WO2021136462A1 - Furan derivatives and application thereof in medicine - Google Patents

Abstract

A class of furan derivatives and an application thereof in medicine; specifically, the furan derivatives relate to pyrimidine derivatives represented by general formula (I'), or stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or eutectic crystals thereof, a pharmaceutical composition comprising same, and a use of the compounds or composition in the preparation of a DNA-PK inhibitor, wherein the definition of each substituent in general formula (I') is the same as the definition thereof in the specification.

Description

Furan derivatives and their applications in medicine Technical field

The present invention relates to furan derivatives, or their stereoisomers, solvates, prodrugs, metabolites, deuterated substances, pharmaceutically acceptable salts or co-crystals, their pharmaceutical compositions and their use in the preparation of DNA-PK inhibitors .

Background technique

DNA-dependent protein kinase (DNA-PK) is a DNA-PK enzyme complex composed of Ku70/Ku80 heterodimer and DNA-dependent protein kinase catalytic subunit (DNA-PKcs). The enzyme complex needs to be activated with the participation of DNA to perform its corresponding functions (George et al., 2019). As a serine/threonine protein kinase, DNA-PK belongs to the PIKK (phosphatidylinositol 3-kinase-related kinase) family. It not only repairs intracellular DNA double-strand breaks (DSBs) and cell DNA recombination Or it plays an important role in the process of antibody DNA rearrangement (V(D)J recombination), and also participates in physiological processes such as chromosome modification, transcription regulation, and telomere maintenance.

In the normal physiological process, a variety of factors may lead to the occurrence of DSBs in DNA: For example, DSBs often appear as intermediate products in the process of somatic DNA recombination. This physiological process is very important for the formation of the functional immune system of all vertebrates; DNA replication is in progress. When the replication fork encounters damaged bases, it may also cause single-strand or double-strand breaks; DNA may also generate DSBs due to the attack of reactive oxygen species (ROS) during normal metabolism (Cannan & Pederson, 2016). In addition, there are a variety of exogenous factors that may also cause DSBs, such as ionizing radiation (IR) and chemotherapeutic agents (such as topoisomerase II inhibitors) (George et al., 2019). If DSBs are not repaired or repaired incorrectly, mutations and/or chromosomal aberrations will occur, eventually leading to cell death. In order to cope with the harm caused by DSBs, eukaryotic cells have evolved a variety of mechanisms to repair damaged DNA to maintain cell viability and genome stability. In eukaryotic cells, the most important way of DNA repair is non-homologous end-joining (NHEJ). This method of directly connecting broken DNA does not require the participation of homologous DNA fragments, and can occur at any stage of the cell cycle. NHEJ is a dynamic process mediated by DNA-PK that requires the participation of multiple proteins and signaling pathways. The basic process is as follows: (1) Ku70/Ku80 heterodimer recognizes and binds to the ends of double-stranded DNA breaks; (2) Recruitment DNA-PKcs, XRCC4-DNA ligase IV complex and other proteins to both sides of the DNA break double-strand; (3) DNA-PKcs autophosphorylate and activate its own kinase activity; (4) DNA-PKcs as an adhesive to connect Break both ends of the DNA to prevent exonuclease from degrading DN A; (5) Process the DNA to remove the unlinkable ends or other forms of damage at the break; (6) XRCC4-DNA ligase IV complex Repair DNA ends (in some cases, DNA polymerase may be required to synthesize new ends before ligation). When DNA-PKcs is phosphorylated, it can induce protein conformation to change and regulate the activity of various proteins in the NHEJ process (such as Artemis, Ku70, Ku80, DNA ligase), which is essential for the repair process of DN A. Therefore, phosphorylated DNA-PKcs (pDNA-PKcs) is often used as a marker of cellular DSBs.

Studies have shown that DNA-PK activity is related to the occurrence and development of a variety of tumors: for example, DNA-PKcs in melanoma can promote angiogenesis and tumor metastasis; DNA-PKcs expression in multiple myeloma is significantly up-regulated; radiotherapy The content of Ku protein in tolerant thyroid tumors is significantly increased (Ihara, Ashizawa, Shichijo, & Kudo, 2019). Therefore, it can be considered to combine DNA-PK inhibitors with anti-tumor therapies that cause DNA damage (such as IR, chemotherapeutic agents, etc.) to improve the effect. The use of DNA-PK inhibitors can interfere with the DNA repair function of normal cells to a certain extent. However, there are many DNA repair pathways in normal cells as a supplement, and tumor cells face strong DNA replication pressure and lack effective DNA repair methods. . By inhibiting the activity of tumor cell DNA-PK, the killing effect of other anti-tumor drugs on tumor cells can be improved.

After years of research, several DNA-PK inhibitors have been discovered. The first compound found to have DNA-PK kinase inhibitory activity is a fungal metabolite-Wortmannin, with an IC50 (DNA-PK) of about 15nM. This compound also plays an important role in the acetylation and phosphorylation of p53 protein ( Sarkaria et al., 1998); The quercetin derivative LY294002 reported later also has DNA-PK inhibitory activity (Maira, Stauffer, Schnell, & Garcia-Echeverria, 2009); later based on the structure of LY294002, NU7026 and NU7441 were developed A generation of DNA-PK inhibitors. Although it has been confirmed that these compounds have a good killing effect on tumor cells, they have problems such as high toxicity and poor selectivity and cannot enter clinical development (Maira et al., 2009). Other DNA-PK inhibitors have also been reported, such as OK1035, SU11752, PP121, KU-0060648 and other small molecule compounds, but these compounds also have defects such as low specificity for DNA-PK (George et al., 2019). Therefore, there is still a need to develop DNA-PK inhibitors with high activity, high specificity, and low toxicity to better meet clinical needs.

Summary of the invention

One or more embodiments of the present application provide furan derivatives, or their stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals, and their pharmaceutical compositions and Preparation of DNA-PK inhibitors; these compounds have high inhibitory activity and high selectivity to DNA-PK, and can be used as chemotherapy and radiotherapy sensitizers to effectively treat cancer, improve the curative effect of the prior art, and reduce toxic and side effects.

One or more embodiments of the present application provide a compound of general formula (I'), or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals:

among them:

R ₀ and R ₁ are each independently selected from H, -OH, cyano, halogen, -NH ₂ , C _1-6 alkyl or C _1-6 alkoxy, the C _1-6 alkyl optionally Further substituted by 1-3 substituents selected from D or halogen;

Or two R ₀ and the atoms to which they are connected form a 3- to 8-membered ring, and the 3- to 8-membered ring optionally contains 1 to 3 heteroatoms selected from N, O or S, and the 3- to 8-membered ring The ring is optionally further substituted with one or more substituents selected from -OH, carboxy, halogen, cyano, =0, C _1-6 alkyl or amino;

R _{2 is} selected from H or C _1-6 alkyl;

R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group, -C _1-6 alkylene-C _4-12 heterocyclic group, C _6-12 spiro compound or C _{6-12 heterospiro} compound, so The C ₃ heterocyclic group and C _4-12 heterocyclic group contain 1 to 3 heteroatoms selected from N, O or S, the C _1-6 alkyl group, C _3-12 carbocyclic group, C ₃ Heterocyclic group, C _4-12 heterocyclic group, C _1-6 alkylene group may be further selected by one or more selected from -OH, -OR ^a1 , carboxy, halogen, cyano, =O, C _{1 -6} alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR ^a1 R ^a2 , -C(=O)OC _1-6 alkyl, -C(= O) NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituent substituted And the C _1-6 alkyl group, C _1-6 heteroalkyl group, C _2-6 alkenyl group or C _2-6 alkynyl group in the substituent is optionally further selected from one or more -OH, carboxyl, cyano, halogen, -OR ^a1 , -NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl Or substituted by a substituent of a _{C 5-12 heteroaryl group;}

R _{4 is} selected from H, C _1-6 alkyl, C _3-12 cycloalkyl, said C _1-6 alkyl and C _3-8 cycloalkyl are optionally further selected from 1-3 from D or Substituted by halogen substituents;

R ^a1 and R ^a2 are each independently selected from H, C _1-6 alkyl, -C(=O)R ^a3 or -C(=O)NR ^a4 R ^a5 , wherein the C _1-6 alkyl is any Optionally, one or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkane Group, C ₃ heterocycloalkyl, or C _4-12 heterocycloalkyl substituent; or ^{Ra4, Ra5} and N atom form a 3- to 8-membered heterocyclic ring, said 3- to 8-membered heterocyclic ring Contain 1 to 3 heteroatoms selected from N, O or S;

R ^{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl;

n is 0, 1, 2 or 3;

p is 0, 1, 2, or 3.

One or more embodiments of the application provide a compound, or its stereoisomer, solvate, prodrug, metabolite, deuterated product, pharmaceutically acceptable salt or co-crystal, wherein the compound is selected from the general formula ( I) The compound shown:

among them:

R ₀ and R _{1 are} selected from H, -OH, cyano, halogen, -NH ₂ , C _1-6 alkyl or C _1-6 alkoxy, and the alkyl group is optionally further selected from 1-3 Substituted by D or halogen substituent;

Or two R ₀ and the atoms to which they are connected can form a 3- to 8-membered ring, the ring may contain 1 to 3 heteroatoms selected from N, O or S, and the ring is optionally further divided by 1 or more Substituted by a substituent selected from -OH, carboxyl, halogen, cyano, =O, C _{1-6 alkyl or amino;}

R _{2 is} selected from H or C _1-6 alkyl;

R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group or -C _1-6 alkylene-C _4-12 heterocyclic group, said heterocyclic group may contain 1 to 3 selected from N, O or S heteroatom, the alkyl group, carbocyclic group, heterocyclic group, alkylene group may be further selected by one or more selected from -OH, carboxyl, halogen, cyano, =0, C _1-6 Alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR _a1 R _a2 , -C(=O)OC _1-6 alkyl, -C(=O) NR _a1 R _a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents are substituted; and The alkyl, heteroalkyl, alkenyl or alkynyl group is optionally further selected by one or more selected from -OH, carboxyl, cyano, halogen, -OR _a1 , -NR _a1 R _a2 , C _3-12 Cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents;

R _a1 and R _{a2 are} selected from H, C _1-6 alkyl, -C(=O)R _a3 or -C(=O)NR _a4 R _a5 , wherein the C _1-6 alkyl is optionally further One or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkyl, C ₃ heterocycloalkyl, or substituted by a substituent of _{C 4-12 heterocycloalkyl; or R a4} and R _a5 and N atoms form a 3- to 8-membered heterocyclic ring, and the ring may contain one or more Heteroatom selected from N, O or S;

R _{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl;

R _a4 and R _{a5 are} selected from H or C _1-6 alkyl;

n and p are each independently selected from 0, 1, 2 or 3.

One or more embodiments of the application provide a compound, or its stereoisomer, solvate, prodrug, metabolite, deuterated product, pharmaceutically acceptable salt or co-crystal, wherein the compound is selected from the general formula ( IA) The compound shown:

among them:

R ₀ and R ₁ are each independently selected from halogen or C _1-6 alkyl, and said C _1-6 alkyl is optionally further substituted with 1-3 substituents selected from D or halogen;

Or two R ₀ and the atoms to which they are connected form a 3- to 8-membered ring, and the 3- to 8-membered ring optionally contains 1 to 3 heteroatoms selected from N, O or S, and the 3- to 8-membered ring The ring is optionally further substituted with one or more substituents selected from -OH, carboxy, halogen, cyano, =0, C _1-6 alkyl or amino;

R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group, -C _1-6 alkylene-C _4-12 heterocyclic group, C _6-12 spiro compound or C _{6-12 heterospiro} compound, so The C ₃ heterocyclic group and C _4-12 heterocyclic group contain 1 to 3 heteroatoms selected from N, O or S, the C _1-6 alkyl group, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, C _1-6 alkylene group may be further selected by one or more selected from -OH, -OR ^a1 , carboxy, halogen, cyano, =O, C _{1 -6} alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR ^a1 R ^a2 , -C(=O)OC _1-6 alkyl, -C(= O) NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituent substituted And the C _1-6 alkyl group, C _1-6 heteroalkyl group, C _2-6 alkenyl group or C _2-6 alkynyl group in the substituent is optionally further selected from one or more -OH, carboxyl, cyano, halogen, -OR ^a1 , -NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl Or substituted by a substituent of a _{C 5-12 heteroaryl group;}

R _{4 is} selected from H, C1-6 alkyl, C3-12 cycloalkyl, said C _1-6 alkyl and C _3-8 cycloalkyl are optionally further selected from 1-3 selected from D or halogen Substituent substituted;

R ^a1 and R ^a2 are each independently selected from H, C _1-6 alkyl, -C(=O)R ^a3 or -C(=O)NR ^a4 R ^a5 , wherein the C _1-6 alkyl is any Optionally, one or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkane Group, C ₃ heterocycloalkyl or C _4-12 heterocycloalkyl substituent; or ^{Ra4, Ra5} and N atom form a 3- to 8-membered heterocyclic ring, and the 3- to 8-membered heterocyclic ring includes 1 to 3 heteroatoms selected from N, O or S;

R ^{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl;

R ^a4 and R ^a5 are each independently selected from H or C _1-6 alkyl;

n is 0, 1, 2 or 3;

p is 0, 1, 2, or 3.

One or more embodiments of the application provide a compound, or its stereoisomer, solvate, prodrug, metabolite, deuterated product, pharmaceutically acceptable salt or co-crystal, wherein the compound is selected from the general formula ( II) The compound shown:

among them:

R ₀ and R _{1 are} selected from halogen or C _1-6 alkyl, and the alkyl is optionally further substituted with 1-3 substituents selected from D or halogen;

Or two R ₀ and the atoms to which they are connected can form a 3- to 8-membered ring, and the ring may contain 1 to 3 heteroatoms selected from N, O or S, and the ring is optionally further divided by 1 or more. Substituted by a substituent selected from -OH, carboxyl, halogen, cyano, =O, C _{1-6 alkyl or amino;}

R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group or -C _1-6 alkylene-C _4-12 heterocyclic group, said heterocyclic group may contain 1 to 3 selected from N, O or S heteroatom, the alkyl group, carbocyclic group, heterocyclic group, alkylene group may be further selected by one or more selected from -OH, carboxyl, halogen, cyano, =0, C _1-6 Alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR _a1 R _a2 , -C(=O)OC _1-6 alkyl, -C(=O) NR _a1 R _a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents are substituted; and The alkyl, heteroalkyl, alkenyl or alkynyl group is optionally further selected by one or more selected from -OH, carboxyl, cyano, halogen, -OR _a1 , -NR _a1 R _a2 , C _3-12 Cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents;

R _a1 and R _{a2 are} selected from H, C _1-6 alkyl, -C(=O)R _a3 or -C(=O)NR _a4 R _a5 , wherein the C _1-6 alkyl is optionally further One or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkyl, C ₃ Heterocycloalkyl or C _4-12 heterocycloalkyl substituents; or R _a4 and R _a5 and N atoms form a 3- to 8-membered heterocyclic ring, the ring may contain one or more options Heteroatoms from N, O or S;

R _{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl;

R _a4 and R _{a5 are} selected from H or C _1-6 alkyl;

n and p are each independently selected from 0, 1, 2 or 3.

One or more embodiments of the application provide a compound, or its stereoisomer, solvate, prodrug, metabolite, deuterated product, pharmaceutically acceptable salt or co-crystal, wherein the compound is selected from the general formula ( I'), (IA), (I) or (II), wherein:

R _{0 is} selected from methyl;

R _{1 is} selected from halogen or C _1-4 alkyl, and the alkyl is optionally further substituted with 1-3 substituents selected from D or halogen;

R _{3 is} selected from C _1-6 alkyl, C _3-6 carbocyclic group, C _4-8 heterocyclic group, -C _1-2 alkylene-C _3-8 carbocyclic group or -C _1-2 alkylene Alkyl-C _4-8 heterocyclic group, the heterocyclic group may contain 1 to 3 heteroatoms selected from N or O, and the alkyl group, carbocyclic group, heterocyclic group, and alkylene group may be any Optionally further substituted by one or more substituents selected from -OH, halogen, cyano, =OC _1-4 alkoxy or C _1-4 alkyl; and the alkyl in the substituents is any Optionally further substituted by one or more substituents selected from -OH, carboxyl, cyano or halogen;

n is selected from 0, 1 or 2;

p is selected from 1.

One or more embodiments of the present application provide a compound, or its stereoisomer, solvate, metabolite, prodrug, deuterium, pharmaceutically acceptable salt or co-crystal, wherein the compound is selected from but not limited to :

或者

or

One or more embodiments of the present application provide an intermediate for preparing the compound of the present invention, and the intermediate is selected from but not limited to:

One or more embodiments of the present application provide a pharmaceutical composition, the pharmaceutical composition comprising:

(1) The compound of the present invention or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals;

(2) Optional one or more other active ingredients; and

(3) A pharmaceutically acceptable carrier and/or excipient.

One or more embodiments of the present application provide that the pharmaceutical composition of the present application, or the compound or its stereoisomers, solvates, prodrugs, metabolites, deuterated substances, pharmaceutically acceptable salts or co-crystals are prepared in Use in medicines for the treatment of cancer.

One or more embodiments of the present application provide that the pharmaceutical composition of the present application, or the compound or its stereoisomers, solvates, prodrugs, metabolites, deuterated substances, pharmaceutically acceptable salts or co-crystals are prepared in Use in DNA-PK inhibitors.

One or more embodiments of the present application provide the compound of the present application for use as a medicine.

One or more embodiments of the present application provide the compound of the present application for use as a DNA-PK inhibitor.

One or more embodiments of the present application provide a compound of the present application for use in a method of treating, preventing, or inhibiting cancer.

One or more embodiments of the present application provide a compound of the present application for use in a method of inhibiting DNA-PK.

One or more embodiments of the present application provide a method of treating, preventing or inhibiting cancer, which comprises administering the compound of the present application to a subject in need.

One or more embodiments of the present application provide a method for inhibiting DNA-PK, which includes administering the compound of the present application to a subject in need.

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

The carbon, hydrogen, oxygen, sulfur, nitrogen or F, Cl, Br, and I involved in the groups and compounds of the present invention include their isotopes, and the carbon involved in the groups and compounds of the present invention , Hydrogen, oxygen, sulfur or nitrogen are optionally further replaced by one or more of their corresponding isotopes, wherein carbon isotopes include ¹² C, ¹³ C and ¹⁴ C, and hydrogen isotopes include protium (H), deuterium (D, Also called heavy hydrogen), tritium (T, also called super heavy hydrogen), oxygen isotopes include ¹⁶ O, ¹⁷ O and ¹⁸ O, sulfur isotopes include ³² S, ³³ S, ³⁴ S and ³⁶ S, and nitrogen isotopes include ¹⁴ N and ¹⁵ N, fluorine isotopes include ¹⁷ F and ¹⁹ F, chlorine isotopes include ³⁵ Cl and ³⁷ Cl, and bromine isotopes include ⁷⁹ Br and ⁸¹ Br.

"Alkyl" refers to a linear or branched saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, preferably 1 to 8 (for example, 1, 2, 3, 4, 5, 6, 7, 8) carbon atoms The alkyl group of is more preferably an alkyl group of 1 to 6 carbon atoms, and still more preferably an alkyl group of 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl And its various branched isomers; when the alkyl group is substituted, it may be optionally further substituted with one or more substituents.

"Alkoxy" refers to a group formed by replacing at least one carbon atom in an alkyl group with an oxygen atom. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropyl Oxy and cyclobutoxy. The definition of the alkyl group is the same as the definition of "alkyl" mentioned above.

"Alkenyl" refers to a straight line consisting of 1 to 10 (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon-carbon double bonds consisting of 2 to 20 carbon atoms. Chain or branched unsaturated aliphatic hydrocarbon group, preferably 2 to 12 (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) carbon atoms alkenyl group, more preferably 2 to The alkenyl group of 8 carbon atoms is more preferably the alkenyl group of 2 to 6 carbon atoms. Non-limiting examples include vinyl, propen-2-yl, buten-2-yl, buten-2-yl, penten-2-yl, penten-4-yl, hexen-2-yl, Hexen-3-yl, hepten-2-yl, hepten-3-yl, hepten-4-yl, octen-3-yl, nonen-3-yl, decen-4-yl and undecenyl En-3-yl. The alkenyl group may be further substituted with one or more substituents.

"Alkynyl" refers to those containing 1 to 10 (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) carbon-carbon triple bonds, consisting of 2 to 20 carbon atoms Straight or branched chain unsaturated aliphatic hydrocarbon group, preferably 2 to 12 (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12) carbon atom alkynyl group, more preferably 2 An alkynyl group having to 8 carbon atoms, and an alkynyl group having 2 to 6 carbon atoms is more preferable. Non-limiting examples include ethynyl, propyn-1-yl, propyn-2-yl, butyn-1-yl, butyn-2-yl, butyn-3-yl, 3,3-dimethyl Butyn-2-yl, pentyn-1-yl, pentyn-2-yl, hexyn-1-yl, 1-heptyn-1-yl, heptyn-3-yl, heptyn-4- Base, octyn-3-yl, nonyn-3-yl, decyn-4-yl, undecyn-3-yl, dodecyn-4-yl. The alkynyl group may be optionally further substituted with one to more substituents.

"Aryl" refers to a substituted or unsubstituted aromatic ring, which can be a 5- to 8-membered (e.g., 5, 6, 7, 8-membered) monocyclic ring, 5 to 12-membered (e.g., 5, 6, 7 , 8, 9, 10, 11, 12 membered) bicyclic or 10 to 15 membered (for example, 10, 11, 12, 13, 14, 15 membered) tricyclic ring system, which can be bridged or spiro ring, non-limiting implementation Examples include phenyl and naphthyl. The aryl group may be further substituted with one or more substituents.

"Heteroaryl" refers to a substituted or unsubstituted aromatic ring, which can be 3 to 8 membered (e.g. 3, 4, 5, 6, 7, 8 membered) monocyclic, 5 to 12 membered (e.g. 5, 6, 7, 8, 9, 10, 11, 12 membered) bicyclic or 10 to 15 membered (e.g. 10, 11, 12, 13, 14, 15 membered) tricyclic ring system, and contains 1 to 6 (e.g. 1, 2, 3, 4, 5, 6) heteroatoms selected from N, O or S, preferably 5 to 8 membered heteroaryl groups, and 1 to 4 (e.g. 1, 2 , 3, 4) N and S can be oxidized into various oxidation states. The heterocyclic group can be attached to a hetero atom or a carbon atom, and the heteroaryl group can be a bridged ring or a spiro ring. Non-limiting examples include cyclopyridyl, furyl, thienyl, pyranyl, pyrrolyl, pyrimidinyl, Pyrazinyl, pyridazinyl, imidazolyl, piperidinyl benzimidazolyl, benzopyridyl, pyrrolopyridyl. The heteroaryl group is optionally further substituted with one or more substituents.

所述的“碳环基”或“碳环”任选进一步被1个或者多个取代基所取代。 "Carbocyclic group" or "carbocyclic ring" refers to a saturated or unsaturated aromatic ring or a non-aromatic ring. When it is an aromatic ring, its definition is the same as the definition of "aryl"above; when it is a non-aromatic ring, it can be 3 to 10 members (for example, 3, 4, 5, 6, 7, 8, 9, 10 Yuan), 4 to 12 yuan (e.g. 4, 5, 6, 7, 8, 9, 10, 11, 12 yuan) bicyclic ring or 10 to 15 yuan (e.g. 10, 11, 12, 13, 14, 15 Member) tricyclic ring system, which can be bridged or spiro ring, non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2 -Alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexenyl, cyclohexadienyl, cyclo Heptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl,

The "carbocyclic group" or "carbocyclic ring" is optionally further substituted with one or more substituents.

"Heterocyclic group" or "heterocyclic ring" refers to a saturated or unsaturated aromatic heterocyclic ring or non-aromatic heterocyclic ring. When it is an aromatic heterocyclic ring, its definition is the same as the definition of "heteroaryl" above; when When it is a non-aromatic heterocyclic ring, it can be a 3- to 10-membered (e.g. 3, 4, 5, 6, 7, 8, 9, 10-membered) monocyclic ring, 4 to 12-membered (e.g. 4, 5, 6, 7, 8, 9, 10, 11, 12 membered) bicyclic or 10 to 15 membered (e.g. 10, 11, 12, 13, 14, 15 membered) tricyclic ring system, and contains 1 to 4 (e.g. 1, 2, 3, 4) heteroatoms selected from N, O or S, preferably 3 to 8 membered heterocyclic groups. One to four (for example, 1, 2, 3, 4) N and S optionally substituted in the "heterocyclic group" or "heterocyclic ring" can be oxidized to various oxidation states; "heterocyclic group" or "Heterocycle" can be attached to a heteroatom or carbon atom; "heterocyclic group" or "heterocycle" can be a bridged ring or a spiro ring. Non-limiting examples of "heterocyclic group" or "heterocyclic ring" include oxirane, glycidyl, aziridinyl, oxetanyl, azetidinyl, thietanyl , 1,3-dioxolane, 1,4-dioxolane, 1,3-dioxanyl, azepanyl, oxepanyl, thiepanyl, oxygen Azepine, diazepine, thiazepine, pyridinyl, piperidinyl, homopiperidinyl, furyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyridine Azinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, thiazinyl, 1,3-dithiazinyl, dihydrofuran Group, dithiopentyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzene Bisimidazolyl, benzopyridyl, pyrrolopyridyl, chromanyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl , Dioxanyl, 1,3-dioxolyl, pyrazolinyl, dithianyl, dithiazolinyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolinyl, 1,2,3,4-Tetrahydroisoquinolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 3H-indolylquinazinyl, N-pyridylurea, 1,1-dioxothiomorpholinyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]non Alkyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptyl. The "heterocyclic group" or "heterocyclic ring" may be optionally further substituted with one or more substituents.

"Cycloalkyl" refers to a saturated cyclic hydrocarbon group whose ring can be 3 to 10 membered (e.g. 3, 4, 5, 6, 7, 8, 9, 10 membered) monocyclic, 4 to 12 membered (e.g. 4 , 5, 6, 7, 8, 9, 10, 11, 12 yuan) bicyclic or 10 to 20 yuan (e.g. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 yuan) more In the ring system, the ring carbon atoms preferably have 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms. Non-limiting examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexyl Alkenyl, cycloheptenyl, 1,5-cyclooctadienyl, 1,4-cyclohexadienyl and cycloheptatrienyl, etc. When the cycloalkyl group is substituted, it may be further substituted with one or more substituents.

"Heterocycloalkyl" refers to a substituted or unsubstituted saturated non-aromatic ring group, which can be 3 to 8 membered (for example, 3, 4, 5, 6, 7, 8 membered) monocyclic, 4 to 12 membered (E.g. 4, 5, 6, 7, 8, 9, 10, 11, 12-membered) bicyclic or 10 to 15-membered (e.g. 10, 11, 12, 13, 14, 15-membered) tricyclic ring system, including 1, 2 or 3 heteroatoms selected from N, O or S, preferably 3 to 8 membered heterocyclic group. The selectively substituted N and S in the "heterocycloalkyl" ring can be oxidized to various oxidation states; the "heterocycloalkyl" can be connected to a heteroatom or a carbon atom; the "heterocycloalkyl" can be a bridge Ring or spiro ring. Non-limiting examples of "heterocycloalkyl" include oxirane ethyl, aziridinyl, oxetanyl, azetidinyl, 1,3-dioxolane, 1,4-dioxolane, Oxolane, 1,3-dioxanyl, azepanyl, piperidinyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl, tetrahydrofuranyl , Tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxa Tricyclic[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptyl.

When the above-mentioned "alkyl", "alkoxy", "alkenyl", "alkynyl", "aryl", "heteroaryl", "carbocyclyl", "carbocyclic", " When "heterocyclyl", "heterocycle", "cycloalkyl", "heterocycloalkyl" or "heterocyclyl" are substituted, they may optionally be further substituted with 0, 1, 2, 3, 4, 5, 6 , 7, 8, 9 or 10 selected from F, Cl, Br, I, hydroxyl, mercapto, nitro, cyano, amino, C _1-6 alkylamino, =0, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl, -NR ^q4 R ^q5 , =NR ^q6 , -C(=O)OC _1-6 alkyl, -OC(=O) C _1-6 alkyl, -C(=O)NR ^q4 R ^q5 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl, C _5-12 heteroaryl, -C(=O)OC _6-12 aryl, -OC(=O)C _6-12 aryl, -OC(=O)C _5-12 heteroaryl, -C( ＝O)OC _5-12 heteroaryl, -OC(=O)C ₃ heterocycloalkyl, -OC(=O)C _4-12 heterocycloalkyl, -C(=O)OC ₃ heterocycloalkane Group, -C(=O)OC _4-12 heterocycloalkyl, -OC(=O)C _3-12 cycloalkyl, -C(=O)OC _3-12 cycloalkyl, -NHC(=O )C ₃ heterocycloalkyl, -NHC(=O)C _4-12 heterocycloalkyl, -NHC(=O)C _6-12 aryl, -NHC(=O)C _5-12 heteroaryl, -NHC(=O)C _3-12 cycloalkyl, -NHC(=O)C _2-6 alkenyl or -NHC(=O)C _2-6 alkynyl substituents, and the said Substituents C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl, C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 Heterocycloalkyl, C _6-12 aryl, C _5-12 heteroaryl, -NHC(=O)C _6-12 aryl, -NHC(=O)C _5-12 heteroaryl, -NHC( =0)C ₃ heterocycloalkyl, -NHC(=O)C _4-12 heterocycloalkyl or -NHC(=O)C _3-12 cycloalkyl is optionally further selected from 1 to 3 OH, F, Cl, Br, I, C _1-6 alkyl, C _1-6 alkoxy, -NR ^q4 R ^q5 or =0 substituents; R ^{q1 is} selected from C _1-6 alkyl, C _{1 -6} alkoxy or C _6-12 aryl; R ^q2 and R ^{q3 are} selected from H or C _1-6 alkyl; wherein R ^q4 and R ^{q5 are} selected from H, C _1-6 alkyl, -NH( C=NR ^q1 )NR ^q2 R ^q3 , -S(=O) ₂ NR ^q2 R ^q3 , -C(=O)R ^q1 or -C(=O)NR ^q2 R ^q3 , Wherein, the C _1-6 alkyl group is optionally further substituted by one or more selected from OH, F, Cl, Br, I, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 Substituted by aryl, C _5-12 heteroaryl, C _3-12 cycloalkyl, C ₃ heterocycloalkyl or C _4-12 heterocycloalkyl substituent; or R ^q4 is formed with R ^q5 and N atom A 3- to 8-membered heterocyclic ring, said heterocyclic ring may contain one or more heteroatoms selected from N, O or S.

"Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" means that the compound of the present invention maintains the biological effectiveness and characteristics of the free acid or free base, and the free acid is combined with a non-toxic inorganic base or An organic base is a salt obtained by reacting the free base with a non-toxic inorganic acid or organic acid.

"Pharmaceutical composition" refers to a mixture of one or more compounds of the present invention, their pharmaceutically acceptable salts or prodrugs, and other chemical components, where "other chemical components" refer to pharmaceutically acceptable compounds. Accepted carriers, excipients and/or one or more other therapeutic agents.

"Carrier" refers to a material that does not cause significant irritation to the organism and does not eliminate the biological activity and characteristics of the administered compound.

"Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate the administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugar, starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, adhesives Agent and disintegrant.

A "prodrug" refers to a compound of the present invention that can be converted into a biologically active compound by metabolism in the body. The prodrug of the present invention is prepared by modifying the amino or carboxyl group in the compound of the present invention, and this modification can be removed by conventional operations or in vivo to obtain the parent compound. When the prodrug of the present invention is administered to a mammalian individual, the prodrug is split to form free amino or carboxyl groups.

"Co-crystal" refers to the crystal formed by the combination of active pharmaceutical ingredient (API) and co-crystal former (CCF) under the action of hydrogen bonds or other non-covalent bonds. The pure state of API and CCF are both at room temperature. Solid, and there is a fixed stoichiometric ratio between the components. A eutectic is a multi-component crystal, which includes both a binary eutectic formed between two neutral solids and a multi-element eutectic formed between a neutral solid and a salt or solvate.

"Stereoisomers" refer to isomers produced by the different arrangements of atoms in a molecule in space, including cis-trans isomers, enantiomers and conformational isomers.

"Optional" or "optionally" or "selective" or "selectively" means that the event or condition described later can but does not necessarily occur, and the description includes the situation in which the event or condition occurs and its failures. What happened. For example, "heterocyclic group optionally substituted by an alkyl group" means that the alkyl group may but does not necessarily exist, and 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. Happening.

Detailed ways

The following embodiments illustrate the technical solutions of the present invention in detail, but the protection scope of the present invention includes but is not limited thereto.

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 is measured with (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic instrument, the solvent is deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD) ), the internal standard is tetramethylsilane (TMS);

For MS measurement (Agilent 6120B (ESI) and Agilent 6120B (APCI));

HPLC determination uses Agilent 1260DAD high pressure liquid chromatograph (Zorbax SB-C18 100×4.6mm, 3.5μM);

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 for thin layer chromatography (TLC) is 0.15mm-0.20mm, and the size used for 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;

The known starting materials of the present invention can be synthesized by or according to methods known in the art, or can be purchased from Titan Technology, Anaiji Chemical, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Bailingwei Technology, etc. the company;

Nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon with a volume of about 1L;

The hydrogen atmosphere refers to the reaction flask connected to a hydrogen balloon with a volume of about 1L;

The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times;

There are no special instructions in the examples, and the reaction is carried out under a nitrogen atmosphere;

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

No special instructions in the examples, the reaction temperature is room temperature, and the most suitable reaction temperature for room temperature is 20°C-30°C;

DCM: dichloromethane;

EA: ethyl acetate;

HCl: hydrochloric acid;

THF: Tetrahydrofuran;

DMF: N,N-dimethylformamide;

PE: petroleum ether;

TLC: thin layer chromatography;

SFC: Supercritical fluid chromatography;

NCS: N-chlorosuccinimide;

Pd(dppf)Cl ₂ : [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride;

DMSO: dimethyl sulfoxide;

DTT: Dithiothreitol;

ATP: Adenosine triphosphate;

DNA: Deoxyribonucleotide;

IC50: refers to the concentration of the compound when the activity of DNA-PK kinase is inhibited by 50%;

X-Phos: 2-Dicyclohexylphosphorus-2,4,6-triisopropylbiphenyl.

