5.5Hz,1H),8.34(d,J＝2.0Hz,1H),7.92(d,J＝5.5Hz,1H),7.57(ddt,J＝11.4,6.3,1.8Hz,4H),7.48–7.32(m,6H),4.37(dq,J＝9.8,3.6Hz,1H),3.89(d,J＝14.0Hz,1H),3.79(d,J＝14.0Hz,1H),2.69(dt,J＝9.8,6.6Hz,2H),1.95(dq,J＝14.3,7.3Hz,1H),1.82–1.62(m,1H),1.23(dd,J＝7.0,4.6Hz,2H),0.98(s,9H).

LCMS found 502:504＝3:1[M+H]

Example 7: synthesis of (R) -3' - ((3- ((3- ((tert-butyldiphenylsilyl) oxy) pyrrol-1-yl) methyl) -1, 7-dinaphthalen-8-yl) amino) -2,2' -dimethyl- [1,1' -diphenyl ] -3-carboxylic acid

Step 1

Into a reaction flask were successively charged Block 1 (0.3 g,0.6mmol,1.0 eq), isopropanol, compound 1 (0.145g, 0.6mmol,1.0 eq) and TosOH-H ₂ O (0.117g, 0.61mmol, 1.0eq), reflux heating for 8h, tlc monitoring (HEP: EA = 3.

LCMS found 721[M+H]

Step 2:

compound 2 (287mg, 0.4mmol) was dissolved in THF/H ₂ O (3.

LCMS found 707[M+H]

(R) -N- (3-bromo-2-methylphenyl) -3- ((3- ((tert-butyldiphenylsilyl) oxy) pyrrol-1-yl) methyl) -1, 7-diazanaphthalen-8-amine

Synthesized according to the method of step 1 of reference example 7.

(R) -N3- (3- ((3- ((tert-butyldiphenylsilyl) oxy) pyrrol-1-yl) methyl) -1, 7-diazanaphthalen-8-yl) -2,2' -dimethyl- [1,1' -diphenyl ] -3,3' -diamine

Synthesized according to the method of step 1 of reference example 7.

Example 8: synthesis of N- (5- (((3S) -cyclopentyl) methyl) -1-methyl-4-oxo-4, 5-dihydro-1H-imidazo [4,5-c ] pyridin-2-yl) -3- ((7 (((R) -3-hydroxypyrrolidin-1-yl) methyl) quinolin-4-yl) oxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3,3' -dicarboxamide.

The synthetic route is as follows:

to a solution of (R) -3' - ((7- ((3-hydroxypyrrol-1-yl) methyl) quinolin-4-yl) oxy) -2,2' -dimethyl- [1,1' -diphenyl ] -3-carboxylic acid (468mg, 1mmol) and 2-amino-5- (((3S) -3-hydroxycyclopentyl) methyl) -1-methyl-1, 5-dihydro-4H-imidazo [4,5-c ] pyridin-4-one (262mg, 1mmol) in DMF (3 mL) at room temperature were added HATU (456 mg) and DIEA (258 mg). The mixture was then heated to 50 ℃ and stirred overnight, then cooled to room temperature. The mixture was purified by prep-HPLC to give the product.

LCMS found 713.3[M+H] ⁺

Example 9: synthesis of N, N '- (2, 2' -dimethyl- [1,1 '-diphenyl ] -3,3' -diyl) bis (5- (2-hydroxyethyl) -1-methyl-4-oxo-4, 5-dihydro-1H-pyrrolo [3,2-c ] pyridine-2-carboxamide).

Step 1:

200mg of Compound 1 are added to 5.6ml of toluene and 4.5ml of methanol, and a solution of trimethylsilylated diazomethane in cyclohexane (2M) is added dropwise until the yellow colour does not disappear anymore (0.25 ml); stir for 1min, quench the reaction with acetic acid (0.2 ml) and adjust pH =8 with aqueous sodium bicarbonate. Spin-drying the solvent; EA extraction, organic phase combination, salt washing, drying, spin-dry mixing and column chromatography to obtain 214mg of white solid. Yield: 100 percent

Step 2

Dissolving 300mg of compound 2 and 149mg of compound 3 in 15ml of THF, performing nitrogen protection, performing ice salt bath, dropwise adding 2.5ml (2M) of NaHMDS THF solution, performing heat preservation reaction for 20min, performing LCMS detection, ensuring that raw materials are basically completely reacted, adding water to quench and react, performing EA extraction, combining organic phases, performing salt washing, drying, performing spin-drying mixing on a sample, and passing through a column to obtain 160mg of light yellow solid. Yield: 37.2 percent

