WO2020151707A1

WO2020151707A1 - Novel small molecule cd73 inhibitor, preparation method therefor and pharmaceutical use thereof

Info

Publication number: WO2020151707A1
Application number: PCT/CN2020/073526
Authority: WO
Inventors: 董平; 李心; 王浩蔚; 刘健; 庄凌航; 张儒民; 陶维康
Original assignee: 江苏恒瑞医药股份有限公司; 上海恒瑞医药有限公司
Priority date: 2019-01-22
Filing date: 2020-01-21
Publication date: 2020-07-30
Also published as: CN112955444B; CN112955444A

Abstract

The present disclosure relates to a novel small molecule CD73 inhibitor, a preparation method therefor and a pharmaceutical use thereof. Particularly, the present disclosure relates to a small molecule CD73 inhibitor represented by general formula (I), a preparation method therefor, a pharmaceutical composition containing the derivative, and use thereof as a therapeutic agent, particularly in the preparation of a medicament for treating CD73-mediated diseases and conditions. The substituents of general formula (I) are as defined in description.

Description

A new type of small molecule CD73 inhibitor, its preparation method and its application in medicine

Technical field

The present disclosure belongs to the field of medicine, and relates to a novel small molecule CD73 inhibitor represented by the general formula (I) or its prodrug, its preparation method and pharmaceutical composition containing the derivative and its use as a therapeutic agent, especially as a preparation Use in medicine for the treatment of CD73-mediated diseases and disorders.

Background technique

CD73 is also called extracellular-5'-nucleotidase (Ecto-5'-Nucleotidase, EC3.1.3.5, ecto-5'-NT), which is anchored to the substance through glycosylphosphatidylinositol (GPI) A glycoprotein of the membrane. It is expressed in different tissues, especially colon, kidney, brain, liver, heart, lung, spleen, lymph nodes and bone marrow (Antonioli, L. et al., Trends Mol. Med., 2013, 19, 355-367). The enzymatic activity of CD73 is to catalyze the hydrolysis of nucleoside monophosphates to their corresponding nucleosides (e.g. 5-AMP to adenosine). CD73 mainly exerts its enzymatic nucleoside products, especially adenosine in the extracellular space, on epithelial ion and fluid transport, tissue barrier function control, hypoxia adaptation, ischemic preconditioning, anti-inflammatory and immunosuppressive signals, etc. Physiological effects (Colgan, SP. et al. Purinergic Signaling, 2006, 2, 351-360).

As a ubiquitous extracellular signal molecule with neuromodulation properties, adenosine produces a wide range of physiological responses in the human body through its interaction with adenosine receptors (receptor subtypes: A1, A2A, A2B, and A3), including in the heart The properties of the vascular system to relax blood vessels and inhibit atrioventricular conduction; play a sedative effect on local neuronal excitability inhibition in the central nervous system, and also have anticonvulsant and neuroprotective effects (Dunwiddie, TV et al., Annu. Rev. Neurosci. 2001, 24, 31-55); the effect of contracting bronchial tubes in the respiratory system (Pauwels, R. et al., Drug Development Research, 1993, 28, 318-321); and mediating immune or inflammatory responses in the immune system (Hasko , G. et al., "A Key Link between Metabolism and Brain Activity", 2013, 233-251).

CD73 is widely expressed in many cancer types (Antonioli, L. et al., Trends in Cancer, 2016, 2(2), 95-109), and is associated with the prediction of the likelihood of occurrence of many cancer types (Allard, D., etc.) Human, Immunotherapy, 2016, 8(2), 145-163). CD73 promotes cancer metastasis (Yang, Q. et al., Pathol. Oncol. Res., 2013, 19, 811-814) and chemoresistance (Loi, S. et al., PNAS, 2013, 110(27), 11091-11096 ). In the immune system, CD73 is found on the surface of macrophages, lymphocytes, regulatory T cells, myeloid-derived suppressor cells (MDSCs) and dendritic cells. Extracellular adenosine, mainly produced by CD73, can accumulate in the tumor microenvironment for a long time, activate adenosine receptors, promote tumor-induced mononuclear phagocytes, relieve anti-tumor T cell responses, expand the MDSCs population, and trigger immunosuppression, which is beneficial Cancer cells escape immune surveillance, thereby promoting cancer transformation and growth (Antonioli, L. et al., Nature Reviews Cancer, 2013, 13, 842-857).

At present, most of the small molecule drug candidates as CD73 inhibitors are in the discovery stage. Published patent applications include WO2015164573, WO2017098421, WO2017120508, WO2017153952, US20170044203, WO2018049145, WO2018067424 and WO2018094148. But no small molecule compound has entered clinical research. Therefore, it is necessary to discover and develop new CD73 inhibitors, provide new treatments for inhibiting tumor development and metastasis, increase the efficacy of anti-cancer treatments, and restore immune cell response by reducing the level of extracellular adenosine in the tumor microenvironment Effective resistance of cancer cells to treat cancer.

Summary of the invention

Therefore, the purpose of the present disclosure is to provide a compound represented by general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, Prodrug, mixture form or its pharmaceutically acceptable salt:

among them:

Y is -O- or -S-;

Z is -O- or -NH-;

W is selected from -O-, -S-, -N(R ¹⁰ )- and -C(R ¹¹ )(R ¹² )-;

M is selected from NH, -O- or -S-;

G ¹ and G ^{2 are the} same or different, and are each independently selected from -N- or -C(R ¹³ )-;

L is an alkylene group, wherein the alkylene group is optionally further selected from alkyl, halogen, amino, nitro, cyano, hydroxy, hydroxyalkyl, alkoxy, cycloalkyl, heterocyclyl, Substituted by one or more substituents in the aryl group and heteroaryl group;

Q is selected from -CH ₂ -OC(R ^s )(R ^t )-, -CH ₂ -N(R ^m )-C(R ^s )(R ^t )-, -CH ₂ -SC(R ^s )(R ^t )-, -CH ₂ -S(O) ₂ -C(R ^s )(R ^t )-, -phenyl-OC(R ^s )(R ^t )-, -CH ₂ -phenyl-OC(R ^s )(R ^t )-, -CH ₂ -heterocyclyl-, -C(R ^m )(R ⁿ ), -CH ₂ -C(R ^m )(R ⁿ )-C(R ^s )(R ^t )-, -C(R ^s )=C(R ^t )-, -C(R ^m )(R ⁿ )-C(R ^s )(R ^t )-, -C(R ^m )(R ⁿ )- C(R ^s )=C(R ^t )-and -C(R ^s )=C(R ^t )-C(R ^m )(R ⁿ );

R ^s , R ^t , R ^m and R ^{n are the} same or different, and are each independently selected from H, D, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl and amino;

Or R ^s and R ^t together form an oxo group;

Or R ^m and R ⁿ together form an oxo group;

R ¹ , R ² , R ³ and R ^{4 are the} same or different, and are each independently selected from hydroxyl, hydrogen, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cyano, amino , Azido and OR ¹⁴ ;

Or R ¹ and R ² together with the carbon atoms to which they are attached form a cycloalkyl or heterocyclic group, wherein the heterocyclic group contains 1 to 3 identical or different heteroatoms selected from N, O and S, wherein The cycloalkyl and heterocyclic groups are optionally further selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and hetero One or more substituents in the cyclic group are substituted;

R ^{5 is} selected from hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro and azido;

R ⁶ and R ^{7 are the} same or different, and are each independently selected from a hydrogen atom, an alkyl group, -C(R ^m )(R ⁿ )-OC(O)OR ^d , -C(R ^m )(R ⁿ )- OC(O)R ^d , -C(R ^m )(R ⁿ )C(O)OR ^d , cycloalkyl, heterocyclic, aryl and heteroaryl, wherein the alkyl group is optionally further selected Substituted by one or more substituents among cycloalkyl, heterocyclyl, aryl and heteroaryl;

R ^{d is} selected from hydrogen atom, alkyl group and alkoxy group;

R ^{8 is} selected from hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and heterocyclic group ；

R ^{9 is} selected from a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, a haloalkyl group, a hydroxyalkyl group, a cycloalkyl group, a heterocyclic group, an aryl group and a heteroaryl group, wherein the alkyl group, alkenyl group, alkynyl group , Cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro , Cycloalkyl, heterocyclyl, aryl and heteroaryl substituted by one or more substituents;

R ¹⁰ is an alkyl group;

R ¹¹ and R ^{12 are the} same or different, and are each independently selected from a hydrogen atom, a halogen, an alkyl group, and an alkenyl group;

R ^{13 is} selected from hydrogen atom, halogen, alkyl, alkoxy, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclic, aryl and heteroaryl;

R ^{14 is} selected from -C(O)R ¹⁵ , -C(O)OR ¹⁵ , -S(O) ₂ R ¹⁵ and -P(O)(OR ⁶ )(OR ⁷ );

R ^{15 is} selected from hydrogen atom, halogen, alkyl, haloalkyl, hydroxy and hydroxyalkyl;

n is 0 or 1.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, which are compounds represented by the general formula (II) or tautomers, mesosomes, racemates, enantiomers Conformers, diastereoisomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them:

R ¹ to R ⁹ , G ¹ , G ² , W, Y, Z, Q, L, M and n are as defined in the general formula (I).

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein W is -O-.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein said R ² , R ⁴ and R ⁵ are hydrogen atoms.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein R ¹ and R ^{3 are the} same or different, and are each independently selected from hydroxyl, hydrogen, halogen, alkyl, alkoxy, haloalkyl , Haloalkoxy and hydroxyalkyl, preferably hydroxy.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, which are compounds represented by general formula (III) or tautomers, mesosomes, racemates, enantiomers Conformers, diastereoisomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them

R ⁶ to R ⁹ , M, Q, L and n are as defined in the general formula (I).

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixtures or pharmaceutically acceptable salts thereof, wherein said G ¹ is -N-; G ² is -CH-.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein said R ⁶ and R ^{7 are the} same or different, and are each independently selected from a hydrogen atom, an alkyl group or -C(R ^m )( R ⁿ )-OC(O)OR ^d ; R ^m , R ⁿ and R ^d are as defined in the general formula (I).

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein said Q is -CH ₂ -O-CH ₂ -.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein said L is -(CH ₂ )s-; s is 1, 2 or 3.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein M is NH.

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, which are compounds represented by general formula (IV) or tautomers, mesosomes, racemates, enantiomers Conformers, diastereoisomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them:

R ⁸ , R ⁹ and n are as defined in the general formula (I).

In some embodiments of the present disclosure, the compound represented by the general formula (I) or its tautomers, mesosomes, racemates, enantiomers, diastereomers Forms, prodrugs, mixtures or pharmaceutically acceptable salts thereof, which are compounds represented by the general formula (IV-1) or tautomers, mesoisomers, racemates, enantiomers Forms, diastereomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them:

R ⁸ and R ⁹ are as defined in the general formula (I).

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, which are compounds represented by general formula (IV-2) or tautomers, mesosomes, racemates, and Enantiomers, diastereomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them:

R ¹⁶ and R ¹⁷ together with the connected nitrogen atom form a heterocyclic group, and the heterocyclic group is preferably a spiro heterocyclic group; wherein the heterocyclic group optionally contains 1 to 1 nitrogen atom. 2 identical or different heteroatoms selected from N, O and S, and the heterocyclic group is optionally further selected from alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl Substituted by one or more substituents in the group, cycloalkyl, heterocyclyl, aryl and heteroaryl;

R ^{8 is} as defined in the general formula (I).