Example

Intermediate 1

6-Methyl-2,3-dihydrobenzofuran-5-amine (Intermediate 1)

6-methyl-2,3-dihydrobenzofuran-5-amine

first step:

1-Bromo-2-(2-bromoethoxy)-4-methylbenzene (1B)

1-bromo-2-(2-bromoethoxy)-4-methylbenzene

Mix 1,2-dibromoethane (100.4g, 534.67mmol) with acetonitrile (100mL), then add 2-bromo-5-methylphenol 1A (25g, 133.67mmol), and finally add potassium carbonate (55.42g) , 401.01mmol), reacted at 80°C for 5h. After the reaction was completed, it was filtered, the filtrate was concentrated, and the silica gel column chromatography (pure petroleum ether) was separated and purified to obtain the title compound 1B (colorless liquid, 34 g, yield 86.51%).

The second step:

6-Methyl-2,3-dihydrobenzofuran (1C)

6-methyl-2,3-dihydrobenzofuran

Add 1-bromo-2-(2-bromoethoxy)-4-methylbenzene 1B (34g, 115.65mmol) into a dry reaction flask, dissolve it with dry tetrahydrofuran (160mL), and then drop at -78℃ Add n-butyllithium (55mL, 138.78mmol), and continue the reaction for 1.5h after the addition is complete. After the reaction is complete, add water (20ml) to the reaction solution for quenching, remove the organic solvent under reduced pressure, extract twice with ethyl acetate, combine the organic phases, dry and concentrate, and use silica gel column chromatography to separate and purify (pure petroleum ether) to obtain The title compound 1C (colorless liquid, 10 g, yield 64.43%).

third step:

6-Methyl-5-nitro-2,3-dihydrobenzofuran (1D)

6-methyl-5-nitro-2,3-dihydrobenzofuran

Dissolve 6-methyl-2,3-dihydrobenzofuran 1C (10g, 74.53mmol) in acetic acid (50mL), add nitric acid (11.8mL, 178.87mmol, 68% purity) dropwise at room temperature, and the addition is complete Then continue to react for 10 minutes. TLC monitored the completion of the reaction. The reaction solution was poured into ice water, extracted three times with ethyl acetate, dried and concentrated the organic phase, separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 15/1) to obtain the title compound 1D (yellow) Solid, 7.0 g, yield 52.43%).

the fourth step:

6-Methyl-2,3-dihydrobenzofuran-5-amine (Intermediate 1)

6-methyl-2,3-dihydrobenzofuran-5-amine

Mix 6-methyl-5-nitro-2,3-dihydrobenzofuran 1D (7.0g, 39.07mmol) with 110mL (ethanol/water=10/1) and add iron powder (10.9g, 195.33). mmol), and finally add dilute hydrochloric acid (9.8mL, 2mol/L), and react at 85°C for 2h. The reaction solution was filtered to remove iron powder, the filtrate was concentrated, and the pH was adjusted to weakly alkaline with saturated sodium bicarbonate solution, and then extracted three times with ethyl acetate. The organic phases were combined, dried and concentrated, and separated and purified by silica gel column chromatography (petroleum ether/ Ethyl acetate = 5/1) to obtain the title compound Intermediate 1 (brown solid, 4.5 g, yield 77.05%).

1H NMR(400MHz DMSO)δ6.52(s,1H),6.38(s,1H),4.36-4.31(t,2H),4.25(s,2H),3.02-2.98(t,2H),1.98(s ,3H).

LC-MS m/z(ESI)=150.10[M+1].

Intermediate 2

6-Chloro-2,3-dihydrobenzofuran-5-amine (Intermediate 2)

6-chloro-2,3-dihydrobenzofuran-5-amine

first step:

1-Bromo-2-(2-bromoethoxy)-4-chlorobenzene (2B)

1-bromo-2-(2-bromoethoxy)-4-chlorobenzene

Mix 1,2-dibromoethane (109.2g, 581.28mmol) with acetonitrile (120mL), then add 2-bromo-5-chlorophenol 2A (30g, 144.61mmol), and finally add potassium carbonate (60g, 434.12 mmol), react at 80°C for 5h. After the reaction was completed, it was filtered, the filtrate was concentrated, and the silica gel column chromatography was separated and purified (petroleum ether/ethyl acetate=200/1) to obtain the title compound 2B (white solid, 31 g, yield 68.19%).

The second step:

6-Chloro-2,3-dihydrobenzofuran (2C)

6-chloro-2,3-dihydrobenzofuran

Add 1-bromo-2-(2-bromoethoxy)-4-chlorobenzene 2B (31g, 98.60mmol) into a dry reaction flask, dissolve with dry tetrahydrofuran (160mL), and then add dropwise at -78℃ N-Butyllithium (45.5mL, 118.32mmol), the reaction was continued for 1.5h after the addition was completed. After the reaction was completed, the reaction solution was quenched by adding water (20 mL), removing the organic solvent under reduced pressure, extracting twice with ethyl acetate, combining the organic phases, drying and concentrating, and using silica gel column chromatography to separate and purify (pure petroleum ether) to obtain The title compound 2C (colorless liquid, 15g, yield 98.42%)

1H NMR (400MHz CDCl3) δ 6.98-6.96 (dt, 1H), 6.72-6.69 (dd, 1H), 6.67 (d, 1H), 4.50-4.46 (t, 2H), 3.08-3.03 (t, 2H) .

third step:

6-chloro-5-nitro-2,3-dihydrobenzofuran (2D)

6-chloro-5-nitro-2,3-dihydrobenzofuran

6-Chloro-2,3-dihydrobenzofuran 2C (15g, 97.03mmol) was dissolved in acetic acid (110mL), and nitric acid (15.5mL, 232.87mmol, 68% purity) was added dropwise at 70°C. After completion, the reaction was continued for 30 minutes. TLC monitored the completion of the reaction, the reaction solution was poured into ice water, extracted with ethyl acetate three times, dried and concentrated the organic phase, separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 20/1) to obtain the title compound 2D (yellow) Solid, 12.5 g, yield 64.55%).

LC-MS m/z(ESI)=200.00[M+1].

the fourth step:

6-Chloro-2,3-dihydrobenzofuran-5-amine (Intermediate 2)

6-chloro-2,3-dihydrobenzofuran-5-amine

Mix 6-chloro-5-nitro-2,3-dihydrobenzofuran 2D (12.5g, 62.63mmol) with 110mL (ethanol/water=10/1), add iron powder (17.8g, 318.77mmol) ), finally add dilute hydrochloric acid (16.5ml, 2mol/L), and react at 85°C for 2h. The reaction solution was filtered to remove iron powder, the filtrate was concentrated, and the pH was adjusted to weakly alkaline with saturated sodium bicarbonate solution, and then extracted three times with ethyl acetate. The organic phases were combined, dried and concentrated, and separated and purified by silica gel column chromatography (petroleum ether/ Ethyl acetate = 15/1) to obtain the title compound Intermediate 2 (yellow solid, 7.0 g, yield 65.91%).

1H NMR (400MHz DMSO) δ 6.72 (s, 1H), 6.64 (s, 1H), 4.73 (s, 2H), 4.43-4.39 (t, 2H), 3.07-3.02 (t, 2H).

LC-MS m/z(ESI)=170.00[M+1].

Example 1

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9 -Dihydro-8H-purin-8-one (Compound 1)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

first step:

Ethyl 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate (1b)

ethyl 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate

2,4-Dichloropyrimidine-5-carboxylic acid ethyl ester 1a (30.00g, 136.4mmol), tetrahydro-2H-pyran-4-amine hydrochloride (18.66g, 136.4mmol) were dissolved in acetonitrile (600mL ), adding potassium carbonate (46.92g, 340.9mmol) several times with stirring, and stirring at room temperature for 4h. After the reaction was monitored by TLC, it was filtered. The residue was washed with ethyl acetate (300 mL). The filtrate was concentrated to obtain a crude product. The crude product was separated and purified by column (n-hexane: ethyl acetate (v/v) = 1:1) to obtain the title compound 2- Ethyl chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate (1b) (white solid, 30.0 g, yield 77.4%).

1H NMR(400MHz DMSO)δ8.62(s,1H),8.32(d,1H),4.30(q,2H),4.21-4.16(m,1H),3.86-3.83(m,2H),3.48-3.42 (m, 2H), 1.88-1.85 (m, 2H), 1.62-1.53 (m, 2H), 1.31 (t, 3H).

LC-MS m/z(ESI)=286.10[M+1].

The second step:

2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid (1c)

2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 1b (30g, 104.99mmol) in tetrahydrofuran/water (200mL/200mL), Lithium hydroxide (5.03 g, 209.99 mmol) was added, and the mixture was stirred at room temperature for 1 h. TLC monitored the reaction to complete, concentrated to remove tetrahydrofuran, adjusted the pH to 5 with 6N hydrochloric acid, a white solid precipitated out, filtered, the filter cake was washed twice with petroleum ether, and the solid was collected to obtain the title compound 2-chloro-4-((tetrahydro-2H -Pyran-4-yl)amino)pyrimidine-5-carboxylic acid (1c) (white solid, 15.0 g, yield 55.44%).

1H NMR(400MHz DMSO) δ8.60(s,1H), 8.54(d,1H), 4.20-4.15(m,1H), 3.86-3.83(m,2H), 3.48-3.42(m,2H), 1.89 -1.86 (m, 2H), 1.60-1.50 (m, 2H).

LC-MS m/z(ESI)=258.10[M+1].

third step:

2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (1d)

2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid 1c (15g, 58.21mmol) in dimethylacetamide (150mL) and add triethyl Amine (7.38mL, 58.21mmol), diphenyl azide phosphate (12.06mL, 58.21mmol), and then gradually heated to 120°C and stirred for 1.5h. TLC monitors the completion of the reaction, the reaction solution is poured into ice water, the solid is collected by filtration, washed with water 3 times, and concentrated and dried in vacuo to obtain the title compound 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9 -Dihydro-8H-purin-8-one (1d) (white solid, 13.0 g, yield 87.69%).

1H NMR (400MHz DMSO) δ 11.63 (s, 1H), 8.11 (s, 1H), 4.43-4.37 (m, 1H), 3.98-3.94 (m, 2H), 2.59-2.38 (m, 2H), 1.73 -1.65 (m, 2H).

LC-MS m/z(ESI)=255.10[M+1].

the fourth step:

2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (1e)

2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1d (5g, 19.63mmol) in dimethylformamide ( 50mL), add dimethyl sulfate (2.48g, 19.63mmol) and cesium carbonate (9.5g, 29.445mmol) at 0°C, and stir at 0°C for 1h. TLC monitors the end of the reaction. Pour the reaction solution into ice water. A solid precipitates out. Filter and collect the solid to obtain the title compound 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7 , 9-dihydro-8H-purin-8-one (1e) (white solid, 3.0 g, yield 56.87%).

1HNMR(400MHz DMSO)δ8.36(s,1H), 4.50-4.41(m,1H), 3.99-3.95(m,2H), 3.48-3.42(m,2H), 3.34(s,3H), 2.47- 2.38 (m, 2H), 1.70 to 1.66 (m, 2H).

LC-MS m/z(ESI)=268.10[M+1].

the fifth step:

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9 -Dihydro-8H-purin-8-one (Compound 1)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1e (126mg, 0.47mmol), 6- Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (70mg, 0.47mmol), cesium carbonate (305mg, 0.94mmol), tris(dibenzylideneacetone) two palladium (42mg, 0.047mmol) ) 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (58mg, 0.094mmol) was dissolved in dioxane (10mL), protected with nitrogen and ventilated, and stirred at 100°C for 4h. TLC monitors the end of the reaction, the reaction solution is poured into ice water, the solid is collected, and the solid is separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v)=30:1) to obtain the title compound 7-methyl-2 -((6-Methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H- Purine-8-one (Compound 1) (white solid, 40 mg, yield 22.35%).

1H NMR (400MHz DMSO) δ 8.25 (s, 1H), 7.95 (s, 1H), 7.23 (s, 1H), 6.6 (s, 1H), 4.48 (t, 2H), 4.32-4.50 (m, 1H) ),3.92-3.97(dd,2H),3.40(t,2H),3.27(s,3H),3.11(t,2H),2.45-2.54(m,2H),2.11(s,3H),1.61- 1.64 (m, 2H).

LC-MS m/z(ESI)=382.30[M+1].

Example 2

9-(((1r,4r)-4-methoxycyclohexyl)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino) -7,9-dihydro-8H-purin-8-one (compound 2)

9-((1r,4r)-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8 -one

first step:

2-Chloro-4-(trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester (2a)

ethyl 2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate

Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1a (5.13g, 23.22mmol) was dissolved in acetonitrile (20mL), potassium carbonate (6.42g, 46.44mmol) was added with stirring at 0°C, and then slowly added dropwise ( 1r,4r)-4-methoxycyclohexane-1-amine (2.00g, 15.48mmol) in acetonitrile (10mL) was heated to room temperature and stirred for 1h, monitored by TLC until the reaction was over, filtered through diatomaceous earth and filtered with silica gel Column chromatography separation and purification (petroleum ether/ethyl acetate (v/v)=2:1) to obtain the title compound 2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxy Ethyl acid (2a) (white solid, 2.00 g, yield 41.26%).

LC-MS m/z(ESI)=314.10[M+1].

The second step:

2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid (2b)

2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 2a (2.00g, 6.37mmol) in tetrahydrofuran/water (15mL/15mL) and add Lithium hydroxide monohydrate (0.80g, 19.12mmol), stirring at room temperature for 2h, TLC monitoring until the end of the reaction, concentration and evaporation of tetrahydrofuran, adding 2N HCl to adjust the pH to about 3-4, a white solid precipitated, filter, filter cake Wash twice with water and petroleum ether/ethyl acetate (v/v=10/1) to obtain the title compound 2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid ( 2b) (white solid, 1.67 g, yield 91.70%).

LC-MS m/z(ESI)=286.10[M+1].

third step:

2-chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one (2c)

2-chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-(trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid 2b (1.57g, 5.49mmol) with N,N-dimethylacetamide (16mL) at room temperature Under stirring, triethylamine (0.76 mL, 5.49 mmol) and diphenyl azide phosphate (1.18 mL, 5.49 mmol) were added and reacted for 2 h, and then the temperature was raised to 110° C. and refluxed for 2.5 h. TLC monitors to the end of the reaction, the reaction solution is extracted with water and dichloromethane, and the organic layer is concentrated to obtain the title compound 2-chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purine -8-one (2c) (white solid, 1.70 g, crude product, yield 109.47%).

1H NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.11(s,1H),4.20-4.10(m,1H),3.25(s,3H),3.22-3.14(m,1H), 2.30–2.18(m,2H), 2.15–2.06(m,2H), 1.81–1.73(d,2H), 1.31–1.18(m,2H).

LC-MS m/z(ESI)=283.00[M+1].

the fourth step:

2-Chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (2d)

2-chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Use 2-chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one 2c (1.70g, 6.01mmol) with N,N-dimethylformaldehyde The amide (20 mL) was dissolved, and dimethyl sulfate (0.57 mL, 6.01 mmol) and cesium carbonate (3.92 g, 12.03 mmol) were added under stirring at 0°C to react for 2 h. TLC monitored until the reaction was over, the reaction solution was slowly added dropwise to ice water and stirred, a white solid precipitated out, washed with water and petroleum ether three times and filtered to obtain the title compound 2-chloro-9-(trans-4-methoxycyclohexyl)- 7-Methyl-7,9-dihydro-8H-purin-8-one (2d) (white solid, 1.2 g, yield 67.25%).

1H NMR (400MHz, DMSO-d6) δ 8.34 (s, 1H), 4.26-4.15 (m, 1H), 3.35 (s, 3H), 3.27 (s, 3H), 3.25-3.16 (m, 1H), 2.31–2.19(m,2H), 2.15–2.08(m,2H), 1.83–1.75(m,2H), 1.34–1.21(m,2H).

LC-MS m/z(ESI)=297.10[M+1].

the fifth step:

9-(trans-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-di Hydrogen-8H-purin-8-one (Compound 2)

9-(trans-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 2d (100mg, 0.34mmol), 6-methyl -2,3-Dihydrobenzofuran-5-amine intermediate 1 (101mg, 0.68mmol), cesium carbonate (240mg, 0.68mmol) and Brettphos G3 Pd (31mg, 0.034mmol) were added to the dry reaction flask and replaced with nitrogen After three times, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(trans-4-methoxycyclohexyl)- 7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 2) ( Light pink solid, 65 mg, yield 47.11%).

1H NMR(400MHz,DMSO-d6)δ8.20(s,1H),7.95(s,1H),7.23(s,1H),6.62(s,1H),4.49(t,2H),4.16-4.07( m, 1H), 3.25 (s, 6H), 3.12 (t, 2H), 3.09-3.02 (m, 1H), 2.34-2.22 (m, 2H), 2.14-2.04 (m, 5H), 1.71 (d, 2H), 1.27–1.15 (m, 2H).

LC-MS m/z(ESI)=410.20[M+1].

Example 3

9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7, 9-dihydro-8H-purin-8-one (compound 3)

9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydro-benzofuran-5-yl)amino)-7,9-dihydro-8H-purin -8-one

first step:

Ethyl 2-chloro-4-(trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate (3a)

ethyl 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate

Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1a (5.00g, 30.18mmol) was dissolved in acetonitrile (30mL), potassium carbonate (12.51g, 90.55mmol) was added with stirring at 0°C, and the reaction was slowly added dropwise. The acetonitrile solution (10 mL) of -4-amino-1-methylcyclohexanol hydrochloride (10.01 g, 45.27 mmol) was heated to room temperature and the reaction was stirred for 1 h, monitored by TLC until the reaction was over, filtered through diatomaceous earth, and chromatographed on a silica gel column. Separation and purification (petroleum ether/ethyl acetate (v/v)=2:1) to obtain the title compound 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5- Ethyl carboxylate (3a) (white solid, 4.70 g, yield 49.63%).

1H NMR(400MHz,Chloroform-d)δ8.66(s,1H), 4.36(q,2H), 4.30-4.21(m,1H), 2.10-2.00(m,2H), 1.74-1.63(m,5H) ), 1.62–1.50 (m, 3H), 1.40 (t, 3H), 1.31 (s, 3H).

LC-MS m/z(ESI)=314.10[M+1].

The second step:

2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (3b)

2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 3a (4.70g, 14.98mmol) in tetrahydrofuran/water (40mL/20mL) Add lithium hydroxide monohydrate (1.89g, 44.94mmol), stir the reaction at room temperature for 2h, monitor by TLC until the reaction is over, concentrate and evaporate the tetrahydrofuran, add 2N HCl to adjust the pH to about 3-4, a white solid precipitates out, filter , The filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v=10/1) to obtain the title compound 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine -5-carboxylic acid (3b) (white solid, 4.20 g, yield 98.15%).

1H NMR(400MHz,DMSO-d6)δ8.68(d,1H),8.56(s,1H),4.02(s,1H),3.16(s,1H),1.91-1.83(m,2H),1.51-- 1.45 (m, 6H), 1.15 (s, 3H).

LC-MS m/z(ESI)=314.10[M+1].

third step:

2-chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (3c)

2-chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one

Use 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 3b (4.34g, 15.19mmol) with N,N-dimethylacetamide (30mL ) Was dissolved, and triethylamine (2.11 mL, 15.19 mmol) and diphenyl azide phosphate (3.27 mL, 15.19 mmol) were added under stirring at room temperature and reacted for 2 hours, and then heated to 110° C. and refluxed for 2.5 hours. TLC monitoring to the end of the reaction, the reaction solution was added with water and a white solid precipitated, filtered, the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v=10/1) to obtain the title compound 2-chloro-9-(anti- -4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (3c) (white solid, 1.82 g, yield 42.36%).

LC-MS m/z(ESI)=283.00[M+1].

the fourth step:

2-chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (3d)

2-chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Use 2-chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 3c (1.80g, 6.37mmol) with N,N-two Methylformamide (20 mL) was dissolved, and dimethyl sulfate (0.60 mL, 6.37 mmol) and cesium carbonate (3.11 g, 9.55 mmol) were added under stirring at 0°C to react for 2 h. The reaction was monitored by TLC until the reaction was over. The reaction solution was slowly added dropwise to ice water and stirred, a white solid precipitated out, washed with water and petroleum ether three times, and filtered to obtain the title compound 2-chloro-9-(trans-4-hydroxy-4-methyl ring Hexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (3d) (white solid, 0.80 g, yield 42.34%).

1H NMR(400MHz,DMSO-d6)δ8.35(s,1H),4.48(s,1H),4.23-4.12(m,1H),3.35(s,3H),2.42-2.25(m,2H), 1.65 (d, 4H), 1.58-1.47 (m, 2H), 1.26 (s, 3H).

LC-MS m/z(ESI)=297.10[M+1].

the fifth step:

9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7, 9-dihydro-8H-purin-8-one (compound 3)

9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydro-benzofuran-5-yl)amino)-7,9-dihydro-8H-purin -8-one

The 2-chloro-9-((1r,4r)-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 3d (100mg, 0.34 mmol), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (101mg, 0.68mmol), cesium carbonate (240mg, 0.68mmol) and Brettphos G3 Pd (31mg, 0.034mmol) were added Dry the reaction flask, replace it with nitrogen three times, add dry 1,4-dioxane (1 mL), and react at 110°C for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(trans-4-hydroxy-4-methyl ring Hexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 3) (Light pink solid, 42 mg, yield 30.44%).

1H NMR (400MHz, DMSO-d6) δ 8.22 (s, 1H), 7.97 (s, 1H), 7.10 (s, 1H), 6.60 (s, 1H), 4.48 (t, 2H), 4.35 (s, 1H), 4.10–3.99(m,1H), 3.26(s,3H), 3.10(t,2H), 2.30–2.19(m,2H), 2.09(s,3H), 1.58–1.39(m,6H) ,0.98(s,3H).

LC-MS m/z(ESI)=410.20[M+1].

Example 4

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9 -Dihydro-8H-purin-8-one (Compound 4)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one

first step:

4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (4a)

ethyl 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5g, 22.6mmol), potassium carbonate (6.2g, 45.2mmol) in acetonitrile (20mL), add 4-aminopiperidine- Tert-butyl 1-carboxylate (4.5g, 22.6mmol), stirred at room temperature for 20h, monitored by TLC until the reaction was complete, added water and ethyl acetate for extraction three times, dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ Ethyl acetate (v/v)=10:)), and concentrated to obtain the title compound 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid Ethyl ester (4a) (white solid, 8.2 g, yield 95%).

1H NMR(400MHz DMSO) δ8.63(s,1H), 8.32(d,1H), 4.33-4.28(m,2H), 4.17-4.14(m,1H), 3.85(d,2H), 2.94(s , 2H), 1.88-1.80 (m, 2H), 1.51-1.44 (m, 2H), 1.41 (s, 9H), 1.32-1.29 (m, 3H).

LC-MS m/z(ESI)=385.10[M+1].

The second step:

4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid (4b)

4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxy-lic acid

Dissolve 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 4a (8.2g, 21.3mmol) in tetrahydrofuran/water (10mL /5mL), lithium hydroxide (1.8g, 42.7mmol) was added, and the mixture was stirred at room temperature for 1h. TLC monitors the reaction until the reaction is complete, concentrates and evaporates the tetrahydrofuran, adjusts the pH to 4-5, and a white solid precipitates out. It is filtered. The filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1) and concentrated to obtain the title Compound 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid (4b) (white solid, 7g, yield 86%).

1H NMR(400MHz DMSO)δ8.59(s,1H), 8.54(d,1H), 4.20-4.10(m,1H), 3.87-3.84(m,2H), 2.96(s,3H), 1.91-1.73 (m, 3H), 1.41 (s, 9H).

LC-MS m/z(ESI)=357.10[M+1].

third step

Tert-Butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate (4c)

tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate

Dissolve 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid 4b (7g, 19.6mmol) in dimethylacetamide (10mL) , Add triethylamine (1.96g, 19.6mmol) and diphenyl azide phosphate (5.4g, 19.6mmol), then gradually increase the temperature to 110°C and stir for 1.5h. TLC monitors until the reaction is complete. The reaction solution is concentrated and the residue is used Separation and purification by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) gave the title compound 4-(2-chloro-8-oxo-7,8-dihydro-9H-purine-9) -Yl)piperidine-1-carboxylic acid tert-butyl ester (4c) (white solid, 6.4 g, yield 87%).

1H NMR(400MHz DMSO)δ8.14(s,1H),4.41-4.33(m,1H),4.06(d,2H),2.88(s,2H), 2.30-2.20(m,2H),1.84-1.71 (m, 2H), 1.43 (s, 9H).

LC-MS m/z(ESI)=354.10[M+1].

the fourth step

Tert-Butyl 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate (4d)

tert-butyl4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate

Dissolve tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate 6c (6.4g, 18.1mmol) in dimethyl Formamide (10 mL), dimethyl sulfate (2.28 g, 18.1 mmol) and cesium carbonate (8.5 g, 27.1 mmol) were added at 0°C, and stirred at 0°C for 0.5 h. TLC monitors until the reaction is complete, water is added to the reaction solution, a solid precipitates out, filtered to obtain the title compound 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purine-9 -Yl)piperidine-1-carboxylic acid tert-butyl ester (4d) (white solid, 5.4 g, yield 79%).

1H NMR (400MHz DMSO) δ 8.36 (s, 1H), 4.46-4.37 (m, 1H), 4.05 (d, 2H), 3.35 (s, 3H), 2.87 (s, 2H), 2.33-2.19 (m , 2H), 1.74-1.72 (m, 2H), 1.43 (s, 9H).

LC-MS m/z(ESI)=368.10[M+1].

the fifth step:

2-chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one (4e)

2-chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one

Add tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate 4d (2g, 5.4mmol) into the reaction flask, 2M hydrochloric acid-ethyl acetate solution (10 mL) was added with stirring at room temperature, and stirred at room temperature for 4 hours. TLC monitoring until the reaction is complete, the reaction solution is concentrated to obtain the title compound 2-chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one hydrochloric acid Salt (4e) (white solid, 1.4 g, yield 91%).

1H NMR (400MHz DMSO) δ 8.39 (s, 1H), 6.42 (s, 1H), 4.59-4.53 (m, 1H), 3.39 (s, 2H), 3.36 (s, 3H), 3.16-3.04 (m , 2H), 2.62-2.50 (m, 2H), 2.07-1.93 (m, 2H).

LC-MS m/z(ESI)=268.10[M+1].

The sixth step:

2-chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one (4f)

2-chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one 4e (1.4g, 5.4mmol) in methanol (20mL), 4A molecular sieve (100mg) was added, then paraformaldehyde (783mg, 27mmol) was added, the reaction was stirred at room temperature for 6h, and then sodium cyanoborohydride (1g, 16.2mmol) was added. TLC monitored until the reaction was complete, filtered and concentrated to obtain the title compound 2-chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one (4f) (white solid, 600 mg, yield 62%).

1H NMR(400MHz DMSO)δ8.35(s,1H), 4.21-4.13(m,1H), 3.35(s,3H), 2.92(d,2H), 2.45-2.41(m,2H), 2.23(s , 3H), 2.09-2.03 (m, 2H), 1.70-1.67 (m, 2H).

LC-MS m/z(ESI)=282.10[M+1].

The seventh step:

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9 -Dihydro-8H-purin-8-one (Compound 4)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one

The 2-chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one 4f (100mg, 0.35mmol), 6-methyl 2-,3-dihydrobenzofuran-5-amine intermediate 1 (106mg, 0.71mmol), cesium carbonate (231mg, 0.71mmol) and Brettphos G3 Pd (32mg, 0.035mmol) into the dry reaction flask, nitrogen After three replacements, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-(((6-methyl -2,3-Dihydrobenzofuran-5-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one (compound 4 ) (White solid, 35 mg, yield 25.00%).

1H NMR(400MHz,DMSO-d6)δ8.20(s,1H),7.94(s,1H),7.25(s,1H),6.60(s,1H),4.49(t,2H),4.13-4.03( m, 1H), 3.26 (s, 3H), 3.12 (t, 2H), 2.86 (d, 2H), 2.59-2.47 (m, 2H), 2.19 (s, 3H), 2.12 (s, 3H), 1.94 (t, 2H), 1.61 (d, 2H).

LC-MS m/z(ESI)=395.20[M+1].

Example 5

9-Cyclohexyl-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purine-8- Ketone (Compound 5)

9-cyclohexyl-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Ethyl 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylate (5a)

ethyl 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5.0g, 22.6mmol), potassium carbonate (6.2g, 45.2mmol) in acetonitrile (50mL), add cyclohexylamine (2.2 g, 22.6mmol), stirred at room temperature for 20h. TLC monitors the completion of the reaction, adds water and ethyl acetate to extract three times, dry with anhydrous sodium sulfate and separate and purify by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=10:1), and concentrate to obtain the title compound 2-chloro- Ethyl 4-(cyclohexylamino)pyrimidine-5-carboxylate (5a) (white solid, 4.1 g, yield 64%).

1H NMR(400MHz,Chloroform-d)δ8.64(s,1H), 8.53-8.23(m,1H), 4.35(q,2H), 4.25-4.05(m,1H), 2.01-1.95(m,2H) ),1.77-1.72(m,2H),1.64-1.61(m,1H),1.50-1.41(m,3H),1.39(t,3H),1.35-1.18(m,2H).

LC-MS m/z(ESI)=284.10[M+1].

The second step:

2-Chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid (5b)

2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid ethyl ester 5a (4.1g, 14.4mmol) in tetrahydrofuran/water (20mL/20mL), add lithium hydroxide (691mg, 28.8mmol) , Stir at room temperature for 1h. The reaction was monitored by TLC, the tetrahydrofuran was spin-dried, and the pH was adjusted to 4-5. A white solid precipitated out. The filter cake was washed twice with petroleum ether/ethyl acetate (v/v=10/1) and concentrated to obtain the title compound 2. -Chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid (5b) (white solid, 3.0 g, yield 81%).

1H NMR(400MHz,DMSO-d6)δ13.77(s,1H),8.76-8.45(m,2H),4.01-3.92(m,1H),2.05-1.79(m,2H),1.79-1.65(m ,2H),1.60-1.54(m,1H),1.42-1.19(m,5H).

LC-MS m/z(ESI)=256.10[M+1].

third step:

2-chloro-9-cyclohexyl-7,9-dihydro-8H-purin-8-one (5c)

2-chloro-9-cyclohexyl-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid 5b (3.0g, 11.7mmol) in dimethylacetamide (30mL), add triethylamine (1.18g, 11.7mmol), stack Diphenyl nitrogen phosphate (3.22g, 11.7mmol), then gradually heated to 110°C and stirred for 1.5h. The end of the reaction was monitored by TLC, the reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound 2-chloro-9-cyclohexyl-7,9- Dihydro-8H-purin-8-one (5c) (white solid, 2.2 g, yield 74%).

1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),8.12(s,1H),4.19-4.11(m,1H),1.92-1.57(m,6H),1.52-0.93(m,4H) ).

LC-MS m/z(ESI)=253.10[M+1].

the fourth step:

2-chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one (5d)

2-chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-cyclohexyl-7,9-dihydro-8H-purin-8-one 5c (2.2g, 8.7mmol) in dimethylformamide (15mL), add two sulfuric acid at 0℃ Methyl ester (1.1g, 8.7mmol) and cesium carbonate (4.25g, 13mmol) were stirred and reacted for 0.5h. The end of the reaction was monitored by TLC, water was added to the reaction solution, a solid precipitated out, and the title compound 2-chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one (5d ) (Yellow solid, 1.1 g, yield 48%).

1H NMR(400MHz,DMSO-d6)δ8.34(s,1H),4.22-4.41(m,1H),3.35(s,3H),2.22-2.12(m,2H),1.91-160(m,5H) ), 1.43-1.33 (m, 2H), 1.27-1.14 (m, 1H).

LC-MS m/z(ESI)=267.10[M+1].

the fifth step:

9-Cyclohexyl-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purine-8- Ketone (Compound 5)

9-cyclohexyl-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one 5d (100mg, 0.37mmol), 6-methyl-2,3-dihydrobenzo Furan-5-amine Intermediate 1 (112mg, 0.75mmol), Cesium Carbonate (244mg, 0.75mmol) and Brettphos G3 Pd (34mg, 0.037mmol) were added to the dry reaction flask, replaced with nitrogen three times and then added to the dry 1,4- Dioxane (1 mL) was reacted at 110°C for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-cyclohexyl-7-methyl-2-(( (6-Methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 5) (pale yellow solid, 34mg, yield 23.90%).

1H NMR(400MHz,DMSO-d6)δ8.18(s,1H),7.95(s,1H),7.26(s,1H),6.61(s,1H),4.49(t,2H),4.16-4.05( m, 1H), 3.26 (s, 3H), 3.11 (t, 2H), 2.27-2.15 (m, 2H), 2.12 (s, 3H), 1.83-1.76 (m, 2H), 1.71-1.65 (m, 2H), 1.38–1.22 (m, 3H), 1.17–1.04 (m, 1H).

LC-MS m/z(ESI)=380.20[M+1].