Step 3

Dissolving 160mg of Compound 4 in 2ml of methanol, adding 100mg 10% Pd/C, introducing hydrogen, reacting at room temperature for 30min, sampling, detecting by TLC, the raw materials reacting substantially completely, filtering out 10% Pd/C, and spin-drying to obtain 140mg of an off-white solid crude product; the crude product is directly put into the next step. Yield: 91.5 percent

Step 4

Dissolving 55mg of compound 2 and 75mg of compound 5 in 3.8ml of THF, performing nitrogen protection, performing ice salt bath, dropwise adding 0.45ml of (2M) NaHMDS THF solution, and reacting for 1h under the condition of keeping temperature after dropwise adding; adding water to quench and react, extracting with EA, combining organic phases, washing with salt, drying, mixing with a rotary drying sample, and passing through a column to obtain 53mg yellow solid. Yield: 41.9 percent

Step 5

53mg of Compound 6 was dissolved in 2.5ml of THF, 2.5ml of triethylamine trihydrofluoride was added thereto, the mixture was reacted at room temperature for 2 hours, TLC was sampled, the reaction of the starting material was substantially completed (. About.30% of the product), THF was distilled off under reduced pressure, and preparative TLC separation was carried out to obtain 6.1mg of THF. Yield: 20.3 percent of

¹ H NMR(400MHz,DMSO-d ₆ )δ9.85(s,1H),7.55(s,1H),7.43(d,J＝7.5Hz,1H),7.39(d,J＝7.9Hz,1H),7.30(t,J＝7.8Hz,1H),7.02(d,J＝7.4Hz,1H),6.62(d,J＝7.5Hz,1H),4.00(t,J＝5.7Hz,2H),3.95(s,3H),3.62(t,J＝5.6Hz,2H),1.96(s,4H),1.24(s,1H).

Example 10: synthesis of (1r, 4r) -4- ((2, 2' -dimethyl-3 ' - (4-oxo-4, 5,6, 7-tetrahydrothiazolo [4,5-c ] pyridopyridin-2-yl) - [1,1' -diphenyl ] -3-yl) carbamoyl) -1-methyl-1, 4,6, 7-tetrahydro-5H-imidazo [4,5-c ] pyridopyridin-5-yl) methyl) cyclohexane-1-carboxylic acid.

The synthesis route is as follows:

step 1:

dissolve 1 (318 mg) in dioxane (15 mL) at room temperature, add 2 (323 mg) and 1.5mL Na ₂ CO ₃ Aqueous (2N), vacuum degassed, and Pd (dppf) Cl added ₂ (73.2 mg), degassed again and reacted at 90 degrees overnight. After cooling to room temperature, the reaction was extracted with water and EtOAc, and the organic phase was washed with saturated NaCl, dried, spun-dried, and passed through a column (HEP: etOAc =5, 1-2.

Step 2:

compound 3 (150 mg) was dissolved in DMF (2 ml) at room temperature, HATU (196.7 mg) and DIEA (113 mg) were added, and the mixture was stirred for 10 minutes, followed by addition of compound 4 (138.7 mg), and the reaction was continued at room temperature overnight. The reaction was extracted with EtOAc after addition of water, and the organic phase was washed with saturated NaCl, dried, spun-dried, and passed through a column (HEP: etOAc =5, 1-1.

And step 3:

compound 5 (100 mg) was dissolved in 4N HCl/dioxane (2 ml) at room temperature, stirred for 30 minutes, TLC detected disappearance of starting material and reaction was concentrated and used directly for the next reaction.

And 4, step 4:

compound 6 (100 mg, N) was dissolved in MeOH/H at room temperature ₂ To O (1ml, 1) ₂ O (12.2 mg), and stirred for 2 hours. TLC detects the disappearance of the raw materials, the reaction is adjusted to pH = 6-7 by 1N HCl, and the organic phase is prepared by high performance liquid chromatography to obtain (1r, 4r) -4- ((2, 2 '-dimethyl-3' - (4-oxo-4, 5,6, 7-tetrahydrothiazole [4, 5-c))]And pyridin-2-yl) - [1,1' -Diphenyl]-3-yl) carbamoyl) -1-methyl-1, 4,6, 7-tetrahydro-5H-imidazo [4,5-c]And pyridin-5-yl) methyl) cyclohexane-1-carboxylic acid. LC-MS found 653.3 (M + H)