In some embodiments of the present disclosure, the compound represented by the general formula (I) according to the present disclosure, or its tautomer, meso, racemate, enantiomer, or non-pair Enantiomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, wherein R ⁸ is halogen.

In some embodiments of the present disclosure, the compound represented by the general formula (I) or its tautomers, mesosomes, racemates, enantiomers, diastereomers In the form of a prodrug, a prodrug, a mixture or a pharmaceutically acceptable salt thereof, wherein said R ^{9 is} selected from alkyl, cycloalkyl and heterocyclic group, wherein said alkyl, cycloalkyl and heterocyclic group are optionally One selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl Or more substituents, the cycloalkyl group and heterocyclic group include cycloalkyl and heterocyclic group condensed aryl and heteroaryl; preferably selected from C _1-6 alkyl, C _3-12 Cycloalkyl, 4- to 12-membered heterocyclyl and C _1-6 alkyl substituted by phenyl; more preferably selected from propyl,

In some embodiments of the present disclosure, the compound represented by the general formula (I) or its tautomers, mesosomes, racemates, enantiomers, diastereomers In the form of a prodrug, a mixture, or a pharmaceutically acceptable salt thereof, wherein -(M) _n -R ^{9 is} selected from:

Typical compounds of the present disclosure include but are not limited to:

Or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its pharmaceutically acceptable salt.

Another aspect of the present disclosure relates to a compound represented by general formula (IA):

Or its tautomer, meso, racemate, enantiomer, diastereoisomer, prodrug, mixture form or its pharmaceutically acceptable salt,

among them:

Y is -O- or -S-;

Z is O;

R ^x is an alkyl group;

W is selected from -O-, -S-, -N(R ¹⁰ )- and -C(R ¹¹ )(R ¹² )-;

M is selected from -NH-, -O- or -S-;

Q is selected from -CH ₂ -OC(R ^s )(R ^t )-, -CH ₂ -N(R ^m )-C(R ^s )(R ^t )-, -CH ₂ -SC(R ^s )(R ^t )-, -CH ₂ -S(O) ₂ -C(R ^s )(R ^t )-, -phenyl-OC(R ^s )(R ^t )-, -CH ₂ -phenyl-OC(R ^s )(R ^t )-, -CH ₂ -heterocyclyl-, -C(R ^m )(R ⁿ )-, -CH ₂ -C(R ^m )(R ⁿ )-C(R ^s )(R ^t )-, -C(R ^s )=C(R ^t )-, -C(R ^m )(R ⁿ )-C(R ^s )(R ^t )-, -C(R ^m )(R ⁿ ) -C(R ^s )=C(R ^t )-and -C(R ^s )=C(R ^t )-C(R ^m )(R ⁿ );

Or R ^s and R ^t together form an oxo group;

Or R ^m and R ⁿ together form an oxo group;

R ² and R ^{4 are the} same or different, and are each independently selected from hydroxyl, hydrogen, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cyano, amino, azido and OR ¹⁴ ;

R ¹⁰ is an alkyl group;

n is 0 or 1.

Another aspect of the present disclosure relates to a compound represented by the general formula (IIIA):

Or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its pharmaceutically acceptable salt,

among them:

R ^x is an alkyl group;

M is selected from -NH-, -O- or -S-;

Or R ^s and R ^t together form an oxo group;

Or R ^m and R ⁿ together form an oxo group;

R ^{9 is} selected from the group consisting of a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, a haloalkyl group, a hydroxyalkyl group, a cycloalkyl group, a heterocyclic group, an aryl group and a heteroaryl group, wherein the alkyl group, alkenyl group and alkynyl group , Cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro , Cycloalkyl, heterocyclyl, aryl and heteroaryl substituted by one or more substituents;

n is 0 or 1.

Typical compounds of general formula (IA) of the present disclosure include but are not limited to:

Another aspect of the present disclosure relates to a method for preparing a compound represented by general formula (I), which comprises the following steps:

The protective group of the compound of general formula (IA) is removed to obtain the compound of general formula (I),

among them:

Z is -O-;

R ^x is an alkyl group;

R ⁶ and R ⁷ are hydrogen atoms;

R ¹ and R ³ are hydroxyl groups;

R ² , R ⁴ , R ⁵ , R ⁸ , R ⁹ , Q, L, Y, W, G ¹ , G ² , M and n are as defined in the general formula (I).

Another aspect of the present disclosure relates to a method for preparing a compound represented by general formula (II), which includes the following steps:

The protective group of the compound of general formula (IIA) is removed to obtain the compound of general formula (II),

among them:

Z is -O-;

R ^x is an alkyl group;

R ⁶ and R ⁷ are hydrogen atoms;

R ¹ and R ³ are hydroxyl groups;

R ² , R ⁴ , R ⁵ , R ⁸ , R ⁹ , Q, L, Y, W, G ¹ , G ² , M and n are as defined in the general formula (II).

Another aspect of the present disclosure relates to a method for preparing a compound represented by general formula (III), which includes the following steps:

The protective group of the compound of general formula (IIIA) is removed to obtain the compound of general formula (III),

among them:

R ^x is an alkyl group;

R ⁶ and R ⁷ are hydrogen atoms;

R ⁸ , R ⁹ , Q, L, G ¹ , G ² , M and n are as defined in the general formula (III).

Another aspect of the present disclosure relates to a method for preparing a compound represented by general formula (IV), which includes the following steps:

The protective group of the compound of general formula (IVA) is removed to obtain the compound of general formula (IV),

among them:

R ⁸ , R ⁹ and n are as defined in the general formula (IV).

Another aspect of the present disclosure relates to a pharmaceutical composition containing a therapeutically effective amount of the compound represented by the general formula (I) of the present disclosure or its tautomer, mesosome, racemic Isomers, enantiomers, diastereomers, prodrugs, mixtures or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

The present disclosure further relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or The use of its pharmaceutically acceptable salt or a pharmaceutical composition containing it in the preparation of a medicament for inhibiting CD73.

The present disclosure further relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or The use of its pharmaceutically acceptable salt or a pharmaceutical composition containing it in the preparation of a medicament for treating or preventing CD73-mediated diseases.

The present disclosure further relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or The use of its pharmaceutically acceptable salt or a pharmaceutical composition containing it in the preparation of a medicine for treating or preventing diseases mediated by adenosine and adenosine receptors.

The present disclosure further relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or Its pharmaceutically acceptable salt, or pharmaceutical composition containing it is prepared for the treatment of tumors, autoimmune diseases, immune system diseases, inflammatory diseases, nervous system diseases, neurodegenerative diseases and central nervous system diseases, depression, Use in medicines for Parkinson's disease, cerebral and cardiac ischemic diseases, sleep disorders or fibrosis; wherein the tumor is preferably selected from melanoma, brain tumor, esophageal cancer, stomach cancer, liver cancer, pancreatic cancer, colorectal cancer , Lung cancer (such as non-small cell lung cancer or small cell lung cancer), kidney cancer, breast cancer, ovarian cancer, uterine cancer, endometriosis, prostate cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, Bone cancer, seminoma, testicular tumor, uterine cancer, head and neck cancer, myeloma (such as multiple myeloma), lymphoma, leukemia (such as chronic myelogenous leukemia, chronic lymphocytic leukemia, lymphoblastic leukemia, or acute myeloid leukemia) Leukemia), thyroid cancer, ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, chorioepithelial carcinoma and pediatric tumors.

The present disclosure also relates to a method for inhibiting CD73, which comprises administering to a patient a therapeutically effective amount of a compound represented by general formula (I) or its tautomers, mesosomes, racemates, and enantiomers. Isomers, diastereomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.

The present disclosure also relates to a method for the treatment or prevention of CD73-mediated diseases, which comprises administering to a desired patient a therapeutically effective amount of a compound represented by general formula (I) or its tautomer, mesosome, or exoisomer. Racemates, enantiomers, diastereomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.

The present disclosure also relates to a treatment for tumors, autoimmune diseases, immune system diseases, inflammatory diseases, nervous system diseases, neurodegenerative diseases and central nervous system diseases, depression, Parkinson's disease, cerebral and cardiac hemostatic diseases, A method for sleep disorders or fibrosis, which comprises administering a therapeutically effective amount of a compound represented by general formula (I) or its tautomers, mesosomes, racemates, or enantiomers to a desired patient , Diastereoisomers, prodrugs, mixture forms or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them; wherein the tumor is preferably selected from melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreas Cancer, colorectal cancer, lung cancer (such as non-small cell lung cancer or small cell lung cancer), kidney cancer, breast cancer, ovarian cancer, uterine cancer, endometriosis, prostate cancer, skin cancer, neuroblastoma, sarcoma , Osteochondroma, bone cancer, seminoma, testicular tumor, uterine cancer, head and neck cancer, myeloma (such as multiple myeloma), lymphoma, leukemia (such as chronic myelogenous leukemia, chronic lymphocytic leukemia, lymphoblast Cell leukemia or acute myeloid leukemia), thyroid cancer, ureteral tumors, bladder cancer, gallbladder cancer, duct cancer, chorioepithelial cancer and pediatric tumors

The present disclosure further relates to a compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture Form or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same, which is used as a medicine.

The present disclosure also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or Its pharmaceutically acceptable salt or pharmaceutical composition containing it is used as a CD73 inhibitor.

The present disclosure also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or Its pharmaceutically acceptable salt or pharmaceutical composition containing it is used for the treatment or prevention of CD73-mediated diseases.

The present disclosure also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or Its pharmaceutically acceptable salt or pharmaceutical composition containing it is used for the treatment of tumors, autoimmune diseases, immune system diseases, inflammatory diseases, nervous system diseases, neurodegenerative diseases and central nervous system diseases, depression, Pa Kinsen’s disease, confirmed hematological diseases of the brain and heart, sleep disorders or fibrosis; wherein the tumor is preferably selected from melanoma, brain tumor, esophageal cancer, stomach cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer (such as non-small cell Lung cancer or small cell lung cancer), kidney cancer, breast cancer, ovarian cancer, uterine cancer, endometriosis, prostate cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, bone cancer, seminoma , Testicular cancer, uterine cancer, head and neck cancer, myeloma (such as multiple myeloma), lymphoma, leukemia (such as chronic myelogenous leukemia, chronic lymphocytic leukemia, lymphoblastic leukemia or acute myeloid leukemia), thyroid cancer, Ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, choriocarcinoma and pediatric tumors.

The active compound can be formulated into a form suitable for administration by any appropriate route, and the active compound is preferably in a unit dose form, or a form in which the patient can self-administer in a single dose. The unit dosage of the compound or composition of the present disclosure can be tablet, capsule, cachet, bottled syrup, powder, granule, lozenge, suppository, rejuvenated powder or liquid preparation.

The dosage of the compound or composition used in the treatment methods of the present disclosure will generally vary with the severity of the disease, the weight of the patient, and the relative efficacy of the compound. However, as a general guide, a suitable unit dose may be 0.1-1000 mg.

In addition to the active compound, the pharmaceutical composition of the present disclosure may contain one or more auxiliary materials selected from the following ingredients: fillers (diluents), binders, wetting agents, disintegrants or excipients Wait. Depending on the method of administration, the composition may contain 0.1 to 99% by weight of the active compound.

The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as tablets, dragees, lozenges, water or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or Elixirs. The oral composition may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives, In order to provide pleasing and delicious medicinal preparations. The tablet contains the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for the preparation of tablets for mixing.