Example 6

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7, 9-dihydro-8H-purin-8-one (compound 6)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8 -one

first step:

Ethyl 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylate (6a)

ethyl 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5.0g, 22.6mmol), potassium carbonate (6.2g, 45.2mmol) in acetonitrile (50mL), add tetrahydro-2H- at 0℃ Pyran-3-amine (2.3g, 22.6mmol), stirred at room temperature for 20h. Add water and ethyl acetate to extract three times, dry with anhydrous sodium sulfate and separate and purify by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=10:1), and concentrate to obtain the title compound 2-chloro-4-((四Hydrogen-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (6a) (white solid, 4.1 g, yield 64%).

1H NMR(400MHz,Chloroform-d)δ8.69(s,1H),8.68(s,1H),4.37(q,2H),4.34-4.24(m,1H),3.89-3.84(m,1H), 3.77-3.66(m,2H),3.58-3.54(m,1H),2.01-1.95(m,1H),1.86-1.65(m,2H),1.69-1.63(m,1H),1.40(t,3H) ).

LC-MS m/z(ESI)=286.10[M+1].

The second step:

2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid (6b)

2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 6a (4.1g, 14.3mmol) in tetrahydrofuran/water (20mL/20mL) , Lithium hydroxide (686 mg, 28.6 mmol) was added, and the reaction was stirred at room temperature for 1 h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 4-5, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title compound 2-Chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid (6b) (white solid, 3.5 g, yield 95%).

1H NMR(400MHz,DMSO-d6)δ13.83(s,1H),8.75(d,1H),8.59(s,1H),4.14-4.07(m,1H),3.76-3.72(m,1H), 3.59 (t, 1H), 3.47-3.42 (m, 2H), 1.93-1.86 (m, 1H), 1.79-1.62 (m, 2H), 1.58-1.49 (m, 1H).

LC-MS m/z(ESI)=258.10[M+1].

third step:

2-chloro-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one (6c)

2-chloro-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid 6b (3.5g, 13.6mmol) in dimethylacetamide (30mL), and then Triethylamine (1.37g, 13.6mmol) and diphenyl azide phosphate (3.74g, 13.6mmol) were added and then gradually heated to 110°C and stirred for 1.5h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound 2-chloro-9-(tetrahydro-2H- Pyran-3-yl)-7,9-dihydro-8H-purin-8-one (6c) (white solid, 2.2 g, yield 63%).

1H NMR(400MHz,DMSO-d6)δ11.67(s,1H),8.13(s,1H),4.29-4.21(m,1H),3.86-3.77(m,2H),3.37-3.29(m,2H) ), 2.04–1.39 (m, 4H).

LC-MS m/z(ESI)=255.10[M+1].

the fourth step:

2-chloro-7-methyl-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one (6d)

2-chloro-7-methyl-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one 6c (2.2g, 8.7mmol) in dimethylformamide (15mL), add dimethyl sulfate (1.1g, 8.7mmol) and cesium carbonate (4.25g, 13mmol) at 0°C and stir for 0.5h. TLC monitored to the end of the reaction, water was added to the reaction solution, a solid precipitated out, filtered to obtain the title compound 2-chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one ( 6d) (yellow solid, 1.7 g, yield 73%).

1H NMR(400MHz,DMSO-d6)δ8.36(s,1H), 4.35-4.23(m,1H), 3.91-3.77(m,2H), 3.35(s,3H),3.34-3.33(m,2H) ), 2.50-2.44 (m, 1H), 1.98-1.89 (m, 1H), 1.83-1.62 (m, 2H).

LC-MS m/z(ESI)=269.10[M+1].

the fifth step:

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7, 9-dihydro-8H-purin-8-one (compound 6)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one 6d (100mg, 0.37mmol), 6-methyl-2,3-dihydrobenzo Furan-5-amine intermediate 1 (112mg, 0.75mmol), cesium carbonate (242mg, 0.75mmol) and Brettphos G3 Pd (34mg, 0.037mmol) were added to the dry reaction flask, replaced with nitrogen three times and then added to the dry 1,4- Dioxane (1 mL) was reacted at 110°C for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-(((6-methyl -2,3-Dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one (compound 6) (Light pink solid, 42 mg, yield 29.59%).

1H NMR (400MHz, DMSO-d6) δ 8.25 (s, 1H), 7.97 (s, 1H), 7.22 (s, 1H), 6.62 (s, 1H), 4.49 (t, 2H), 4.27-4.16 ( m,1H),3.94-3.82(m,2H), 3.75(dd,1H), 3.26(s,3H), 3.23-3.16(m,1H), 3.11(t,2H), 2.49-2.40(m, 1H), 2.12 (s, 3H), 1.85 (d, 1H), 1.75-1.61 (m, 2H).

LC-MS m/z(ESI)=382.20[M+1].

Example 7

(S)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9- Dihydro-8H-purin-8-one (Compound 7)

(S)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-furan-3-yl)-7,9-dihydro-8H-purin -8-one

first step:

(S)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (7a)

ethyl(S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5g, 5.35mmol) and potassium carbonate (1.4g, 22.6mmol) in acetonitrile (20mL), add (S)-tetrahydrofuran- at 0°C 3-amine (660mg, 5.35mmol), stirred at room temperature for 20h. Add water and ethyl acetate to extract three times, dry with anhydrous sodium sulfate and separate and purify by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=10:1), and concentrate to obtain the title compound (S)-2-chloro-4 -((Tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (7a) (white solid, 2.2 g, yield 36%).

1H NMR(400MHz DMSO)δ8.68(s,1H), 8.58(d,1H), 4.79-4.77(m,1H), 4.39-4.33(m,2H), 4.04-3.98(m,2H), 3.90 -3.87 (m, 1H), 3.76-3.73 (m, 1H), 2.43-2.33 (m, 1H), 1.95-1.88 (m, 1H), 1.39 (t, 3H).

LC-MS m/z(ESI)=272.00[M+1].

The second step:

(S)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid (7b)

(S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid

Dissolve (S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 7a (2.2g, 8.1mmol) in tetrahydrofuran/water (5mL/5mL) and add Lithium hydroxide (681 mg, 16.2 mmol) was stirred at room temperature for 1 h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 4-5, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title compound (S)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid (7b) (white solid, 1.7 g, yield 89%).

1H NMR (400MHz DMSO) δ 13.83 (s, 1H), 8.65 (s, 1H), 8.63 (d, 1H), 4.61 (s, 1H), 3.89-3.80 (m, 2H), 3.76-3.70 (m , 1H), 3.65-3.63 (m, 1H), 2.50-2.25 (m, 1H), 1.87-1.86 (m, 1H).

LC-MS m/z(ESI)=244.20[M+1].

third step:

(S)-2-chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (7c)

(S)-2-chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one

(S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid 7b (1.7g, 7mmol) was dissolved in dimethylacetamide (10mL), and triethylamine ( 707 mg, 7 mmol), diphenyl azide phosphate (1.9 g, 7 mmol), and then gradually heated to 110° C. and stirred for 1.5 h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:6) to obtain the title compound (S)-2-chloro-9-(tetrahydrofuran) -3-yl)-7,9-dihydro-8H-purin-8-one (7c) (white solid, 1.4g, yield 87%).

1H NMR(400MHz DMSO)δ8.13(s,1H), 5.00-4.93(m,1H), 4.12-4.06(dd,1H), 3.97(t,1H), 3.88-3.87(m,2H), 3.33 (s, 3H), 2.43-2.37 (m, 1H), 2.25-2.20 (m, 1H).

LC-MS m/z(ESI)=241.20[M+1].

the fourth step:

(S)-2-Chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (7d)

(S)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one

Dissolve (S)-2-chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 7c (1.4g, 5.8mmol) in dimethylformamide (10mL ), add dimethyl sulfate (735 mg, 5.8 mmol) and cesium carbonate (2.85 g, 8.7 mmol) at 0° C. and stir and react for 0.5 h. TLC monitors to the end of the reaction, water is added to the reaction solution, a solid is precipitated, and the title compound (S)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro is obtained by filtration. -8H-purin-8-one (7d) (white solid, 1.2g, yield 85%).

1H NMR(400MHz DMSO)δ8.35(s,1H),5.04-4.97(m,1H),4.13-4.07(dd,1H),3.98(t,1H),3.90-3.84(m,2H), 3.35 (s,3H),2.44-2.37(m,1H),2.28-2.27(m,1H).

LC-MS m/z(ESI)=255.20[M+1].

the fifth step:

(S)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9- Dihydro-8H-purin-8-one (Compound 7)

(S)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-furan-3-yl)-7,9-dihydro-8H-purin -8-one

Add (S)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 7d (100mg, 0.39mmol), 6-methyl -2,3-Dihydrobenzofuran-5-amine intermediate 1 (117mg, 0.79mmol), cesium carbonate (256mg, 0.75mmol) and Brettphos G3 Pd (36mg, 0.039mmol) were added to the dry reaction flask and replaced with nitrogen After three times, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound (S)-7-methyl-2-((( 6-Methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (compound 7) (Light yellow solid, 43 mg, yield 20.79%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),7.98(s,1H),7.19(s,1H),6.61(s,1H),4.93-4.83(m,1H),4.49( t, 2H), 3.96--3.87(m, 2H), 3.82--3.75(m, 2H), 3.27(s, 3H), 3.11(t, 2H), 2.41--2.32(m, 1H), 2.21-2.12( m, 1H), 2.11 (s, 3H).

LC-MS m/z(ESI)=368.20[M+1].

Example 8

(R)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9- Dihydro-8H-purin-8-one (Compound 8)

(R)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-furan-3-yl)-7,9-dihydro-8H-purin -8-one

first step:

(R)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (8a)

ethyl(R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5g, 5.35mmol) and potassium carbonate (1.4g, 22.6mmol) in acetonitrile (20mL), add (R)-tetrahydrofuran- at 0°C 3-amine (660mg, 5.35mmol), stirred at room temperature for 20h. Add water and ethyl acetate to extract three times, dry with anhydrous sodium sulfate and separate and purify by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=10:1), and concentrate to obtain the title compound (R)-2-chloro-4 -((Tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (8a) (white solid, 4.3 g, yield 65%).

1H NMR(400MHz DMSO)δ8.68(s,1H),8.58(d,1H),4.83-4.77(m,1H),4.39-4.33(m,2H),4.02-3.98(m,2H),3.90 -3.86 (m, 1H), 3.76-3.73 (m, 1H), 2.43-2.34 (m, 1H), 1.98-1.88 (m, 1H), 1.39 (t, 3H).

LC-MS m/z(ESI)=272.00[M+1].

The second step:

(R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid (8b)

(R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid

(R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 8a (4.3g, 15.8mmol) was dissolved in tetrahydrofuran/water (5mL/5mL) and added Lithium hydroxide (1.3g, 31.7mmol) was stirred at room temperature for 1h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 4-5, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title compound (R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid (8b) (white solid, 2.9 g, yield 87%).

1H NMR(400MHz DMSO)δ13.83(s,1H),8.65(s,1H),8.59(d,1H),4.62-4.60(m,1H),3.89-3.80(m,2H),3.76-3.70 (m, 1H), 3.64-3.56 (m, 1H), 2.33-2.23 (m, 1H), 1.88-1.83 (m, 1H).

LC-MS m/z(ESI)=244.20[M+1].

third step:

(R)-2-chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (8c)

(R)-2-chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one

Dissolve (R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid 8b (2.9g, 11.9mmol) in dimethylacetamide (20mL) and add triethylamine (1.2g, 11.9mmol), diphenyl azide phosphate (3.3g, 11.9mmol), and then gradually heated to 110°C and stirred for 1.5h. The reaction was monitored by TLC and the reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:6) to obtain the title compound (R)-2-chloro-9-(tetrahydrofuran) -3-yl)-7,9-dihydro-8H-purin-8-one (8c) (white solid, 2.2 g, yield 68%).

1H NMR(400MHz DMSO)δ8.36(s,1H),5.04-4.96(m,1H),4.13-4.07(dd,1H),3.97(t,1H),3.87-3.84(m,2H),3.33 (s, 3H), 2.40-2.36 (m, 1H), 2.29-2.27 (m, 1H).

LC-MS m/z(ESI)=241.20[M+1].

the fourth step:

(R)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (8d)

(R)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one

(R)-2-chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 8c (2.2g, 9.1mmol) was dissolved in dimethylformamide (10mL ), add dimethyl sulfate (1.1 g, 9.1 mmol) and cesium carbonate (4.5 g, 13.7 mmol) at 0° C. and stir and react for 0.5 h. TLC monitors to the end of the reaction, water is added to the reaction solution, a solid is precipitated, and the title compound (R)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro is obtained by filtration. -8H-purin-8-one (8d) (white solid, 1.7 g, yield 82%).

1H NMR (400MHz DMSO) δ8.41 (s, 1H), 5.05-4.97 (m, 1H), 4.13-4.08 (dd, 1H), 3.98 (t, 1H), 3.91-3.84 (m, 2H), 3.33 (s, 3H), 2.45-2.27 (m, 1H), 2.26-2.20 (m, 1H).

LC-MS m/z(ESI)=255.20[M+1].

the fifth step:

(R)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9- Dihydro-8H-purin-8-one (Compound 8)

(R)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-furan-3-yl)-7,9-dihydro-8H-purin -8-one

Add (R)-2-chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 8d (100mg, 0.39mmol), 6-methyl -2,3-Dihydrobenzofuran-5-amine intermediate 1 (117mg, 0.79mmol), cesium carbonate (256mg, 0.75mmol) and Brettphos G3 Pd (36mg, 0.039mmol) were added to the dry reaction flask and replaced with nitrogen After three times, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound (R)-7-methyl-2-(( 6-Methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (compound 8) (Light yellow solid, 60 mg, yield 41.59%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),7.98(s,1H),7.19(s,1H),6.61(s,1H),4.93-4.82(m,1H),4.49( t, 2H), 3.95-3.88(m, 2H), 3.81-3.75(m, 2H), 3.27(s, 3H), 3.11(t, 2H), 2.40-2.32(m, 1H), 2.18-2.12( m, 1H), 2.11 (s, 3H).

LC-MS m/z(ESI)=368.20[M+1].

Example 9

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(((tetrahydrofuran-3-yl)methyl)-7,9 -Dihydro-8H-purin-8-one (compound 9)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8- one

first step:

Ethyl 2-chloro-4-((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylate (9a)

ethyl 2-chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (6.2g, 22.6mmol), potassium carbonate (6.2g, 44.8mmol) in acetonitrile (20mL), add (tetrahydrofuran-3- Methyl) formamide (2.3g, 22.6mmol), stirred at room temperature for 20h. Add water and ethyl acetate to extract three times, dry with anhydrous sodium sulfate and separate and purify by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=10:1), and concentrate to obtain the title compound 2-chloro-4-((tetrahydrofuran) -3-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester (9a) (white solid, 4.4 g, yield 85%).

1H NMR (400MHz DMSO) δ 8.58-8.61 (m, 2H), 4.31 (q, 2H), 3.78-3.73 (m, 1H), 3.69-3.58 (m, 2H), 3.48-3.44 (m, 3H) , 2.57-2.53 (m, 1H), 1.98-1.91 (m, 1H), 1.62-1.58 (m, 1H), 1.31 (t, 3H).

LC-MS m/z(ESI)=286.20[M+1].

The second step:

2-Chloro-4-((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylic acid (9b)

2-chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylate

Dissolve 2-chloro-4-((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester 9a (4.4g, 15.3mmol) in tetrahydrofuran/water (10mL/5mL), add hydrogen Lithium oxide (736 mg, 30.6 mmol) was stirred at room temperature for 1 h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 4-5, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title compound 2-Chloro-4-((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylic acid (9b) (white solid, 3.4 g, yield 80%).

1H NMR (400MHz DMSO) δ 13.75 (s, 1H), 8.75 (s, 1H), 8.57 (s, 1H), 3.78-3.73 (m, 3H), 3.47-3.43 (m, 3H), 2.58-2.52 (m, 1H), 1.98-1.89 (m, 3H), 1.63-1.55 (m, 1H).

LC-MS m/z(ESI)=259.20[M+1].

third step:

2-Chloro-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one (9c)

2-chloro-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylic acid 9b (3.4g, 13.0mmol) in dimethylacetamide (20mL), add triethylamine ( 1.58g, 13.0mmol), diphenyl azide phosphate (3.6g, 13.0mmol), and then gradually heated to 110°C and stirred for 1.5h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound 2-chloro-9-((tetrahydrofuran-3- (Yl)methyl)-7,9-dihydro-8H-purin-8-one (9c) (white solid, 1.46 g, yield 50%).

1H NMR (400MHz DMSO) δ 11.65 (s, 1H), 8.14 (s, 1H), 3.80-3.74 (m, 3H), 3.66-3.57 (m, 2H), 3.52-3.49 (m, 1H), 2.73 -2.69 (m, 1H), 1.98-1.89 (m, 1H), 1.67-1.60 (m, 1H).

LC-MS m/z(ESI)=255.20[M+1].

the fourth step:

2-Chloro-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one (9d)

2-chloro-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one 9c (1.6g, 5.2mmol) in dimethylformamide (10mL ), dimethyl sulfate (663 mg, 5.2 mmol) and cesium carbonate (2.4 g, 7.7 mmol) were added at 0° C. and the reaction was stirred for 1 h. TLC monitors to the end of the reaction, water is added to the reaction solution, a solid is precipitated, and the title compound 2-chloro-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro is obtained by filtration. -8H-purin-8-one (9d) (white solid, 1.2 g, yield 80%).

1H NMR(400MHz DMSO)δ8.35(s,1H),3.81-3.75(m,3H),3.65-3.57(m,2H),3.52-3.49(m,1H),3.37(s,3H),2.73 -2.70 (m, 1H), 1.96-1.91 (m, 1H), 1.67-1.61 (m, 1H).

LC-MS m/z(ESI)=268.20[M+1].

the fifth step:

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(((tetrahydrofuran-3-yl)methyl)-7,9 -Dihydro-8H-purin-8-one (compound 9)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8- one

The 2-chloro-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one 9d (100mg, 0.37mmol), 6-methyl -2,3-Dihydrobenzofuran-5-amine intermediate 1 (112mg, 0.74mmol), cesium carbonate (242mg, 0.74mmol) and Brettphos G3 Pd (34mg, 0.037mmol) were added to the dry reaction flask and replaced with nitrogen After three times, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-(((6-methyl -2,3-Dihydrobenzofuran-5-yl)amino)-9-(((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one (compound 9 ) (Off-white solid, 22 mg, yield 15.50%).

1H NMR (400MHz, DMSO-d6) δ 8.25 (s, 1H), 7.95 (s, 1H), 7.18 (s, 1H), 6.60 (s, 1H), 4.49 (t, 2H), 4.24 (p, 1H), 3.81-3.72 (m, 2H), 3.66-3.57 (m, 2H), 3.28 (s, 3H), 3.11 (t, 2H), 2.10 (s, 3H), 1.93-1.77 (m, 3H) ,1.70-1.60(m,1H).

LC-MS m/z(ESI)=382.20[M+1].

Example 10

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H -Purin-8-one (Compound 10)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one

first step:

4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (10a)

ethyl 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5g, 22.6mmol), potassium carbonate (6.2g, 45.2mmol) in acetonitrile (20mL), add 4-aminopiperidine- Tert-Butyl 1-carboxylate (4.5g, 22.6mmol), stirred at room temperature for 20h, TLC monitored until the reaction was complete, added water and ethyl acetate for three times, dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ Ethyl acetate (v/v)=10:)), and concentrated to obtain the title compound 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid Ethyl ester (10a) (white solid, 8.2 g, yield 95%).

1H NMR(400MHz DMSO) δ8.63(s,1H), 8.32(d,1H), 4.33-4.28(m,2H), 4.17-4.14(m,1H), 3.85(d,2H), 2.94(s , 2H), 1.88-1.80 (m, 2H), 1.51-1.44 (m, 2H), 1.41 (s, 9H), 1.32-1.29 (m, 3H).

LC-MS m/z(ESI)=385.10[M+1].

The second step:

4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid (10b)

4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxy-lic acid

Dissolve 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 10a (8.2g, 21.3mmol) in tetrahydrofuran/water (10mL /5mL), lithium hydroxide (1.8g, 42.7mmol) was added, and the mixture was stirred at room temperature for 1h. TLC monitors the reaction until the reaction is complete, concentrates and evaporates the tetrahydrofuran, adjusts the pH to 4-5, and a white solid precipitates out, filtered, the filter cake was washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title Compound 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid (10b) (white solid, 7g, yield 86%).

1H NMR(400MHz DMSO)δ8.59(s,1H), 8.54(d,1H), 4.20-4.10(m,1H), 3.87-3.84(m,2H), 2.96(s,3H), 1.91-1.73 (m, 3H), 1.41 (s, 9H).

LC-MS m/z(ESI)=357.10[M+1].

third step

Tert-Butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate (10c)

tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate

Dissolve 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid 10b (7g, 19.6mmol) in dimethylacetamide (10mL) , Add triethylamine (1.96g, 19.6mmol) and diphenyl azide phosphate (5.4g, 19.6mmol), then gradually increase the temperature to 110°C and stir for 1.5h. TLC monitors until the reaction is complete. The reaction solution is concentrated and the residue is used Separation and purification by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) gave the title compound 4-(2-chloro-8-oxo-7,8-dihydro-9H-purine-9) -Yl)piperidine-1-carboxylic acid tert-butyl ester (10c) (white solid, 6.4 g, yield 87%).

1H NMR(400MHz DMSO)δ8.14(s,1H),4.41-4.33(m,1H),4.06(d,2H),2.88(s,2H), 2.30-2.20(m,2H),1.84-1.71 (m, 2H), 1.43 (s, 9H).

LC-MS m/z(ESI)=354.10[M+1].

the fourth step:

Tert-Butyl 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate (10d)

tert-butyl4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate

Dissolve tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate 10c (6.4g, 18.1mmol) in dimethyl Formamide (10 mL), dimethyl sulfate (2.28 g, 18.1 mmol) and cesium carbonate (8.5 g, 27.1 mmol) were added at 0°C, and stirred at 0°C for 0.5 h. TLC monitors until the reaction is complete, water is added to the reaction solution, a solid is precipitated, and the title compound 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purine-9 is obtained by filtration. -Yl)piperidine-1-carboxylic acid tert-butyl ester (10d) (white solid, 5.4 g, yield 79%).

1H NMR (400MHz DMSO) δ 8.36 (s, 1H), 4.46-4.37 (m, 1H), 4.05 (d, 2H), 3.35 (s, 3H), 2.87 (s, 2H), 2.33-2.19 (m , 2H), 1.74-1.72 (m, 2H), 1.43 (s, 9H).

LC-MS m/z(ESI)=368.10[M+1].

the fifth step:

4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purine- 9-yl) piperidine-1-carboxylic acid tert-butyl ester (10e)

tert-butyl 4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine -1-carboxylate

The 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl) piperidine-1-carboxylic acid tert-butyl ester 10d (200mg, 0.54mmol), 6-Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (161mg, 1.08mmol), cesium carbonate (381mg, 1.08mmol) and Brettphos G3 Pd (49mg, 0.054mmol) were added to the dry reaction After replacing the flask with nitrogen three times, add dry 1,4-dioxane (1 mL), and react at 110°C for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was filtered with celite. The filtrate was concentrated to obtain crude 4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl) )Amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylic acid tert-butyl ester (10e), directly cast to the next step without purification.

The sixth step:

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H -Purin-8-one (Compound 10)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one

The crude 4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H- Purin-9-yl)piperidine-1-carboxylic acid tert-butyl ester 10e was dissolved in dichloromethane (2 mL), and then trifluoroacetic acid (0.19 mL, 2.5 mmol) was added, and the reaction was carried out at room temperature for 1 h. TLC monitors to the end of the reaction, adjust the pH to 8-9 with saturated sodium bicarbonate solution, add dichloromethane for extraction three times, concentrate the organic phase, and use silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v)=40: 1)) Purified to obtain the title compound 7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(piperidin-4-yl)-7 , 9-dihydro-8H-purin-8-one (compound 10) (white solid, 40 mg, yield 19.38%).

1H NMR(400MHz,DMSO-d6)δ8.18(s,1H),7.93(s,1H),7.24(s,1H),6.60(s,1H),4.48(t,2H),4.22-4.12( m, 1H), 3.26 (s, 3H), 3.12 (t, 2H), 3.03 (d, 2H), 2.55-2.48 (m, 2H), 2.38-2.27 (m, 2H), 2.12 (s, 3H) ,1.60(d,2H).

LC-MS m/z(ESI)=381.20[M+1].

Example 11

9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7 ,9-Dihydro-8H-purin-8-one (Compound 11)

9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8 -one

first step:

Ethyl 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate (11a)

ethyl 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5.0g, 22.6mmol) and potassium carbonate (6.2g, 45.2mmol) in acetonitrile (50mL), add (1s, 4s) at 0℃ -4-amino-1-methylcyclohexan-1-ol (2.9g, 22.6mmol), stirred at room temperature for 20h. Add water and ethyl acetate to extract three times, dry with anhydrous sodium sulfate and separate and purify by silica gel column chromatography (n-hexane/ethyl acetate (v/v) = 10:1), and concentrate to obtain the title compound 2-chloro-4-((cis Ethyl 4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate (11a) (white solid, 4.1 g, yield 58%).

1H NMR(400MHz,Chloroform-d)δ8.65(s,1H),8.37(d,1H),4.35(q,2H),4.09-4.04(m,1H),1.94-1.82(m,2H), 1.82-1.45 (m, 6H), 1.38 (t, 3H), 1.28 (s, 3H).

LC-MS m/z(ESI)=314.10[M+1].

The second step:

2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (13b)

2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 11a (4.1g, 13.1mmol) in tetrahydrofuran/water (20mL/20mL) Lithium hydroxide (629 mg, 26.2 mmol) was added to the mixture, and the reaction was stirred at room temperature for 1 h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 4-5, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title compound 2-Chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (11b) (white solid, 3.5 g, yield 93%).

1H NMR(400MHz,DMSO-d6)δ13.77(s,1H),8.56(s,1H),8.49(d,1H),3.93-3.83(m,2H),1.78-1.50(m,6H), 1.41 (td, 2H), 1.11 (s, 3H).

LC-MS m/z(ESI)=286.10[M+1].

third step:

2-chloro-9-cis-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (11c)

2-chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 11b (3.5g, 12.2mmol) in dimethylacetamide (30mL) and add Triethylamine (1.24g, 12.2mmol), diphenyl azide phosphate (3.36g, 12.2mmol), then gradually heated to 110°C and stirred for 1.5h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound 2-chloro-9-cis-4-hydroxy- 4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (11c) (white solid, 1.4g, yield 41%).

1H NMR(400MHz,DMSO-d6)δ11.58(s,1H),8.11(s,1H),4.22(s,1H),4.22-4.02(m,1H),2.65-2.56(m,2H), 1.83-1.60 (m, 2H), 1.47-1.43 (m, 4H), 1.18 (s, 3H).

LC-MS m/z(ESI)=283.10[M+1].

the fourth step:

2-Chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (11d)

2-chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-cis-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 11c (1.4g, 4.9mmol) in dimethylformamide (15mL), add dimethyl sulfate (618mg, 4.9mmol) and cesium carbonate (3.2g, 9.8mmol) at 0°C and stir and react for 0.5h. TLC monitored to the end of the reaction, water was added to the reaction solution, a solid precipitated out, filtered to obtain the title compound 2-chloro-9-cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9- Dihydro-8H-purin-8-one (11d) (yellow solid, 502 mg, yield 34%).

LC-MS m/z(ESI)=297.10[M+1].

the fifth step:

9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7 ,9-Dihydro-8H-purin-8-one (Compound 11)

9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-9-cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 11d (80mg, 0.27mmol), 6- Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (80mg, 0.54mmol), cesium carbonate (176mg, 0.54mmol) and Brettphos G3 Pd (24mg, 0.027mmol) were added to the dry reaction flask, After nitrogen replacement three times, dry 1,4-dioxane (1 mL) was added, and the reaction was carried out at 110° C. for 5 h. The reaction was monitored by TLC until the reaction was over. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(cis-4-hydroxy-4-methyl ring Hexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 11) (Off-white solid, 46 mg, yield 41.67%).

1H NMR(400MHz,DMSO-d6)δ8.09(s,1H),7.92(s,1H),7.27(s,1H),6.59(s,1H),4.48(t,2H),4.14-4.04( m, 1H), 4.02 (s, 1H), 3.25 (s, 3H), 3.13 (t, 2H), 2.63 (q, 2H), 2.12 (s, 3H), 1.66 (d, 2H), 1.46-1.34 (m, 4H), 1.14 (s, 3H).

LC-MS m/z(ESI)=410.20[M+1].

Example 12

9-(cis-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-di Hydrogen-8H-purin-8-one (Compound 12)

9-(cis-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Ethyl 2-chloro-4-(cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate (12a)

ethyl 2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate

Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1a (5.13g, 23.22mmol) was dissolved in acetonitrile (20mL), potassium carbonate (6.42g, 46.44mmol) was added with stirring at 0°C, and then slowly added dropwise ( 1s,4s)-4-methoxycyclohexane-1-amine (2.00g, 15.48mmol) in acetonitrile (10mL) was heated to room temperature and stirred for 1h, monitored by TLC until the reaction was over, filtered through diatomaceous earth and filtered with silica gel Column chromatography separation and purification (petroleum ether/ethyl acetate (v/v)=2:1) was purified to obtain the title compound 2-chloro-4-((1s,4s)-4-methoxycyclohexyl)amino)pyrimidine- Ethyl 5-carboxylate (12a) (white solid, 2.00 g, yield 41.26%).

LC-MS m/z(ESI)=314.10[M+1].

The second step:

2-Chloro-4-(cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid (12b)

2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 12a (2.00g, 6.37mmol) in tetrahydrofuran/water (15mL/15mL) and add Lithium hydroxide monohydrate (0.80g, 19.12mmol), stirred at room temperature for 2h, monitored by TLC until the end of the reaction, concentrated and evaporated off the tetrahydrofuran, then added 2N HCl to adjust the pH to about 3-4, a white solid precipitated, filtered, filter cake Wash twice with water and petroleum ether/ethyl acetate (v/v=10/1) to obtain the title compound 2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid ( 12b) (white solid, 1.67 g, yield 91.70%).

LC-MS m/z(ESI)=286.10[M+1].

third step:

2-Chloro-9-(cis-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one (12c)

2-chloro-9-(cis-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid 12b (1.57g, 5.49mmol) with N,N-dimethylacetamide (16mL), Triethylamine (0.76 mL, 5.49 mmol) and diphenyl azide phosphate (1.18 mL, 5.49 mmol) were added under stirring at room temperature and reacted for 2 hours, then the temperature was raised to 110° C. and the reaction was refluxed for 2.5 hours. TLC monitors to the end of the reaction, the reaction solution is extracted with water and dichloromethane, and the organic layer is concentrated to obtain the title compound 2-chloro-9-(cis-4-methoxycyclohexyl)-7,9-dihydro-8H-purine -8-one (12c) (white solid, 1.70 g, crude product, yield 109.47%).

LC-MS m/z(ESI)=283.00[M+1].

the fourth step:

2-Chloro-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (12d)

2-chloro-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Use 2-chloro-9-(cis-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one 12c (1.70g, 6.01mmol) with N,N-dimethylformaldehyde The amide (20 mL) was dissolved, and dimethyl sulfate (0.57 mL, 6.01 mmol) and cesium carbonate (3.92 g, 12.03 mmol) were added under stirring at 0°C to react for 2 h. TLC monitored until the reaction was over, the reaction solution was slowly added dropwise to ice water and stirred, a white solid precipitated out, washed with water and petroleum ether three times and filtered to obtain the title compound 2-chloro-9-(cis-4-methoxycyclohexyl)- 7-Methyl-7,9-dihydro-8H-purin-8-one (12d) (white solid, 1.2 g, yield 67.25%).

LC-MS m/z(ESI)=297.10[M+1].

the fifth step:

9-(cis-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-di Hydrogen-8H-purin-8-one (Compound 12)

9-(cis-4-methoxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 12d (100mg, 0.34mmol), 6-methyl -2,3-Dihydrobenzofuran-5-amine intermediate 1 (100mg, 0.68mmol), cesium carbonate (220mg, 0.68mmol) and Brettphos G3 Pd (30mg, 0.034mmol) were added to the dry reaction flask and replaced with nitrogen After three times, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(cis-4-methoxycyclohexyl)- 7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 12) ( Light yellow solid, 47 mg, yield 34.06%).

1H NMR(400MHz,DMSO-d6)δ8.16(s,1H),7.93(s,1H), 7.18(s,1H), 6.60(s,1H), 4.48(t,2H), 4.18-4.09( m, 1H), 3.39(s, 1H), 3.25(s, 3H), 3.19(s, 3H), 3.12(t, 2H), 2.48-2.39(m, 2H), 2.11(s, 3H), 2.01 –1.95(m,2H),1.50-1.39(m,4H).

LC-MS m/z(ESI)=410.20[M+1].

Example 13

9-(1,1-Dioxytetrahydro-2H-thiopyran-4-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl) Amino)-7,9-dihydro-8H-purin-8-one (compound 13)

9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

first step:

2-chloro-4-((1,1-dioxytetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (13a)

ethyl 2-chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (8g, 36mmol) and potassium carbonate (9.9g, 72mmol) in acetonitrile (30mL), and slowly add 4-aminotetrahydro- 2H-thiopyran 1,1-dioxide (6.6g, 36mmol), heated to room temperature and stirred for 20h. TLC monitors to the end of the reaction, water is added to the reaction solution, a solid is precipitated, and the title compound 2-chloro-4-((1,1-dioxytetrahydro-2H-thiopyran-4-yl)amino is obtained by filtration. ) Ethyl pyrimidine-5-carboxylate (13a) (brown solid, 6.3 g, yield 76%).