Example 11: synthesis of (R) -5- (2-hydroxyethyl) -N- (3 ' - ((3- ((3-hydroxypyrrolin-1-yl) methyl) -1, 7-dinaphthalen-8-yl) amino) -2,2' -dimethyl- [1,1' -diphenyl ] -3-yl) -1-methyl-4-oxo-4, 5-dihydro-1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

The synthetic route is as follows:

step 1:

compound 1 (120 mg) and compound 2 (86.5 mg) were dissolved in THF (2 ml) under ice-bath, and N was added ₂ A2M solution of NaHMDS (0.7 mL THF) was added with protection and the ice bath was continued for 1 hour. TLC showed complete reaction of starting material, saturated NH was added ₄ And quenching the reaction by using a Cl aqueous solution, extracting by using EA, and carrying out column chromatography separation to obtain 80mg of yellow solid.

Step 2:

compound 3 (90 mg) was added to a mixture of THF (1 mL) and TEA/3HF (1 mL) and reacted for 2 hours. TLC detects the disappearance of the raw material, and saturated NaHCO is added into the reaction liquid ₃ The aqueous solution was adjusted to pH =9, extracted with etoac, spin dried, and preparative isolated to give 12.4mg yellow solid.

¹ H NMR(400MHz,DMSO-d ₆ )δ10.98(d,J＝94.5Hz,1H),9.88(s,1H),9.09(s,1H),8.52(s,1H),8.18(d,J＝8.3Hz,1H),8.05(d,J＝5.9Hz,1H),7.56(s,1H),7.38(ddd,J＝30.9,13.6,7.3Hz,4H),7.26–7.17(m,1H),7.08(t,J＝6.5Hz,1H),6.97(d,J＝7.6Hz,1H),6.62(d,J＝7.4Hz,1H),4.67(s,2H),4.47(s,1H),4.01(s,2H),3.95(s,3H),3.71–3.60(m,4H),2.93(m,1H),2.34-2.32(s,1H),2.07(s,3H),2.00(s,3H),1.95-1.85(m,1H),1.21-1.15(m,3H).

LCMS found 658[M+H] ⁺

Example 12: synthesis of (R) -7-cyano-5- (2-hydroxyethyl) -N- (3 ' - ((3- ((3-hydroxypyrrolin-1-yl) methyl) -1, 7-dinaphthalen-8-yl) amino) -2,2' -dimethyl- [1,1' -diphenyl ] -3-yl) -1-methyl-4-oxo-4, 5-dihydro-1H-pyrrolo [3,2-c ] pyridine-2-carboxamide

The synthesis route is as follows:

step 1:

compound 1 (300 mg) was dissolved in toluene (8.4 mL)/methanol (6) at room temperature6 mL) of the mixture, 2M cyclohexane solution (1.08 mL) of trimethylsilyl diazomethane was added dropwise until the yellow color did not disappear, TLC was used to detect the completion of the reaction of the raw materials, 0.3mL of AcOH was added to quench the reaction, and saturated NaHCO was added ₃ The pH was adjusted to =9, the solvent was evaporated under reduced pressure, extracted with EtOAc and passed through a column to give 240mg of a white solid

Step 2:

compound 3 (136 mg) and compound 4 (150 mg) were dissolved in THF (3 mL) in an ice bath, naHMDS (2M in THF, 0.88 mL) was added under N2 protection, and the reaction was continued for 1 hour in an ice bath. TLC shows that the raw material reacts completely, saturated NH4Cl is added for quenching reaction, EA extraction and column chromatography separation are carried out to obtain 80mg of yellow solid.

And step 3:

compound 5 (40 mg) was dissolved in THF (0.5 mL) at room temperature, and TBAF/THF (0.1 mL) was added to react for 3 hours. TLC showed the starting material was reacted completely, the solvent was distilled off under reduced pressure and preparative isolation gave 6.3mg of a yellow solid.