Aqueous suspensions contain the active substance and excipients suitable for the preparation of aqueous suspensions for mixing. The aqueous suspension may also contain one or more preservatives such as ethyl paraben or n-propyl paraben, one or more coloring agents, one or more flavoring agents and one or more sweeteners. Flavoring agent.

Oil suspensions can be formulated by suspending active ingredients in vegetable oils. The oil suspension may contain thickeners. The above-mentioned sweeteners and flavoring agents can be added to provide a palatable preparation.

By adding water, dispersible powders and granules suitable for preparing aqueous suspensions can be provided with active ingredients and dispersing or wetting agents for mixing, suspending agents or one or more preservatives. Suitable dispersing or wetting agents and suspending agents can illustrate the above examples. Other excipients such as sweeteners, flavoring agents and coloring agents may also be added. These compositions are preserved by adding antioxidants such as ascorbic acid.

The pharmaceutical composition of the present disclosure may also be in the form of an oil-in-water emulsion.

The pharmaceutical composition may be in the form of a sterile injectable aqueous solution. Acceptable solvents or solvents that can be used include water, Ringer's solution and isotonic sodium chloride solution. The sterile injection preparation may be a sterile injection oil-in-water microemulsion in which the active ingredient is dissolved in an oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. Then the oil solution is added to a mixture of water and glycerin to form a microemulsion. The injection or microemulsion can be injected into the patient's bloodstream by local large-scale injection. Alternatively, it is best to administer the solution and microemulsion in a manner that maintains a constant circulating concentration of the compound of the present disclosure. To maintain this constant concentration, a continuous intravenous delivery device can be used. An example of such a device is the Deltec CADD-PLUS.TM. 5400 intravenous pump.

The pharmaceutical composition may be in the form of a sterile injection water or oil suspension for intramuscular and subcutaneous administration. The suspension can be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents mentioned above. The sterile injection preparation can also be a sterile injection solution or suspension prepared in a parenterally acceptable non-toxic diluent or solvent. In addition, sterile fixed oil can be conveniently used as a solvent or suspension medium.

The compounds of the present disclosure can be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid in the rectum and thus will melt in the rectum to release the drug. Such substances include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycol.

As is well known to those skilled in the art, the dosage of the drug depends on many factors, including but not limited to the following factors: the activity of the specific compound used, the age of the patient, the weight of the patient, the health of the patient, and the behavior of the patient , The patient’s diet, time of administration, mode of administration, rate of excretion, combination of drugs, etc.; in addition, the best mode of treatment such as the mode of treatment, the daily dosage of the compound (I) or the amount of pharmaceutically acceptable salt The type can be verified according to the traditional treatment plan.

Detailed description of the invention

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably containing 1 to 6 carbon atoms Atom of the alkyl group. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1 ,2-Dimethylpropyl, 2,2-Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2- Methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3 -Dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2 -Methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethyl Pentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2 ,5-Dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethyl 2-methylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethyl Hexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers. More preferred are lower alkyl groups containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and sec-butyl. Group, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Butyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl Group, 2,3-dimethylbutyl, etc. Alkyl groups can be substituted or unsubstituted. When substituted, substituents can be substituted at any available point of attachment. The substituents are independently optionally selected from halogen, alkyl, haloalkyl, alkoxy Group, haloalkoxy, alkylthio, alkylamino, alkenyl, alkynyl, mercapto, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, heteroaryl , Cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo group substituted by one or more substituents.

The term "alkylene" refers to a saturated linear or branched aliphatic hydrocarbon group, which has two residues derived from the removal of two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkane, which is A straight or branched chain group containing 1 to 20 carbon atoms, preferably containing 1 to 12 carbon atoms, more preferably an alkylene group containing 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene Group (-CH2CH2CH2CH2-) and so on. The alkylene group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably independently optionally selected from alkyl, alkenyl, alkynyl , Alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy Substituted by one or more substituents among thiol, cycloalkylthio, heterocycloalkylthio and oxo.

The term "alkenyl" refers to an alkyl compound containing a carbon-carbon double bond in the molecule, wherein the definition of the alkyl group is as described above. Alkenyl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from hydrogen atoms, alkyl groups, alkoxy groups, halogens, halogenated alkyl groups, hydroxyl groups, It is substituted by one or more substituents among hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl.

The term "alkynyl" refers to an alkyl compound containing a carbon-carbon triple bond in the molecule, wherein the definition of the alkyl group is as described above. The alkynyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from hydrogen atoms, alkyl groups, alkoxy groups, halogens, halogenated alkyl groups, hydroxyl groups, It is substituted by one or more substituents among hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl and heteroaryl.

The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where alkyl and cycloalkyl are as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is independently optionally selected from halogen, alkyl, haloalkyl , Alkoxy, haloalkoxy, alkylthio, alkylamino, alkenyl, alkynyl, mercapto, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, One or more substituents among heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo groups are substituted.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 Carbon atoms (e.g. 3, 4, 5, 6, 7 or 8 carbon atoms), most preferably 5 to 7 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Groups, cyclooctyl groups, etc.; polycyclic cycloalkyls include spiro, fused, and bridged cycloalkyls.

The term "spirocycloalkyl" refers to a polycyclic group that shares one carbon atom (called a spiro atom) between 5- to 20-membered monocyclic rings, which may contain one or more double bonds, but none of the rings have complete conjugate Π electronic system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of shared spiro atoms between the ring and the ring, the spirocycloalkyl group is classified into a single spirocycloalkyl group, a bispirocycloalkyl group or a polyspirocycloalkyl group, preferably a single spirocycloalkyl group and a bispirocycloalkyl group. More preferably, it is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, or 5-membered/6-membered monospirocycloalkyl. Non-limiting examples of spirocycloalkyl groups include:

The term "fused cycloalkyl" refers to a 5- to 20-membered all-carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more rings may contain one or Multiple double bonds, but none of the rings have a fully conjugated π-electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyls, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic alkyl. Non-limiting examples of fused cycloalkyl groups include:

The term "bridged cycloalkyl" refers to a 5- to 20-membered, all-carbon polycyclic group with any two rings sharing two carbon atoms that are not directly connected. It may contain one or more double bonds, but no ring has complete Conjugated π electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

The cycloalkyl ring includes the above-mentioned cycloalkyl group fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples include indanyl (Phenocyclopentyl), tetrahydronaphthyl, phenylcyclopentyl, benzocycloheptyl, etc.; preferably indanyl and tetrahydronaphthyl. Cycloalkyl groups can be optionally substituted or unsubstituted. When substituted, the substituents can be substituted at any available point of attachment. The substituents are independently optionally selected from halogen, alkyl, haloalkyl , Alkoxy, haloalkoxy, alkylthio, alkylamino, alkenyl, alkynyl, mercapto, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, One or more substituents among heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo groups are substituted.

The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent which contains 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) _m (where m is an integer of 0 to 2) heteroatoms, but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably contains 5 to 6 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrazole Hydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably tetrahydropyranyl, piperidinyl, and pyrrolidinyl. Polycyclic heterocyclic groups include spiro, fused, and bridged heterocyclic groups.

The term "spiroheterocyclic group" refers to a polycyclic heterocyclic group that shares one atom (called a spiro atom) between 5- to 20-membered monocyclic rings, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O ) _m (where m is an integer of 0 to 2) heteroatoms, and the remaining ring atoms are carbon. It can contain one or more double bonds, but none of the rings have a fully conjugated π-electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the ring and the ring, the spiro heterocyclic group is classified into a single spiro heterocyclic group, a dispiro heterocyclic group or a polyspiro heterocyclic group, preferably a single spiro heterocyclic group and a dispiro heterocyclic group. More preferably, it is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monospiro heterocyclic group. Non-limiting examples of spiroheterocyclic groups include:

The term "fused heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system. One or more rings may contain one or more Double bond, but none of the rings have a fully conjugated π-electron system, one or more of the ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (where m is an integer from 0 to 2), and the rest of the ring The atom is carbon. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

The term "bridged heterocyclic group" refers to a 5- to 14-membered polycyclic heterocyclic group with any two rings sharing two atoms that are not directly connected. It may contain one or more double bonds, but none of the rings has a complete common A conjugated π-electron system in which one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O) _m (where m is an integer of 0 to 2), and the remaining ring atoms are carbon. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan (for example, 7, 8, 9 or 10 yuan). According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

The heterocyclic ring includes the above heterocyclic groups (including monocyclic and polycyclic heterocyclic groups, such as spiro heterocyclic groups, fused heterocyclic groups and bridged heterocyclic groups) fused to aryl, heteroaryl or cycloalkane On the base ring, the ring connected to the parent structure is a heterocyclic group, non-limiting examples of which include:

The heterocyclic group may be optionally substituted or unsubstituted. When substituted, the substituent may be substituted at any available attachment point. The substituent is independently optionally selected from halogen, alkyl, haloalkyl , Alkoxy, haloalkoxy, alkylthio, alkylamino, alkenyl, alkynyl, mercapto, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, One or more substituents among heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo groups are substituted.

The term "aryl" refers to a 6 to 14-membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) with a conjugated π-electron system, preferably 6 to 10-membered, more preferably 5 to 6 members, such as phenyl and naphthyl. The aryl ring includes the above-mentioned aryl group fused on a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and non-limiting examples thereof include:

Aryl groups can be substituted or unsubstituted. When substituted, the substituents can be substituted at any available point of attachment. The substituents are independently optionally selected from halogen, alkyl, haloalkyl, and alkoxy. Group, haloalkoxy, alkylthio, alkylamino, alkenyl, alkynyl, mercapto, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, heteroaryl , Cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo group substituted by one or more substituents.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, where the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl groups are preferably 5 to 10 members, containing 1 to 3 heteroatoms; more preferably 5 or 6 members, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furyl, thienyl, thiazolyl, pyridine Azolyl, oxazolyl, pyrrolyl, 1H-1,2,3-triazolyl, 4H-1,2,4-triazolyl, 4H-1,2,3-triazolyl, 1H-tetrazole Group, 2H-tetrazolyl, 5H-tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably pyrazolyl or Imidazolyl. The heteroaryl ring includes the above-mentioned heteroaryl group fused on an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples thereof include:

Heteroaryl groups can be optionally substituted or unsubstituted. When substituted, the substituents can be substituted at any available point of attachment. The substituents are independently optionally selected from halogen, alkyl, haloalkyl , Alkoxy, haloalkoxy, alkylthio, alkylamino, alkenyl, alkynyl, mercapto, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, One or more substituents among heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo groups are substituted.

The term "amino protecting group" is to keep the amino group unchanged when other parts of the molecule react, and to protect the amino group with a group that is easy to remove. Non-limiting examples include tert-butoxycarbonyl, acetyl, benzyl, allyl, 2,4-dimethoxybenzyl, p-methoxybenzyl, and the like. These groups may be optionally substituted with 1-3 substituents selected from halogen, alkoxy or nitro. The amino protecting group is preferably 2,4-dimethoxybenzyl.

The term "cycloalkyloxy" refers to cycloalkyl-O-, where cycloalkyl is as defined above.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, where the alkyl group is as defined above.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, where alkoxy is as defined above.

The term "deuterated alkyl" refers to an alkyl group substituted with one or more deuterium atoms, where the alkyl group is as defined above.

The term "hydroxy" refers to the -OH group.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, where the alkyl group is as defined above.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "hydroxy" refers to the -OH group.

The term "amino" refers to -NH ₂ .

The term "cyano" refers to -CN.

The term "azido" refers to a -N = N ⁺ = N ^-.