1H NMR(400MHz DMSO)δ8.64(s,1H),8.33(d,1H),4.34-4.30(m,3H),3.42-3.35(m,1H),3.08(d,1H),2.24-2.06 (m, 4H), 1.31 (t, 3H).

LC-MS m/z(ESI)=334.20[M+1].

The second step:

2-chloro-4-((1,1-dioxytetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5-carboxylic acid (13b)

2-chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((1,1-dioxytetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 13a (3g, 9mmol) in tetrahydrofuran/ Lithium hydroxide monohydrate (756mg, 18mmol) was added to water (15mL/15mL), and the reaction was stirred at room temperature for 2h. TLC monitored until the reaction was over, concentrated and evaporated off the tetrahydrofuran, then added 2N HCl to adjust the pH to about 3-4, white color The solid precipitated out, filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v=10/1) to obtain the title compound 2-chloro-4-((1,1-dioxytetrahydro-2H- Thiopyran-4-yl)amino)pyrimidine-5-carboxylic acid (13b) (brown solid, 2.3 g, yield 84%).

1H NMR (400MHz DMSO) δ 13.80 (s, 1H), 8.60 (s, 1H), 8.57 (d, 1H), 4.37-4.29 (m, 1H), 3.50-3.27 (m, 2H), 3.07 (d ,2H),2.25-2.21(m,2H),2.08-1.98(m,2H).

LC-MS m/z(ESI)=306.20[M+1].

third step:

2-chloro-9-(1,1-dioxytetrahydro-2H-thiopyran-4-yl)-7,9-dihydro-8H-purin-8-one (13c)

2-chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((1,1-dioxytetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5-carboxylic acid 13b (2.3g, 7.5mmol) in dimethyl Acetamide (10 mL), triethylamine (757 mg, 7.5 mmol) and diphenyl azide phosphate (2 g, 7.5 mmol) were added, and then gradually heated to 120° C. and stirred for 1.5 h. TLC monitors to the end of the reaction, the reaction solution is poured into water and a solid precipitates out, and it is filtered to obtain the title compound 2-chloro-9-(1,1-dioxytetrahydro-2H-thiopyran-4-yl)- 7,9-dihydro-8H-purin-8-one (13c) (white solid, 1.7 g, yield 58%).

1H NMR (400MHz DMSO) δ 11.67 (s, 1H), 8.14 (s, 1H), 4.69-4.61 (m, 1H), 3.55-3.45 (m, 2H), 3.13 (d, 2H), 2.94-2.77 (m,2H),2.14-2.02(m,2H).

LC-MS m/z(ESI)=303.20[M+1].

the fourth step:

2-chloro-9-(1,1-dioxytetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (13d )

2-chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-(1,1-dioxytetrahydro-2H-thiopyran-4-yl)-7,9-dihydro-8H-purin-8-one 13c (1.7g, 5.6 mmol) was dissolved in dimethylformamide (10 mL), dimethyl sulfate (709 mg, 4.9 mmol) and cesium carbonate (2.7 g, 9.8 mmol) were added at 0° C. and the reaction was stirred for 1 h. TLC monitored to the end of the reaction, water was added to the reaction solution, a solid precipitated out, filtered to obtain the title compound 2-chloro-9-(1,1-dioxytetrahydro-2H-thiopyran-4-yl)- 7-Methyl-7,9-dihydro-8H-purin-8-one (13d) (white solid, 1.3 g, yield 82%).

1H NMR (400MHz DMSO) δ 8.33 (s, 1H), 4.26 (s, 1H), 3.33 (s, 3H), 2.99 (s, 2H), 2.11-2.04 (m, 2H), 1.80-1.61 (m ,2H),1.44(d,2H).

LC-MS m/z(ESI)=317.20[M+1].

the fifth step:

9-(1,1-Dioxytetrahydro-2H-thiopyran-4-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl) Amino)-7,9-dihydro-8H-purin-8-one (compound 13)

9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

Combine 2-chloro-9-(1,1-dioxytetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 13d (100mg, 0.32mmol), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (94mg, 0.63mmol), cesium carbonate (206mg, 0.63mmol) and Brettphos G3 Pd (29mg, 0.032mmol) was added to a dry reaction flask, replaced with nitrogen three times and then dry 1,4-dioxane (1mL) was added, and reacted at 110°C for 5h. TLC monitors until the reaction is complete. After cooling to room temperature, it is separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(((1s,4s)-4-methoxy Cyclohexyl)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purine-8- Ketone (Compound 13) (off-white solid, 28 mg, yield 20.65%).

1H NMR(400MHz,DMSO-d6)δ8.14(s,1H),7.98(s,1H),7.25(s,1H),6.59(s,1H),4.61-4.52(m,1H), 4.48( t, 2H), 3.47-3.39 (m, 2H), 3.27 (s, 3H), 3.20-3.08 (m, 4H), 2.98-2.86 (m, 2H), 2.12 (s, 3H), 2.04 (d, 2H).

LC-MS m/z(ESI)=430.10[M+1].

Example 14

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydrofuran-2-yl)methyl)-7,9-bis Hydrogen-8H-purin-8-one (Compound 14)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8- one

first step:

Ethyl 2-chloro-4-((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylate (14a)

ethyl 2-chloro-4-(((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5g, 22.6mmol) and potassium carbonate (6.2g, 44.8mmol) in acetonitrile (20mL), and slowly add 4-aminotetramethylene dropwise with stirring at 0°C. Hydrogen-2H-thiopyran 1,1-dioxide (2.3g, 22.6mmol), heated to room temperature and stirred for 20h. TLC monitoring until the end of the reaction, water and ethyl acetate were added to the reaction solution for extraction, and the organic layer was concentrated and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v) = 10:1) to obtain the title compound 2. -Chloro-4-((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester (14a) (white solid, 4.7 g, yield 87%).

1H NMR(400MHz DMSO)δ8.62(s,1H),8.57(s,1H),4.33-4.28(m,2H),4.04-4.01(m,1H),3.82-3.77(m,1H), 3.68 -3.58 (m, 1H), 3.47-3.42 (m, 1H), 1.96-1.81 (m, 2H), 1.59-1.54 (m, 1H), 1.32-1.29 (m, 3H).

LC-MS m/z(ESI)=286.20[M+1].

The second step:

2-Chloro-4-((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylic acid (14b)

2-chloro-4-(((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester 14a (4.7g, 16.4mmol) in tetrahydrofuran/water (15mL/15mL), add hydrogen Lithium oxide monohydrate (788mg, 32.8mmol), stirred at room temperature for 1h. TLC monitors to the end of the reaction, concentrates and evaporates the tetrahydrofuran, adds 2N HCl to adjust the pH to about 3-4, a white solid is precipitated, filtered, and the filter cake is washed with water and petroleum ether/ethyl acetate (v/v=10/1) The title compound 2-chloro-4-((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylic acid (14b) (white solid, 3.8 g, yield 83%) was obtained in the second time.

1H NMR (400MHz DMSO) δ 13.75 (s, 1H), 8.74 (t, 1H), 8.58 (s, 1H), 4.40-4.01 (m, 1H), 3.82-3.76 (m, 1H), 3.69-3.58 (m, 2H), 3.46-3.40 (m, 1H), 1.95-1.90 (m, 1H), 1.86-1.81 (m, 1H), 1.58-1.53 (m, 1H).

LC-MS m/z(ESI)=259.2[M+1].

third step:

2-Chloro-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one (14c)

2-chloro-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((tetrahydrofuran-2-yl)methyl)amino)pyrimidine-5-carboxylic acid 14b (3.8g, 14.7mmol) in dimethylacetamide (20mL), add triethylamine ( 1.8g, 17.6mmol), diphenyl azide phosphate (4.4g, 16.2mmol), and then gradually heated to 120°C and stirred for 1.5h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound 2-chloro-9-((tetrahydrofuran-2- (Yl)methyl)-7,9-dihydro-8H-purin-8-one (14c) (white solid, 1.3 g, yield 46%).

LC-MS m/z(ESI)=255.20[M+1].

the fourth step:

2-Chloro-7-methyl-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one (14d)

2-chloro-7-methyl-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one 14c (1.3g, 5.1mmol) in dimethylformamide (10mL ), dimethyl sulfate (709 mg, 4.9 mmol) and cesium carbonate (2.7 g, 9.8 mmol) were added at 0° C. and the reaction was stirred for 1 h. TLC monitors to the end of the reaction, water is added to the reaction solution, a solid is precipitated, and the title compound 2-chloro-7-methyl-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro is obtained by filtration. -8H-purin-8-one (14d) (white solid, 600 mg, yield 45%).

1H NMR (400MHz DMSO) δ 8.35 (s, 1H), 4.24-4.20 (m, 1H), 3.90-3.85 (m, 1H), 3.79-3.73 (m, 2H), 3.60 (q, 1H), 3.37 (s, 3H), 1.98-1.77 (m, 3H), 1.70-1.63 (m, 1H).

LC-MS m/z(ESI)=268.20[M+1].

the fifth step:

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(((tetrahydrofuran-2-yl)methyl)-7,9 -Dihydro-8H-purin-8-one (Compound 14)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8- one

The 2-chloro-7-methyl-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one 14d (50mg, 0.19mmol), 6-methyl -2,3-Dihydrobenzofuran-5-amine intermediate 1 (56mg, 0.37mmol), cesium carbonate (121mg, 0.37mmol) and Brettphos G3 Pd (17mg, 0.019mmol) were added to the dry reaction flask and replaced with nitrogen After three times, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-((6-methyl- 2,3-Dihydrobenzofuran-5-yl)amino)-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8-one (compound 14) ( Lilac solid, 64 mg, yield 90.14%).

1H NMR(400MHz,DMSO-d6)δ8.26(s,1H),7.95(s,1H),7.19(s,1H),6.60(s,1H),4.49(t,2H),3.76-3.68( m,3H),3.66-3.59(m,2H),3.55-3.50(m,1H), 3.28(s,3H), 3.11(t,2H), 2.78-2.67(m,1H), 2.11(s, 3H), 1.96-1.85 (m, 1H), 1.68-1.58 (m, 1H).

LC-MS m/z(ESI)=382.20[M+1].

Example 15

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydro-2H-pyran-4-yl)methyl) -7,9-dihydro-8H-purin-8-one (Compound 15)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H- purin-8-one

first step:

2-chloro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester (15a)

ethyl 2-chloro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carbo-xylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (5g, 22.6mmol), potassium carbonate (6.2g, 44.8mmol) in acetonitrile (20mL), slowly add dropwise (tetrahydro- 2H-pyran-4-yl)formamide (2.6g, 2.6mmol) was heated to room temperature and stirred for 20h. TLC monitoring until the end of the reaction, water and ethyl acetate were added to the reaction solution for extraction, and the organic layer was concentrated and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v) = 10:1) to obtain the title compound 2. -Chloro-4-((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester (15a) (white solid, 3.6 g, yield 70%).

1H NMR(400MHz DMSO)δ8.60(s,1H),8.54(t,1H),4.31(q,2H), 3.84(dd,2H), 3.38(t,2H), 3.29-3.23(m,2H) ), 1.87-1.82 (m, 1H), 1.55 (dd, 2H), 1.30 (t, 3H), 1.25-1.17 (m, 2H).

LC-MS m/z(ESI)=301.20[M+1].

The second step:

2-chloro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid (15b)

2-chloro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid

2-Chloro-4-((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid ethyl ester 15a (4.6g, 15.3mmol) was dissolved in tetrahydrofuran/water (15mL/ 15mL), lithium hydroxide monohydrate (788mg, 32.8mmol) was added, and the reaction was stirred at room temperature for 1h. TLC monitors to the end of the reaction, concentrates and evaporates the tetrahydrofuran, adds 2N HCl to adjust the pH to about 3-4, a white solid is precipitated, filtered, and the filter cake is washed with water and petroleum ether/ethyl acetate (v/v=10/1) The title compound 2-chloro-4-((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid (15b) (white solid, 4.0g, yield 83%) .

1H NMR (400MHz DMSO) δ 13.72 (s, 1H), 8.71 (s, 1H), 8.56 (s, 1H), 3.84 (dd, 2H), 3.37 (t, 2H), 3.29-3.23 (m, 2H) ), 1.87-1.80 (m, 1H), 1.54 (dd, 2H), 1.26-1.17 (m, 2H).

LC-MS m/z(ESI)=273.20[M+1].

third step:

2-chloro-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one (15c)

2-chloro-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((tetrahydro-2H-pyran-4-yl)methyl)amino)pyrimidine-5-carboxylic acid 15b (4g, 14.6mmol) in dimethylacetamide (20mL), Triethylamine (2.9 g, 29 mmol) and diphenyl azide phosphate (5.2 g, 19 mmol) were added, and then gradually heated to 120° C. and stirred for 1.5 h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound 2-chloro-9-((tetrahydro-2H -Pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one (15c) (white solid, 1.46 g, yield 50%).

1H NMR(400MHz DMSO)δ11.63(s,1H),8.13(s,1H),3.81(dd,2H),3.66(d,2H),3.25-3.15(m,2H),2.07-2.00(m ,1H), 1.50 (dd, 2H), 1.29-1.21 (m, 2H).

LC-MS m/z(ESI)=269.20[M+1].

the fourth step:

2-chloro-7-methyl-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one (15d)

2-chloro-7-methyl-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one 15c (1.4g, 5.4mmol) was dissolved in two Methylformamide (10 mL), dimethyl sulfate (688 mg, 5.4 mmol) and cesium carbonate (2.6 g, 8.0 mmol) were added at 0° C. and the reaction was stirred for 1 h. TLC monitored to the end of the reaction, water was added to the reaction solution, a solid precipitated out, filtered to obtain the title compound 2-chloro-7-methyl-9-((tetrahydro-2H-pyran-4-yl)methyl)- 7,9-dihydro-8H-purin-8-one (15d) (white solid, 1.2g, yield 80%).

1H NMR (400MHz DMSO) δ 8.34 (s, 1H), 3.81 (dd, 2H), 3.70 (d, 2H), 3.36 (s, 3H), 3.25-3.19 (m, 2H), 2.07-2.01 (m ,1H),1.51(dd,2H),1.29-1.19(m,2H).

LC-MS m/z(ESI)=283.20[M+1].

the fifth step:

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydro-2H-pyran-4-yl)methyl) -7,9-dihydro-8H-purin-8-one (Compound 15)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H- purin-8-one

The 2-chloro-7-methyl-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one 15d (100mg, 0.35mmol ), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (106mg, 0.71mmol), cesium carbonate (230mg, 0.71mmol) and Brettphos G3 Pd (32mg, 0.035mmol) were added to the dry After replacing the reaction flask with nitrogen for three times, add dry 1,4-dioxane (1 mL), and react at 110°C for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-(((6-methyl -2,3-Dihydrobenzofuran-5-yl)amino)-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H-purine-8 -Ketone (Compound 15) (yellow solid, 22 mg, yield 90.14%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),7.95(s,1H),7.20(s,1H),6.61(s,1H),4.49(t,2H),3.85-3.80( m, 2H), 3.60 (d, 2H), 3.28 (s, 3H), 3.26-3.20 (m, 2H), 3.11 (t, 2H), 2.11 (s, 3H), 2.09-2.02 (m, 1H) ,1.50–1.45(m,2H),1.25–1.22(m,2H).

LC-MS m/z(ESI)=396.20[M+1].

Example 16

9-((trans-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-di Hydrogen-8H-purin-8-one (Compound 16)

9-(trans-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Ethyl 2-chloro-4-((trans-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate (16a)

ethyl 2-chloro-4-((trans-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate

Dissolve (trans-3-aminocyclobutane-1-ol hydrochloride (3.00g, 34.48mmol) in acetonitrile (25mL), add potassium carbonate (14.30g, 103.45mmol), 2,4 -Dichloro-5-pyrimidinecarboxylic acid ethyl ester 1a (11.43g, 51.72mmol), heated to room temperature and stirred for 1h, monitored by TLC until the reaction is over, filtered through diatomaceous earth and purified by silica gel column chromatography (petroleum ether/ethyl acetate) (v/v)=2:1) Purified to obtain the title compound 2-chloro-4-((trans-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid ethyl ester (16a) (white solid, 2.9g , The yield is 30.95%).

1H NMR(400MHz,DMSO-d6)δ8.61(s,1H), 8.48(d,1H), 5.17-5.11(m,1H), 4.61-4.51(m,1H), 4.36-4.27(m,3H) ), 2.32-2.20 (m, 4H), 1.32 (t, 3H).

LC-MS m/z(ESI)=272.00[M+1].

The second step:

4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (16b)

ethyl 4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2-chloro-4-((trans-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid ethyl ester 18a (3.10g, 11.41mmol) in dichloromethane (20mL), add tert-butyl Diphenylchlorosilane (4.45mL, 17.11mmol), imidazole (1.94g, 28.52mmol), protected by nitrogen and ventilated, stirred at room temperature for 2h, monitored by TLC until the end of the reaction, and purified by silica gel column chromatography (petroleum ether/ethyl acetate) Ester (v/v)=10:1) was purified to obtain the title compound 4-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5- Ethyl carboxylate (16b) (colorless liquid, 4.18 g, yield 64.26%).

LC-MS m/z(ESI)=510.20[M+1].

third step:

4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid (16c)

4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid

Dissolve 4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 16b (4.18g, 8.19mmol) In tetrahydrofuran/water (30mL/15mL), lithium hydroxide monohydrate (1.03g, 24.58mmol) was added, the reaction was stirred at room temperature for 2h, TLC monitored until the reaction was over, concentrated and evaporated off the tetrahydrofuran, and then 2N HCl was added to adjust the pH to 3 4. Add water and ethyl acetate to extract twice, concentrate the organic layer and use silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v)=20:1) to obtain the title compound 4-((反-3-(( Tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid (16c) (white solid, 3.87 g, yield 97.97%).

LC-MS m/z(ESI)=482.20[M+1].

the fourth step:

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9-dihydro-8H-purin-8-one (18d)

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9-dihydro-8H-purin-8-one

Use 4-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid 16c (3.87g, 8.03mmol) with N, N -Dimethylacetamide (38mL) was dissolved, triethylamine (1.11mL, 8.03mmol) and diphenyl azide phosphate (1.73mL, 8.03mmol) were added under stirring at room temperature, and then reacted for 2h, then heated to 110℃ and refluxed for 2.5 h. TLC monitoring until the end of the reaction, the reaction solution was added with water and ethyl acetate for extraction, and the organic layer was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1) to obtain the title compound 9-( Trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9-dihydro-8H-purin-8-one (16d) (white solid, 1.81g , The yield is 47.06%).

1H NMR (400MHz, DMSO-d6) δ 11.59 (s, 1H), 8.09 (s, 1H), 7.65-7.62 (m, 2H), 7.62-7.60 (m, 2H), 7.47-7.39 (m, 6H) ), 5.05–4.97(m,1H), 4.96–4.89(m,1H), 2.98-2.84(m,2H), 2.54-2.46(m,2H), 1.02(s,9H).

LC-MS m/z(ESI)=479.20[M+1].

the fifth step:

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one( 18e)

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9-dihydro-8H-purine-8-one 16d (1.81g, 3.78mmol) was dissolved in N,N-dimethylformamide (20mL), dimethyl sulfate (0.36mL, 3.78mmol) and cesium carbonate (2.47g, 7.57mmol) were added and reacted for 2h under stirring at 0°C. TLC monitors until the reaction is over, the reaction solution is slowly added dropwise to ice water and stirred, ethyl acetate is added for extraction, the concentrated organic layer is separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 3:1) Purified to obtain the title compound 9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl-7,9-dihydro-8H-purine- 8-ketone (16e) (white solid, 1.80 g, yield 96.62%).

LC-MS m/z(ESI)=493.20[M+1].

The sixth step:

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran -5-yl)amino)-7,9-dihydro-8H-purin-8-one (16f)

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one 16e (200mg, 0.41mmol), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1, (121mg, 0.82mmol), cesium carbonate (264mg, 0.82mmol) and Brettphos G3 Pd ( 37 mg, 0.041 mmol) was added to a dry reaction flask, and N _{2 was} replaced three times, then dry 1,4-dioxane (1 mL) was added, and the reaction was carried out at 110° C. for 5 hours. Cool to room temperature, filter, and concentrate the filtrate to obtain crude product (16f), which is directly used for the next step without purification.

The seventh step:

9-((trans-3-hydroxycyclobutyl)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9- Dihydro-8H-purin-8-one (Compound 16)

9-(trans-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The crude product 16f obtained in the sixth step was dissolved in 1,4-dioxane (5 mL), then concentrated hydrochloric acid (1 mL) was added dropwise, and the reaction was carried out at room temperature for 2 hours. TLC monitors to the end of the reaction, adjusts the pH to about 7 with saturated sodium bicarbonate solution, adds dichloromethane for extraction three times, combines the organic phases, concentrates, and uses silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v=40: 1)) Purification to obtain the title compound 9-((trans-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl) Amino)-7,9-dihydro-8H-purin-8-one (compound 16) (white solid, 54 mg, yield 36.23%).

1H NMR(400MHz,DMSO-d6)δ8.24(s,1H),7.97(s,1H),7.22(s,1H),6.61(s,1H),5.07(d,1H),4.99(p, 1H), 4.49 (t, 2H), 4.40--4.33 (m, 1H), 3.25 (s, 3H), 3.11 (t, 2H), 3.02-2.92 (m, 2H), 2.19-2.13 (m, 2H) , 2.12 (s, 3H).

LC-MS m/z(ESI)=368.10[M+1].

Example 17

9-(6-(Hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5- (Yl)amino)-7,9-dihydro-8H-purin-8-one (compound 17)

9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro- 8H-purin-8-one

first step:

Tert-butyl((3,4-dihydro-2H-pyran-2-yl)methoxy)diphenylsilane (17b)

tert-butyl((3,4-dihydro-2H-pyran-2-yl)methoxy)diphenylsilane

Dissolve (3,4-dihydro-2H-pyran-2-yl)methanol 17a (20.0g, 175.22mmol) in dichloromethane (400mL), add tert-butyldiphenylchlorosilane (68.35mL, 262.84mmol), imidazole (29.82g, 438.06mmol), protected by nitrogen and ventilated, stirred overnight at room temperature, monitored by TLC until the end of the reaction, washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure on silica gel Column chromatography separation and purification (petroleum ether/ethyl acetate (v/v) = 100:1) to obtain the title compound tert-butyl ((3,4-dihydro-2H-pyran-2-yl)methoxy) Diphenylsilane (17b) (colorless liquid, 42.78 g, yield 75.00%).

¹ H NMR (400MHz, CDCl ₃ ): δ7.71-7.68 (m, 4H), 7.45-7.37 (m, 6H), 6.37-6.35 (d, 1H), 4.68-4.65 (m, 1H), 3.96 3.93(m,1H),3.83-3.77(m,1H),3.71-3.67(m,1H),2.08-2.00(m,1H),1.99-1.92(m,2H),1.74-1.71(m,1H) ), 1.02(s, 9H).

LC-MS m/z(ESI)=353.20[M+1].

The second step:

6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol(17c)

6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol

Add tert-butyl((3,4-dihydro-2H-pyran-2-yl)methoxy)diphenylsilane 17b (23.5g, 66.7mmol) to tetrahydrofuran (200mL), under nitrogen protection, After cooling to -78°C, borane dimethyl sulfide complex (100 mL, 2M, 200.00 mmol) was added dropwise to the reaction solution, and after the addition was completed, it was naturally warmed to room temperature and stirred overnight. After slowly adding 1N sodium hydroxide aqueous solution to the reaction system until no borane gas was released, 30% hydrogen peroxide (90 mL) was added to the reaction solution, and the reaction solution was stirred at 45° C. for 2 h. Water and ethyl acetate were added to the reaction system for extraction, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by reverse phase column chromatography (acetonitrile/water (v/v) = 5:95) The title compound 6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol (17c) (colorless liquid, 18.03g, yield 73.00%) .

LC-MS m/z(ESI)=371.30[M+1].

third step:

2-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3-dione (17d)

2-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3-dione

Dissolve 6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol 17c (13.0g, 35.1mmol) in tetrahydrofuran (150mL), stir at room temperature Add phthalimide (5.16g, 35.1mmol) and triphenylphosphine (13.8g, 52.7mmol), then slowly add diisopropyl azodicarboxylate (10.7g, 52.7mmol) dropwise and increase the temperature The reaction was refluxed at 70°C for 1 hour, monitored by TLC until the reaction was over, the reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 100:1) to obtain the title 2-(6-((( Tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3-dione (17d) (white solid, 9.0g, yield 51.34%).

LC-MS m/z(ESI)=500.20[M+1].

the fourth step:

6-(((tert-butyldiphenylsilyl)oxy)methyltetrahydro-2H-pyran-3-amine(17e)

6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-amine

The 2-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3-dione 19d(9.0 g, 18.0mmol) was dissolved in methanol (50mL), and then hydrazine hydrate (1.8g, 36.0mmol) was added. The reaction solution was refluxed for 2h. After the reaction was completed, dichloromethane (50mL) was added to precipitate a large amount of solids, filtered and rotated. After drying, the viscous liquid solid obtained was washed twice with dichloromethane and filtered. The organic phases were combined and concentrated under reduced pressure to obtain the title compound 6-((tert-butyldiphenylsilyl)oxy)methyltetrahydro- 2H-pyran-3-amine (17e) (colorless liquid, 6.3 g, yield 94.10%).

LC-MS m/z(ESI)=370.20[M+1].

the fifth step:

4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (19f)

ethyl 4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2,4-dichloro-5-pyrimidine ethyl carboxylate 1a (6.94g, 31.40mmol) in acetonitrile (25mL), add potassium carbonate (8.68g, 62.80mmol) while stirring at room temperature, and slowly add 6- ((Tert-butyldiphenylsilyl)oxy)methyltetrahydro-2H-pyran-3-amine 17e (7.40g, 20.93mmol) in acetonitrile (15mL), the reaction was stirred for 2h, TLC monitored until the reaction At the end, after filtration through diatomaceous earth, it was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=20:1) to obtain the title compound 4-((6-(((tert-butyldiphenylsilane) (Yl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (17f) (light yellow solid, 5.20g, yield 46.86%) .

LC-MS m/z(ESI)=554.30[M+1].

The sixth step:

4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid (17g )

4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylate

4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl Ester 17f (5.20g, 9.383mmol) was dissolved in tetrahydrofuran/water (30mL/15mL), lithium hydroxide monohydrate (1.18g, 28.15mmol) was added, the reaction was stirred at room temperature for 3 hours, and then heated to 50°C for 2 hours, monitored by TLC To the end of the reaction, concentrate and evaporate the tetrahydrofuran, add 2N HCl to adjust the pH to 3-4, add water and ethyl acetate for extraction twice, concentrate the organic layer and separate and purify by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 4:1) Purified to obtain the title compound 4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloro Pyrimidine-5-carboxylic acid (17 g) (white solid, 2.57 g, yield 52.06%).

LC-MS m/z(ESI)=526.20[M+1].

The seventh step:

9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9-dihydro-8H-purine -8-ketone (17h)

9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one

The 4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid 17g (2.57g, 4.88mmol) dissolved in N,N-dimethylacetamide (24mL), add triethylamine (0.68mL, 4.88mmol) and diphenyl azide phosphate (1.05mL, 4.88mmol) under stirring at room temperature After reacting for 2h, the temperature was raised to 110°C and refluxed for 2.5h. TLC monitoring until the end of the reaction, the reaction solution was added with water and ethyl acetate for extraction, and the organic layer was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1) to obtain the title compound 9-( 6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9-dihydro-8H-purine-8- Ketone (17h) (light pink solid, 1.60 g, yield 62.66%).

1H NMR (400MHz, DMSO-d6) δ 11.65 (s, 1H), 8.12 (s, 1H), 7.67-7.62 (m, 4H), 7.46-7.39 (m, 6H), 4.32-4.24 (m, 1H) ), 4.13-4.06 (m, 1H), 4.00-3.93 (m, 1H), 3.91-3.81 (m, 2H), 3.56 (dd, 1H), 2.53-2.47 (m, 1H), 2.07-1.98 (m ,1H),1.85-1.72(m,2H),1.01(s,9H).

LC-MS m/z(ESI)=523.20[M+1].

The eighth step:

9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7-methyl-7,9-bis Hydrogen-8H-purin-8-one (17i)

9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one

Add 9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9-dihydro-8H- Purine-8-one 17h (1.60g, 3.06mmol) was dissolved with N,N-dimethylformamide (15mL), and dimethyl sulfate (0.29mL, 3.06mmol) and cesium carbonate (2.00g, 6.13mmol) reacted for 1h, monitored by TLC to the end of the reaction, added water and ethyl acetate for extraction, concentrated the organic layer and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=3:2) to obtain the title Compound 9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7-methyl-7,9- Dihydro-8H-purin-8-one (17i) (light pink liquid, 1.6 g, yield 97.21%).

1H NMR(400MHz,DMSO-d6)δ8.35(s,1H),7.67-7.62(m,4H),7.49-7.39(m,6H),4.36-4.28(m,1H),4.14-4.07(m ,1H),4.00-3.93(m,1H),3.90-3.81(m,2H),3.57(dd,1H),3.35(s,3H),2.55-2.44(m,1H),2.09-1.98(m ,1H),1.87-1.73(m,2H),1.01(s,9H).

LC-MS m/z(ESI)=537.30[M+1].

Step 9:

9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-(((6-methyl (2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (17j)

9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino )-7,9-dihydro-8H-purin-8-one

Add 9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7-methyl-7,9- Dihydro-8H-purin-8-one 17i (150mg, 0.28mmol), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (83mg, 0.56mmol), cesium carbonate (182mg , 0.56mmol) and Brettphos G3 Pd (25mg, 0.028mmol) were added to a dry reaction flask, replaced with N2 three times, and then added dry 1,4-dioxane (1mL), and reacted at 110°C for 5h. TLC monitored to the end of the reaction, cooled to room temperature and filtered, and the filtrate was concentrated to obtain the crude product 17j, which was directly used for the next step without purification.

The tenth step:

9-(6-(Hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl )Amino)-7,9-dihydro-8H-purin-8-one (compound 17)

9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro- 8H-purin-8-one

The crude product 17j obtained in the ninth step was dissolved with 1,4-dioxane (5 mL), then concentrated hydrochloric acid (1 mL) was added dropwise, and the reaction was carried out at room temperature for 2 h. TLC monitors until the reaction is over, adjust the pH to about 7 with saturated sodium bicarbonate solution, add DCM for extraction three times, combine the organic phases, concentrate and use silica gel column chromatography for separation and purification (dichloromethane/methanol (v/v=40:1) ) To obtain the title compound 9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzene P-furan-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 17) (white solid, 33 mg, yield 28.72%).

1H NMR(400MHz,DMSO-d6)δ8.25(s,1H),7.97(s,1H),7.13(s,1H),6.62(s,1H),4.55-4.46(m,3H), 4.24-- 4.15 (m, 1H), 4.07 (t, 1H), 3.68-3.60 (m, 1H), 3.59-3.52 (m, 1H), 3.44 (dd, 1H), 3.38-3.31 (m, 2H), 3.25 ( s, 3H), 3.13 (t, 2H), 2.09 (s, 3H), 1.85-1.76 (m, 1H), 1.71-1.58 (m, 2H).

LC-MS m/z(ESI)=412.20[M+1].

Example 18

9-(3-Hydroxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H- Purine-8-one (Compound 18)

9-(3-hydroxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Tert-Butyl (3-hydroxycyclohexyl) carbamate (18b)

tert-butyl(3-hydroxycyclohexyl)carbamate

Dissolve tert-butyl (3-oxocyclohexyl) carbamate 18a (8.20g, 38.45mmol) in tetrahydrofuran (40mL), add sodium borohydride (4.36g, 115.35mmol) under stirring at 0°C and react for 3h, TLC monitors to the end of the reaction, slowly add saturated sodium carbonate solution to the reaction solution, stir at room temperature for 3 hours, add ethyl acetate for extraction, concentrate the organic layer to obtain the title compound tert-butyl (3-hydroxycyclohexyl) carbamate (18b) (Yellow liquid, 8.00 g, yield 96.95%).

LC-MS m/z(ESI)=216.20[M+1].

The second step:

3-Aminocyclohexane-1-ol (20c)

3-aminocyclohexan-1-ol hydrochloride

Dissolve tert-butyl (3-hydroxycyclohexyl) carbamate 18b (8.00g, 37.16mmol) in 2M hydrogen chloride-ethyl acetate (35mL), stir the reaction at room temperature for 4h, monitor by TLC until the reaction is complete, concentrate and evaporate The solvent was used to obtain the title compound 3-aminocyclohexane-1-ol hydrochloride (18c) (yellow liquid, crude product, 4.00 g, yield 71.05%).

LC-MS m/z(ESI)=116.10[M+1].

third step:

Ethyl 2-chloro-4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate (18d)

ethyl 2-chloro-4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 3-aminocyclohexane-1-ol hydrochloride 18c (4.00g, 26.38mmol) in acetonitrile (30mL), add 2,4-dichloro-5-pyrimidinecarboxylic acid ethyl ester 1a( 8.75g, 39.57mmol), potassium carbonate (10.94g, 79.14mmol), heated to room temperature and stirred for 2h, monitored by TLC until the reaction was over, filtered through diatomaceous earth and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v /v)=2:1) Purified to obtain the title compound 2-chloro-4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester (18d) (light yellow solid, 1.40g, yield 17.71 %).

LC-MS m/z(ESI)=300.10[M+1].

the fourth step:

4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (18e)

ethyl 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2-chloro-4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 18d (1.10g, 3.67mmol) in dichloromethane (20mL), add tert-butyldiphenyl Chlorosilane (1.43mL, 5.50mmol), imidazole (0.62g, 9.17mmol), protected by nitrogen and ventilated, stirred at room temperature for 2h, monitored by TLC until the end of the reaction, and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v /v)=8:1) was purified to obtain the title compound 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester ( 18e) (colorless liquid, 1.90 g, yield 96.21%).