1H NMR(400MHz,DMSO-d6)δ10.02(s,1H),9.31(s,1H),8.87(s,1H),8.43(d,J＝8.3Hz,1H),8.34(s,1H),8.19(s,1H),8.06(d,J＝5.7Hz,1H),7.61(s,1H),7.40(d,J＝8.1Hz,1H),7.38–7.26(m,2H),7.18(d,J＝5.8Hz,1H),7.09(d,J＝7.5Hz,1H),6.88(d,J＝7.6Hz,1H),6.64(s,1H),5.33(t,J＝4.8Hz,1H),4.92(t,J＝5.6Hz,1H),4.72(d,J＝4.6Hz,1H),4.24(s,3H),4.07(d,J＝5.7Hz,1H),3.83(d,J＝10.4Hz,2H),3.65(d,J＝5.7Hz,2H),2.74(d,J＝6.1Hz,1H),2.68(s,1H),2.41(s,1H),2.10(s,3H),2.01-1.90(m,5H),1.46-1.40(m,2H).

LCMS found 683[M+H] ⁺

Biological assay

Example A: PD-1/PD-L1 homogeneous time-resolved fluorescence (HTRF) binding assays

The assay was performed in standard black 384 well polystyrene plates with a final volume of 20. Mu.L. Inhibitors were first serially diluted in DMSO and added to the well, followed by the addition of other reaction components. The final concentration of DMSO in the assay was 1%. The assay was carried out in PBS buffer (pH7.4) containing 0.05% Tween-20 and 0.1% BSA at 25 ℃. Recombinant human PD-L1 protein (19-238) with a His tag at the C-terminus was purchased from Acrobiosystems (PD 1-H5229). Recombinant human P with Fc tag at C-terminusD-1 protein (25-167) was also purchased from Acrobiosystems (PD 1-H5257). PD-L1 and PD-1 proteins were diluted in assay buffer and 0.1. Mu.l of the solution was extracted and added to the well of the plate. The plates were centrifuged and the proteins were preincubated with inhibitors for 40 minutes. After incubation, 0.1. Mu.l HTRF detection buffer containing europium-blocked labeled anti-human IgG (PerkinElmer-AD 0212) Fc specific and anti-His

Allophycocyanin (APC, perkinelmer-AD 0059H) -conjugated antibody. After centrifugation, the plates were incubated at 25 ℃ for 60 minutes. The data were read (665 nm/620nm ratio) in a PHERAstar FS reader. The final concentrations in the assay were-3 nM 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 give IC50 values for the inhibitors.

The IC50 values of the compounds exemplified in the examples are expressed in the following manner: + = IC50 ≦ 100nM; + =100nM woven fabric IC50 is less than or equal to 1000nM; + ++=1000nM Once-constructed IC50

Data for the 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.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes or modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the appended claims of the present application.

Claims

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

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

L ₁ selected from the group consisting of: a bond, substituted or unsubstituted C1-C4 alkylene-S-, -O-, substituted or unsubstituted-NH-,

substituted or unsubstituted->

L ₂ Is a chemical bond;

L ₃ selected from the group consisting of: chemical bond, substituted or unsubstituted C1-C4 alkylene, substituted or unsubstituted

Is a group having the structure shown below:

Wherein,

Z ¹ selected from the group consisting of: o, NRf, N-O-Rh; wherein said Rf is selected from the group consisting of: H. -C (= O) -NRdRe, -C (= O) -substituted or unsubstituted C ₁ -C ₆ Alkyl, -C (= O) -substituted or unsubstituted C ₃ -C ₁₀ A cycloalkyl group; the Rh is selected from the following group: H. substituted or unsubstituted C ₁ -C ₆ An alkyl group;

Z ² 、Z ³ each independently selected from the group consisting of: CH. CH (CH) ₂ ；

Y ¹ 、Y ² Each independently selected from the group consisting of: CH. NH, N, O, S;

is a single bond or a double bond;

and is

Is an aromatic fragment;

r is H, substituted or unsubstituted C ₁ -C ₆ Alkyl, or- (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s ；

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 A cycloalkyl group;

selected from the group consisting of: 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 said heterocyclyl has 1-3 heteroatoms;

selected from the group consisting of: a substituted or unsubstituted C6 arylene, or a substituted or unsubstituted 5-7 membered heteroarylene; or said>

Is none;

selected from the group consisting of: a substituted or unsubstituted C6 arylene, or a substituted or unsubstituted 5-7 membered heteroarylene;

R ₁ selected from the group consisting of: H. halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, oxo, -CN, hydroxy, NRdRe, substituted or unsubstituted C1-C6 amino; or (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s ；

R ₄ Selected from the group consisting of: H. halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, oxo, -CN, hydroxy, substituted or unsubstituted C1-C6 amino;