The term "nitro" refers to -NO ₂ .

The term "carbonyl" refers to C=O.

The term "carboxy" refers to -C(O)OH.

The term "carboxylate group" refers to -C(O)O(alkyl) or -C(O)O(cycloalkyl), where alkyl and cycloalkyl are as defined above.

The present disclosure also includes compounds of formula (I) in various deuterated forms. Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize the compound of formula (I) in the deuterated form with reference to relevant literature. When preparing the deuterated form of the compound of formula (I), commercially available deuterated starting materials can be used, or they can be synthesized using conventional techniques using deuterated reagents. Deuterated reagents include, but are not limited to, deuterated borane and tri-deuterated. Borane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated ethyl iodide and deuterated methyl iodide, etc.

"Optional" or "optionally" means that the event or environment described later can but does not have to occur, and the description includes the occasion where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted by an alkyl group" means that an alkyl group may but does not have to be present. The description includes the case where the heterocyclic group is substituted by an alkyl group and the case where the heterocyclic group is not substituted by an alkyl group. .

"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms independently of each other substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without too much effort. For example, an amino group or a hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (eg, olefinic) bond.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, and other components such as physiological/pharmaceutically acceptable carriers And excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and thus the biological activity.

"Pharmaceutically acceptable salt" refers to the salt of the compound of the present disclosure. Such salt is safe and effective when used in mammals, and has due biological activity.

Synthetic method of the compound of the present disclosure

In order to accomplish the purpose of the present disclosure, the present disclosure adopts the following technical solutions:

Option One

The compound represented by general formula (I) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its The preparation method of medicinal salt includes the following steps:

among them:

Z is -O-;

R ^x is an alkyl group;

R ⁶ and R ⁷ are hydrogen atoms;

R ¹ and R ³ are hydroxyl groups;

Reagents for deprotection include but are not limited to trifluoroacetic acid, trimethylsilyl bromide, and hydrochloric acid.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, trifluoroacetic acid, methanol, ethanol, n-butanol, tert-butanol, toluene, tetrahydrofuran, methylene chloride, petroleum ether, ethyl acetate, n-hexane , Dimethyl sulfoxide, 1,4-dioxane, ethylene glycol dimethyl ether, water or N,N-dimethylformamide and mixtures thereof.

Option II

The compound represented by general formula (II) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its The preparation method of medicinal salt includes the following steps:

among them:

Z is -O-;

R ^x is an alkyl group;

R ⁶ and R ⁷ are hydrogen atoms;

R ¹ and R ³ are hydroxyl groups;

third solution

The compound represented by general formula (III) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its The preparation method of medicinal salt includes the following steps:

among them:

R ^x is an alkyl group;

R ⁶ and R ⁷ are hydrogen atoms;

Option Four

The compound represented by general formula (IV) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its The preparation method of medicinal salt includes the following steps:

among them:

R ⁸ , R ⁹ and n are as defined in the general formula (IV).

detailed description

The present disclosure is further described below in conjunction with embodiments, but these embodiments do not limit the scope of the present disclosure.

Example

The structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 ^-6 (ppm). NMR was measured with Bruker AVANCE-400 and 500 Ultrashield nuclear magnetometers, and the solvents were deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), and deuterated methanol (CD ₃ OD). Labeled as tetramethylsilane (TMS).

The MS is measured with FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantageMAX) and Shimadzu LCMS-2020 liquid chromatography-mass spectrometer.

HPLC determination uses Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18 150×4.6mm column), Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150×4.6mm column) and Shimadzu OPTIONBOX-L high pressure liquid chromatography Instrument (Gimini 5um NX-C18 100x21.2mm chromatographic column).

Chiral HPLC analysis and determination use LC-10A vp (Shimadzu) or SFC-analytical (Berger Instruments Inc.).

The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm～0.2mm, and the size of thin layer chromatography separation and purification products is 0.4mm ~0.5mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

Chiral preparative column chromatography uses Prep Star SD-1 (Varian Instruments Inc.) or SFC-multigram (Berger Instruments Inc.).

CombiFlash rapid preparation instrument uses Combiflash Rf200 (TELEDYNE ISCO).

The average value of ₅₀ measured kinase inhibition rate and IC NovoStar using a microplate reader (BMG, Germany).

The known starting materials of the present disclosure can be synthesized by or according to methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Darui Chemicals, Shanghai Bi De Pharmaceutical Technology Co., Ltd. and other companies.

There is no special description in the examples, and the reaction can be carried out in an argon atmosphere or a nitrogen atmosphere.

The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1L.

The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.

The pressure hydrogenation reaction uses Parr 3916EKX hydrogenator and Qinglan QL-500 hydrogen generator or HC2-SS hydrogenator.

The hydrogenation reaction is usually evacuated, filled with hydrogen, and repeated three times.

The microwave reaction uses CEM Discover-S 908860 type microwave reactor.

There is no special description in the examples, and the solution refers to an aqueous solution.

There are no special instructions in the examples, and the reaction temperature is room temperature, which is 20°C to 30°C.

The monitoring of the reaction progress in the examples adopts thin layer chromatography (TLC), the developing reagent used in the monitoring reaction, the eluent system of column chromatography and the developing reagent system of thin layer chromatography used to purify the compound include: A: Dichloromethane/methanol system, B: n-hexane/ethyl acetate system, C: dichloromethane/ethyl acetate system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triacetate can also be added Adjust with alkaline or acidic reagents such as ethylamine and acetic acid.

Example 1

((((2R,3S,4R,5R)-5-((6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl )-3,4-Dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

6-Chloro-N-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine 1b

The compound 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine 1a (1.75g, 9.26mmol, finished) was added to methanol solvent (45.0mL), and triethylamine ( 1.40g, 1.49mmol) and cyclopentylamine (790mg, 9.28mmol) were stirred and reacted for 6 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1b (690 mg, yield: 31%).

MS m/z(ESI): 238.1[M+1].

Second step

(Diethoxyphosphono)methyl 2-nitrobenzenesulfonate 1d

Add diethyl (hydroxymethyl) phosphonate 1c (1.00g, 5.95mmol, Sinopharm) and pyridine (941mg, 11.90mmol) into dichloromethane solvent (40.0mL), add 2-nitrobenzenesulfonyl chloride ( 1.58g, 7.14mmol), the reaction was stirred for 2 hours. Ethyl acetate (50 mL) and water (50 mL) were added, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1d (1.50 g, yield: 71%).

¹ H NMR (400MHz, CDCl ₃ ) δ 8.16-8.18 (m, 1H), 7.80-7.87 (m, 3H), 4.45-4.47 (m, 2H), 4.17-4.24 (m, 4H), 1.33-1.36 (m,6H).

third step

Tert-butyl(((3aR,4R,6S,6aR)-6-(iodomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol (Alk-4-yl)methoxy)dimethylsilane 1f

Add ((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][ 1,3] Dioxolane-4-yl)methanol 1e (4.00g, 12.56mmol, prepared by the method disclosed in the document "Tetrahedron, 2012, 68, 1540-1546"), iodine (7.01g, 27.63mmol), triphenylphosphine (6.59g, 25.12mmol) and imidazole (2.57g, 37.68mmol) were added to toluene solvent (50.0mL), and the mixture was heated at 110°C for 3 hours. After cooling to room temperature, ethyl acetate (50 mL) and water (50 mL) were added, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1f (5.10 g, yield: 94%).

MS m/z(ESI): 429.1[M+1].

the fourth step

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-N-cyclopentyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine 1g

Cesium carbonate (10.04g, 30.81mmol) was added to a solution of compound 1b (2.44g, 10.27mmol) and compound 1f (4.40g, 10.27mmol) in N,N-dimethylformamide (35.0mL), the mixture The reaction was heated at 70°C for 36 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain 1 g of the title compound (1.00 g, yield: 18%).

MS m/z(ESI): 538.3[M+1].

the fifth step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)- 2,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methanol 1h

N-Tetrabutylammonium fluoride (628 mg, 2.79 mmol) was added to a tetrahydrofuran (20.0 mL) solution of compound 1 g (1.00 g, 1.86 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1h (610 mg, yield: 77%).

MS m/z(ESI): 424.2[M+1].

Sixth step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl )-2,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxolane-4-yl)methoxy)methyl)diethyl phosphonate 1i

Mix compound 1h (200mg, 0.47mmol), compound 1d (250mg, 0.71mmol) and dry N,N-dimethylformamide solvent (5.0mL) together, add magnesium tert-butoxide (241mg, 1.41mmol) . The mixture was heated at 70°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 1i (200 mg, yield: 74%).

MS m/z(ESI): 574.2[M+1].

Seventh step

Compound 1i (800mg, 1.39mmol) was added to dry dichloromethane solvent (40.0mL), trimethylsilyl bromide (2.13g, 13.94mmol) was added to the solution, the reaction was stirred for 4 hours, and then at 0°C Add saturated sodium bicarbonate solution (2.0 mL) and stir for 10 minutes. The reaction solution was concentrated under reduced pressure, and the residue obtained was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18 100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 1 (400 mg, yield: 60%).

MS m/z(ESI):478.1[M+1].

¹ H NMR (400MHz, CD ₃ OD) δ8.06 (s, 1H), 4.50-4.58 (m, 4H), 4.16-4.18 (m, 2H), 4.14-4.15 (m, 1H), 3.56-3.65 ( m, 4H), 2.07-2.10 (m, 2H), 1.29-1.68 (m, 6H).

Example 2

((((2R,3S,4R,5R)-5-((6-chloro-4-(((S)-1-phenethyl)amino)-1H-pyrazolo[3,4-d] (Pyrimidine-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

(Di-tert-Butoxyphosphono)methyl 2-nitrobenzenesulfonate 2b

Di-tert-butyl (hydroxymethyl) phosphonate 2a (1.00g, 4.46mmol, prepared by the method disclosed in the patent application "US20140088314") and pyridine (706mg, 8.92mmol) were added to the dichloromethane solvent (40.0mL ), add 2-nitrobenzenesulfonyl chloride (1.58 g, 7.14 mmol), stir and react for 2 hours. Ethyl acetate (50 mL) and water (50 mL) were added, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2b (1.20 g, yield: 66%).

¹ H NMR (400MHz, CDCl ₃ ) δ 8.15-8.17 (m, 1H), 7.82-7.83 (m, 3H), 4.26-4.29 (m, 2H), 1.51 (s, 18H).

Second step

(S)-6-chloro-N-(1-phenethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine 2c

Compound 1a (2.00 g, 10.58 mmol) was added to acetonitrile solvent (20.0 mL), and sodium bicarbonate (1.78 g, 21.16 mmol) and (S)-1-phenylethylamine (1.28 g, 10.58 mmol) were added to the solution , The reaction was stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2c (1.00 g, yield: 35%).

MS m/z(ESI): 274.0[M+1].

third step

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-N-((S)-1-phenethyl)-1H-pyrazolo[3,4-d ]Pyrimidine-4-amine 2d

Cesium carbonate (1.43g, 4.38mmol) was added to the N,N-dimethylformamide (10.0mL) solution of compound 2c (400mg, 1.46mmol) and compound 1f (688mg, 1.61mmol), and the mixture was heated at 70 The reaction was heated at °C for 36 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2d (260 mg, yield: 31%).

MS m/z(ESI): 574.3[M+1].

the fourth step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(((S)-1-phenethyl)amino)-1H-pyrazolo[3,4-d]pyrimidine- 1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methanol 2e

N-Tetrabutylammonium fluoride (95 mg, 0.42 mmol) was added to the tetrahydrofuran (3.0 mL) solution of compound 2d (160 mg, 0.28 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2e (120 mg, yield: 94%).