LC-MS m/z(ESI)=538.20[M+1].

the fifth step:

4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid (18f)

4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylate

4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 18e (1.90g, 3.53mmol) was dissolved in tetrahydrofuran/ In water (20mL/20mL), add lithium hydroxide monohydrate (0.44g, 10.59mmol), stir the reaction at room temperature for 2h, monitor by TLC until the reaction is over, concentrate and evaporate the tetrahydrofuran, add 2N HCl to adjust the pH to 3-4, add water It was extracted twice with ethyl acetate, and the organic layer was concentrated to obtain the title compound 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid 18f (White solid, 1.10g, yield 61.08%).

LC-MS m/z(ESI)=510.20[M+1].

The sixth step:

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9-dihydro-8H-purin-8-one (18g)

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9-dihydro-8H-purin-8-one

Use 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid 18f (1.10g, 2.16mmol) with N,N-di Methyl acetamide (23 mL) was dissolved, triethylamine (0.30 mL, 2.16 mmol) and diphenyl azide phosphate (0.46 mL, 2.16 mmol) were added under stirring at room temperature and reacted for 2 h, then the temperature was raised to 110° C. and refluxed for 2.5 h. TLC monitoring until the end of the reaction, the reaction solution was added with water and ethyl acetate for extraction, and the organic layer was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1) to obtain the title compound 9-( 3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9-dihydro-8H-purin-8-one 18g (white solid, 0.31g, yield 28.35% ).

LC-MS m/z(ESI)=507.20[M+1].

The seventh step:

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one (18h)

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9-dihydro-8H-purin-8-one 18g (0.30g, 0.59mmol) Dissolve with N,N-dimethylformamide (10mL), add dimethyl sulfate (0.06mL, 0.59mmol) and cesium carbonate (0.39g, 1.18mmol) under stirring at 0°C and react for 1h. TLC monitors until the reaction is over, the reaction solution is slowly added dropwise to ice water and stirred, ethyl acetate is added for extraction, the organic layer is concentrated to obtain the title compound 9-(3-((tert-butyldiphenylsilyl)oxy) Hexyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one (18h) (colorless liquid, 0.30 g, yield 97.31%).

LC-MS m/z(ESI)=521.20[M+1].

Step 8:

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5- (Yl)amino)-7,9-dihydro-8H-purin-8-one (20i)

9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin -8-one

Add 9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one 18h (200mg , 0.38mmol), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (115mg, 0.76mmol), cesium carbonate (251mg, 0.76mmol) and Brettphos G3 Pd (35mg, 0.038mmol) ) Add a dry reaction flask, _{replace with N 2} three times, then add dry 1,4-dioxane (1 mL), and react at 110° C. for 5 hours. TLC monitored to the end of the reaction, cooled to room temperature and filtered. The filtrate was concentrated to obtain the crude product 18i, which was directly used in the next step without purification.

Step 9:

9-(3-Hydroxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H- Purine-8-one (Compound 18)

9-(3-hydroxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The crude product 18i obtained in the eighth step was dissolved with 1,4-dioxane (5 mL), then concentrated hydrochloric acid (1 mL) was added dropwise, and the reaction was carried out at room temperature for 2 h. TLC monitors to the end of the reaction, adjust the pH to about 7 with saturated sodium bicarbonate solution, add dichloromethane for extraction three times, combine the organic phases, concentrate and use silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v=40: 1)) Purification to obtain the title compound 9-(3-hydroxycyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7 , 9-dihydro-8H-purin-8-one (Compound 18) (white solid, 22 mg, yield 14.50%).

1H NMR (400MHz, DMSO-d6) δ 8.20 (s, 1H), 7.96 (s, 1H), 7.26 (s, 1H), 6.60 (s, 1H), 4.78 (d, 1H), 4.48 (t, 2H), 4.17-4.08(m, 1H), 3.52-3.42(m, 1H), 3.26(s, 3H), 3.21-3.05(m, 2H), 2.20-2.03(m, 5H), 1.90-1.79( m, 2H), 1.78-1.70 (m, 1H), 1.59 (d, 1H), 1.34-1.21 (m, 1H), 1.11-0.99 (m, 1H).

LC-MS m/z(ESI)=396.20[M+1].

Example 19

Trans-4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H- Purin-9-yl)cyclohexane-1-carbonitrile (Compound 19)

trans-4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane- 1-carbonitrile

first step:

Tert-Butyl (trans-4-carbamoylcyclohexyl) carbamate (19b)

tert-butyl(trans-4-carbamoylcyclohexyl)carbamate

Trans-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid 19a (5.0g, 20.5mmol), O-(7-azabenzotriazole)-N,N,N ',N'-Tetramethyluronium hexafluorophosphate (9.4g, 24.7mmol) was dissolved in dichloromethane (15mL), stirred at 0℃ for 20min, and N,N diisopropylethylamine (10.5g , 82mmol) and ammonium chloride (3.3g, 61.5mmol), stirred at room temperature for 4h. TLC monitors to the end of the reaction, add 10 mL of water to the reaction solution, separate the organic phase, wash with saturated brine once, dry with anhydrous sodium sulfate and mix the sample with silica gel, pass the product with a normal phase column passer, and concentrate to obtain the title compound tert-butyl Group (trans-4-carbamoylcyclohexyl) carbamate (19b) (white solid, 4.4 g, yield 83%).

LC-MS m/z(ESI)=243.30[M+1].

The second step:

Tert-butyl (trans-4-cyanocyclohexyl) carbamate (19c)

tert-butyl(trans-4-cyanocyclohexyl)carbamate

Tert-butyl (trans-4-carbamoylcyclohexyl) carbamate 19b (4.4g, 18.0mmol) was dissolved in 70mL of pyridine, cooled in an ice bath, and then phosphorus oxychloride (7.7mL) was added dropwise to the reaction In the solution, stir in an ice bath for 1 hour. TLC monitors the completion of the reaction, adds 20 mL of water in an ice bath, extracts 4 times with ethyl acetate, then washes the organic phase 7 times with acid water, and finally washes twice with saturated brine, dried with anhydrous sodium sulfate, filtered and concentrated to dryness. The compound tert-butyl (trans-4-cyanocyclohexyl) carbamate (19c) (yellow solid, 1.2 g, yield 30%).

1H NMR(400MHz DMSO)δ6.80(m,1H),3.24-3.22(m,1H),2.63-2.55(m,1H),1.99-1.95(m,2H),1.77-1.73(m,2H) , 1.56-1.46 (m, 2H), 1.36 (s, 9H), 1.21-1.11 (m, 2H).

LC-MS m/z(ESI)=225.30[M+1].

third step:

Trans-4-aminocyclohexane-1-carbonitrile (19d)

trans-4-aminocyclohexane-1-carbonitrile

Dissolve tert-butyl (trans-4-cyanocyclohexyl) carbamate 19c (1.2 g, 9.6 mmol) in hydrochloric acid ethyl acetate solution (20 mL), heat to 45° C., and stir for 2 h. TLC monitored to the end of the reaction, a white solid precipitated, concentrated and dried to obtain the title compound trans-4-aminocyclohexane-1-carbonitrile (19d) (white solid, 660 mg, yield 60%).

LC-MS m/z(ESI)=125.30[M+1].

the fourth step:

2-Chloro-4-(trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester (19e)

ethyl 2-chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (1.4g, 5.35mmol), potassium carbonate (1.4g, 10.7mmol) in acetonitrile (10mL), add (1r, 4r) at 0℃ -4-Aminocyclohexane-1-carbonitrile 19d (660 mg, 5.35 mmol) was stirred at room temperature for 20 h. TLC monitors until the reaction is complete, the reaction solution is added with water and ethyl acetate for extraction three times, washed with saturated brine once, dried with anhydrous sodium sulfate and separated by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=10: 1) Purification to obtain the title compound 2-chloro-4-((1r,4r)-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester (19e) (white solid, 810g, yield 62% ).

1H NMR (400MHz DMSO) δ8.62 (s, 1H), 8.27 (d, 1H), 4.30 (q, 2H), 4.04-3.96 (m, 1H), 4.04-3.67 (m, 1H), 2.76-2.70 (m, 1H), 2.04-2.00 (m, 2H), 1.96-1.92 (m, 2H), 1.72-1.62 (m, 2H), 1.47-1.37 (m, 2H), 1.30 (t, 3H).

LC-MS m/z(ESI)=310.20[M+1].

the fifth step:

2-chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid (19f)

2-chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 19e (810mg, 2.6mmol) in tetrahydrofuran/water (4mL/4mL) and add hydroxide Lithium (247mg, 5.2mmol), stirred at room temperature for 1h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 4-5, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether/ethyl acetate (v/v=10/1), and concentrated to obtain the title compound 2-Chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid (19f) (white solid, 790mg, yield 86%)

1H NMR(400MHz DMSO)δ8.58(s,1H),8.49(d,1H),4.01-3.87(m,1H),2.76-2.71(m,1H),2.03-1.93(m,4H),1.72 -1.63 (m, 2H), 1.44-1.35 (m, 2H).

LC-MS m/z(ESI)=282.30[M+1].

The sixth step:

Trans-4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (19g)

trans-4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile

2-Chloro-4-(trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid 19f (730mg, 2.6mmol) was dissolved in dimethylacetamide (10mL), and triethylamine (263mg, 2.6mmol), diphenyl azide phosphate (715mg, 2.6mmol), then gradually heated to 110°C and stirred for 1.5h. The reaction was monitored by TLC until the end of the reaction. The reaction solution was concentrated. The residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:10) to obtain the title compound trans-4-(2-chloro-8-oxygen). Di-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (19 g) (white solid, 553 mg, yield 68%).

1H NMR (400MHz DMSO) δ 11.63 (s, 1H), 8.12 (s, 1H), 4.23-4.16 (m, 1H), 2.78-2.72 (m, 1H), 2.25-2.12 (m, 4H), 1.82 -1.68(m,4H).

LC-MS m/z(ESI)=278.30[M+1].

The seventh step:

Trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (19h)

trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile

Dissolve 19 g (553 mg, 2 mmol) of trans-4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile in dimethylform Amide (5 mL), dimethyl sulfate (252 mg, 2 mmol) and cesium carbonate (977 mg, 3 mmol) were added at 0°C, and stirred at 0°C for 30 min. TLC monitored to the end of the reaction, 10mL of water was added to the reaction solution, a solid precipitated out, filtered to obtain the title compound trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H- Purin-9-yl)cyclohexane-1-carbonitrile (19h) (yellow solid, 420 mg, yield 72%).

1H NMR (400MHz DMSO) δ8.34 (s, 1H), 4.28-4.21 (m, 1H), 2.79-2.72 (m, 1H), 2.22-2.13 (m, 4H), 1.82-1.70 (m, 4H) .

LC-MS m/z(ESI)=292.30[M+1].

The eighth step:

Trans-4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H- Purin-9-yl)cyclohexane-1-carbonitrile (Compound 19)

trans-4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane- 1-carbonitrile

Trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile 19h (100mg, 0.34mmol) , 6-Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (102mg, 0.68mmol), cesium carbonate (223mg, 0.68mmol) and Brettphos G3 Pd (31mg, 0.034mmol) were added to the dry In the reaction tube, _{replace with N 2} three times, then add dry 1,4-dioxane (1 mL), and react at 110° C. for 5 h. TLC monitors until the reaction is complete, cools to room temperature, and uses silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v=40:1)) to obtain the title compound trans-4-(7-methyl-2-( (6-Methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (Compound 19) (pale yellow solid, 34 mg, yield 24.52%).

1H NMR(400MHz,DMSO-d6)δ8.18(s,1H),7.94(s,1H),7.21(s,1H),6.62(s,1H),4.50(t,2H),4.21-4.12( m, 1H), 3.25 (s, 3H), 3.12 (t, 2H), 2.59-2.53 (m, 1H), 2.31-2.19 (m, 2H), 2.18-2.09 (m, 5H), 1.80-1.73 ( m,2H),1.71-1.61(m,2H).

LC-MS m/z(ESI)=405.20[M+1].

Example 20

9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino) -7,9-dihydro-8H-purin-8one (Compound 20)

9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin -8-one

first step:

Ethyl 2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylate (20a)

2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylate

Dissolve 2-isopropyl-4-methylpyridine-3-amine (21.61g, 143.92mmol), 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (26.51g, 119.93mmol) in acetonitrile (400mL), add potassium carbonate (33.15g, 239.86mmol) under stirring at 0°C, and then gradually increase the temperature to 90°C and stir for 72h. The reaction was monitored by TLC until the end of the reaction. Water and ethyl acetate were added to the reaction solution for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the title compound 2-chloro-4-((2-isopropyl-4- Methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (20a) (brown solid, 44.23 g, yield 91.95%).

1H NMR(400MHz,DMSO-d6)δ9.83(s,1H), 8.77(s,1H), 8.39(d,1H), 7.20(d,1H), 4.39(q,2H), 3.10-3.03( m, 1H), 2.13 (s, 3H), 1.36 (t, 3H), 1.12 (d, 6H).

LC-MS m/z(ESI)=335.10[M+1].

The second step:

2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid (20b)

2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid

The 2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 20a (44.23g, 131.42mmol) was mixed with tetrahydrofuran/water (200mL /200mL) was dissolved, and then lithium hydroxide (11.03g, 262.84mmol) was added and stirred at room temperature for 3h. TLC monitors to the end of the reaction, spin-dry the tetrahydrofuran, adjust the pH to 1-2 with 2N hydrochloric acid, precipitate a solid, filter, and wash the filter cake with water to dry to obtain the title compound 2-chloro-4-((2-isopropyl-4- Methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid (20b) (brown solid, 29.29 g, yield 72.66%).

1H NMR(400MHz,DMSO-d6)δ10.23(s,1H),8.72(s,1H),8.38(d,1H),7.20(d,1H),3.11-3.04(m,1H),2.13( s, 3H), 1.26 (d, 1H), 1.12 (d, 6H).

LC-MS m/z(ESI)=307.09[M+1].

third step:

2-Chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin-8-one (20c)

2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid 20b (25.00g, 81.5mmol) in tetrahydrofuran, and then lower the temperature To 0°C, add diphenyl azide phosphate (24.6 g, 89.5 mmol) and triethylamine (9.8 g, 97.75 mmol). After stirring at room temperature for 1 hour, the temperature was gradually increased to 120°C for 2 hours. The reaction was monitored by TLC until the end of the reaction. Water and ethyl acetate were added to the reaction solution for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated and separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1/ 1) Purified to obtain the title compound 2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin-8-one (20c) (white solid , 18g, yield 75%).

1H NMR(400MHz,DMSO-d6)δ12.06(s,1H),8.59(d,1H),8.33(s,1H),7.35(dd,1H),2.80-2.73(m,1H),2.05( s, 3H), 1.09 (dd, 6H).

LC-MS m/z(ESI)=304.10[M+1].

the fourth step:

2-Chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (20d)

2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin-8-one 20c (500mg, 1.65mmol) in N , N-Dimethylformamide (10mL), add cesium carbonate (536mg, 1.65mmol) and dimethyl sulfate (269mg, 2.14mmol) at 0°C, and stir for 1.5h at 0°C. TLC monitors to the end of the reaction, adds water and ethyl acetate to the reaction solution to extract 3 times, the organic phase is dried over anhydrous sodium sulfate, concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1/1) Obtain the title compound 2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (20d) (white solid, 258 mg, yield 49.32%).

1H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.56(s,1H),7.36(d,1H),3.49(s,3H),2.71-2.78(m,1H),2.04( s, 3H), 1.10 (d, 3H), 1.06 (d, 3H).

LC-MS m/z(ESI)=318.10[M+1].

the fifth step:

9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino) -7,9-dihydro-8H-purin-8one (Compound 20)

9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin -8-one

The 2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 20d (80mg, 0.25mmol ), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (75mg, 0.50mmol), cesium carbonate (164mg, 0.50mmol) and Brettphos G3 Pd (23mg, 0.025mmol) were added to the dry The reaction tube was _{replaced with N 2} three times, then dry 1,4-dioxane (1 mL) was added, and the reaction was carried out at 110°C for 5 hours. TLC monitors until the reaction is complete, cools to room temperature, uses silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to separate and purify to obtain the title compound 9-(2-isopropyl-4-methyl) Pyridin-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purine-8 Ketone (Compound 20) (light pink solid, 32 mg, yield 29.52%).

1H NMR(600MHz,DMSO-d6)δ8.53(d,1H), 8.37(s,1H), 8.08(s,1H), 7.31(d,1H), 7.07(s,1H), 6.56(s, 1H), 4.46(t, 2H), 3.39(s, 3H), 3.06(t, 2H), 2.83-2.75(m, 1H), 2.06(s, 3H), 2.03(s, 3H), 1.14-1.08 (m,6H).

LC-MS m/z(ESI)=431.20[M+1].

Example 21

9-(cis-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro -8H-purin-8-one (Compound 21)

9-(cis-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Ethyl 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate (21a)

ethyl 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate

(1s, 3s)-3-aminocyclobutane-1-ol hydrochloride (3.00g, 34.48mmol) was dissolved in acetonitrile (25mL), and potassium carbonate (14.30g, 103.45mmol) was added while stirring at 0°C, Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1a (11.43g, 51.72mmol), heated to room temperature and stirred for 1h, monitored by TLC until the end of the reaction, filtered through diatomaceous earth and purified by silica gel column chromatography (petroleum ether/ Ethyl acetate (v/v)=2:1) was purified to obtain the title compound 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid ethyl ester (21a) (white solid) , 3.3g, yield 32.91%).

1H NMR(400MHz,DMSO-d6)δ8.60(s,1H),8.41(d,1H),4.96(s,1H),4.31(q,2H),4.08--3.97(m,1H),3.94-- 3.84 (m, 1H), 2.71-2.63 (m, 2H), 1.88-1.80 (m, 2H), 1.33-1.29 (m, 3H).

LC-MS m/z(ESI)=272.00[M+1].

The second step:

2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid (21b)

2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate

Dissolve 2-chloro-4-(((1s,3s)-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid ethyl ester 21a (3.30g, 12.14mmol) in tetrahydrofuran/water (30mL/15mL) Add lithium hydroxide monohydrate (1.53g, 36.44mmol), stir the reaction at room temperature for 2h, monitor by TLC until the reaction is over, concentrate and evaporate the tetrahydrofuran, add 2N HCl to adjust the pH to about 3-4, white solid precipitates out, filter , The filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v=10/1) to obtain the title compound 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxy Acid (21b) (white solid, 2.44 g, yield 82.45%).

1H NMR(400MHz,DMSO-d6)δ8.61(d,1H),8.57(s,1H),4.43-4.20(m,1H),4.07-3.96(m,1H),3.93-3.83(m,1H) ), 2.72-2.63 (m, 2H), 1.87-1.77 (m, 2H).

LC-MS m/z(ESI)=244.00[M+1].

third step:

2-Chloro-9-(cis-3-hydroxycyclobutyl)-7,9-dihydro-8H-purin-8-one (21c)

2-chloro-9-(cis-3-hydroxycyclobutyl)-7,9-dihydro-8H-purin-8-one

Use 2-chloro-4-(((1s,3s)-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid 21b (2.44g, 10.01mmol) with N,N-dimethylacetamide (20mL ) Was dissolved, triethylamine (1.39mL, 10.01mmol) and diphenyl azide phosphate (2.16mL, 10.01mmol) were added under stirring at room temperature and reacted for 2h, and the temperature was raised to 110°C and refluxed for 2.5h. TLC monitors to the end of the reaction, the reaction solution is extracted with water and dichloromethane, and the organic layer is concentrated to obtain the title compound 2-chloro-9-(cis-3-hydroxycyclobutyl)-7,9-dihydro-8H-purine -8-one (21c) (white solid, 0.90 g, yield 37.35%).

1H NMR(400MHz,DMSO-d6)δ11.59(s,1H), 8.12(s,1H), 4.31-4.20(m,1H), 4.00-3.92(m,1H), 3.80-3.54(m,1H) ), 2.83–2.73(m,2H), 2.58–2.51(m,2H).

LC-MS m/z(ESI)=241.00[M+1].

the fourth step:

2-Chloro-9-(cis-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one (21d)

2-chloro-9-(cis-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Use 2-chloro-9-((1s,3s)-3-hydroxycyclobutyl)-7,9-dihydro-8H-purin-8-one 21c (0.70g, 1.25mmol) with N,N-two Methylformamide (10 mL) was dissolved, and dimethyl sulfate (0.28 mL, 1.25 mmol) and cesium carbonate (1.42 g, 4.36 mmol) were added under stirring at 0°C to react for 2 h. TLC monitors until the reaction is over, the reaction solution is slowly added dropwise to ice water and stirred, ethyl acetate is added for extraction, the concentrated organic layer is separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:1) Purified to obtain the title compound 2-chloro-9-(cis-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one (21d) (white solid, 0.32g, Yield 41.20%).

1H NMR(400MHz,DMSO-d6)δ8.33(s,1H),5.31(d,1H),4.34-4.23(m,1H),4.02-3.91(m,1H),3.34(s,3H), 2.81-2.72 (m, 2H), 2.58-2.51 (m, 2H).

LC-MS m/z(ESI)=255.00[M+1].

the fifth step:

9-(cis-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro -8H-purin-8-one (Compound 21)

9-(cis-3-hydroxycyclobutyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-((1s,3s)-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one 21d (50mg, 0.20mmol), 6 -Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (59mg, 0.40mmol), cesium carbonate (128mg, 0.4mmol) and Brettphos G3 Pd (19mg, 0.020mmol) were added to the dry reaction tube , N _{2 was} replaced three times, and then dry 1,4-dioxane (1 mL) was added, and the reaction was carried out at 110° C. for 5 h. TLC monitors until the reaction is complete, cool to room temperature, use silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to separate and purify to obtain the title compound 9-((1s, 3s)-3-hydroxy Butyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one( Compound 21) (light yellow solid, 22 mg, yield 30.50%).

1H NMR (400MHz, DMSO-d6) δ 8.14 (s, 1H), 7.95 (s, 1H), 7.26 (s, 1H), 6.60 (s, 1H), 5.11 (d, 1H), 4.48 (t, 2H), 4.23 - 4.14 (m, 1H), 3.94-3.88 (m, 1H), 3.25 (s, 3H), 3.13 (t, 2H), 2.82 - 2.73 (m, 2H), 2.54-2.44 (m, 2H), 2.12(s, 3H).

LC-MS m/z(ESI)=368.10[M+1].

Example 22

9-(4,4-Difluorocyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro -8H-purin-8-one (Compound 22)

9-(4,4-difluorocyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Ethyl 2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylate (22a)

ethyl-2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylate

2,4-Dichloropyrimidine-5-carboxylic acid ethyl ester 1a (6.5g, 29.41mmol), 4,4-difluorocyclohexyl-1-amine hydrochloride (5.0g, 29.41mmol) were dissolved in acetonitrile ( 100mL), potassium carbonate (10.16g, 73.52mmol) was added under stirring at room temperature to react for 4h. After the reaction was monitored by TLC, it was filtered and the residue was washed with ethyl acetate. The filtrate was concentrated and separated and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)=1:1) to obtain the title compound 2-chloro-4- ((Tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (22a) (white solid, 8.0 g, yield 85.10%).

1H NMR(400MHz,DMSO-d6)δ8.85(s,1H),8.11(d,1H),4.34-4.25(m,2H),4.21-4.10(m,1H),2.11-1.91(m,6H) ), 1.71-1.57 (m, 2H), 1.23 (t, 3H).

LCMS m/z(ESI)=320.10[M+1].

The second step:

2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylic acid (22b)

2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 22a (4g, 12.47mmol) in tetrahydrofuran/water (50mL/50mL) and add hydroxide Lithium (597.22mg, 24.94mmol), stirred at room temperature for 1h. TLC monitors until the reaction is complete, spin dry the tetrahydrofuran, adjust the pH to 5 with 2N hydrochloric acid, a white solid precipitates out, filter, wash the filter cake twice with petroleum ether, collect the solid to obtain the title compound 2-chloro-4-((4,4 -Difluorocyclohexyl)amino)pyrimidine-5-carboxylic acid (22b) (white solid, 3.4g, yield 91.53%).

1H NMR(400MHz,DMSO-d6)δ13.79(s,1H),8.60(s,1H),8.55(d,1H),4.22-4.06(m,1H),2.12-1.89(m,6H), 1.72-1.56 (m, 2H).

LCMS m/z(ESI)=292.00[M+1].

third step:

2-chloro-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one (22c)

2-chloro-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid 22b (3.34g, 11.45mmol) in dimethylacetamide (50mL), add three Ethylamine (1.6mL, 11.45mmol), diphenyl azide phosphate (1.22mL, 11.45mmol), then gradually heated to 120°C and stirred for 1.5h. TLC monitors the completion of the reaction, the reaction solution is poured into ice water, the solid is collected by filtration, washed with water 3 times, concentrated and dried in vacuo to obtain the title compound 2-chloro-9-(4,4-difluorocyclohexyl)-7,9-dihydro -8H-purin-8-one (22c) (white solid, 2.2 g, yield 78.5%).

1H NMR(400MHz,DMSO-d6)δ11.66(s,1H),8.13(s,1H),4.46-4.36(m,1H),2.17-1.94(m,6H),1.87-1.80(m,2H) ).

LCMS m/z(ESI)=289.10[M+1].

the fourth step:

2-chloro-7-methyl-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one (22d)

2-chloro-9-(4,4-difluorocyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one 22c (2.62g, 9.08mmol) in dimethylformamide (50mL) Add dimethyl sulfate (1.14g, 9.08mmol) and cesium carbonate (4.44g, 13.61mmol) at 0°C, and stir at 0°C for 1h. TLC monitors the end of the reaction. Pour the reaction solution into ice water. A solid precipitates out. The solid is filtered and collected to obtain the title compound 2-chloro-7-methyl-9-(4,4-difluorocyclohexyl)-7,9- Dihydro-8H-purin-8-one (22d) (white solid, 2.2 g, yield 80%).

1H NMR(400MHz,DMSO-d6)δ8.37(s,1H),4.55-4.39(m,1H),3.33(s,3H),2.21-2.01(m,6H),1.90-1.79(m,2H) ).

LCMS m/z(ESI)=303.10[M+1].

the fifth step:

9-(4,4-Difluorocyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro -8H-purin-8-one (Compound 22)

9-(4,4-difluorocyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The 2-chloro-7-methyl-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one 22d (100mg, 0.33mmol), 6-methyl- 2,3-Dihydrobenzofuran-5-amine intermediate 1 (99mg, 0.66mmol), cesium carbonate (215mg, 0.66mmol) and Brettphos G3 Pd (30mg, 0.033mmol) were added to the dry reaction tube and replaced _{with N 2} Three times, then dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. Cooled to room temperature, and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(4,4-difluorocyclohexyl)-7-methyl-2 -((6-Methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (Compound 22) (off-white solid, 62mg, product Rate 45.18%).

1H NMR(400MHz,DMSO-d6)δ8.18(s,1H),7.98(s,1H), 7.24(s,1H), 6.60(s,1H), 4.48(t,2H), 4.38-4.28( m, 1H), 3.27 (s, 3H), 3.11 (t, 2H), 2.56-2.43 (m, 2H), 2.12 (s, 3H), 2.11-1.92 (m, 4H), 1.77 (d, 2H) .

LC-MS m/z(ESI)=416.20[M+1].

Example 23

4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo 7,8-dihydro-9H-purine-9 -Yl)bicyclo[2.2.2]octane-1-carbonitrile (compound 23)

4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2. 2]octane-1-carbonitrile

first step:

Methyl 4-carbamoylbicyclo[2.2.2]octane-1-carboxylate (23b)

methyl 4-carbamoylbicyclo[2.2.2]octane-1-carboxylate

Dissolve 4-(methoxycarbonyl)bicyclo[2.2.2]octane-1-carboxylic acid 23a (10.0g, 47.12mmol) in anhydrous dichloromethane (200mL), add 2-(7-nitrogen under ice bath) Heterobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (18.81g, 49.47mmol), keep the temperature for 30min, add N,N-diisopropylethylamine (24.62mL, 141.35mmol), then slowly added ammonium chloride solid (3.78g, 70.67mmol) several times. The reaction gradually returned to room temperature and reacted overnight. TLC monitors the end of the reaction, the reaction solution is directly used with 0.5N hydrochloric acid solution 100mL, after separation, the organic phase is successively 100mL with water and saturated brine, dried, and concentrated to obtain the target compound 4-carbamoylbicyclo[2.2.2]octane- Methyl 1-formate (23b) (white solid, 21 g, crude product).

1H NMR (400MHz, DMSO-d6) δ 6.94 (s, 1H), 6.73 (s, 1H), 3.57 (s, 3H), 1.72-1.63 (m, 12H).

LC-MS m/z(ESI)=212.10[M+1].

The second step:

4-cyanobicyclo[2.2.2]octane-1-carboxylic acid methyl ester (23c)

methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate

Dissolve 4-carbamoyl bicyclo[2.2.2]octane-1-carboxylic acid methyl ester 23b (21g, 99.40mmol) in dichloromethane (200mL), add pyridine (16.02mL, 198.81mmol) in an ice bath, trifluoro Acetic anhydride (21.00ml, 149.10mmol), keep the temperature and continue the reaction for 1h. TLC monitors the end of the reaction, directly filter the reaction solution, wash the filter cake with 100ml of dichloromethane, combine the organic phases, then use 1N hydrochloric acid solution, water, and saturated brine each with 100ml, dry, concentrate and separate by column chromatography to obtain the target The product 4-cyanobicyclo[2.2.2]octane-1-carboxylic acid methyl ester (23c) (white solid, 5.3g, 60mg, two-step yield 58.21%).

1H NMR (400MHz, CDCl3-d6) δ 3.66 (s, 3H), 1.98-1.94 (m, 6H), 1.86-1.82 (m, 6H).

LC-MS m/z(ESI)=194.10[M+1].

third step:

4-cyanobicyclo[2.2.2]octane-1-carboxylic acid (23d)

4-cyanobicyclo[2.2.2]octane-1-carboxylic acid

Dissolve 4-cyanobicyclo[2.2.2]octane-1-carboxylic acid methyl ester 23c (5.3g, 27.43mmol) in tetrahydrofuran 50mL, water 50mL, add lithium hydroxide (1.73g, 41.14mmol), room temperature Stir for 1h. TLC monitors that the reaction is complete, concentrates to remove tetrahydrofuran, adjusts the pH to 5 with 2N hydrochloric acid, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether, and the solid is collected to obtain the title compound 4-cyanobicyclo[2.2.2]octane -1-carboxylic acid (23d) (white solid, 5.0 g, crude product).

1H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 1.89-1.85 (m, 6H), 1.72-1.68 (m, 6H).

LC-MS m/z(ESI)=180.10[M+1].

the fourth step:

4-aminobicyclo[2.2.2]octane-1-carbonitrile hydrochloride (23e)

4-aminobicyclo[2.2.2]octane-1-carbonitrile hydrochloride

Dissolve 4-cyanobicyclo[2.2.2]octane-1-carboxylic acid 23d (5.0g, 27.90mmol) in toluene (60mL), add diphenyl azide phosphate (6.01mL, 27.90mmol) in an ice bath And triethylamine (3.88mL, 27.90mmol), the reaction solution was stirred at room temperature for 1h, then the temperature was raised to 90°C and the reaction was continued for 3h. TLC monitors until the reaction is complete, cool to room temperature, slowly pour 100mL 1N hydrochloric acid solution, a large amount of solids appear, filter, collect the solids, beat with ethyl acetate (150mL), and dry in vacuo to obtain the title compound 4-aminobicyclo[2.2.2 ] Octane-1-nitrile hydrochloride (23e) (white solid, 7.3 g, crude product, yield 80%).

1H NMR(401MHz,DMSO-d6)δ8.45(s,2H),2.01–1.97(m,6H),1.81-1.76(m,6H).

LC-MS m/z(ESI)=151.20[M+1].

the fifth step:

2-chloro-4-((4-cyanobicyclo[2.2.2]octane-1-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (23f)

ethyl 2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylate

The 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (8.74g, 39.53mmol), 4-aminobicyclo[2.2.2]octane-1-carbonitrile hydrochloride 23e (7.38g, 39.53mmol, 60% purity), potassium carbonate (21.85g, 158.13mmol) was dissolved in acetonitrile (200mL), and the reaction solution was reacted at room temperature for 16h. The end of the reaction was monitored by TLC, filtered, and the solid was washed with a small amount of acetonitrile. The filtrate was combined and concentrated. The crude product was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=1:1) to obtain the title compound 2-chloro- 4-((4-Cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (23f) (white solid, 4.0 g, yield 30.23%).

1H NMR(400MHz,DMSO-d6)δ8.63(s,1H), 8.29(s,1H), 4.30(q,2H), 2.10–2.00(m,12H), 1.30(t,3H).

LC-MS m/z(ESI)=335.10[M+1].

The sixth step:

2-chloro-4-((4-cyanobicyclo[2.2.2]octane-1-yl)amino)pyrimidine-5-carboxylic acid (23g)

2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 23f (4g, 11.95mmol) in tetrahydrofuran 50mL, water 50mL Lithium hydroxide (1.01 g, 23.90 mmol) was added to the mixture, and stirred at room temperature for 1 h. TLC monitored the reaction to complete, concentrated to remove tetrahydrofuran, adjusted the pH to 5 with 2N hydrochloric acid, a white solid precipitated out, filtered, the filter cake was washed twice with petroleum ether, the solid was collected to obtain the title compound 2-chloro-4-((4-cyano Bicyclo[2.2.2]octane-1-yl)amino)pyrimidine-5-carboxylic acid (23g) (3.6g, white solid, yield 98.23%), proceed directly to the next experiment.