R ₂ and R ₃ Each independently selected from the group consisting of: H. halogen, substituted or unsubstituted C1-C6 alkyl, -CN;

each L is _1a Each independently is a group selected from the group consisting of; chemical bond, substituted or unsubstituted C ₁ -C ₇ An alkylene group;

L _2a selected from the group consisting of: substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-10 membered heterocyclyl having 1-3 heteroatoms;

L _3a selected from the group consisting of: hydroxyl, amino, carboxyl;

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

each s is 0 or 1;

unless otherwise specified, the term "substituted" means substituted with one or more substituents selected from the group consisting of: halogen, -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 unsubstituted or substituted with one or more substituents selected from the group consisting of: C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, C3-C8 cycloalkyl having 1-3 substituents selected from N, S and5-10 membered heteroaryl of a heteroatom of O, 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; the substituents are selected from the following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

in each of the above formulae, any of the heteroatoms is selected from the group consisting of: n, S and O.

2. The compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof

A bicyclic structure of 5-7 membered ring and 5-6 membered ring.

3. The compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof

Is equal to->

4. The compound of claim 1, or an optical isomer, a pharmaceutically acceptable salt thereof, wherein said ring is

And/or->

Having a substituent represented by formula IV below:

wherein, L is ₄ Is substituted or unsubstitutedC1-C4 alkylene;

selected from the group consisting of: a substituted or unsubstituted 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O;

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

5. The compound of claim 4, or an optical isomer, or a pharmaceutically acceptable salt thereof, wherein

Is a 5-7 membered nitrogen containing heterocyclic group.

6. The compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I has the structure of formula Ia:

substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

Y ¹ 、Y ² Each independently selected from the group consisting of: CH. NH, N, O or S;

is a single or double bond;

and is provided with

Is an aromatic fragment;

r is H, substituted or unsubstituted C ₁ -C ₆ Alkyl, or- (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s Wherein, in the process,

L _3a selected from the group consisting of: hydroxyl, amino, carboxyl;

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

each s is 0 or 1;

unless otherwise specified, "substituted" means substituted with one or more substituents selected from the group consisting of: halogen-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 unsubstituted or substituted with one or more substituents selected from the group consisting of: C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, C3-C8 cycloalkyl, 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 following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

7. The compound of claim 6, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of compounds 1-125 of the following table:

。

8. the compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I has the structure shown in formula Ib:

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

substituted or unsubstituted>

Is a group having the structure shown below:

Wherein,

Z ¹ selected from the group consisting of: o, NRf, N-O-Rh; wherein, said Rf is selected from the group consisting of: H. -C (= O) -NRdRe, -C (= O) -substituted or unsubstituted C ₁ -C ₆ Alkyl, -C (= O) -substituted or unsubstituted C ₃ -C ₁₀ A cycloalkyl group; the Rh is selected from the following group: H. substituted or unsubstitutedC of (A) ₁ -C ₆ An alkyl group;

is a single or double bond;

and is provided with

Is an aromatic fragment;

each L _1a Each independently is a group selected from the group consisting of; chemical bond, substituted or unsubstituted C ₁ -C ₇ An alkylene group;

L _2a selected from the group consisting of: substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-10 member heterocyclyl having 1-3 heteroatoms;

L _3a selected from the group consisting of: hydroxyl, amino, carboxyl;

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

each s is 0 or 1;

unless otherwise specified, "substituted" means substituted with one or more substituents selected from the group consisting of: halogen, -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 unsubstituted or substituted with one or more substituents selected from the group consisting of: C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, C3-C8 cycloalkyl, 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; the substituent is selected from the following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

9. The compound of claim 8, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of compounds 126-202:

。

10. the compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I has the structure of formula Ic:

substituted or unsubstituted->

Is a group having the structure shown below:

Z ¹ Selected from the group consisting of: o, NRf, N-O-Rh; wherein, said Rf is selected from the group consisting of: H. -C (= O) -NRdRe, -C (= O) -substituted or unsubstituted C ₁ -C ₆ Alkyl, -C (= O) -substituted or unsubstituted C ₃ -C ₁₀ A cycloalkyl group; the Rh is selected from the following group: H. substituted or unsubstituted C ₁ -C ₆ An alkyl group;

is a single or double bond;

and is

Is an aromatic fragment;

L _3a selected from the group consisting of: hydroxyl, amino, carboxyl;