MS m/z(ESI): 457.9[M-1].

the fifth step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(((S)-1-phenethyl)amino)-1H-pyrazolo[3,4-d] Pyrimidine-1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)methyl)phosphine Di-tert-butyl ester 2f

Mix compound 2e (100mg, 0.22mmol), compound 2b (160mg, 0.39mmol) and dry N,N-dimethylformamide solvent (3.0mL) together, add magnesium tert-butoxide (112mg, 0.66mmol) . The mixture was heated at 70°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 2f (100 mg, yield: 69%).

MS m/z(ESI): 665.9[M+1].

Sixth step

((((2R,3S,4R,5R)-5-((6-chloro-4-(((S)-1-phenethyl)amino)-1H-pyrazolo[3,4-d] (Pyrimidine-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid 2

Compound 2f (22 mg, 0.033 mmol) was added to the trifluoroacetic acid solvent (2.0 mL), water (0.5 mL) was added to the solution, and the reaction was stirred for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 2 (12 mg, yield: 71%).

MS m/z(ESI): 513.8[M+1].

¹ H NMR (400MHz, CD ₃ OD) δ8.11 (s, 1H), 7.42-7.44 (m, 2H), 7.31-7.35 (m, 2H), 7.24-7.26 (m, 1H), 5.53-5.54 ( m, 1H), 4.45-4.48 (m, 3H), 4.06-4.16 (m, 3H), 3.65-3.69 (m, 4H), 1.61-1.62 (m, 3H).

Example 3

((((2R,3S,4R,5R)-5-((6-chloro-4-(((R)-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyridine Azolo[3,4-d]pyrimidin-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

(R)-6-chloro-N-(2,3-dihydro-1H-inden-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine 3a

Compound 1a (2.22g, 11.74mmol) was added to acetonitrile solvent (50.0mL), to this solution was added triethylamine (3.56g, 35.24mmol) and (R)-2,3-dihydro-1H-indene-1 -Amine hydrochloride (2.0g, 11.78mmol, finished), stirred and reacted for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 3a (2.00 g, yield: 59%).

MS m/z(ESI): 286.1[M+1].

Second step

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-N-((R)-2,3-dihydro-1H-inden-1-yl)-1H- Pyrazolo[3,4-d]pyrimidin-4-amine 3b

Cesium carbonate (3.42g, 10.49mmol) was added to the N,N-dimethylformamide (20.0mL) solution of compound 3a (1.00g, 3.49mmol) and compound 1f (1.49g, 3.47mmol), and the mixture The reaction was heated at 70°C for 16 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 3b (600 mg, yield: 29%).

MS m/z(ESI): 586.3[M+1].

third step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(((R)-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyrazolo [3,4-d]pyrimidin-1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxolane-4-yl)methanol 3c

N-Tetrabutylammonium fluoride (144 mg, 0.64 mmol) was added to the tetrahydrofuran (30.0 mL) solution of compound 3b (250 mg, 0.43 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 3c (150 mg, yield: 75%).

MS m/z(ESI): 470.1[M-1].

the fourth step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(((R)-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyridine Azolo[3,4-d]pyrimidin-1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxolane-4-yl )Methoxy)methyl)di-tert-butylphosphonate 3d

Compound 3c (100 mg, 0.21 mmol) and compound 2b (104 mg, 0.25 mmol) were dissolved in dry N,N-dimethylformamide solvent (10.0 mL), and magnesium tert-butoxide (108 mg, 0.63 mmol) was added. The mixture was heated and reacted at 60°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 3d (100 mg, yield: 70%).

MS m/z(ESI): 677.9[M+1].

the fifth step

((((2R,3S,4R,5R)-5-((6-chloro-4-(((R)-2,3-dihydro-1H-inden-1-yl)amino)-1H-pyridine Azolo[3,4-d]pyrimidin-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid 3

Compound 3d (100 mg, 0.15 mmol) was added to dichloromethane (5.0 mL), trifluoroacetic acid (1.0 mL) was added, water (0.2 mL) was added to the solution, and the reaction was stirred for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18 100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 3 (20 mg, yield: 26%).

MS m/z(ESI): 524.2[M-1].

¹ H NMR (400MHz, DMSO-d ₆ ) δ9.11-9.13 (m, 1H), 8.16 (s, 1H), 7.16-7.31 (m, 4H), 5.79-5.85 (m, 1H), 4.42 to 4.51 (m, 4H), 4.27-4.31 (m, 1H), 3.99-4.05 (m, 2H), 3.88 (s, 2H), 3.47-3.50 (m, 3H), 3.26-3.35 (m, 2H), 2.87 -3.32(m,3H),1.94-2.00(m,1H).

Example 4

(((((2R,3S,4R,5R)-5-((6-chloro-4-(propylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl) -3,4-Dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

6-chloro-N-n-propyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine 4a

Compound 1a (2.0 g, 10.58 mmol) was added to acetonitrile solvent (50.0 mL), triethylamine (3.21 g, 31.72 mmol) and n-propylamine (625 mg, 10.57 mmol) were added to the solution, and the reaction was stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 4a (1.5 g, yield: 67%).

MS m/z(ESI): 212.0[M+1].

Second step

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-N-n-propyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine 4b

Cesium carbonate (2.47g, 7.57mmol) was added to compound 4a (535mg, 2.53mmol) and compound 1f (2.16g, 5.04mmol) in dimethyl sulfoxide (20.0mL) solution, and the mixture was heated at 80°C for reaction 16 hours. After cooling to ambient temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 4b (220 mg, yield: 17%).

MS m/z(ESI): 512.3[M+1].

third step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(propylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2 ,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methanol 4c

N-Tetrabutylammonium fluoride (145 mg, 0.64 mmol) was added to the tetrahydrofuran (20.0 mL) solution of compound 4b (220 mg, 0.43 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 4c (60 mg, yield: 35%).

MS m/z(ESI): 398.1[M+1].

the fourth step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(propylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl) -2,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxolane-4-yl)methoxy)methyl)di-tert-butyl phosphonate 4d

Mix compound 4c (60mg, 0.15mmol), compound 2b (92mg, 0.22mmol) and dry N,N-dimethylformamide solvent (10.0mL) together, add magnesium tert-butoxide (76mg, 0.44mmol) . The mixture was heated and reacted at 60°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 4d (50 mg, yield: 55%).

MS m/z(ESI): 604.1[M+1].

the fifth step

(((((2R,3S,4R,5R)-5-((6-chloro-4-(propylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl) -3,4-Dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid 4

Compound 4d (50 mg, 0.082 mmol) was added to dichloromethane (5.0 mL), trifluoroacetic acid (1.0 mL) was added, water (0.2 mL) was added to the solution, and the reaction was stirred for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18 100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 4 (10 mg, yield: 27%).

MS m/z(ESI): 452.1[M+1].

¹ H NMR (400MHz, CD ₃ OD) δ 8.06 (s, 1H), 4.51-4.54 (m, 2H), 4.43-4.44 (m, 1H), 4.11-4.17 (m, 3H), 3.49-3.71 ( m, 6H), 1.67-1.73 (m, 2H), 0.98-1.02 (m, 3H).

Example 5

((((2R,3S,4R,5R)-5-((6-chloro-4-(cycloheptylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl )-3,4-Dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

6-Chloro-N-cycloheptyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine 5a

Compound 1a (2.00 g, 10.58 mmol) was added to acetonitrile solvent (20.0 mL), sodium bicarbonate (1.78 g, 21.16 mmol) and cycloheptylamine (1.45 g, 12.81 mmol) were added to the solution, and the reaction was stirred for 16 hours. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 5a (2.00 g, yield: 71%).

MS m/z(ESI): 266.2[M+1].

Second step

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-N-cycloheptyl-1H-pyrazolo[3,4-d]pyrimidin-4-amine 5b

Cesium carbonate (1.43g, 4.38mmol) was added to the N,N-dimethylformamide (5.0mL) solution of compound 5a (200mg, 0.753mmol) and compound 1f (387mg, 0.903mmol), and the mixture was heated at 70 The reaction was heated at °C for 12 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 5b (220 mg, yield: 52.0%).

MS m/z(ESI): 566.2[M+1].

third step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(cycloheptylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)- 2,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methanol 5c

N-Tetrabutylammonium fluoride (600 mg, 2.66 mmol) was added to the tetrahydrofuran (5.0 mL) solution of compound 5b (300 mg, 0.28 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 5c (60 mg, yield: 25%).

MS m/z(ESI): 452.2[M+1].

the fourth step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(cycloheptylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl )-2,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)methyl)di-tert-butyl phosphonate 5d

Mix compound 5c (55mg, 0.122mmol), compound 2b (75mg, 0.18mmol) and dry N,N-dimethylformamide solvent (3.0mL) together, add magnesium tert-butoxide (52mg, 0.30mmol) . The mixture was heated at 70°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 5d (35 mg, yield: 44%).

MS m/z(ESI): 657.9[M+1].

the fifth step

((((2R,3S,4R,5R)-5-((6-chloro-4-(cycloheptylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl )-3,4-Dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid 5

Compound 5d (35 mg, 0.053 mmol) was added to trifluoroacetic acid solvent (2.0 mL), water (0.2 mL) was added to the solution, and the reaction was stirred for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 5. (15 mg, yield: 56%).

MS m/z(ESI): 506.2[M+1].

¹ H NMR (400MHz, CD ₃ OD) δ8.06 (s, 1H), 4.51-4.53 (m, 2H), 4.42-4.47 (m, 1H), 4.15-4.17 (m, 1H), 4.14-4.15 ( m, 2H), 4.07 (br, 1H), 3.57-3.67 (m, 4H), 2.01-2.04 (m, 2H), 1.28-1.76 (m, 12H).

Example 6

((((2R,3S,4R,5R)-5-((6-chloro-4-(2-azaspiro[3.5]non-2-yl)-1H-pyrazolo[3,4-d ]Pyrimidin-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

6-chloro-4-(2-azaspiro[3.5]non-2-yl)-1H-pyrazolo[3,4-d]pyrimidine 6a

Compound 1a (1.50g, 7.94mmol) was added to acetonitrile solvent (20.0mL), and sodium bicarbonate (1.65g, 11.94mmol) and 2-azaspiro[3.5]nonane (1.00g, 7.99mmol) were added to the solution. , Shaoyuan), the reaction was stirred for 16 hours. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 6a (2.00 g, yield: 91%).

MS m/z(ESI): 278.2[M+1].

Second step

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-4-(2-azaspiro[3.5]non-2-yl)-1H-pyrazolo[3 ,4-d]pyrimidine 6b

Cesium carbonate (1.20g, 3.69mmol) was added to compound 6a (500mg, 1.80mmol) and compound 1f (0.78mg, 1.82mmol) in N,N-dimethylformamide (10.0mL) solution, the mixture was added to The reaction was heated at 70°C for 12 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 6b (800 mg, yield: 77%).

MS m/z(ESI): 578.2[M+1].

third step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(2-azaspiro[3.5]non-2-yl)-1H-pyrazolo[3,4-d]pyrimidine -1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxolane-4-yl)methanol 6c

N-Tetrabutylammonium fluoride (153 mg, 0.68 mmol) was added to the tetrahydrofuran (5.0 mL) solution of compound 6b (250 mg, 0.43 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 6c (180 mg, yield: 90%).