1H NMR (600MHz, DMSO-d6) δ 13.85 (s, 1H), 8.59 (s, 1H), 8.56 (s, 1H), 2.07-1.98 (m, 12H).

LCMS m/z(ESI)=307.10[M+1].

The seventh step:

4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile (23h)

4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile

Dissolve 23g (3.8g, 12.39mmol) of 2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid in dimethylacetamide ( 50mL), triethylamine (1.72mL, 12.39mmol) and diphenyl azide phosphate (2.67mL, 12.39mmol) were added, and then gradually heated to 120°C and stirred for 1.5h. TLC monitors the completion of the reaction, pour the reaction solution into ice water, filter to collect the solid, wash 3 times with water, concentrate and dry in vacuo to obtain 4-(2-chloro-8-oxo-7,8-dihydro-9H-purine-9- Yl)bicyclo[2.2.2]octane-1-carbonitrile (23h) (3.3g, white solid, yield 87.7%).

1H NMR(400MHz,DMSO-d6)δ11.61(s,1H), 8.09(s,1H), 2.48–2.40(m,6H), 2.09–2.03(m,6H).

LC-MS m/z(ESI)=304.20[M+1].

The eighth step:

4-(2-Chloro-7-methyl-8-oxo 7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile (23i)

4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile

Dissolve 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-nitrile 23h (3.3g, 11.43mmol) in Dimethylformamide (50mL), dimethyl sulfate (1.44g, 11.43mmol) and cesium carbonate (5.59g, 17.15mmol) were added at 0°C, and stirred at 0°C for 1h. TLC monitors the end of the reaction. Pour the reaction solution into ice water. A solid precipitates out. The solid is filtered and collected to obtain the title compound 4-(2-chloro-7-methyl-8-oxo7,8-dihydro-9H-purine -9-yl)bicyclo[2.2.2]octane-1-carbonitrile (23i) (3.1 g, white solid, yield 89.59%).

1H NMR(401MHz,DMSO-d6)δ8.33(s,1H), 3.29(s,3H), 2.48–2.41(m,6H), 2.10–2.04(m,6H).

LC-MS m/z(ESI)=318.20[M+1].

Step 9:

4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo 7,8-dihydro-9H-purine-9 -Yl)bicyclo[2.2.2]octane-1-carbonitrile (compound 23)

4-(7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2. 2]octane-1-carbonitrile

The 4-(2-chloro-7-methyl-8-oxo 7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile 23i (100mg, 0.31mmol ), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (94mg, 0.62mmol), cesium carbonate (205mg, 0.62mmol) and Brettphos G3 Pd (28mg, 0.031mmol) were added to the dry The reaction tube was _{replaced with N 2} three times, then dry 1,4-dioxane (1 mL) was added, and the reaction was carried out at 110°C for 5 hours. After cooling to room temperature, silica gel column chromatography was used for separation and purification (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 4-(7-methyl-2-((6-methyl-2, 3-Dihydrobenzofuran-5-yl)amino)-8-oxo7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octan-1-one (compound 23) (Off-white solid, 53 mg, yield 39.12%).

1H NMR (400MHz, DMSO-d6) δ 8.16 (s, 1H), 7.95 (s, 1H), 7.23 (s, 1H), 6.61 (s, 1H), 4.50 (t, 2H), 3.20 (s, 3H), 3.12(t,2H), 2.45–2.37(M,6H), 2.12(s,3H), 2.04–1.96(m,6H).

LC-MS m/z(ESI)=431.20[M+1].

Example 24

9-(8-oxabicyclo[3.2.1]octan-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino )-7,9-dihydro-8H-purin-8-one

9-(8-oxabicyclo[3.2.1]octan-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

first step:

8-oxabicyclo[3.2.1]octan-3-one oxime (24b)

8-oxabicyclo[3.2.1]octan-3-one oxime

Dissolve 8-oxabicyclo[3.2.1]octan-3-one 24a (1.4g, 11.1mmol), hydroxylamine hydrochloride (925mg, 13.3mmol) and potassium carbonate (3.1g, 22.2mmol) in ethanol/water ( 10mL/5mL) in a mixed solvent, react at 80°C for 2h. After the reaction, the reaction solution was directly concentrated, quenched with water, extracted with ethyl acetate, dried with anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure to obtain the title compound 8-oxabicyclo[3.2.1]oct-3- Ketone oxime (24b) (light yellow solid, 1.56 g, yield 100%).

LC-MS m/z(ESI)=142.10[M+1].

The second step:

8-oxabicyclo[3.2.1]oct-3-amine(24c)

8-oxabicyclo[3.2.1]octan-3-amine

Dissolve 8-oxabicyclo[3.2.1]octan-3-one oxime 24b (1.5g, 10.64mmol) in methanol (30mL), add nickel chloride hexahydrate (2.53g, 10.64mmol) at room temperature, React at room temperature for 0.5h. The reaction solution was cooled to -30°C, and sodium borohydride (6.0g, 159.6mmol) was slowly added. After the addition, it was slowly raised to room temperature and reacted overnight; the reaction was completed, quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered The organic solvent was removed under reduced pressure to obtain the title compound 8-oxabicyclo[3.2.1]oct-3-amine (24c) (light yellow oil, 639 mg, yield 45%).

LC-MS m/z(ESI)=128.10[M+1].

third step:

4-((8-oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (24d)

ethyl 4-((8-oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (1.1g, 5.03mmol), potassium carbonate (1.74g, 12.58mmol) in acetonitrile (20mL), add 8-oxabicyclo at 0℃ [3.2.1] Octan-3-amine 24c (639 mg, 5.03 mmol), stirred at room temperature for 20 h. Add 30 mL of water, a solid precipitated out, filtered and washed with water 3 times, and concentrated to obtain the title compound 4-((8-oxabicyclo[3.2.1]octane-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid Ethyl ester (24d) (white solid, 1.01 g, yield 64.3%).

1H NMR(400MHz,DMSO)δ8.91(d,1H),8.64(s,1H), 4.38-4.31(m,4H), 4.30-4.24(m,1H), 2.20-2.08(m,2H), 2.02–1.90 (m, 4H), 1.66 (d, 2H), 1.32 (t, 3H).

LC-MS m/z(ESI)=312.10[M+1].

the fourth step:

4-((8-oxabicyclo[3.2.1]oct-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid (24e)

4-((8-oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid

Dissolve 4-((8-oxabicyclo[3.2.1]octane-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 24d (1.0g, 3.21mmol) in 10mL of tetrahydrofuran, water In 5 mL, lithium hydroxide (308 mg, 12.83 mmol) was added, and the mixture was stirred at room temperature for 1 h. The tetrahydrofuran was spin-dried and the pH was adjusted to 4-5. A white solid precipitated out and filtered. The filter cake was washed twice with petroleum ether/ethyl acetate (v/v=10/1) and concentrated to obtain the title compound 4-((8 -Oxabicyclo[3.2.1]oct-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid (24e) (white solid, 808 mg, yield 87.9%).

1H NMR (400MHz, DMSO) δ 13.83 (s, 1H), 9.17 (d, 1H), 8.59 (s, 1H), 4.32 (s, 2H), 4.29-4.22 (m, 1H), 2.17-2.07 ( m, 2H), 2.00–1.89 (m, 4H), 1.65 (d, 2H).

LC-MS m/z(ESI)=284.20[M+1].

the fifth step:

9-(8-oxabicyclo[3.2.1]oct-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one (24f)

9-(8-oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one

Dissolve 4-((8-oxabicyclo[3.2.1]oct-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid 24e (808mg, 2.85mmol) in dimethylacetamide (20mL) Add triethylamine (288mg, 2.85mmol) and diphenyl azide phosphate (784mg, 2.85mmol), then gradually increase the temperature to 120°C and stir for 1.5h. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=5:1～1:10) to obtain the title compound 9-(8-oxabicyclo[3.2.1 ]Oct-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one (24f) (white solid, 653 mg, yield 71%).

1H NMR (600MHz, DMSO) δ 11.64 (s, 1H), 8.12 (s, 1H), 4.47-4.44 (m, 2H), 4.42-4.40 (m, 1H), 2.28-2.20 (m, 2H), 2.07–2.01(m,2H), 1.96–1.90(m,2H), 1.81–1.75(m,2H).

LC-MS m/z(ESI)=281.10[M+1].

The sixth step:

9-(8-oxabicyclo[3.2.1]oct-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one (24g)

9-(8-oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one

Dissolve 9-(8-oxabicyclo[3.2.1]oct-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one 24f (653mg, 2.33mmol) in dimethyl Add dimethyl sulfate (293 mg, 2.33 mmol) and cesium carbonate (1.52 g, 4.65 mmol) to methyl formamide (10 mL) at 0°C, and stir at 0°C for 1 h. Then 10mL of water was added, extracted with ethyl acetate 3 times, the organic phase was dried over anhydrous sodium sulfate, concentrated, a solid precipitated out, filtered to obtain the title compound 9-(8-oxabicyclo[3.2.1]oct-3-yl )-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one (24g) (white solid, 533mg, yield 81.6%).

1H NMR (400MHz, DMSO) δ 8.36 - 8.34 (m, 1H), 4.51-4.47 (m, 2H), 4.45 - 4.42 (m, 1H), 3.34 (s, 3H), 2.31 - 2.22 (m, 2H) ), 2.07–1.98(m,2H), 1.98–1.90(m,2H), 1.79(t,2H).

LC-MS m/z(ESI)=295.20[M+1].

The seventh step:

9-(8-oxabicyclo[3.2.1]octane-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino )-7,9-dihydro-8H-purin-8-one (Compound 24)

9-(8-oxabicyclo[3.2.1]octan-3-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

The 9-(8-oxabicyclo[3.2.1]oct-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one 24g (100mg, 0.34mmol ), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (101mg, 0.68mmol), cesium carbonate (221mg, 0.68mmol) and Brettphos G3 Pd (31mg, 0.034mmol) were added to the dry The reaction tube was _{replaced with N 2} three times, then dry 1,4-dioxane (1 mL) was added, and the reaction was carried out at 110°C for 5 hours. Cooled to room temperature, and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(8-oxabicyclo[3.2.1]octan-3-yl )-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 24 ) (Off-white solid, 60 mg, yield 43.40%).

1H NMR(400MHz,DMSO-d6)δ8.27(s,1H),7.98(s,1H), 7.09(s,1H), 6.64(s,1H), 4.49(t,2H), 4.38-4.29( m, 3H), 3.25 (s, 3H), 3.11 (t, 2H), 2.15-2.05 (m, 5H), 1.95 (t, 2H), 1.81-1.74 (m, 2H), 1.50-1.40 (d, 2H).

LC-MS m/z(ESI)=408.20[M+1].

Example 25

7-methyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.5]nonyl-7-yl) -7,9-dihydro-8H-purin-8-one

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin -8-one

first step:

2-oxaspiro[3.5]non-7-one oxime (25b)

2-oxaspiro[3.5]nonan-7-one oxime

Dissolve 2-oxaspiro[3.5]nonyl-7-one 25a (1.5g, 10.7mmol), hydroxylamine hydrochloride (744mg, 10.7mmol) and potassium carbonate (2.95g, 21.4mmol) in ethanol/water (10mL/ 5mL) in a mixed solvent, react at 80°C for 2h. After the reaction, the reaction solution was directly concentrated, quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure to obtain the title compound 2-oxaspiro[3.5]non-7-one oxime (25b) (Light yellow solid, 1.45 g, yield 89%).

LC-MS m/z(ESI)=156.10[M+1].

The second step:

2-oxaspiro[3.5]non-7-amine (25c)

2-oxaspiro[3.5]nonan-7-amine

Dissolve 2-oxaspiro[3.5]non-7-one oxime 25b (1.45g, 9.34mmol) in methanol (30mL) and add nickel chloride hexahydrate (2.22g, 9.34mmol) at room temperature. Reaction for 0.5h. The reaction solution was cooled to -30°C, and sodium borohydride (5.3g, 140.1mmol) was slowly added. After the addition, the temperature was slowly raised to room temperature and reacted overnight; the reaction was completed, quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered The organic solvent was removed under reduced pressure to obtain the title compound 2-oxaspiro[3.5]non-7-amine (25c) (light yellow oil, 481 mg, yield 37.6%).

LC-MS m/z(ESI)=142.20[M+1].

third step:

Ethyl 4-((2-oxaspiro[3.5]non-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (25d)

ethyl 4-((2-oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylate

Dissolve 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester 1a (753mg, 3.41mmol), potassium carbonate (940mg, 6.81mmol) in acetonitrile (20mL), add 8-oxabicyclo[3.2 .1] Octan-3-amine 25c (481mg, 3.41mmol), stirred at room temperature for 20h. Add 30 mL of water, a solid precipitated out, filtered and washed with water 3 times, and concentrated to obtain the title compound ethyl 4-((2-oxaspiro[3.5]non-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (25d) (white solid, 640 mg, yield 57.6%).

1H NMR (400MHz, DMSO) δ8.61 (s, 1H), 8.29 (d, 1H), 4.34 - 4.29 (m, 4H), 4.24 (s, 2H), 3.98 - 3.89 (m, 1H), 2.05 - 1.92(m,2H), 1.86–1.75(m,2H), 1.65–1.56(m,2H), 1.45–1.35(m,2H), 1.34–1.27(m,3H).

LC-MS m/z(ESI)=326.20[M+1].

the fourth step:

4-((2-oxaspiro[3.5]nonyl-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid (25e)

4-((2-oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid

Ethyl 4-((2-oxaspiro[3.5]non-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 25d (640mg, 1.96mmol) was dissolved in tetrahydrofuran 10mL, water 5mL, Lithium hydroxide (189 mg, 7.86 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The tetrahydrofuran was spin-dried and adjusted to pH 4-5. A white solid precipitated out. Filtered. The filter cake was washed twice with petroleum ether/ethyl acetate (v/v=10/1) and concentrated to obtain the title compound 4-((2 -Oxaspiro[3.5]nonyl-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid (25e) (white solid, 518 mg, yield 88.5%).

1H NMR (400MHz, DMSO) δ 13.74 (s, 1H), 8.57 (s, 1H), 8.48 (d, 1H), 4.32 (s, 2H), 4.24 (s, 2H), 3.97-3.84 (m, 1H), 1.99(d,2H), 1.86–1.76(m,2H), 1.66–1.54(m,2H), 1.42–1.26(m,2H).

LC-MS m/z(ESI)=298.10[M+1].

the fifth step:

2-Chloro-9-(2-oxaspiro[3.5]non-7-yl)-7,9-dihydro-8H-purin-8-one (25f)

2-chloro-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 4-((2-oxaspiro[3.5]nonyl-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid 25e (518mg, 1.74mmol) in dimethylacetamide (20mL) Add triethylamine (175 mg, 1.74 mmol) and diphenyl azide phosphate (479 mg, 1.74 mmol), then gradually increase the temperature to 120° C. and stir for 1.5 h. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=5:1～1:10) to obtain the title compound 2-chloro-9-(2-oxaspiro [3.5] Non-7-yl)-7,9-dihydro-8H-purin-8-one (25f) (white solid, 353 mg, yield 72.8%).

1H NMR (600MHz, DMSO) δ 11.61 (s, 1H), 8.11 (s, 1H), 4.41 (s, 2H), 4.24 (s, 2H), 4.15-4.08 (m, 1H), 2.24-2.08 ( m, 4H), 1.69 (d, 2H), 1.64-1.51 (m, 2H).

LC-MS m/z(ESI)=295.10[M+1].

The sixth step:

2-Chloro-7-methyl-9-(2-oxaspiro[3.5]non-7-yl)-7,9-dihydro-8H-purin-8-one (25g)

2-chloro-7-methyl-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(2-oxaspiro[3.5]non-7-yl)-7,9-dihydro-8H-purin-8-one 25f (353mg, 1.18mmol) in dimethylform To the amide (10 mL), dimethyl sulfate (150 mg, 1.18 mmol) and cesium carbonate (464 mg, 3.36 mmol) were added at 0°C, and the mixture was stirred at 0°C for 1 h. Then 10 mL of water was added, extracted with ethyl acetate 3 times, the organic phase was dried over anhydrous sodium sulfate, concentrated, a solid precipitated out, filtered to obtain the title compound 2-chloro-7-methyl-9-(2-oxaspiro[ 3.5] Non-7-yl)-7,9-dihydro-8H-purin-8-one (25 g) (white solid, 313 mg, yield 85.5%).

1H NMR (400MHz, DMSO) δ 8.34 (s, 1H), 4.41 (s, 2H), 4.25 (s, 2H), 4.22-4.08 (m, 1H), 3.34 (s, 3H), 2.22-2.05 ( m, 4H), 1.69 (d, 2H), 1.64-1.50 (m, 2H).

LC-MS m/z(ESI)=309.20[M+1].

The seventh step:

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.5]nonyl-7-yl)- 7,9-dihydro-8H-purin-8one (Compound 25)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin -8-one

25g (100mg, 0.32mmol) of 2-chloro-7-methyl-9-(2-oxaspiro[3.5]non-7-yl)-7,9-dihydro-8H-purin-8-one, 6-Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (97mg, 0.64mmol), cesium carbonate (211mg, 0.64mmol) and Brettphos G3 Pd (29mg, 0.032mmol) were added to the dry reaction Replace the tube with N2 three times, then add dry 1,4-dioxane (1 mL), and react at 110°C for 5 hours. Cool to room temperature, use silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-((6-methyl-2,3-di Hydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.5]nonyl-7-yl)-7,9-dihydro-8H-purin-8one (compound 25) (off-white Solid, 50 mg, yield 36.63%).

1H NMR (400MHz, DMSO-d6) δ 8.24 (s, 1H), 7.96 (s, 1H), 7.07 (s, 1H), 6.63 (s, 1H), 4.47 (t, 2H), 4.19 (s, 2H), 4.10 (s, 2H), 4.06-3.96 (m, 1H), 3.25 (s, 3H), 3.12 (t, 2H), 2.13-1.99 (m, 7H), 1.55 (d, 2H), 1.52 –1.41(m,2H).

LC-MS m/z(ESI)=422.20[M+1].

Example 26

9-(3-Hydroxybicyclo[3.2.1]octyl-8-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino) -7,9-dihydro-8H-purin-8-one

9-(3-hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

first step:

8-aminobicyclo[3.2.1]octyl-3-one hydrochloride (26b)

8-aminobicyclo[3.2.1]octan-3-one hydrochloride

Dissolve tert-butyl 3-oxobicyclo[3.2.1]oct-8-yl)carbamate 26a (1.0 g, 4.18 mmol) in 4N dioxane hydrochloride (40 mL), and react at room temperature for 1 h. TLC monitored the completion of the reaction, and the reaction solution was directly concentrated to dryness to obtain the target compound 8-aminobicyclo[3.2.1]octan-3-one hydrochloride (26b) (white solid, 30mg, yield 99.42%).

1H NMR (401MHz, DMSO-d6) δ8.77 (s, 2H), 3.36 (m, 2H), 2.80-2.77 (m, 2H), 2.16-2.13 (m, 2H), 1.87-1.85 (m, 2H) ), 1.43-1.42 (m, 2H).

LC-MS m/z(ESI)=176.20[M+1].

The second step:

Ethyl 2-chloro-4-(((3-oxobicyclo[3.2.1]oct-8-yl)amino)pyrimidine-5-carboxylate (26c)

ethyl 2-chloro-4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylate

Ethyl 2,4-dichloropyrimidine-5-carboxylate 1a (1.38g, 6.26mmol), (1R, 5S, 8s)-8-aminobicyclo[3.2.1]octan-3-one hydrochloride 26b (730mg, 4.17mmol), potassium carbonate (1.73g, 12.52mmol) was dissolved in acetonitrile (20mL), and the reaction solution was reacted at room temperature for 16h. The end of the reaction was monitored by TLC, filtered, and the solid was washed with a small amount of acetonitrile. The filtrate was combined and concentrated. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain the target compound 2-chloro-4-((( 3-Oxybicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (26c) (white solid, 1.0 g, yield 74.01%).

1H NMR(400MHz,DMSO-d6)δ8.86(d,2H),8.68(s,1H),4.36-4.30(q,2H),4.20-4.16(m,1H),2.66-2.65(m,2H) ), 2.56-2.55 (m, 2H), 2.21-2.17 (m, 2H), 1.97-1.93 (m, 2H), 1.53-1.33 (m, 2H), 1.31 (t, 3H).

LC-MS m/z(ESI)=324.10[M+1].

third step:

2-chloro-4-((3--3-oxobicyclo[3.2.1]oct-8-yl)amino]pyrimidine-5-carboxylic acid (26d)

2-chloro-4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((3-oxobicyclo[3.2.1]oct-8-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 26c (1.0g, 3.09mmol) in 20mL of tetrahydrofuran and 20mL of water , Lithium hydroxide (259mg, 6.18mmol) was added and stirred at room temperature for 1h. TLC monitored the reaction to complete, concentrated to remove tetrahydrofuran, adjusted the pH to 5 with 2N hydrochloric acid, a white solid precipitated out, filtered, the filter cake was washed twice with petroleum ether, the solid was collected to obtain the title compound 2-chloro-4-((3-3- Oxybicyclo[3.2.1]octan-8-yl)amino]pyrimidine-5-carboxylic acid (26d) (white solid, 800mg, yield 87.59%).

1H NMR(400MHz,DMSO-d6)δ13.93(s,1H),9.20(d,2H),8.64(s,1H),4.20-4.16(m,1H),2.66-2.64(m,2H), 2.54-2.53 (m, 2H), 2.21-2.16 (m, 2H), 1.96-1.93 (m, 2H), 1.53-1.47 (m, 2H).

LC-MS m/z(ESI)=296.10[M+1].

the fourth step:

2-chloro-9-(3-oxobicyclo[3.2.1]oct-8-yl)-7,9-dihydro-8H-purin-8-one (26e)

2-chloro-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((3-3-oxobicyclo[3.2.1]oct-8-yl)amino)pyrimidine-5-carboxylic acid 26d (800mg, 2.71mmol) in dimethylacetamide (20mL ), add triethylamine (0.37mL, 2.7mmol) and diphenyl azide phosphate (0.58mL, 2.7mmol), then gradually increase the temperature to 90°C and stir for 2h. TLC monitors the completion of the reaction, the reaction solution is poured into ice water, the solid is collected by filtration, washed with water 3 times, and concentrated and dried in vacuo to obtain the title compound 2-chloro-9-(3-oxobicyclo[3.2.1]oct-8-yl) -7,9-dihydro-8H-purin-8-one (26e) (white solid, 630 mg, yield 94.71%).

1H NMR(400MHz,DMSO-d6)δ11.66(s,1H),8.13(s,1H),4.01-3.99(m,1H),3.65-3.64(m,2H),2.66-2.61(m,2H) ), 2.12-2.08 (m, 2H), 1.89-1.86 (m, 2H), 1.54-1.49 (m, 2H).

LC-MS m/z(ESI)=293.00[M+1].

the fifth step:

2-chloro-7-methyl-9-(3-oxobicyclo[3.2.1]oct-8-yl)-7,9-dihydro-8H-purin-8-one(26f)

2-chloro-7-methyl-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(3-oxobicyclo[3.2.1]oct-8-yl)-7,9-dihydro-8H-purin-8-one 26e (630mg, 2.15mmol) in dimethyl Methyl formamide (10 mL), dimethyl sulfate (0.2 mL, 2.15 mmol) and cesium carbonate (1.4 g, 4.3 mmol) were added at 0°C, and the mixture was stirred at 0°C for 1 h. TLC monitors the end of the reaction. Pour the reaction liquid into ice water. A solid precipitates out. The solid is filtered and collected to obtain the title compound 2-chloro-7-methyl-9-(3-oxobicyclo[3.2.1]oct-8- Yl)-7,9-dihydro-8H-purin-8-one (26f) (white solid, 490 mg, yield 74.22%).

1H NMR(400MHz,DMSO-d6)δ8.37(s,1H),4.03-4.01(m,1H),3.66-3.65(m,2H),3.36(s,3H),2.65-2.59(m,2H) ), 2.13-2.08 (m, 2H), 1.89-1.87 (m, 2H), 1.55-1.50 (m, 2H).

LC-MS m/z(ESI)=307.10[M+1].

The sixth step:

2-chloro-9-(3-hydroxybicyclo[3.2.1]oct-8-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (26g)

2-chloro-9-(3-hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-7,9-dihydro-8H-purin-8-one

The 2-chloro-7-methyl-9-(3-oxobicyclo[3.2.1]oct-8-yl)-7,9-dihydro-8H-purin-8-one 26f (490mg, 1.6mmol ) Was dissolved in tetrahydrofuran (30 mL) and methanol (30 mL), and sodium borohydride (181.30 mg, 4.79 mmol) was added at 0°C. TLC monitors the end of the reaction. Pour the reaction solution into ice water. A solid precipitates out. The solid is filtered and collected to obtain the title compound 2-chloro-9-(3-hydroxybicyclo[3.2.1]oct-8-yl)-7-methyl Glyc-7,9-dihydro-8H-purin-8-one (26 g) (white solid, 380 mg, yield 77.04%).

1H NMR(400MHz,DMSO-d6)δ8.37(s,1H),4.28-4.20(m,1H),3.74-3.66(m,2H),3.45-3.43(m,2H),3.37(s,3H) ), 2.14-2.07 (m, 1H), 1.74-1.50 (m, 5H), 1.28-1.22 (m, 2H).

LC-MS m/z(ESI)=309.10[M+1].

The seventh step:

9-(3-Hydroxybicyclo[3.2.1]octyl-8-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino) -7,9-dihydro-8H-purin-8-one (compound 26)

9-(3-hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H -purin-8-one

The 2-chloro-9-(3-hydroxybicyclo[3.2.1]oct-8-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 26g (50mg, 0.16mmol) , 6-Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (49mg, 0.32mmol), cesium carbonate (106mg, 0.32mmol) and Brettphos G3 Pd (15mg, 0.016mmol) were added to the dry In the reaction tube, _{replace with N 2} three times, then add dry 1,4-dioxane (1 mL), and react at 110° C. for 5 h. Cooled to room temperature, and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(3-hydroxybicyclo[3.2.1]octyl-8-yl) -7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 26) (Off-white solid, 25mg, yield 36.63%).

1H NMR (400MHz, DMSO-d6) δ 8.18 (s, 1H), 8.00 (s, 1H), 7.20 (s, 1H), 6.60 (s, 1H), 4.49 (t, 2H), 4.24 (d, 1H), 3.74--3.63(m,1H), 3.53--3.49(m,1H), 3.45--3.39(m,2H), 3.28(s,3H), 3.09(t,2H), 2.12(s,3H) ,1.66-1.53(m,4H), 1.52-1.46(m,2H),1.33-1.25(m,2H).

LC-MS m/z(ESI)=422.20[M+1].

Example 27

9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9- Dihydro-8H-purin-8-one (Compound 27)

9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

first step:

Tert-Butyl (3-hydroxy-3-methylcyclohexyl) carbamate (27b)

tert-butyl(3-hydroxy-3-methylcyclohexyl)carbamate

Dissolve tert-butyl (3-oxocyclohexyl) carbamate 27a (5.00g, 23.4mmol) in tetrahydrofuran (80mL) and pre-cool at -78℃, then slowly add 1.6M methyl lithium dropwise over 45min. Ether solution (61.5mL, 98.28mmol), the reaction solution was stirred at -78°C for 1h, then the remaining 1.6M methyl lithium-ether solution (61.5mL, 98.28mmol) was added, and the reaction was stirred at -78°C for 1h. TLC monitors to the end of the reaction, adds saturated ammonium chloride solution to quench the reaction, adds water and ethyl acetate for extraction, concentrates the organic layer and uses silica gel column chromatography to separate and purify (petroleum ether/ethyl acetate (v/v)=8:1 ) Purified to obtain the title compound tert-butyl (3-hydroxy-3-methylcyclohexyl) carbamate (27b) (white solid, 2.07 g, yield 38.53%).

1H NMR (400MHz, Chloroform-d) δ 4.30 (s, 1H), 3.68 (s, 1H), 1.90 (d, 3H), 1.67 (q, 1H), 1.59-1.48 (m, 2H), 1.37 ( s,9H),1.19–1.12(m,4H),1.06(t,1H),0.96–0.84(m,1H).

LC-MS m/z(ESI)=230.20[M+1].

The second step:

3-amino-1-methylcyclohexane-1-ol hydrochloride (27c)

3-amino-1-methylcyclohexan-1-ol hydrochloride

Tert-butyl (3-hydroxy-3-methylcyclohexyl) carbamate 27b (2.07 g, 9.03 mmol) was dissolved in 4M hydrogen chloride-1,4-dioxane (10 mL), and the reaction was stirred at room temperature. TLC monitors to the end of the reaction, concentrates and evaporates the 1,4-dioxane solution to obtain the title compound 3-amino-1-methylcyclohexane-1-ol hydrochloride (27c) (light yellow solid, crude product, 1.49) g, yield 99.60%).

LC-MS m/z(ESI)=130.20[M+1].

third step:

Ethyl 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylate (27d)

ethyl 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxyl-ate

2,4-Dichloropyrimidine-5-carboxylic acid ethyl ester 1a (2.99g, 13.54mmol), 3-amino-1-methylcyclohexane-1-ol hydrochloride 27c (1.49g, 9.03mmol) Dissolved in acetonitrile (20 mL), stirred and added potassium carbonate (3.74 g, 27.08 mmol), and stirred at room temperature for 4 h. TLC monitors to the end of the reaction, concentrates and purifies by column separation (petroleum ether: ethyl acetate (v/v)=4:1) to obtain the title compound 2-chloro-4-((3-hydroxy-3-methylcyclohexyl) Amino)pyrimidine-5-carboxylic acid ethyl ester (27d) (white solid, 2.23 g, yield 78.74%).

1H NMR(400MHz,Chloroform-d)δ8.57(s,1H), 8.19(d,1H), 4.43-4.32(m,1H), 4.27(q,2H), 2.07-1.95(m,2H), 1.83–1.69(m,2H), 1.64–1.56(m,2H), 1.31(t,4H), 1.26(d,1H), 1.21(s,3H), 1.10–1.05(m,1H).

LC-MS m/z(ESI)=314.10[M+1].

the fourth step:

2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (27e)

2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylate

Dissolve 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid ethyl ester 27d (2.23g, 7.11mmol) in tetrahydrofuran/water (15mL/15mL), Lithium hydroxide (895 mg, 21.32 mmol) was added, and the mixture was stirred at room temperature for 1 h. TLC monitors until the reaction is complete, concentrates to remove tetrahydrofuran, adjusts the pH to 3-4 with 2N hydrochloric acid, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether, and the solid is collected to obtain the title compound 2-chloro-4-((3 -Hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (27e) (white solid, 1.6g, yield 78.79%).

LC-MS m/z(ESI)=286.10[M+1].

the fifth step:

2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (27e)

2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 27d (1.38g, 4.83mmol) in dimethylacetamide (20mL) and add triethyl Amine (0.67mL, 4.83mmol), diphenyl azide phosphate (1.04mL, 4.83mmol), then gradually heated to 90°C and stirred for 2h. TLC monitors to the end of the reaction, the reaction solution is poured into ice water, the solid is collected by filtration, washed with water 3 times, and concentrated and dried in vacuo to obtain the target compound 2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7,9 -Dihydro-8H-purin-8-one (27e) (white solid, 1.35 g, yield 98.86%).

1H NMR (400MHz, DMSO-d6) δ 11.60 (s, 1H), 8.10 (s, 1H), 4.62-4.54 (m, 1H), 4.35 (s, 1H), 2.19 (t, 1H), 2.10- 1.99 (m, 1H), 1.70 to 1.53 (m, 5H), 1.30 to 1.22 (m, 1H), 1.16 (s, 3H).

LC-MS m/z(ESI)=283.10[M+1].

The sixth step:

2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (27f)

2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 27e (1.35g, 4.77mmol) in dimethylformamide ( 10 mL), dimethyl sulfate (0.45 mL, 4.77 mmol) and cesium carbonate (1.56 g, 4.77 mmol) were added at 0°C, and the reaction was stirred for 1 h. TLC monitors to the end of the reaction, the reaction solution is poured into ice water, a solid precipitates out, filtered and collected the solid to obtain the title compound 2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7 , 9-dihydro-8H-purin-8-one (27f) (white solid, 873 mg, yield 61.61%).

1H NMR (400MHz, DMSO-d6) δ 8.34 (s, 1H), 4.66-4.58 (m, 1H), 4.37 (s, 1H), 3.35 (s, 3H), 2.20 (t, 1H), 2.11 2.00 (m, 1H), 1.72-1.54 (m, 5H), 1.32-1.24 (m, 1H), 1.17 (s, 3H).

LC-MS m/z(ESI)=297.10[M+1].

The seventh step:

9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9- Dihydro-8H-purin-8-one (Compound 27)

9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one

The 2-chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 27f (100mg, 0.34mmol), 6-methyl 2-,3-dihydrobenzofuran-5-amine intermediate 1 (100mg, 0.67mmol), cesium carbonate (220mg, 0.67mmol), Brettphos G3 Pd (30mg, 0.034mmol) into the dry reaction flask, nitrogen After three replacements, dry 1,4-dioxane (1 mL) was added and reacted at 110° C. for 5 h. TLC monitors until the reaction is complete. After cooling to room temperature, it is separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(3-hydroxy-3-methylcyclohexyl) -7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purin-8-one (compound 27) (Light pink solid, 80 mg, yield 58.01%).

1H NMR(400MHz,DMSO-d6)δ8.21(s,1H),7.94(s,1H),7.21(s,1H),6.60(s,1H),4.58-4.50(m,1H), 4.48( t, 2H), 4.27(s, 1H), 3.25(s, 3H), 3.10(t, 2H), 2.26(t, 1H), 2.11(s, 3H), 2.04-1.96(m, 1H), 1.74 –1.60(m,2H), 1.58–1.49(m,3H), 1.19–1.13(m,1H), 1.12(s,3H).