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

each s is 0 or 1;

rd, re are each independently selected from the group consisting of: H. substituted or unsubstitutedC ₁ -C ₆ Alkyl, substituted or unsubstituted C ₃ -C ₁₀ A cycloalkyl group;

selected from the group consisting of: 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 said heterocyclyl has 1-3 heteroatoms; (ii) a

Unless otherwise specified, the term "substituted" means substituted with one or more substituents selected from the group consisting of: halogen, -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 unsubstituted or substituted with one or more substituents selected from the group consisting of: C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, C3-C8 cycloalkyl, 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 following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

11. The compound of claim 10, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of compounds 203-313;

。

12. the compound of claim 1, or an optical isomer, a pharmaceutically acceptable salt thereof, wherein the compound of formula I is selected from the group consisting of formulas Id-1 and Id-2, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof:

substituted or unsubstituted->

Rf is selected from the group consisting of: H. -C (= O) -NRdRe, -C (= O) -substituted or unsubstituted C ₁ -C ₆ Alkyl, -C (= O) -substituted or unsubstituted C ₃ -C ₁₀ A cycloalkyl group; the Rh is selected from the following group: H. substituted or unsubstituted C ₁ -C ₆ An alkyl group;

Z ² and Z ³ Each independently selected from the group consisting of: CH. CH (CH) ₂ ；

Y ¹ And Y ² Each independently selected from the group consisting of: CH. NH, N, O, S;

is a single bond or a double bond;

and is provided with

Is an aromatic fragment;

r is H, substituted or unsubstituted C ₁ -C ₆ Alkyl, or- (L) _1a ) _r -(L _2a ) _s -(L _3a ) _s Wherein

L _3a selected from the group consisting of: hydroxyl, amino, carboxyl;

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

each s is 0 or 1;

unless otherwise specified, the term "substituted" means substituted with one or more substituents selected from the group consisting of: halogen, -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 unsubstituted or substituted with one or more substituents selected from the group consisting of: C1-C6 alkyl, C1-C6 alkoxy, C6-C10 aryl, C3-C8 cycloalkyl, 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, 5-10 membered heterocyclyl having 1-3 heteroatoms selected from N, S and O; the substituent is selected from the following group: halogen, hydroxy, carboxy, cyano, C1-C6 alkoxy, C1-C6 alkylamino;

13. A compound, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, selected from the group consisting of compounds 314-349:

。

14. a process for the preparation of a compound of formula I according to claim 1, wherein said compound is 1-7, 2-7 or 3-7 and said process comprises a step selected from the group consisting of those depicted in schemes 1,2 or 3:

synthesis scheme 1

(a) Taking a halide 1-1 and a proper coupling reagent 1-2 as basic raw materials, and obtaining an intermediate compound 1-3 through a coupling reaction catalyzed by palladium or copper;

(b) Taking the intermediate 1-3 as a raw material, and reacting with carboxylic acid 1-4 under the action of a condensing agent to obtain an amide intermediate 1-5;

(c) Taking the intermediate 1-5 as a raw material, and removing a protecting group under an acidic condition to obtain an intermediate 1-6;

synthesis scheme 2:

(a) Carboxylic ester 2-1 is taken as a raw material, and is subjected to aminolysis reaction with amine 2-2 under the catalysis of Lewis acid to obtain an intermediate compound 2-3;

(b) Taking the intermediate 2-3 and a proper coupling reagent 2-4 as basic raw materials, and obtaining an intermediate compound 2-5 through a coupling reaction catalyzed by palladium or copper;

said Cy is

Is->

n、m、p、q、Y ¹ 、Y ² 、Z ¹ 、Z ² 、Z ³ And R is as defined in claim 1.

15. A pharmaceutical composition comprising (1) a compound of claim 1 or an optical isomer thereof, or a pharmaceutically acceptable salt thereof; (2) a pharmaceutically acceptable carrier.

16. Use of the compound or the optical isomer thereof according to claim 1, or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 15, for the preparation of a pharmaceutical composition for the prevention and/or treatment of diseases associated with the activity or expression amount of PD-1/PD-L1.

17. A PD-1/PD-L1 inhibitor, wherein said inhibitor comprises a compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof.

18. A method for non-therapeutically inhibiting the PD-1/PD-L1 interaction in vitro comprising the steps of: contacting a compound of claim 1, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof, with PD-L1 protein.