MS m/z(ESI): 464.2[M+1].

the fourth step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(2-azaspiro[3.5]non-2-yl)-1H-pyrazolo[3,4-d ]Pyrimidin-1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)methyl) Di-tert-butyl phosphonate 6d

Compound 6c (100 mg, 0.22 mmol), compound 2b (133 mg, 0.32 mmol) and N,N-dimethylformamide solvent (3.0 mL) were mixed together, and magnesium tert-butoxide (74 mg, 0.43 mmol) was added. The mixture was heated at 70°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 6d (110 mg, yield: 76%).

MS m/z(ESI): 669.9[M+1].

the fifth step

((((2R,3S,4R,5R)-5-((6-chloro-4-(2-azaspirocyclo[3.5]non-2-yl)-1H-pyrazolo[3,4- d)pyrimidin-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid 6

Compound 6d (50 mg, 0.075 mmol) was added to trifluoroacetic acid solvent (2.0 mL), water (0.2 mL) was added to the solution, and the reaction was stirred at room temperature for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18 100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 6 (15 mg, yield: 39%).

MS m/z(ESI): 518.6[M+1].

¹ H NMR (400MHz, CD ₃ OD) δ7.98 (s, 1H), 4.57-4.61 (m, 1H), 4.44-4.46 (m, 2H), 4.09-4.14 (m, 4H), 4.05 (br, 1H), 3.95 (s, 2H), 3.71-3.73 (d, 3H), 3.64-3.65 (m, 1H), 1.75-1.77 (m, 4H), 1.52-1.56 (m, 4H), 1.45-1.47 ( m,2H).

Example 7

(((2R,3S,4R,5R)-5-((6-chloro-4-(2',3'-dihydrospiro[azetidine-3,1'-indenyl]-1- Yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid

first step

2,3-Dihydro-1H-indene-1-carboxylic acid methyl ester 7b

2,3-Dihydro-1H-indene-1-carboxylic acid 7a (10.00g, 61.65mmol, finished) was dissolved in methanol solvent (200.0mL), concentrated sulfuric acid (3.0mL) was added dropwise, and heated to 60℃ , Stir for 18 hours. The reaction solution was quenched by adding water (70 mL), extracted with ethyl acetate (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The title compound 7b (8.00 g, yield 73%) was obtained.

Second step

1-(hydroxymethyl)-2,3-dihydro-1H-indene-1-carboxylic acid 7c

Compound 7b (7.00g, 39.72mmol), formaldehyde (9.67g, 119.18mmol, purity 37%) and potassium carbonate (16.47g, 119.17mmol) were added to dimethyl sulfoxide (70.0mL), and the reaction was stirred for 18 hours . 20 mL of water was added to the reaction solution, and extraction was performed with ethyl acetate (70 mL). The pH of the aqueous phase was adjusted to about 3 with 1N hydrochloric acid, extracted with ethyl acetate (70 mL), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The title compound 7c (6 g, yield 78%) was obtained.

MS m/z(ESI): 191.0[M-1].

third step

N-benzyl-1-(hydroxymethyl)-2,3-dihydro-1H-indene-1-carboxamide 7d

Compound 7c (6.00g, 31.21mmol), phenylmethylamine (3.34g, 31.21mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (8.97g, 46.82mmol), 1-hydroxybenzotriazole (7.12g, 46.82mmol), and diisopropylethylamine (8.06g, 62.43mmol) were added to the N,N-dimethylformamide solvent (60.0mL) , The reaction was stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7d (8.00 g, yield 91%).

MS m/z(ESI): 282.0[M+1].

the fourth step

(1-(Benzylcarbamoyl)-2,3-dihydro-1H-inden-1-yl)methanesulfonate 7e

Compound 7d (8.00g, 28.43mmol) and triethylamine (2.87g, 28.43mmol) were dissolved in dichloromethane solvent (60.0mL), and methylsulfonyl chloride (3.25g, 28.44mmol) was added dropwise at 0°C, The reaction was stirred for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7e (8.00 g, yield 78%).

MS m/z(ESI): 360.1[M+1].

the fifth step

1-Benzyl-2',3'-dihydrospiro[azetidine-3,1'-indene]-2-one 7f

Compound 7e (8.00 g, 22.25 mmol) and potassium carbonate (6.14 g, 44.51 mmol) were dissolved in acetonitrile solvent (100.0 mL), and heated to 70° C. and stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7f (5.00 g, yield 85%).

MS m/z(ESI): 264.0[M+1].

Sixth step

1-Benzyl-2',3'-dihydrospiro[azetidine-3,1'-indene] 7g

Aluminum trichloride (5.06 g, 37.98 mmol) was dissolved in tetrahydrofuran solvent (20.0 mL), lithium tetrahydroaluminum (2.16 g, 56.96 mmol) was added in portions at 0°C, and the reaction was stirred at 0°C for 1 hour. A pre-prepared tetrahydrofuran solution of 10 mL of compound 7f (5.00 g, 18.98 mmol) was added dropwise, and the mixture was stirred at room temperature for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7g (4.00g, yield 84%).

MS m/z(ESI): 250.2[M+1].

Seventh step

2',3'-Dihydrospiro[azetidine-3,1'-indene]7h

Compound 7g (4.00g, 16.04mmol), ammonium formate (3.04g, 48.13mmol), palladium hydroxide (1.12g, 1.60mmol, purity 20%) were added to methanol solvent (100.0mL), and heated to Stir at 70°C for 18 hours. Methanol (50 mL) was added, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system A to obtain the title compound 7h (1.90 g, yield 74%).

MS m/z(ESI): 160.2[M+1].

Eighth step

6-chloro-4-(2',3'-dihydrospiro[azetidine-3,1'-indenyl]-1-yl)-1H-pyrazolo[3,4-d]pyrimidine 7i

Compound 1a (660 mg, 3.492 mmol) was added to acetonitrile solvent (10 mL), sodium bicarbonate (317 mg, 3.774 mmol) and compound 7h (500 mg, 3.14 mmol) were added to the solution, and the reaction was stirred for 16 hours. 50 mL of water was added to the reaction solution, extracted with ethyl acetate (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound 7i (800 mg, yield: 81%).

MS m/z(ESI): 312.0[M+1].

Step 9

1-(((3aS,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d ][1,3]Dioxolane-4-yl)methyl)-6-chloro-4-(2',3'-dihydrospiro[azetidine-3,1'-indene Yl]-1-yl)-1H-pyrazolo[3,4-d]pyrimidine 7j

Cesium carbonate (1.195g, 3.654mmol) was added to compound 7i (380mg, 1.218mmol) and compound 1f (523mg, 1.221mmol) in N,N-dimethylformamide (10.0mL) solution, the mixture was heated to 70 Heat at ℃ for 36 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7j (200 mg, yield: 26%).

MS m/z(ESI): 612.3[M+1].

Tenth step

((3aR,4R,6R,6aS)-6-((6-chloro-4-(2',3'-dihydrospiro[azetidine-3,1'-indenyl]-1-yl )1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxolane -4-base) methanol 7k

N-Tetrabutylammonium fluoride (150 mg, 0.666 mmol) was added to the tetrahydrofuran (5.0 mL) solution of compound 7j (200 mg, 0.326 mmol), and the reaction was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7k (100 mg, yield: 61%).

MS m/z(ESI): 497.9[M-1].

Eleventh step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(2',3'-dihydrospiro[azetidine-3,1'-indenyl]-1 -Yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxa Cyclopentane-4-yl)methoxy)methyl)di-tert-butyl phosphonate 7l

Compound 7k (100 mg, 0.2 mmol), compound 2b (99 mg, 0.24 mmol) and N,N-dimethylformamide solvent (3.0 mL) were mixed together, and magnesium tert-butoxide (103 mg, 0.3 mmol) was added. The mixture was heated at 70°C for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 7l (60 mg, yield: 42%).

MS m/z(ESI): 703.9[M+1].

Twelfth step

(((2R,3S,4R,5R)-5-((6-chloro-4-(2',3'-dihydrospiro[azetidine-3,1'-indenyl]-1- Yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphonic acid 7

Compound 7l (60 mg, 0.085 mmol) was added to trifluoroacetic acid (0.5 mL) and dichloromethane (1 mL), water (0.05 mL) was added to the solution, and the reaction was stirred for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18 100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 7 (5 mg, yield: 10%).

MS m/z(ESI): 552.2[M+1].

¹ H NMR (400MHz, CD ₃ OD) δ 8.02 (s, 1H), 7.51 (dd, 2H), 7.24-7.28 (m, 3H), 5.35 (t, 1H), 4.62-4.88 (m, 3H) ,4.47-4.45(m,4H),4.18-4.17(m,2H),4.16-4.15(m,1H),3.63-3.68(m,4H),2.99(t,2H),2.52-2.55(m, 2H), 2.04 (br, 1H), 1.63 (br, 1H).

Example 8

((((((2R,3S,4R,5R)-5-((6-chloro-4-cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl (Yl)-3,4-Dihydroxytetrahydrofuran-2-yl)methoxy)methyl)phosphono)di(oxy))bis(methylene)diisopropyl dicarbonate 8

first step

((((3aR,4R,6R,6aS)-6-((6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl )-2,2-Dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)methyl)phosphonic acid 8a

Compound 1 (100 mg, 0.21 mmol) was added to acetone solvent (30.0 mL), concentrated sulfuric acid (184 mg, 1.88 mmol) was added to the solution, and the reaction was stirred for 1 hour. Solid sodium bicarbonate (1000 mg, 11.90 mmol) was added and stirred for 10 minutes, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound 8a (85 mg, yield: 78%).

MS m/z(ESI): 518.1[M+1].

Second step

((((((3aR,4R,6R,6aS)-6-((6-chloro-4-(cyclopentylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl) (Methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)methyl)phosphonophosphono)bis(oxygen) (Base)) bis(methylene) diisopropyl dicarbonate 8b

Diisopropylethylamine (749mg, 5.80mmol) and tetra-n-butylammonium bromide (177mg, 0.62mmol) were added to compound 8a (200mg, 0.39mmol) and chloromethyl isopropyl carbonate (883mg, In a solution of 5.79 mmol) in N,N-dimethylformamide (10.0 mL), the mixture was heated at 70°C for 24 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 8b (160 mg, yield: 55%).

MS m/z(ESI): 750.2[M+1].

third step

Compound 8b (240 mg, 0.32 mmol) was added to trifluoroacetic acid solvent (2.5 mL), water (0.25 mL) was added to the solution, and the reaction was stirred for 0.5 hour. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (Shimadzu OPTIONBOX-L high pressure liquid chromatography, Gimini 5um NX-C18100x21.2mm column, elution system: acetonitrile, water) to obtain the title compound 8 (140 mg, yield: 62%).

MS m/z(ESI): 710.1[M+1].

¹ H NMR (400MHz, CDCl ₃ ) δ7.91 (s, 1H), 5.68-5.75 (m, 4H), 4.90-4.98 (m, 2H), 4.57-4.61 (m, 2H), 3.74-4.25 (m , 8H), 2.14-2.22 (m, 3H), 1.70 to 1.80 (m, 6H), 1.31-1.33 (m, 12H).

Test case:

Biological evaluation

Test Example 1 The in vitro inhibitory activity of the compound of the present disclosure on CD73 enzyme

CD73 enzyme is an extracellular-5'-nucleotidase, which converts extracellular nucleoside 5'monophosphate into nucleoside, with AMP or CMP as the preferred substrate.