LC-MS m/z(ESI)=410.20[M+1].

Example 28

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxapheno[3.3]heptane-6-yl) -7,9-dihydro-8H-purin-8-one (compound 28)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin -8-one

first step:

4-((2-oxaspiro[3.3]hept-6-6 yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (28a)

ethyl 4-((2-oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylate

2,4-Dichloropyrimidine-5-carboxylic acid ethyl ester 1a (4.43g, 20.05mmol), 2-oxaspiro[3.3]heptan-6-amine hydrochloride (2.0g, 13.37mmol), potassium carbonate (5.54g, 40.10mmol) was dissolved in acetonitrile (60mL), and the reaction solution was reacted at room temperature for 16h. The end of the reaction was monitored by TLC, filtered, and the solid was washed with a small amount of acetonitrile. The filtrate was combined and concentrated. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain the target compound 4-((2-oxaspiro) [3.3]Heptan-6-6yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester (28a) (white solid, 3.0 g, yield 75.38%).

1H NMR(400MHz,DMSO-d6)δ8.60(s,1H), 8.47(d,1H), 4.63(s,2H), 4.49(s,2H), 4.39-4.33(m,1H), 4.30- 4.27 (q, 2H), 2.66-2.61 (m, 2H), 2.31-2.26 (m, 2H), 1.30 (t, 3H).

LC-MS m/z(ESI)=298.10[M+1].

The second step:

4-((2-oxaspiro[3.3]hept-6-6 yl)amino)-2-chloropyrimidine-5-carboxylic acid (28b)

4-((2-oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylic acid

4-((2-oxaspiro[3.3]hept-6-6 yl)amino)-2-chloropyrimidine-5-carboxylic acid ethyl ester 280a (3.0g, 10.08mmol) was dissolved in tetrahydrofuran/water (30mL/ 30 mL), lithium hydroxide (845 mg, 20.15 mmol) was added, and the mixture was stirred at room temperature for 1 h. TLC monitors until the reaction is complete, concentrates to remove tetrahydrofuran, adjusts the pH to 5 with 2N hydrochloric acid, a white solid is precipitated, filtered, the filter cake is washed twice with petroleum ether, and the solid is collected to obtain the title compound 4-((2-oxaspiro[3.3 ]Hept-6-6yl)amino)-2-chloropyrimidine-5-carboxylic acid (28b) (white solid, 1.8g, yield 66.24%).

1H NMR (400MHz, DMSO-d6) δ 13.76 (s, 1H), 8.64 (d, 1H), 8.57 (s, 1H), 4.64 (s, 2H), 4.49 (s, 2H), 4.40-4.30 ( m, 1H), 2.67-2.61 (m, 2H), 2.28-2.23 (m, 2H).

LC-MS m/z(ESI)=270.20[M+1].

third step:

2-Chloro-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one (28c)

2-chloro-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 4-((2-oxaspiro[3.3]hept-6-6 yl)amino)-2-chloropyrimidine-5-carboxylic acid 28b (1.8g, 6.67mmol) in dimethylacetamide (40mL) , Triethylamine (0.92mL, 6.67mmol) and diphenyl azide phosphate (1.4mL, 6.67mmol) were added, and then gradually heated to 90°C and stirred for 2h. TLC monitors to the end of the reaction, the reaction solution is poured into ice water, the solid is collected by filtration, washed 3 times with water, concentrated and dried in vacuo to obtain 2-chloro-9-(2-oxaspiro[3.3]heptan-6-yl)-7, 9-dihydro-8H-purin-8-one (28c) (white solid, 1.3 g, yield 73.03%).

1H NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.11(s,1H),4.71-4.63(m,5H),3.02-2.94(m,2H),2.69-2.66(m,2H) ).

LC-MS m/z(ESI)=267.10[M+1].

the fourth step:

2-Chloro-7-methyl-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one (28d)

2-chloro-7-methyl-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(2-oxaspiro[3.3]hept-6-yl)-7,9-dihydro-8H-purin-8-one 28c (1.3g, 4.87mmol) in dimethyl Formamide (10 mL), dimethyl sulfate (0.46 mL, 4.87 mmol) and cesium carbonate (3.18 g, 9.75 mmol) were added at 0°C, and the reaction was stirred for 1 h. TLC monitors to the end of the reaction, the reaction solution is poured into ice water, a solid precipitates out, filtered and collected the solid to obtain the title compound 2-chloro-7-methyl-9-(2-oxaspiro[3.3]hept-6-yl )-7,9-dihydro-8H-purin-8-one (28d) (white solid, 875mg, yield 63.94%).

1H NMR(400MHz,DMSO-d6)δ8.35(s,1H),4.76-4.69(m,1H),4.66-4.63(m,4H),3.33(s,3H),3.01-2.95(m,2H) ), 2.71-2.66 (m, 2H).

LC-MS m/z(ESI)=281.10[M+1].

the fifth step:

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxapheno[3.3]heptane-6-yl) -7,9-dihydro-8H-purin-8-one (compound 28)

7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin -8-one

The 2-chloro-7-methyl-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one 28d (50mg, 0.18mmol), 6-Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (53mg, 0.36mmol), cesium carbonate (116mg, 0.36mmol), Brettphos G3 Pd (16mg, 0.018mmol) were added to the dry reaction After replacing the flask with nitrogen three times, add dry 1,4-dioxane (1 mL), and react at 110°C for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-methyl-2-((6-methyl- 2,3-Dihydrobenzofuran-5-yl)amino)-9-(2-oxafero[3.3]heptane-6-yl)-7,9-dihydro-8H-purine-8- Ketone (Compound 28) (light pink solid, 37 mg, yield 52.80%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),7.97(s,1H),7.09(s,1H),6.63(s,1H),4.63-4.55(m,3H),4.48( t, 2H), 4.31 (s, 2H), 3.23 (s, 3H), 3.12 (t, 2H), 2.99-2.90 (m, 2H), 2.55-2.48 (m, 2H), 2.08 (s, 3H) .

LC-MS m/z(ESI)=394.20[M+1].

Example 29

9-(6-(Hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl) Amino)-7,9-dihydro-8H-purin-8-one

9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro- 8H-purin-8-one

first step:

(6-Aminospiro[3.3]heptan-2-yl)methanol (29b)

(6-aminospiro[3.3]heptan-2-yl)methanol

Methyl 6-aminospiro[3.3]heptane-2-carboxylate 29a (2.0g, 11.8mmol) was dissolved in tetrahydrofuran (15mL), the mixture was cooled to 0℃, and 1N lithium aluminum tetrahydrogen solution was slowly added dropwise (23.6mL, 1mol/L tetrahydrofuran solution), after the dripping is completed, slowly warm up to room temperature and stir for 2h. The reaction was monitored by TLC until the reaction was over, methanol was added to quench the reaction, concentrated under reduced pressure to evaporate the solvent, acetonitrile (50 mL) was added to make a slurry filter, and the filtrate was spin-dried to obtain the title compound (6-aminospiro[3.3]heptan-2-yl)methanol ( 29b) (white solid, 1.3 g, yield 77.8%).

LC-MS m/z(ESI)=142.10[M+1].

The second step:

Ethyl 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylate (29c)

ethyl 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylate

Ethyl 2,4-dichloropyrimidine-5-carboxylate 1a (2.03g, 9.19mmol), (6-aminospiro[3.3]heptan-2-yl)methanol 29b (1.3g, 9.19mmol), carbonic acid Potassium (1.27g, 9.19mmol) was dissolved in acetonitrile (25mL), and the reaction was stirred at room temperature for 16h. TLC monitoring until the end of the reaction, the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) = 1: 1) to obtain the title compound 2-chloro-4-((6-(hydroxymethyl) Spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid ethyl ester (29c) (yellow solid, 1.56 g, yield 52.2%).

1H NMR(400MHz,Chloroform-d)δ8.65(s,1H),8.54(d,1H),4.59--4.48(m,1H),4.35(q,2H),3.59(dd,2H),2.66-- 2.58 (m, 1H), 2.51-2.38 (m, 2H), 2.27-2.19 (m, 1H), 2.11-2.04 (m, 2H), 2.03-1.91 (m, 2H), 1.91-1.85 (m, 1H) ), 1.84-1.76 (m, 1H), 1.39 (t, 3H).

LCMS m/z(ESI)=326.10[M+1].

third step:

2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid (29d)

2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid

Dissolve 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid ethyl ester 29c (1.07g, 3.28 mmol) in tetrahydrofuran/water (10mL/10mL), lithium hydroxide monohydrate (0.41g, 9.85mmol) was added, and the reaction was stirred at room temperature for 1h. TLC monitors to the end of the reaction, concentrates and evaporates the tetrahydrofuran, adds 2N HCl to adjust the pH to about 3-4, a white solid is precipitated, filtered, and the filter cake is washed twice with water and petroleum ether/ethyl acetate (v/v=10/1) The title compound 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid (29d) (white solid, 918mg, yield 93.88) %).

1H NMR(400MHz,DMSO-d6)δ12.69(s,1H),9.67(d,1H),8.35(s,1H),4.42-4.31(m,1H),3.30(s,1H),2.55-- 2.46 (m, 3H), 2.40-2.31 (m, 1H), 2.28-2.19 (m, 1H), 2.10-2.00 (m, 2H), 2.00-1.91 (m, 2H), 1.83-1.69 (m, 2H) ).

LC-MS m/z(ESI)=298.10[M+1].

the fourth step:

2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one (29e)

2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one

Use 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid 29d (0.91g, 3.06mmol) with N,N-dimethyl Acetamide (12 mL) was dissolved, triethylamine (0.42 mL, 3.06 mmol) and diphenyl azide phosphate (0.66 mL, 10.01 mmol) were added under stirring at room temperature for 2 hours, and the temperature was raised to 110° C. and refluxed for 2.5 hours. TLC monitors to the end of the reaction, the reaction solution is added with water and ethyl acetate for extraction, the organic layer is concentrated and purified by silica gel column chromatography (dichloromethane/methanol (v/v)=20:1) to obtain the title compound 2-chloro- 9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one (29e) (white solid, 516mg, yield 57.28%) .

1H NMR(400MHz,DMSO-d6)δ11.60(s,1H),8.11(s,1H), 4.68-4.57(m,1H), 4.46(t,1H), 3.37-3.32(m,2H), 2.95–2.81(m,2H), 2.42–2.35(m,1H), 2.32–2.19(m,2H), 2.19–2.12(m,1H), 2.06–1.98(m,1H), 1.89–1.77(m ,2H).

LC-MS m/z(ESI)=295.10[M+1].

the fifth step:

2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (29f)

2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one

Use 2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one 29e (516mg, 1.75mmol) with N , N-dimethylformamide (6mL) was dissolved, and dimethyl sulfate (0.17mL, 1.75mmol) and cesium carbonate (1.14g, 3.50mmol) were added under stirring at 0°C and reacted for 1h. TLC monitors until the reaction is over, the reaction solution is slowly added dropwise to ice water and stirred, ethyl acetate is added for extraction, and the organic layer is concentrated to obtain the title compound 2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptane -2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (29f) (white solid, 470 mg, yield 86.95%).

1H NMR(400MHz,DMSO-d6)δ8.33(s,1H), 4.71-4.60(m,1H), 4.10-3.96(m,2H), 3.35(s,1H), 3.33(s,3H), 2.93–2.81(m, 2H), 2.44–2.36(m, 1H), 2.32–2.21(m, 2H), 2.20–2.13(m, 1H), 2.07–1.99(m, 1H), 1.89–1.78(m ,2H).

LC-MS m/z(ESI)=309.10[M+1].

The sixth step:

9-(6-(Hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl) Amino)-7,9-dihydro-8H-purin-8-one (Compound 29)

9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro- 8H-purin-8-one

The 2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 29f (100mg, 0.32mmol), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (97mg, 0.65mmol), cesium carbonate (211mg, 0.65mmol), Brettphos G3 Pd (29mg, 0.032mmol) Add a dry reaction flask, replace with nitrogen three times, add dry 1,4-dioxane (1 mL), and react at 110° C. for 5 hours. The reaction was monitored by TLC until the end of the reaction. After cooling to room temperature, it was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 9-(6-(hydroxymethyl)spiro[3.3] Heptan-2-yl)-7-methyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-7,9-dihydro-8H-purine- 8-ketone (compound 29) (light pink solid, 70 mg, yield 51.28%).

1H NMR(400MHz,DMSO-d6)δ8.22(s,1H),7.96(s,1H),7.14(s,1H),6.61(s,1H),4.61-4.51(m,1H), 4.48( t, 2H), 4.43 (t, 1H), 3.33--3.29 (m, 2H), 3.23 (s, 3H), 3.11 (t, 2H), 2.87 (t, 1H), 2.80 (t, 1H), 2.29 –2.17(m,2H), 2.14–2.06(m,5H), 1.85–1.76(m,2H), 1.67–1.59(m,1H).

LC-MS m/z(ESI)=422.20[M+1].

Example 30

7-Ethyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7, 9-dihydro-8H-purin-8-one (Compound 30)

7-ethyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

first step:

2-chloro-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (30a)

2-chloro-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1d (400mg, 1.57mmol) in N,N-di Methylformamide (8mL), add cesium carbonate (511mg, 1.57mmol) and iodoethane (293mg, 1.88mmol) at 0℃, stir for 1h at 0℃, then add 10mL of water and extract 3 times with ethyl acetate , The organic phase was dried with anhydrous sodium sulfate, concentrated, and the organic solvent was removed by rotary evaporation to obtain the target product 2-chloro-7-ethyl-9-(tetrahydro-2H-pyran-4-yl(-7,9- Dihydro-8H-purin-8-one (30a) (white solid, 290 mg, 65.32%).

1H NMR(400MHz,DMSO-d6)δ8.44(s,1H), 4.50-4.41(m,1H), 3.99-3.95(m,2H), 3.89(q,2H), 3.45(t,2H), 2.46–2.41(m,2H), 1.67–1.71(m,2H), 1.25(t,3H).

LC-MS m/z(ESI)=283.00[M+1].

The second step:

7-Ethyl-2-(((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7, 9-dihydro-8H-purin-8-one (Compound 30)

7-ethyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 30a (100mg, 0.35mmol), 6- Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (106mg, 0.70mmol), cesium carbonate (230mg, 0.70mmol) and Brettphos G3 Pd (32mg, 0.035mmol) were added to the dry reaction tube, Replace with N ₂ three times, then add dry 1,4-dioxane (1 mL), and react at 110° C. for 5 h. Cooled to room temperature, and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-ethyl-2-(((6-methyl-2,3- Dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 30) (light pink Solid, 44 mg, yield 31.46%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H), 8.02(s,1H), 7.24(s,1H), 6.60(s,1H), 4.49(t,2H), 4.41--4.31( m, 1H), 3.95 (dd, 2H), 3.79 (q, 2H), 3.40 (t, 2H), 3.11 (t, 2H), 2.58-2.45 (m, 2H), 2.12 (s, 3H), 1.68 -1.60(m,2H),1.21(t,3H).

LC-MS m/z(ESI)=396.20[M+1].

Example 31

7-isopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7, 9-dihydro-8H-purin-8--one

7-isopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

first step:

2-Chloro-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (31a)

2-chloro-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1d (520mg, 2.04mmol) in N,N-di Methylformamide (10mL), add cesium carbonate (665mg, 2.04mmol) and 2-iodopropane (416mg, 2.45mmol) at 0℃, stir for 2h at 0℃, then add 20mL of water, and extract 3 with ethyl acetate. Next, the organic phase was dried with anhydrous sodium sulfate, concentrated, and the organic solvent was removed by rotary evaporation to obtain the target product 2-chloro-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7, 9-Dihydro-8H-purin-8-one (31a) (white solid, 465 mg, 76.74%).

1H NMR(400MHz,DMSO-d6)δ8.52(s,1H), 4.65-4.56(m,1H), 4.49-4.40(m,1H), 3.99-3.95(m,2H), 3.44(t,2H) ), 2.46–2.42(m,2H), 1.70–1.67(m,2H), 1.43(d,6H).

LC-MS m/z(ESI)=297.10[M+1].

The second step:

7-isopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7, 9-dihydro-8H-purin-8-one (compound 31)

7-isopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 31a (100mg, 0.37mmol), 6 -Methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (101mg, 0.74mmol), cesium carbonate (220mg, 0.74mmol) and Brettphos G3 Pd (30mg, 0.037mmol) were added to the dry reaction tube , N2 was replaced three times, and then dry 1,4-dioxane (1ml) was added and reacted at 110°C for 5h. Cool to room temperature, use silica gel column chromatography to separate and purify (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-isopropyl-2-(((6-methyl-2,3 -Dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 31) (shallow Yellow solid, 41 mg, yield 29.71%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),8.13(s,1H),7.24(s,1H),6.60(s,1H),4.57-4.51(m,1H), 4.49( t, 2H), 4.41--4.32 (m, 1H), 3.95 (dd, 2H), 3.40 (t, 2H), 3.11 (t, 2H), 2.58 - 2.45 (m, 2H), 2.13 (s, 3H) ,1.66-1.60(m,2H),1.38(d,6H).

LC-MS m/z(ESI)=410.20[M+1].

Example 32

7-Cyclopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7, 9-dihydro-8H-purin-8-one (Compound 32)

7-cyclopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

first step:

2-Chloro-7-cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (32a)

2-chloro-7-cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one

Combine 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1d (1g, 3.9mmol), cyclopropylboronic acid (676mg, 7.8mmol), anhydrous copper acetate (715mg, 3.9mmol), potassium carbonate (1.058g, 7.8mmol), 1,10-phenanthroline) 710mg, 3.9mmol) dissolved in 1,2-dichloroethane 12mL , Open reaction, heat to 70 ℃ and stir for 6h. TLC monitors until the reaction is complete, the reaction solution is concentrated, and the residue is separated and purified by silica gel column chromatography) petroleum ether/ethyl acetate (v/v) = 3/1) to obtain the title compound 2-chloro-7-cyclopropyl-9 -(Tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (34a) (white solid, 690 mg, yield 56%).

1H NMR (400MHz CDCI3) δ8.15 (s, 1H), 4.51-4.59 (m, 1H), 4.12 (dd, 2H), 3.49-3.55 (m, 2H), 2.91-2.96 (m, 1H), 2.67 -2.77 (m, 2H), 1.68-1.77 (m, 2H), 1.13-1.26 (m, 2H), 1.00-1.04 (m, 2H).

LC-MS m/z(ESI)=295.2[M+1].

The second step:

7-Cyclopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7, 9-dihydro-8H-purin-8-one (Compound 32)

7-cyclopropyl-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-7-cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 32a (100mg, 0.34mmol), 6 -Methyl-2,3-dihydrobenzofuran-5-amine (102mg, 0.68mmol), cesium carbonate (222mg, 0.68mmol) and Brettphos G3 Pd (32mg, 0.034mmol) were added to the dry reaction tube, N ₂ Replace three times, then add dry 1,4-dioxane (1 mL), and react at 110° C. for 5 h. Cooled to room temperature, and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-cyclopropyl-2-((6-methyl-2,3- Dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 32) (light yellow Solid, 28 mg, yield 20.25%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),7.94(s,1H),7.23(s,1H),6.60(s,1H), 4.48(t,2H), 4.38-4.28( m, 1H), 3.94 (dd, 2H), 3.39 (t, 2H), 3.11 (t, 2H), 2.90-2.84 (m, 1H), 2.56-2.43 (m, 2H), 2.12 (s, 3H) ,1.65-1.57(m,2H),0.99–0.92(m,2H),0.90–0.84(m,2H).

LC-MS m/z(ESI)=408.20[M+1].

Example 33

7-(methyl-d3)-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl) -7,9-dihydro-8H-purin-8-one (Compound 33)

7-(methyl-d3)-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H -purin-8-one

first step:

2-Chloro-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (33a)

2-chloro-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one

Dissolve 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1d (1.0g, 3.93mmol) in dimethyl sulfoxide (20mL), add cesium carbonate (2.03g, 7.86mmol) at room temperature, then add deuterated methyl iodide (0.45g, 3.93mmol) at 0°C, and react at room temperature for 2h. After the reaction, 5 mL of water was added and extracted with ethyl acetate for 3 times. The organic phase was dried with anhydrous sodium sulfate and concentrated. A solid precipitated out. It was separated and purified by silica gel column chromatography (dichloromethane/methanol (v/v=20:1) )) Purification to obtain the compound 2-chloro-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one ( 33a) (white solid, 0.36 g, yield 34.66%).

1HNMR(400MHz DMSO)δ8.36(s,1H), 4.50-4.41(m,1H), 3.99-3.95(m,2H), 3.48-3.42(m,2H), 2.47-2.38(m,2H), 1.70-1.66 (m, 2H).

LC-MS m/z(ESI)=272.10[M+1].

The second step:

7-(methyl-d3)-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl) -7,9-dihydro-8H-purin-8-one (Compound 33)

7-(methyl-d3)-2-((6-methyl-2,3-dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H -purin-8-one

The 2-chloro-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 33a (100mg, 0.37mmol ), 6-methyl-2,3-dihydrobenzofuran-5-amine intermediate 1 (110mg, 0.74mmol), cesium carbonate (240mg, 0.74mmol) and Brettphos G3 Pd (34mg, 0.037mmol) were added to the dry Replace the reaction tube with N2 three times, then add dry 1,4-dioxane (1ml), and react at 110°C for 5h. Cooled to room temperature, and purified by silica gel column chromatography (dichloromethane/methanol (v/v=40:1)) to obtain the title compound 7-(methyl-d3)-2-((6-methyl-2 ,3-Dihydrobenzofuran-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (Compound 33) (Off-white solid, 50 mg, yield 35.34%).

1H NMR (400MHz, DMSO-d6) δ 8.23 (s, 1H), 7.95 (s, 1H), 7.24 (s, 1H), 6.60 (s, 1H), 4.49 (t, 2H), 4.41-4.32 ( m, 1H), 3.95 (dd, 2H), 3.40 (t, 2H), 3.11 (t, 2H), 2.57-2.44 (m, 2H), 2.12 (s, 3H), 1.68-1.59 (m, 2H) .

LC-MS m/z(ESI)=385.20[M+1].

Example 34

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9- Dihydro-8H-purin-8-one (Compound 34)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The synthesis method of compound 34 is the same as that of compound 1, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H- Pyran-4-yl)-7,9-dihydro-8H-purin-8-one (Compound 34) (white solid, 15 mg, yield 10.55%).

1H NMR (400MHz MeOD) δ 7.86 (s, 1H), 7.41 (s, 1H), 6.99 (s, 1H), 4.67 (t, 2H), 4.51-4.59 (m, 1H), 4.03-4.07 (m ,2H),3.52(m,2H),3.38(s,3H),3.25-3.30(m,2H),2.55-2.66(m,2H),1.75-1.78(m,2H)

LC-MS m/z(ESI)=402.20[M+1].

Example 35

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-((trans-4-methoxycyclohexyl)-7-methyl-7,9-di Hydrogen-8H-purin-8-one (Compound 35)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-((trans-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

Refer to the preparation method of compound 2, and prepare the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-((trans-4-methoxycyclohexyl) -7-Methyl-7,9-dihydro-8H-purin-8-one (Compound 35) (off-white solid, 65 mg, yield 44.88%).

1H NMR(400MHz,DMSO-d6)δ8.28(s,1H),8.00(s,1H),7.53(s,1H),6.91(s,1H),4.57(t,2H),4.16-4.08( m, 1H), 3.27 (s, 3H), 3.25 (s, 3H), 3.18 (t, 2H), 3.09-3.02 (m, 1H), 2.34-2.22 (m, 2H), 2.08 (d, 2H) , 1.71 (d, 2H), 1.26-1.20 (m, 2H).

LC-MS m/z(ESI)=430.10[M+1].

Example 36

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9 -Dihydro-8H-purin-8-one (Compound 36)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8 -one

Using the preparation method of compound 3, the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-((trans-4-hydroxy-4-methyl) was prepared. Cyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (Compound 36) (light yellow solid, 58 mg, yield 40.04%).

1H NMR (400MHz, DMSO-d6) δ 8.38 (s, 1H), 8.01 (s, 1H), 7.34 (s, 1H), 6.88 (s, 1H), 4.56 (t, 2H), 4.38 (s, 1H), 4.08-4.01 (m, 1H), 3.27 (s, 3H), 3.15 (t, 2H), 2.32-2.19 (m, 2H), 1.59-1.39 (m, 6H), 0.98 (s, 3H) .

LC-MS m/z(ESI)=430.10[M+1].

Example 37

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-bis Hydrogen-8H-purin-8-one (Compound 37)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 37 is the same as that of compound 4 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(1-methylpiper (Pyridin-4-yl)-7,9-dihydro-8H-purin-8-one (Compound 37) (pale yellow solid, 36 mg, yield 44.45%).

1H NMR(400MHz,DMSO-d6)δ8.25(s,1H),7.99(s,1H),7.59(s,1H),6.89(s,1H),4.57(t,2H),4.13-4.05( m, 1H), 3.27(s, 3H), 3.18(t, 2H), 2.86(d, 2H), 2.58-2.47(m, 2H), 2.19(s, 3H), 1.95(t, 2H), 1.62 (d, 2H).

LC-MS m/z(ESI)=415.10[M+1].

Example 38

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one( Compound 38)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 38 is the same as that of compound 5 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-cyclohexyl-7-methyl-7,9 -Dihydro-8H-purin-8-one (Compound 38) (light yellow solid, 40 mg, yield 26.68%).

1H NMR(400MHz,DMSO-d6)δ8.24(s,1H),8.00(s,1H),7.57(s,1H),6.90(s,1H),4.57(t,J=8.6Hz,2H) ,4.15-4.07(m,1H),3.27(s,3H),3.17(t,2H),2.26-2.16(m,2H),1.80(d,2H),1.68(d,3H),1.36-1.27 (m, 2H), 1.15-1.05 (m, 1H).

LC-MS m/z(ESI)=400.10[M+1].

Example 39

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-3-yl)-7,9- Dihydro-8H-purin-8-one (Compound 39)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-3- yl)-7,9-dihydro-8H-purin-8 -one

The synthesis method of compound 39 is the same as that of compound 6, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H- Pyran-3-yl)-7,9-dihydro-8H-purin-8-one (Compound 39) (pale yellow solid, 49 mg, yield 32.76%).

1H NMR(400MHz,DMSO-d6)δ8.35(s,1H),8.01(s,1H),7.51(s,1H),6.91(s,1H),4.57(t,2H),4.26-4.18( m, 1H), 3.95-3.82 (m, 2H), 3.77-3.73 (m, 1H), 3.27 (s, 3H), 3.17 (t, 3H), 2.48-2.40 (m, 1H), 1.85 (d, 1H), 1.77-1.61 (m, 2H).

LC-MS m/z(ESI)=402.10[M+1].

Example 40

(R)-2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro -8H-purin-8-one (Compound 40)

(R)-2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin- 8-one

The synthesis method of compound 40 is the same as that of compound 7, to obtain the title compound (R)-2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydrofuran) -3-yl)-7,9-dihydro-8H-purin-8-one (Compound 40) (pale yellow solid, 65 mg, yield 42.68%).

1H NMR(400MHz,DMSO-d6)δ8.32(s,1H),8.03(s,1H),7.51(s,1H),6.90(s,1H),4.94-4.85(m,1H),4.57( t,2H),3.95-3.90(m,2H),3.83-3.76(m,2H), 3.28(s,3H), 3.20-3.14(m,2H), 2.40-2.33(m,1H),2.22- 2.12 (m, 1H).

LC-MS m/z(ESI)=388.10[M+1].

Example 41

(S)-2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro -8H-purin-8-one (Compound 41)

(S)-2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8 -one

The synthesis method of compound 41 is the same as that of compound 8, to obtain the title compound (S)-2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydrofuran) -3-yl)-7,9-dihydro-8H-purin-8-one (Compound 41) (light yellow solid, 67 mg, yield 44.00%).

1H NMR(400MHz,DMSO-d6)δ8.32(s,1H),8.03(s,1H),7.52(s,1H),6.90(s,1H),4.93-4.86(m,1H),4.57( t,2H),3.95-3.88(m,2H),3.83-3.76(m,2H), 3.28(s,3H), 3.17(t,2H), 2.40-2.33(m,1H),2.22-2.12( m,1H).

LC-MS m/z(ESI)=388.10[M+1].

Example 42

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(((tetrahydrofuran-3-yl)methyl)-7,9-bis Hydrogen-8H-purin-8-one (Compound 42)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8- one

The synthesis method of compound 42 is the same as that of compound 9, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(((tetrahydrofuran-3 -Yl)methyl)-7,9-dihydro-8H-purin-8-one (Compound 42) (off-white solid, 48 mg, yield 32.10%).

1H NMR(400MHz,DMSO-d6)δ8.34(s,1H),8.01(s,1H),7.50(s,1H),6.90(s,1H),4.57(t,2H),3.78-3.73( m, 1H), 3.71 (d, 2H), 3.65-3.60 (m, 2H), 3.52 (dd, 1H), 3.29 (s, 3H), 3.17 (t, 2H), 2.78-2.68 (m, 1H) ,1.94-1.86(m,1H),1.68-1.60(m,1H).

LC-MS m/z(ESI)=402.10[M+1].

Example 43

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H- Purine-8-one (Compound 43)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 43 is the same as that of compound 10 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(piperidine-4- Yl)-7,9-dihydro-8H-purin-8-one (Compound 43) (white solid, 30 mg, yield 13.76%).

1H NMR(400MHz,DMSO-d6)δ8.24(s,1H),7.99(s,1H),7.59(s,1H),6.90(s,1H),4.57(t,2H),4.18(tt, 1H), 3.27 (s, 3H), 3.19 (t, 2H), 3.04 (d, 2H), 2.55-2.48 (m, 2H), 2.38-2.28 (m, 2H), 1.61 (d, 2H).

LC-MS m/z(ESI)=401.10[M+1].

Example 44

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-((cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7, 9-dihydro-8H-purin-8-one (Compound 44)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8 -one

The synthetic method of compound 44 is the same as that of compound 11 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-((cis-4-hydroxy-4-methyl) Cyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (Compound 44) (off-white solid, 23 mg, yield 19.84%).

1H NMR(400MHz,DMSO-d6)δ8.09(s,1H),8.00(s,1H),7.68(s,1H),6.89(s,1H),4.56(t,2H),4.14-4.07( m, 1H), 4.05 (s, 1H), 3.28 (s, 3H), 3.20 (t, 2H), 2.70-2.59 (m, 2H), 1.66 (d, 2H), 1.46-1.37 (m, 4H) , 1.15 (s, 3H).

LC-MS m/z(ESI)=430.10[M+1].

Example 45

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro -8H-purin-8-one (Compound 45)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 45 is the same as that of compound 12 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(cis-4-methoxycyclohexyl) -7-Methyl-7,9-dihydro-8H-purin-8-one (Compound 45) (pale yellow solid, 33 mg, yield 22.78%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),7.99(s,1H),7.51(s,1H),6.89(s,1H),4.57(t,2H),4.19-4.10( m,1H), 3.40 (s, 1H), 3.27 (s, 3H), 3.25-3.15 (m, 5H), 2.49-2.40 (m, 2H), 1.98 (d, 2H), 1.50-1.40 (m, 4H).

LC-MS m/z(ESI)=430.20[M+1].

Example 46

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(((tetrahydrofuran-2-yl)methyl)-7,9-bis Hydrogen-8H-purin-8-one (Compound 46)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-((tetrahydrofuran-2-yl)methyl)-7,9-dihydro-8H-purin-8- one

The synthesis method of compound 46 is the same as that of compound 14, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(((tetrahydrofuran-2 -Yl)methyl)-7,9-dihydro-8H-purin-8-one (Compound 46) (brown solid, 29 mg, yield 38.78%).

1H NMR(400MHz,DMSO-d6)δ8.32(s,1H),8.00(s,1H),7.51(s,1H),6.90(s,1H),4.57(t,2H),4.28-4.22( m,1H),3.81-3.73(m,2H),3.68-3.58(m,2H), 3.29(s,3H), 3.17(t,2H),1.95-1.75(m,3H),1.71-1.61( m,1H).

LC-MS m/z(ESI)=402.10[M+1].

Example 47

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-((tetrahydro-2H-pyran-4-yl)methyl)- 7,9-dihydro-8H-purin-8-one (Compound 47)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-((tetrahydro-2H-pyran-4-yl)methyl)-7,9-dihydro-8H- purin-8-one

The synthesis method of compound 47 is the same as that of compound 15, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-((tetrahydro-2H -Pyran-4-yl)methyl)-7,9-dihydro-8H-purin-8-one (compound 47) (white solid, 48 mg, yield 32.63%).

1H NMR(400MHz,DMSO-d6)δ8.31(s,1H),8.00(s,1H),7.52(s,1H),6.90(s,1H),4.57(t,2H),3.85-3.80( m, 2H), 3.61 (d, 2H), 3.29 (s, 3H), 3.27-3.14 (m, 4H), 2.12-2.03 (m, 1H), 1.48 (d, 2H), 1.28-1.18 (m, 2H).

LC-MS m/z(ESI)=416.10[M+1].

Example 48

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(trans-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro- 8H-purin-8-one (Compound 48)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(trans-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 48 is the same as that of compound 16, to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(trans-3-hydroxycyclobutyl)- 7-Methyl-7,9-dihydro-8H-purin-8-one (Compound 48) (white solid, 40 mg, yield 25.43%).

1H NMR (400MHz, DMSO-d6) δ 8.34 (s, 1H), 8.02 (s, 1H), 7.50 (s, 1H), 6.91 (s, 1H), 5.09 (d, 1H), 5.03-4.95 ( m, 1H), 4.57 (t, 2H), 4.39-4.33 (m, 1H), 3.26 (s, 3H), 3.17 (t, 2H), 3.03-2.94 (m, 2H), 2.20-2.14 (m, 2H).