In this assay, recombinant human CD73 expressed from the Chinese Hamster Ovary cell line (R&D Systems) was used to convert cytidine monophosphate (CMP) into cytidine and phosphate. Before adding the substrate, the CD73 enzyme was pre-incubated with the compound for 2 hours. Then the amount of phosphate is measured by a malachite green phosphate detection kit.

Experimental materials and instruments

Malachite green phosphate detection kit: R&D Systems

Recombinant human 5′-nucleotidase (CD73): R&D Systems

HEPES buffer: Gibco, cat#15630-080

CMP: Sigma, cat#C1006

NaCl 5M: Boston Bioproducts, BM-244

TECAN plate reader: TECAN

Experimental steps

The compound was first prepared into a 10 mM solution with DMSO, and then diluted 1:3 with DMSO for a total of 12 concentration points (the compounds with high inhibitory properties were diluted 1:3 with the highest concentration of 10 μM). In each well of a 384-well plate, combine 0.34 nM recombinant human 5'-nucleotidase (CD73) and the tested compound in a solution containing 20 mM HEPES buffer (pH 7.4), 137 mM NaCl, and 0.001% Tween 20. Incubate in assay buffer and pre-incubate at 37°C for 2 hours. The final reaction volume in each well is 12 μL. The highest concentration of the compound is 125 μM and the DMSO concentration is 1.25%. After pre-incubation, 3 μL of CMP dissolved in the assay buffer was added to each reaction well, and the final CMP concentration was 45 μM. The reaction was incubated at 37°C for 15 minutes, and then 3 μL of malachite green reagent A was added to each reaction well. Spin the plate briefly for 30 seconds in the centrifuge. After incubating for another 10 minutes at room temperature, add 3 μL of malachite green reagent B to each reaction well. The reaction plate is briefly rotated in the centrifuge for 30 seconds. After incubating for 20 minutes at room temperature, read the signal at OD ₆₂₀ on a TECAN plate reader. A reaction containing CD73 enzyme, substrate CMP and DMSO (no compound) was used as a positive control for the assay, while a reaction containing substrate CMP and DMSO without CD73 enzyme was used as a negative control for the assay. IC ₅₀ values _were calculated by plotting the logarithm of the compound concentration using GraphPad Prism program appropriate and in percent inhibition.

The in vitro inhibition of CD73 enzyme activity by the compounds of the present disclosure was determined by the above test, and the measured IC ₅₀ values are shown in Table 1 below.

Table 1 The IC ₅₀ value of in vitro inhibition of CD73 enzyme activity by the compounds of the present disclosure

实施例编号Example number	IC ₅₀(nM) IC ₅₀ (nM)
11	0.090.09
33	0.170.17
44	0.250.25
55	2.92.9
66	0.30.3
77	3.43.4

Conclusion: The compound of the present disclosure has a significant inhibitory effect on CD73 enzyme activity in vitro.

Test Example 2 The inhibitory effect of the compound of the present disclosure on the membrane-bound CD73 enzyme activity on human melanoma A375 cells.

In this assay, the membrane-bound CD73 enzyme activity on the surface of human melanoma A375 cells (ATCCCRL-1619) was used to convert cytidine-phosphate (CMP) into cytidine and phosphate in the presence of compound and CMP. Then the amount of phosphate is measured by a malachite green phosphate detection kit.

Experimental materials and instruments

Malachite green phosphate detection kit: R&D Systems, Cat#DY996

A375 cell line: ATCC, Cat#CRL-1619

DMEM: ATCC, Cat#, 30-2002

Trypsin-EDTA 0.25%: Gibco, Cat#25-200-056

FBS: Gibco, Cat#16-140-071

Penicillin-Streptomycin: Gibco, Cat#15-140-122

CMP: Sigma, Cat#C1006

DMSO: Fisher Chemical, Cat#D128-1

HEPES: Gibco, Cat#15630-080

NaCl 5M: Boston Bioproducts, Cat#BM-244

KCl 2M: Ambion, Cat#AM9640G

CaCl ₂ 2M: Fisher, Cat#BP9742

NaHCO ₃ 7.5%: Gibco, Cat#25080-094

Glucose: Gibco, Cat#A2494001

TECAN plate reader: TECAN

Experimental steps

A375 cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin. On the day before the assay, A375 cells were harvested with trypsin and counted. Cells were seeded into 96-well plates in 100 μL of medium (2500 cells/well). The next day, prepare an assay buffer containing 20 mM HEPES, 137 mM NaCl, 5.4 mM KCl, 1.3 mM CaCl ₂ , 4.2 mM NaHCO ₃ and 1 mg/mL glucose. Heat the buffer in a 37°C water bath. In the assay buffer containing 50 μM CMP, start with the highest concentration of 10 μM to prepare 3.16-fold serial dilutions of compounds, and make sure that each dilution contains the same amount of DMSO (0.1%).

Gently remove the medium from the cell culture plate, gently wash the cell layer once with the assay buffer, and then transfer 200 μL of the serially diluted compound solution to the corresponding well. Add 200 μL of assay buffer to the negative control wells, and add 200 μL of DMSO/CMP/assay buffer to the positive control wells.

After incubating at 37°C and 5% CO ₂ for 4 hours, 100 μL of supernatant was transferred from each well to a transparent flat-bottom 96-well plate. Add 20 μL of reagent A from the malachite green phosphate detection kit to each well. Incubate at room temperature for 10 minutes. Add 10 μL of reagent B from the kit to each well, click on the plate to aid mixing. Incubate for 20 minutes at room temperature. Then use TECAN plate reader to read the data on OD620.

data processing

NC is a negative control; PC is a positive control.

IC ₅₀ values _were calculated by plotting the logarithm of the compound concentration using GraphPad Prism program appropriate and in percent inhibition.

The biochemical inhibitory effect of the compounds of the present disclosure on the membrane-bound CD73 enzyme activity on human melanoma A375 cells was determined by the above-mentioned assay method, and the obtained IC ₅₀ values are shown in Table 2 below.

Table 2 The IC ₅₀ value of the compounds of the present disclosure on the inhibition of membrane-bound CD73 enzyme activity on human melanoma A375 cells

实施例编号Example number	IC ₅₀(nM) IC ₅₀ (nM)
11	2.52.5
22	5959
33	9.19.1
55	2020
66	8080
77	9898

Conclusion: The compound of the present disclosure has a significant inhibitory effect on the CD73 enzyme activity in A375 cells.

Pharmacokinetic evaluation

Test Example 3 Mouse pharmacokinetic test of the compound of the present disclosure

1 Overview

Using mice as the test animal, the LC/MS/MS method was used to determine the drug concentration in plasma at different times after the mice were injected with the compound of Example 1 and gavage with the compound of Example 8. To study the pharmacokinetic behavior of the compound of the present disclosure in mice and evaluate its pharmacokinetic characteristics.

2. Test plan

2.1 Experimental drugs

The compound of Example 1 and the compound of Example 8.

2.2 Experimental animals

18 C57 mice, females, were equally divided into 2 groups, purchased from Shanghai Jiesjie Experimental Animal Co., Ltd., animal production license number: SCXK (Shanghai) 2013-0006.

2.3 Drug preparation

Weigh the compound of Example 1, add 5% volume of DMSO and 5% Tween 80 (Shanghai Titan Technology Co., Ltd.) to dissolve, and then add 90% normal saline to prepare a colorless and clear solution of 0.1 mg/ml.

Weigh the compound of Example 8, add 1.5% by volume of DMSO and 98.5% HBSS (Life Technologies) to prepare a 0.5 mg/ml white uniform suspension.

2.4 Administration

A group of C57 mice were fasted overnight and then injected into the administration Example 1. The administration dose was 1 mg/kg, and the administration volume was 0.1 ml/10 g.

Another group of C57 mice were fasted overnight and then administered by gavage in Example 8. The dosage was 10 mg/kg, and the dosage was 0.2 ml/10 g.

3. Operation

The compound of Example 1 was administered to mice by injection, and 0.1ml of blood was collected at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, 24.0 hours before and after administration (3 animals at each time point), and placed In a heparinized test tube (source?), the plasma was separated after centrifugation at 3500 rpm for 10 minutes, and stored at -20°C.

Mice were intragastrically administered the compound of Example 8, and 0.2ml of blood was collected at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, 24.0 hours before and after administration (3 animals at each time point), Place it in a centrifuge tube pre-added with heparinized sodium (Chemical Reagent Co., Ltd. of Sinopharm Group), centrifuge at 3500 rpm for 10 minutes, separate the plasma, and store at -20°C.

To determine the content of the test compound in mouse plasma after drug injection of different concentrations: Take 25μl of mouse plasma at each time after administration and add 50μl (100ng/mL) of the internal standard solution camptothecin (China Institute for the Control of Biological Products) ) And 175μl of acetonitrile, vortexed for 5 minutes, centrifuged for 10 minutes (3700 rpm), plasma sample took 1μl of supernatant for LC/MS/MS (API4000 Triple Quadrupole Tandem Mass Spectrometer (No. 3), USA Applied Biosystems; Shimadzu LC-30AD ultra-high performance liquid chromatography system, Shimadzu, Japan.) analysis.

4. Results of pharmacokinetic parameters

The pharmacokinetic parameters of the compounds of the present disclosure are shown in Table 3 below. The concentration of Example 8 in mouse plasma was lower than the quantitative lower limit, but the product Example 1 was detected.

Table 3 Pharmacokinetic parameters of the compounds of the present disclosure

Conclusion: The compound of the present disclosure has good pharmacokinetic absorption and has pharmacokinetic advantages; Example 8 is the prodrug of Example 1.

Claims

A compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its Medicinal salt:

among them:

Y is -O- or -S-;

Z is -O- or -NH-;

W is selected from -O-, -S-, -N(R 10 )- and -C(R 11 )(R 12 )-;

M is selected from -NH-, -O- or -S-;

G 1 and G 2 are the same or different, and are each independently selected from -N- or -C(R 13 )-;

L is an alkylene group, wherein the alkylene group is optionally further selected from alkyl, halogen, amino, nitro, cyano, hydroxy, hydroxyalkyl, alkoxy, cycloalkyl, heterocyclyl, Substituted by one or more substituents in the aryl group and heteroaryl group;

Q is selected from -CH 2 -OC(R s )(R t )-, -CH 2 -N(R m )-C(R s )(R t )-, -CH 2 -SC(R s )(R t )-, -CH 2 -S(O) 2 -C(R s )(R t )-, -phenyl-OC(R s )(R t )-, -CH 2 -phenyl-OC(R s )(R t )-, -CH 2 -heterocyclyl-, -C(R m )(R n )-, -CH 2 -C(R m )(R n )-C(R s )(R t )-, -C(R s )=C(R t )-, -C(R m )(R n )-C(R s )(R t )-, -C(R m )(R n ) -C(R s )=C(R t )-and -C(R s )=C(R t )-C(R m )(R n );

R s , R t , R m and R n are the same or different, and are each independently selected from H, D, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl and amino;

Or R s and R t together form an oxo group;

Or R m and R n together form an oxo group;

R 1 , R 2 , R 3 and R 4 are the same or different, and are each independently selected from hydroxyl, hydrogen, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cyano, amino , Azido and OR 14 ;

Or R 1 and R 2 together with the carbon atoms to which they are attached form a cycloalkyl or heterocyclic group, wherein the heterocyclic group contains 1 to 3 identical or different heteroatoms selected from N, O and S, wherein The cycloalkyl and heterocyclic groups are optionally further selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and One or more substituents in the heterocyclic group are substituted;

R 5 is selected from hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro and azido;

R 6 and R 7 are the same or different, and are each independently selected from a hydrogen atom, an alkyl group, -C(R m )(R n )-OC(O)OR d , -C(R m )(R n )- OC(O)R d , -C(R m )(R n )C(O)OR d , cycloalkyl, heterocyclic, aryl and heteroaryl, wherein the alkyl group is optionally further selected Substituted by one or more substituents among cycloalkyl, heterocyclyl, aryl and heteroaryl;

R d is selected from hydrogen atom, alkyl group and alkoxy group;

R 8 is selected from hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and heterocyclic group ；

R 9 is selected from a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, a haloalkyl group, a hydroxyalkyl group, a cycloalkyl group, a heterocyclic group, an aryl group and a heteroaryl group, wherein the alkyl group, alkenyl group, alkynyl group , Cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro , Cycloalkyl, heterocyclyl, aryl and heteroaryl substituted by one or more substituents;

R 10 is an alkyl group;

R 11 and R 12 are the same or different, and are each independently selected from a hydrogen atom, a halogen, an alkyl group, and an alkenyl group;

R 13 is selected from hydrogen atom, halogen, alkyl, alkoxy, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclic, aryl and heteroaryl;

R 14 is selected from -C(O)R 15 , -C(O)OR 15 , -S(O) 2 R 15 and -P(O)(OR 6 )(OR 7 );

R 15 is selected from hydrogen atom, halogen, alkyl, haloalkyl, hydroxy and hydroxyalkyl;

n is 0 or 1.
The compound represented by the general formula (I) according to claim 1 or its tautomer, mesoisomer, racemate, enantiomer, diastereomer, prodrug, In the form of a mixture or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (II) or its tautomer, meso, racemate, enantiomer, or diastereomer Body, prodrug, mixture form or its pharmaceutically acceptable salt:

among them:

R 1 to R 9 , G 1 , G 2 , W, Y, Z, Q, L, M, and n are as defined in claim 1.
The compound represented by the general formula (I) according to claim 1 or 2 or its tautomer, meso, racemate, enantiomer, diastereomer, pro In the form of a medicine, a mixture or a pharmaceutically acceptable salt thereof, wherein W is -O-.
The compound represented by the general formula (I) according to any one of claims 1 to 3, or its tautomer, meso, racemate, enantiomer, diastereomer Structure, prodrug, mixture form or pharmaceutically acceptable salt thereof, wherein R 2 , R 4 and R 5 are hydrogen atoms.
The compound represented by the general formula (I) according to any one of claims 1 to 4, or its tautomer, meso, racemate, enantiomer, diastereomer Construct, prodrug, mixture form or pharmaceutically acceptable salt thereof, wherein R 1 and R 3 are the same or different, and are each independently selected from hydroxyl, hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, haloalkyl Oxy and hydroxyalkyl, preferably hydroxy.
The compound represented by the general formula (I) according to any one of claims 1 to 5 or its tautomer, meso, racemate, enantiomer, diastereomer Conformer, prodrug, mixture form or its pharmaceutically acceptable salt, which is a compound represented by the general formula (III) or its tautomer, meso, racemate, enantiomer , Diastereomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them:

G 1 , G 2 , R 6 to R 9 , M, Q, L, and n are as defined in claim 1.
The compound represented by the general formula (I) according to any one of claims 1 to 6 or its tautomer, meso, racemate, enantiomer, diastereomer In the form of a conformer, prodrug, mixture or a pharmaceutically acceptable salt thereof, G 1 is -N-; G 2 is -CH-.
The compound represented by the general formula (I) according to any one of claims 1 to 7 or its tautomer, meso, racemate, enantiomer, diastereomer Conformer, prodrug, mixture form or pharmaceutically acceptable salt thereof, wherein R 6 and R 7 are the same or different, and are each independently selected from a hydrogen atom, an alkyl group, or -C(R m )(R n )-OC (O)OR d ;

R m , R n and R d are as defined in claim 1.
The compound represented by the general formula (I) according to any one of claims 1 to 8 or its tautomer, meso, racemate, enantiomer, diastereomer The structure, prodrug, mixture form or pharmaceutically acceptable salt thereof, wherein Q is -CH 2 -O-CH 2 -.
The compound represented by the general formula (I) according to any one of claims 1 to 9 or its tautomer, meso, racemate, enantiomer, diastereomer A structure, prodrug, mixture form or a pharmaceutically acceptable salt thereof, wherein L is -(CH 2 )s-; s is 1, 2 or 3.
The compound represented by the general formula (I) according to any one of claims 1 to 10, or its tautomer, meso, racemate, enantiomer, diastereomer A structure, prodrug, mixture form or a pharmaceutically acceptable salt thereof, wherein M is NH.
The compound represented by the general formula (I) according to any one of claims 1 to 11 or its tautomer, meso, racemate, enantiomer, diastereomer Conformer, prodrug, mixture form or its pharmaceutically acceptable salt, which is a compound represented by general formula (IV) or its tautomer, meso, racemate, enantiomer , Diastereomers, prodrugs, mixture forms or their pharmaceutically acceptable salts:

among them:

R 8 , R 9 and n are as defined in claim 1.
The compound represented by the general formula (I) according to any one of claims 1 to 12 or a tautomer, meso, racemate, enantiomer, or diastereomer thereof A structure, prodrug, mixture form or a pharmaceutically acceptable salt thereof, wherein R 8 is halogen.
The compound represented by the general formula (I) according to any one of claims 1 to 13 or its tautomer, meso, racemate, enantiomer, diastereomer A structure, prodrug, mixture form or a pharmaceutically acceptable salt thereof, wherein: R 9 is selected from alkyl, cycloalkyl and heterocyclyl, wherein said alkyl, cycloalkyl and heterocyclyl are optionally One or one selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl Multiple substituents are substituted.
The compound represented by the general formula (I) according to any one of claims 1 to 14 or its tautomer, meso, racemate, enantiomer, diastereomer Conform, prodrug, mixture form or pharmaceutically acceptable salt thereof, wherein -(M) n -R 9 is selected from:
The compound represented by the general formula (I) according to any one of claims 1 to 15 or its tautomer, meso, racemate, enantiomer, diastereomer Construct, prodrug, mixture form or pharmaceutically acceptable salt thereof, which is selected from:
A compound represented by the general formula (IA) or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, mixture form or its Medicinal salt:

among them:

Y is -O- or -S-;

Z is -O-;

R x is an alkyl group;

W is selected from -O-, -S-, -N(R 10 )- and -C(R 11 )(R 12 )-;

M is selected from -NH-, -O- or -S-;

G 1 and G 2 are the same or different, and are each independently selected from -N- or -C(R 13 )-;

L is an alkylene group, wherein the alkylene group is optionally further selected from alkyl, halogen, amino, nitro, cyano, hydroxy, hydroxyalkyl, alkoxy, cycloalkyl, heterocyclyl, Substituted by one or more substituents in the aryl group and heteroaryl group;

Q is selected from -CH 2 -OC(R s )(R t )-, -CH 2 -N(R m )-C(R s )(R t )-, -CH 2 -SC(R s )(R t )-, -CH 2 -S(O) 2 -C(R s )(R t )-, -phenyl-OC(R s )(R t )-, -CH 2 -phenyl-OC(R s )(R t )-, -CH 2 -heterocyclyl-, -C(R m )(R n )-, -CH 2 -C(R m )(R n )-C(R s )(R t )-, -C(R s )=C(R t )-, -C(R m )(R n )-C(R s )(R t )-, -C(R m )(R n ) -C(R s )=C(R t )-and -C(R s )=C(R t )-C(R m )(R n );

R s , R t , R m and R n are the same or different, and are each independently selected from H, D, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl and amino;

Or R s and R t together form an oxo group;

Or R m and R n together form an oxo group;

R 2 and R 4 are the same or different, and are each independently selected from hydroxyl, hydrogen, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cyano, amino, azido and OR 14 ;

R 5 is selected from hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro and azido;

R 8 is selected from hydrogen atom, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and heterocyclic group ；

R 9 is selected from a hydrogen atom, an alkyl group, an alkenyl group, an alkynyl group, a haloalkyl group, a hydroxyalkyl group, a cycloalkyl group, a heterocyclic group, an aryl group and a heteroaryl group, wherein the alkyl group, alkenyl group, alkynyl group , Cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro , Cycloalkyl, heterocyclyl, aryl and heteroaryl substituted by one or more substituents;

R 10 is an alkyl group;

R 11 and R 12 are the same or different, and are each independently selected from a hydrogen atom, a halogen, an alkyl group, and an alkenyl group;

R 13 is selected from hydrogen atom, halogen, alkyl, alkoxy, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclic, aryl and heteroaryl;

R 14 is selected from -C(O)R 15 , -C(O)OR 15 , -S(O) 2 R 15 and -P(O)(OR 6 )(OR 7 );

R 15 is selected from hydrogen atom, halogen, alkyl, haloalkyl, hydroxy and hydroxyalkyl;

n is 0 or 1.
The compound represented by the general formula (IA) according to claim 17 or its tautomer, meso, racemate, enantiomer, diastereomer, prodrug, In the form of a mixture or a pharmaceutically acceptable salt thereof, which is selected from:
A method for preparing the compound represented by the general formula (I) according to claim 1, which comprises the following steps:

The protective group of the compound of general formula (IA) is removed to obtain the compound of general formula (I),

among them:

Z is -O-;

R x is an alkyl group;

R 6 and R 7 are hydrogen atoms;

R 1 and R 3 are hydroxyl groups;

R 2 , R 4 , R 5 , R 8 , R 9 , Q, L, Y, W, G 1 , G 2 , M, and n are as defined in claim 1.
A pharmaceutical composition containing a therapeutically effective amount of the compound represented by the general formula (I) according to any one of claims 1 to 16 or its tautomers, mesosomes, racemates Isomers, enantiomers, diastereomers, prodrugs, mixtures or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.
The compound represented by the general formula (I) according to any one of claims 1 to 16 or its tautomer, meso, racemate, enantiomer, diastereomer Use of the construct, prodrug, mixture form or pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 20 in the preparation of a drug for inhibiting CD73.
The compound represented by the general formula (I) according to any one of claims 1 to 16 or its tautomer, meso, racemate, enantiomer, diastereomer The use of the structure, prodrug, mixture form or pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 20 in the preparation of a medicament for the treatment or prevention of CD73-mediated diseases.
The compound represented by the general formula (I) according to any one of claims 1 to 16 or its tautomer, meso, racemate, enantiomer, diastereomer The use of the construct, prodrug, mixture form or pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 20 in the preparation of a medicine for the treatment or prevention of adenosine and adenosine receptor-mediated diseases.
The compound represented by the general formula (I) according to any one of claims 1 to 16 or its tautomer, meso, racemate, enantiomer, diastereomer The structure, prodrug, mixture form or its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 20 is prepared for the treatment of tumors, autoimmune diseases, immune system diseases, inflammatory diseases, nervous system diseases, Neurodegenerative diseases and central nervous system diseases, depression, Parkinson's disease, cerebral and cardiac ischemic diseases, sleep disorders or fibrosis, preferably tumors.
The use according to claim 24, wherein the tumor is preferably selected from melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, breast cancer, ovarian cancer, uterine cancer, Endometriosis, prostate cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, bone cancer, seminoma, testicular tumor, uterine cancer, head and neck cancer, multiple myeloma, lymphoma, leukemia , Thyroid cancer, ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, choriocarcinoma and pediatric tumors.