LC-MS m/z(ESI)=388.10[M+1].

Example 49

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7- Methyl-7,9-dihydro-8H-purin-8-one (Compound 49)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-7,9-dihydro- 8H-purin-8-one

The synthesis method of compound 49 is the same as that of compound 17, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(6-(hydroxymethyl)tetrahydro- 2H-pyran-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (compound 49) (white solid, 25 mg, yield 20.73%).

1H NMR (400MHz, DMSO-d6) δ 8.38 (s, 1H), 8.02 (s, 1H), 7.39 (s, 1H), 6.91 (s, 1H), 4.58 (t, 2H), 4.53 (t, 1H), 4.24-4.16 (m, 1H), 4.08 (t, 1H), 3.69-3.61 (m, 1H), 3.58-3.52 (m, 1H), 3.44 (dd, 1H), 3.38-3.31 (m, 1H), 3.27 (s, 3H), 3.19 (t, 2H), 2.56-2.44 (m, 1H), 1.84-1.78 (m, 1H), 1.69-1.60 (m, 2H).

LC-MS m/z(ESI)=432.20[M+1].

Example 50

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxycyclohexyl)-7-methyl-7,9-dihydro-8H-purine -8-one (Compound 50)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 50 is the same as that of compound 18 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxycyclohexyl)-7-methyl Cyclo-7,9-dihydro-8H-purin-8-one (Compound 50) (white solid, 30 mg, yield 18.80%).

1H NMR (400MHz, DMSO-d6) δ 8.24 (s, 1H), 8.01 (s, 1H), 7.58 (s, 1H), 6.90 (s, 1H), 4.79 (d, 1H), 4.57 (t, 2H), 4.18-4.10 (m, 1H), 3.55-3.43 (m, 1H), 3.27 (s, 3H), 3.25-3.14 (m, 2H), 2.21-2.02 (m, 2H), 1.93-1.79 ( m, 2H), 1.79-1.71 (m, 1H), 1.69-1.57 (m, 1H), 1.35-1.26 (m, 1H), 1.12-1.00 (m, 1H).

LC-MS m/z(ESI)=416.10[M+1].

Example 51

Trans-4-(2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-8-oxo-7,8-dihydro-9H-purine -9-yl)cyclohexane-1-nitrile (Compound 51)

trans-4-(2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane- 1-carbonitrile

The synthesis method of compound 51 is the same as that of compound 19 to obtain the title compound trans-4-(2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-8-oxy (Dihydro-7,8-dihydro-9H-purine-9-yl)cyclohexane-1-carbonitrile compound 51 (white solid, 60 mg, yield 41.20%).

1H NMR(400MHz,DMSO-d6)δ8.25(s,1H),8.00(s,1H),7.54(s,1H),6.92(s,1H),4.58(t,2H),4.22-4.14( m, 1H), 3.27 (s, 3H), 3.18 (t, 2H), 2.60-2.52 (m, 1H), 2.31-2.20 (m, 2H), 2.15 (d, 2H), 1.77 (d, 2H) ,1.73-1.61(m,2H).

LC-MS m/z(ESI)=425.20[M+1].

Example 52

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl- 7,9-dihydro-8H-purin-8-one (Compound 52)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-isopropyl-4-methylpyridin-3-yl)- 7-methyl-7,9-dihydro-8H- purin-8-one

The synthesis method of compound 52 is the same as that of compound 20 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(2-isopropyl-4-methyl Pyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (Compound 52) (white solid, 20 mg, yield 17.62%).

1H NMR(400MHz,DMSO-d6)δ8.54(d,1H),8.50(s,1H),8.13(s,1H),7.32(d,2H),6.85(s,1H),4.55(t, 2H), 3.40 (s, 3H), 3.11 (t, 2H), 2.83-2.75 (m, 1H), 2.05 (s, 3H), 1.15-1.08 (m, 6H).

LC-MS m/z(ESI)=451.10[M+1].

Example 53

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(4,4-difluorocyclohexyl)-7-methyl-7,9-dihydro- 8H-purine-8-one (Compound 53)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(4,4-difluorocyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one

The synthesis method of compound 53 is the same as that of compound 22 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(4,4-difluorocyclohexyl)- 7-Methyl-7,9-dihydro-8H-purin-8-one (Compound 53) (light yellow solid, 25 mg, yield 17.36%).

1H NMR(400MHz,DMSO-d6)δ8.22(s,1H), 8.04(s,1H), 7.56(s,1H), 6.89(s,1H), 4.56(t,2H), 4.37-4.29( m, 1H), 3.29 (s, 3H), 3.17 (t, 2H), 2.56-2.43 (m, 2H), 2.10-1.95 (m, 4H), 1.77 (d, 2H).

LC-MS m/z(ESI)=436.10[M+1].

Example 54

4-(2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-8-oxo-7,8-dihydro-9H-purine-9 -Yl)bicyclo[2.2.2]octane-1-carbonitrile (compound 54)

4-(2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2. 2]octane-1-carbonitrile

The synthetic method of compound 54 is the same as that of compound 23 to obtain the title compound 4-(2-(((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-8-oxo -7,8-Dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile (Compound 54) (white solid, 60 mg, yield 42.28%).

1H NMR(400MHz,DMSO-d6)δ8.23(s,1H),8.00(s,1H),7.51(s,1H),6.91(s,1H), 4.58(t,2H),3.22(s, 3H), 3.18 (t, 2H), 2.44-2.39 (m, 6H), 2.013-1.98 (m, 6H).

LC-MS m/z(ESI)=451.10[M+1].

Example 55

9-(8-oxabicyclo[3.2.1]octyl-3-yl)-2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl -7,9-dihydro-8H-purin-8-one (Compound 55)

9-(8-oxabicyclo[3.2.1]octan-3-yl)-2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-7,9-dihydro-8H -purin-8-one

The synthesis method of compound 55 is the same as that of compound 24 to obtain the title compound 9-(8-oxabicyclo[3.2.1]octyl-3-yl)-2-((6-chloro-2,3-dihydrobenzofuran) -5-yl)amino)-7-methyl-7,9-dihydro-8H-purin-8-one (compound 55) (pale yellow solid, 28 mg, yield 19.29%).

1H NMR(400MHz,DMSO-d6)δ8.43(s,1H), 8.02(s,1H), 7.34(s,1H), 6.93(s,1H), 4.57(t,2H), 4.39-4.27( m, 3H), 3.26 (s, 3H), 3.17 (t, 2H), 2.15-2.05 (m, 2H), 1.97 (t, 2H), 1.82-1.76 (m, 2H), 1.48-1.43 (m, 2H).

LC-MS m/z(ESI)=428.10[M+1].

Example 56

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(2-oxaspiro[3.5]nonyl-7-yl)-7 ,9-Dihydro-8H-purin-8-one (Compound 56)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(2-oxaspiro[3.5].nonan-7-yl)-7,9-dihydro-8H- purin-8-one

The synthesis method of compound 56 is the same as that of compound 25 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(2-oxaspiro [3.5] Nonyl-7-yl)-7,9-dihydro-8H-purin-8-one (Compound 56) (white solid, 38 mg, yield 26.55%).

1H NMR(400MHz,DMSO-d6)δ8.38(s,1H),8.00(s,1H),7.33(s,1H),6.92(s,1H),4.56(t,2H),4.20(s, 2H), 4.12 (s, 2H), 4.08-3.98 (m, 1H), 3.26 (s, 3H), 3.18 (t, 2H), 2.13-2.04 (m, 4H), 1.60-1.43 (m, 4H) .

LC-MS m/z(ESI)=442.20[M+1].

Example 57

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxybicyclo[3.2.1]octyl-8-yl)-7-methyl- 7,9-dihydro-8H-purin-8-one (Compound 57)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-7,9-dihydro-8H -purin-8-one

The synthesis method of compound 57 is the same as that of compound 26 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxybicyclo[3.2.1]octane (Yl-8-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (Compound 57) (white solid, 22 mg, yield 30.74%).

1H NMR (400MHz, DMSO-d6) δ 8.23 (s, 1H), 8.04 (s, 1H), 7.46 (s, 1H), 6.89 (s, 1H), 4.57 (t, 2H), 4.24 (d, 1H), 3.73-3.63(m, 1H), 3.54-3.52(m, 1H), 3.42(s, 2H), 3.29(s, 3H), 3.15(t, 2H), 1.65-1.54(m, 4H) , 1.52-1.48 (m, 2H), 1.32-1.26 (m, 2H).

LC-MS m/z(ESI)=442.10[M+1].

Example 58

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-di Hydrogen-8H-purin-8-one (Compound 58)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8- one

The synthesis method of compound 58 is the same as that of compound 27 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(3-hydroxy-3-methylcyclohexyl) )-7-methyl-7,9-dihydro-8H-purin-8-one (Compound 58) (gray solid, 20 mg, yield 13.81%).

1H NMR (400MHz, DMSO-d6) δ 8.28 (s, 1H), 7.99 (s, 1H), 7.51 (s, 1H), 6.90 (s, 1H), 4.60-4.49 (m, 3H), 4.28 ( s, 1H), 3.26 (s, 3H), 3.16 (t, 2H), 2.26 (t, 1H), 2.09-1.96 (m, 1H), 1.72-1.59 (m, 2H), 1.54 (d, 3H) , 1.19-1.13 (m, 1H), 1.12 (s, 3H).

LC-MS m/z(ESI)=430.20[M+1].

Example 59

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl Gyl-7,9-dihydro-8H-purin-8-one (Compound 59)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro- 8H-purin-8-one

The synthesis method of compound 59 is the same as that of compound 29 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-9-(6-(hydroxymethyl)spiro[3.3 ]Heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (Compound 59) (gray solid, 40 mg, yield 27.93%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.34 (s, 1H), 8.00 (s, 1H), 7.42 (s, 1H), 6.89 (s, 1H), 4.56 (t, 3H), 4.45 ( t,1H),3.33–3.29(m,2H), 3.24(s,3H), 3.17(t,2H), 2.89-2.79(m,2H), 2.29–2.17(m,2H),2.13-2.07( m, 2H), 1.82-1.77 (m, 2H), 1.65-1.59 (m, 1H).

LC-MS m/z(ESI)=416.10[M+1].

Example 60

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9- Dihydro-8H-purin-8-one (Compound 60)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The synthesis method of compound 60 is the same as that of compound 30, and the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-ethyl-9-(tetrahydro-2H- Pyran-4-yl)-7,9-dihydro-8H-purin-8-one (Compound 60) (pale yellow solid, 31 mg, yield 21.08%).

1H NMR (400MHz, DMSO-d6) δ 8.30 (s, 1H), 8.07 (s, 1H), 7.55 (s, 1H), 6.89 (s, 1H), 4.57 (t, 2H), 4.38 (tt, 1H), 3.95 (dd, 2H), 3.80 (q, 2H), 3.41 (t, 2H), 3.17 (t, 2H), 2.57-2.44 (m, 2H), 1.69-1.60 (m, 2H), 1.21 (t, 3H).

LC-MS m/z(ESI)=416.10[M+1].

Example 61

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9 -Dihydro-8H-purin-8-one (Compound 61)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The synthesis method of compound 61 is the same as that of compound 31 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-isopropyl-9-(tetrahydro-2H -Pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 61) (white solid, 15 mg, yield 10.35%).

1H NMR(400MHz,DMSO-d6)δ8.31(s,1H), 8.19(s,1H), 7.55(s,1H), 6.90(s,1H), 4.61-4.50(m,3H), 4.38( tt,1H),3.96(dd,2H),3.41(t,2H),3.17(t,2H),2.51(d,6H),1.68-1.60(m,2H),1.45-1.36(m,6H) .

LC-MS m/z(ESI)=430.20[M+1].

Example 62

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9 -Dihydro-8H-purin-8-one (Compound 62)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The synthesis method of compound 62 is the same as that of compound 32 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-cyclopropyl-9-(tetrahydro-2H -Pyran-4-yl)-7,9-dihydro-8H-purin-8-one (Compound 62) (off-white solid, 14 mg, yield 9.64%).

1H NMR(400MHz,DMSO-d6)δ8.31(s,1H),7.99(s,1H),7.54(s,1H),6.90(s,1H),4.57(t,2H), 4.34(tt, 1H), 3.95(dd, 2H), 3.40(t, 2H), 3.17(t, 2H), 2.89(tt, 1H), 2.55-2.46(s, 2H), 1.66-1.58(m, 2H), 1.00 -0.93 (m, 2H), 0.91-0.85 (m, 2H).

LC-MS m/z(ESI)=428.20[M+1].

Example 63

2-((6-Chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)- 7,9-dihydro-8H-purin-8-one (Compound 63)

2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H -purin-8-one

The synthesis method of compound 63 is the same as that of compound 33 to obtain the title compound 2-((6-chloro-2,3-dihydrobenzofuran-5-yl)amino)-7-(methyl-d3)-9-(tetra Hydrogen-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (Compound 63) (white solid, 45 mg, yield 30.20%).

1H NMR (400MHz, DMSO-d6) δ 8.29 (s, 1H), 8.01 (s, 1H), 7.56 (s, 1H), 6.89 (s, 1H), 4.57 (t, 2H), 4.38 (tt, 1H), 3.96 (dd, 2H), 3.41 (t, 2H), 3.20-3.16 (m, 2H), 2.56-2.44 (m, 2H), 1.67-1.61 (m, 2H).

LC-MS m/z(ESI)=405.10[M+1].

Example 64

2-((6-Fluoro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9- Dihydro-8H-purin-8-one (Compound 64)

2-((6-fluoro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

first step:

6-Fluoro-5-nitro-2,3-dihydrobenzofuran (64b)

6-fluoro-5-nitro-2,3-dihydrobenzofuran

6-Fluoro-2,3-dihydrobenzofuran 64a (0.8g, 5.79mmol) was dissolved in 30mL of acetic acid, nitric acid (660.6mg, 69% purity) was added dropwise under ice bath, and the reaction was stirred at 70°C for 1h. TLC monitors that the reaction is complete, the reaction solution is poured into ice water, extracted, and the organic phase is concentrated to obtain a yellow oily crude product. The crude product is separated by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) = 10/1) to obtain colorless The title compound as a clear solid, 6-fluoro-5-nitro-2,3-dihydrobenzofuran (64b) (white solid, 0.5 g, yield 47.14%).

¹ H NMR (400MHz CDCl ₃ ) δ 7.97 (d, 1H), 6.62 (d, 1H), 4.77 (t, 2H), 3.27 (t, 2H).

The second step:

6-Fluoro-2,3-dihydrobenzofuran-5-amine (64c)

6-fluoro-2,3-dihydrobenzofuran-5-amine

Pd/C (10%, wet carrier) (50mg) was added to a solution of 6-fluoro-5-nitro-2,3-dihydrobenzofuran 64b (0.5g, 2.73mmol) in methanol (100mL), React overnight at room temperature under hydrogen atmosphere. It was filtered and concentrated to obtain the title compound 6-fluoro-2,3-dihydrobenzofuran-5-amine (64c) (colorless solid, 0.4g, yield 95.66%).

¹ H NMR (400MHz CDCl ₃ ) δ 6.66 (d, 1H), 6.50 (d, 1H), 4.52 (t, 2H), 3.1 (t, 2H).

third step:

2-((6-Fluoro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9- Dihydro-8H-purin-8-one (Compound 64)

2-((6-fluoro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one

The 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 3e (315mg, 1.18mmol, refer to the middle of WO2018114999) Body 20 method preparation), 6-fluoro-2,3-dihydrobenzofuran-5-amine 36c (150mg, 0.979mmol), cesium carbonate (638.21g, 1.96mmol), three (dibenzylidene acetone) two Palladium (89.57mg, 0.09mmol) and 2,2'-bis(diphenylphosphino)-1,1'-binaphthalene (121.84mg, 0.18mmol) were dissolved in dioxane, protected by nitrogen and ventilated, Stir at 100°C for 4h. TLC monitors the end of the reaction, the reaction solution is poured into ice water, the solid is collected, and the solid is separated and purified by silica gel column chromatography (dichloromethane: methanol (v/v) = 30/1) to obtain the title compound, 2-((6 -Fluoro-2,3-dihydrobenzofuran-5-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H- Purin-8-one (Compound 64) (white solid, 140 mg, yield 39.74%).

¹ H NMR(400MHz DMSO)δ8.50(s,1H),8.0(s,1H),7.43(d,1H),6.80(d,2H),4.56(t,2H),4.33-4.39(m, 1H), 3.93-3.97 (m, 2H), 3.37-3.45 (m, 2H), 3.27 (s, 3H), 3.15 (t, 2H), 1.64 (t, 2H).

LC-MS m/z(ESI)=386.10[M+1].

Biological test

1. DNA-PK kinase inhibition test

The DNA-PK kinase assay kit (purchased from Promega, product number: V4107, batch number: 0000366495) was used to detect the inhibitory activity of the compound against DNA-PK kinase. Using chemiluminescence to quantify the results, the specific experimental plan is as follows:

i. Construct different concentrations of ADP-fluorescence standard curve according to the kit instructions;

ii. Prepare a 5μL reaction system in a 384-well white plate, and add 1μL of compound to each well (set concentration gradients of 1μM, 200nM, 40nM, 8nM, 1.6nM, 0.32nM, 0.064nM, 0.013nM), 20units DNA- PK kinase, 0.2μg/μL substrate, 10μg/μL DNA, 50μM ATP, 1% DMSO;

iii. Mix well, centrifuge (1000rpm, 30s), and incubate at 37°C for 60min;

iv. Add 5μL ADP-Glo ^TM Reagent to terminate the reaction, mix well, centrifuge (1000rpm, 30s), and incubate at room temperature for 40min;

v. Add 10μL Kinase Detection Reagent, shake and mix, centrifuge (1000rpm, 30s), and incubate at room temperature for 30min;

vi. Measure the fluorescence value using a microplate reader (Thermo fisher, Varioskan LUX). Using GraphPad Pris m 8 to _{calculate IC 50} , the results are shown in Table 1.

Table 1 DNA-PK kinase inhibitory activity of the compounds of the present invention

Compound number IC ₅₀ (nM) Compound 1 19.70 Compound 2 0.91 Compound 3 0.97 Compound 4 47.60 Compound 5 0.01 Compound 6 1.68 Compound 7 11.34 Compound 8 3.35 Compound 9 24.59 Compound 11 0.89 Compound 12 4.63 Compound 13 3.80 Compound 14 11.39 Compound 15 12.33 Compound 16 1.81 Compound 17 5.10 Compound 18 2.14 Compound 19 2.93 Compound 21 4.68 Compound 22 0.01 Compound 23 0.04 Compound 24 1.45 Compound 25 6.12 Compound 26 2.88 Compound 27 12.30 Compound 28 3.40 Compound 29 6.80 Compound 30 5.48 Compound 32 1.68 Compound 33 0.92 Compound 34 0.91 Compound 35 18.77 Compound 36 3.90 Compound 38 28.65 Compound 39 3.83 Compound 40 42.69 Compound 41 12.46 Compound 42 37.87

Compound 44 6.81 Compound 45 63.72 Compound 46 27.80 Compound 47 11.25 Compound 48 4.05 Compound 49 23.80 Compound 50 1.13 Compound 51 35.98 Compound 53 33.03 Compound 54 8.90 Compound 55 3.31 Compound 56 43.41 Compound 57 32.52 Compound 58 23.0 Compound 59 31.30 Compound 60 24.37 Compound 62 14.89 Compound 63 3.80 Compound 64 19.72 Control example 100.2

Note: The comparative example refers to compound 3 of J. Med. Chem (2020), 63(7), 3461-3471, and the comparative example is prepared according to its preparation method.

Conclusion: The results show that, compared with the control example, the compound of the present invention has a more significant inhibitory effect on DNA-PK kinase.

2. Proliferation inhibition experiment

2.1. Experimental materials

A549 cells (purchased from ATCC), Doxorubicin hydrochloride (purchased from Shanghai Taosu Biochemical Technology Co., Ltd., catalog number: T1020), DMEM (10% FBS) medium, 96-well white board, cell viability Detection kit (purchased from Promega, article number: G9241).

2.2. Experimental steps

The A549 cells were resuspended and counted, and then spread in a 96-well plate, 500 cells/well (80 μL); cultured overnight.

Configure 10× drug concentration gradient (1:5): the initial concentration is 100μM, diluted with medium 1:5 to 20μM, 4μM, 800nM, 160nM, 32nM, 6.4nM, 1.3nM (final concentration 10μM, 2μM, 0.4 μM, 80nM, 16nM, 3.2nM, 0.64nM, 0.13nM).

3. Add 10 μL of the test compound to each well, do 2 replicates for each concentration gradient, set up the single-drug group (test compound or Doxorubicin), combination group, and medium control group:

Single-drug group: add the test compound (final concentration 10μM, 2μM, 0.4μM, 80nM, 16nM, 3.2nM, 0.64nM, 0.13nM), or only add doxorubicin (final concentration 10μM, 2μM, 0.4μM, 80nM, 16nM, 3.2nM, 0.64nM, 0.13nM), continue to incubate for 120h in a 37°C incubator;

Combination group: add test compound (final concentration 10μM, 2μM, 0.4μM, 80nM, 16nM, 3.2nM, 0.64nM, 0.13nM), incubate at 37℃ for 1h, add 10nM Doxorubicin (10μL/well) ), continue to incubate for 120h in 37℃ incubator;

Control group: add 20μL of culture medium and continue to incubate for 120h in 37℃ incubator.

4. Take out the cell culture plate and add an equal volume of cell culture medium to the test solution (100 μL medium, add 100 μL test solution).

5. Vibrate on the shaker for 2 minutes to lyse the cells.

6. Leave at room temperature for 10 minutes to stabilize the signal.

7. Microplate reader (Thermo fisher; Varioskan LUX) measures the chemiluminescence value.

Conclusion: The combination of the compound of the present invention and Doxorubicin has a more significant inhibitory effect on the proliferation of A549 cells and reduces cytotoxicity.

3. The transplantation tumor inhibition experiment

3.1 Experimental materials: A549 cells (purchased from ATCC); Doxorubicin liposomes (Dox) (Lipo Doxorubicin, trade name "Libaoduo", purchased from Shanghai Fudan Zhangjiang Biomedical Co., Ltd.); 6-week-old female nude mice ( Body weight 18-20g) (Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.), 10 rats in each group.

3.2 Determination of the inhibitory effect of Doxorubicin in combination with the compound to be screened on A549 xenograft tumors:

3.2.1 Collect the A549 cells in the logarithmic growth phase, wash them twice with pre-cooled PBS and use them for later use;

3.2.2 Balb/c nude mice were adapted to the laboratory environment for 3 days, and A549 cells were subcutaneously inoculated on the right ribs with a cell amount of 5×10 ⁶ per mouse. When the tumor grows to ^{about 200 mm 3} , the efficacy experiment will be carried out;

3.2.3 Randomly group the mice that have successfully grown tumors, and set up a single Doxorubicin (Dox) group, a test compound and Dox combined group, and a control group (Vehicle) for 21 days; mice are given intragastric administration (ig ), twice a day (BID, administration volume 5mL/kg; solvent: 5% DMSO+30% 2-hydroxypropyl-β-cyclodextrin). After intragastric administration in the morning for 1 hour, Lipo Doxorubicin (2.5mg/kg) was injected from the tail vein; once a week (QW, administration volume 5mL/kg;);

3.2.4 Weigh the weight of the mice twice a week, and measure the tumor volume at the same time: the tumor volume (V) is calculated as: V = 1/2 × L _long × L _short ² , and calculate the tumor inhibition rate and tumor inhibition rate (%)=(D21 tumor volume (Vehicle)-D21 tumor volume (administration group))/D21 tumor volume (Vehicle)×100;

3.2.5 After 21 days of administration, the tumor was isolated and weighed, and the weight change rate was calculated. The weight change rate (%)=(D21 weight-D0 weight)/D0 weight×100.

Conclusion: The experimental results show that the combination of the compound of the present invention and Doxorubicin can significantly improve the tumor suppressive effect of Doxorubicin, and will not cause significant weight loss.

The specification of the present invention describes the specific embodiments in detail. Those skilled in the art should realize that the above-mentioned embodiments are exemplary and should not be construed as limiting the present invention. For those skilled in the art, the principle of the present invention is not deviated from Under the premise of making several improvements and modifications to the present invention, the technical solutions obtained by these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims (10)

The compound represented by the general formula (I'), or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals:

among them: R ₀ and R ₁ are each independently selected from H, -OH, cyano, halogen, -NH ₂ , C _1-6 alkyl or C _1-6 alkoxy, the C _1-6 alkyl optionally Further substituted by 1-3 substituents selected from D or halogen; Or two R ₀ and the atoms to which they are connected form a 3- to 8-membered ring, and the 3- to 8-membered ring optionally contains 1 to 3 heteroatoms selected from N, O or S, and the 3- to 8-membered ring The ring is optionally further substituted with one or more substituents selected from -OH, carboxy, halogen, cyano, =0, C _1-6 alkyl or amino; R _{2 is} selected from H or C _1-6 alkyl; R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group, -C _1-6 alkylene-C _4-12 heterocyclic group, C _6-12 spiro compound or C _{6-12 heterospiro} compound, so The C ₃ heterocyclic group and C _4-12 heterocyclic group contain 1 to 3 heteroatoms selected from N, O or S, the C _1-6 alkyl group, C _3-12 carbocyclic group, C ₃ Heterocyclic group, C _4-12 heterocyclic group, C _1-6 alkylene group may be further selected by one or more selected from -OH, -OR ^a1 , carboxy, halogen, cyano, =O, C _{1 -6} alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR ^a1 R ^a2 , -C(=O)OC _1-6 alkyl, -C(= O) NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituent substituted And the C _1-6 alkyl group, C _1-6 heteroalkyl group, C _2-6 alkenyl group or C _2-6 alkynyl group in the substituent is optionally further selected from one or more -OH, carboxyl, cyano, halogen, -OR ^a1 , -NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl Or substituted by a substituent of a _{C 5-12 heteroaryl group;} R _{4 is} selected from H, C _1-6 alkyl, C _3-8 cycloalkyl, said C _1-6 alkyl and C _3-8 cycloalkyl are optionally further selected from 1-3 from D or Substituted by halogen substituents; R ^a1 and R ^a2 are each independently selected from H, C _1-6 alkyl, -C(=O)R ^a3 or -C(=O)NR ^a4 R ^a5 , wherein the C _1-6 alkyl is any Optionally, one or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkane Group, C ₃ heterocycloalkyl, or C _4-12 heterocycloalkyl substituent; or ^{Ra4, Ra5} and N atom form a 3- to 8-membered heterocyclic ring, said 3- to 8-membered heterocyclic ring Contain 1 to 3 heteroatoms selected from N, O or S; R ^{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl; R ^a4 and R ^a5 are each independently selected from H or C _1-6 alkyl; n is 0, 1, 2 or 3; p is 0, 1, 2, or 3. The compound according to claim 1, or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals, wherein the compound is selected from the group consisting of formula (I) compound of:

among them: R ₀ and R _{1 are} selected from H, -OH, cyano, halogen, -NH ₂ , C _1-6 alkyl or C _1-6 alkoxy, and the alkyl group is optionally further selected from 1-3 Substituted by D or halogen substituent; Or two R ₀ and the atoms to which they are connected can form a 3- to 8-membered ring, and the ring may contain 1 to 3 heteroatoms selected from N, O or S, and the ring is optionally further divided by 1 or more. Substituted by a substituent selected from -OH, carboxyl, halogen, cyano, =O, C _{1-6 alkyl or amino;} R _{2 is} selected from H or C _1-6 alkyl; R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group or -C _1-6 alkylene-C _4-12 heterocyclic group, said heterocyclic group may contain 1 to 3 selected from N, O or S heteroatom, the alkyl group, carbocyclic group, heterocyclic group, alkylene group may be further selected by one or more selected from -OH, carboxyl, halogen, cyano, =0, C _1-6 Alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR _a1 R _a2 , -C(=O)OC _1-6 alkyl, -C(=O) NR _a1 R _a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents are substituted; and The alkyl, heteroalkyl, alkenyl or alkynyl group is optionally further selected by one or more selected from -OH, carboxyl, cyano, halogen, -OR _a1 , -NR _a1 R _a2 , C _3-12 Cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents; R _a1 and R _{a2 are} selected from H, C _1-6 alkyl, -C(=O)R _a3 or -C(=O)NR _a4 R _a5 , wherein the C _1-6 alkyl is optionally further One or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkyl, C ₃ heterocycloalkyl, or substituted by a substituent of _{C 4-12 heterocycloalkyl; or R a4} and R _a5 and N atoms form a 3- to 8-membered heterocyclic ring, and the ring may contain one or more Heteroatom selected from N, O or S; R _{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl; R _a4 and R _{a5 are} selected from H or C _1-6 alkyl; n is selected from 0, 1, 2 or 3; p is selected from 0, 1, 2 or 3. The compound according to claim 1, or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals, wherein the compound is selected from general formula (IA) compound of:

among them: R ₀ and R ₁ are each independently selected from halogen or C _1-6 alkyl, and said C _1-6 alkyl is optionally further substituted with 1-3 substituents selected from D or halogen; Or two R ₀ and the atoms to which they are connected form a 3- to 8-membered ring, and the 3- to 8-membered ring optionally contains 1 to 3 heteroatoms selected from N, O or S, and the 3- to 8-membered ring The ring is optionally further substituted with one or more substituents selected from -OH, carboxy, halogen, cyano, =0, C _1-6 alkyl or amino; R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group or -C _1-6 alkylene-C _4-12 heterocyclic group, said C ₃ heterocyclic group and C _4-12 heterocyclic group include 1 Up to 3 heteroatoms selected from N, O or S, the C _1-6 alkyl group, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, C _1-6 The alkylene group is optionally further selected by one or more selected from -OH, carboxyl, halogen, cyano, =0, C _1-6 alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR ^a1 R ^a2 , -C(=O)OC _1-6 alkyl, -C(=O)NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl , C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents; and the C _1-6 alkyl, C _{1- 6} heteroalkyl, C _2-6 alkenyl or C _2-6 alkynyl is optionally further selected by one or more selected from -OH, carboxyl, cyano, halogen, -OR ^a1 , -NR ^a1 R ^a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl group substituents; R _{4 is} selected from H, C _1-6 alkyl, C _3-12 cycloalkyl, said C _1-6 alkyl and C _3-8 cycloalkyl are optionally further selected from 1-3 from D or Substituted by halogen substituents; R ^a1 and R ^a2 are each independently selected from H, C _1-6 alkyl, -C(=O)R ^a3 or -C(=O)NR ^a4 R ^a5 , wherein the C _1-6 alkyl is any Optionally, one or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkane Group, C ₃ heterocycloalkyl or C _4-12 heterocycloalkyl substituent; or ^{Ra4, Ra5} and N atom form a 3- to 8-membered heterocyclic ring, and the 3- to 8-membered heterocyclic ring includes 1 to 3 heteroatoms selected from N, O or S; R ^{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl; R ^a4 and R ^a5 are each independently selected from H or C _1-6 alkyl; n is 0, 1, 2 or 3; p is 0, 1, 2, or 3. The compound according to claim 1, or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals, wherein the compound is selected from general formula (II) compound of:

among them: R ₀ and R _{1 are} selected from halogen or C _1-6 alkyl, and the alkyl is optionally further substituted with 1-3 substituents selected from D or halogen; Or two R ₀ and the atoms to which they are connected can form a 3- to 8-membered ring, and the ring may contain 1 to 3 heteroatoms selected from N, O or S, and the ring is optionally further divided by 1 or more. Substituted by a substituent selected from -OH, carboxyl, halogen, cyano, =O, C _{1-6 alkyl or amino;} R _{3 is} selected from C _1-6 alkyl, C _3-12 carbocyclic group, C ₃ heterocyclic group, C _4-12 heterocyclic group, -C _1-6 alkylene-C _3-12 carbocyclic group, -C _1-6 alkylene-C ₃ heterocyclic group or -C _1-6 alkylene-C _4-12 heterocyclic group, said heterocyclic group may contain 1 to 3 selected from N, O or S heteroatom, the alkyl group, carbocyclic group, heterocyclic group, alkylene group may be further selected by one or more selected from -OH, carboxyl, halogen, cyano, =0, C _1-6 Alkyl, C _1-6 heteroalkyl, C _2-6 alkenyl, C _2-6 alkynyl, -NR _a1 R _a2 , -C(=O)OC _1-6 alkyl, -C(=O) NR _a1 R _a2 , C _3-12 cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents are substituted; and The alkyl, heteroalkyl, alkenyl or alkynyl group is optionally further selected by one or more selected from -OH, carboxyl, cyano, halogen, -OR _a1 , -NR _a1 R _a2 , C _3-12 Cycloalkyl, C ₃ heterocycloalkyl, C _4-12 heterocycloalkyl, C _6-12 aryl or C _5-12 heteroaryl substituents; R _a1 and R _{a2 are} selected from H, C _1-6 alkyl, -C(=O)R _a3 or -C(=O)NR _a4 R _a5 , wherein the C _1-6 alkyl is optionally further One or more selected from OH, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _6-12 aryl, C _5-12 heteroaryl, C _3-12 cycloalkyl, C ₃ Heterocycloalkyl or C _4-12 heterocycloalkyl substituents; or R _a4 and R _a5 and N atoms form a 3- to 8-membered heterocyclic ring, the ring may contain one or more options Heteroatoms from N, O or S; R _{a3 is} selected from C _1-6 alkyl, C _1-6 alkoxy or C _6-12 aryl; R _a4 and R _{a5 are} selected from H or C _1-6 alkyl; n is selected from 0, 1, 2 or 3; p is selected from 0, 1, 2 or 3. The compound according to any one of claims 1 to 4, or its stereoisomer, solvate, prodrug, metabolite, deuterated product, pharmaceutically acceptable salt or co-crystal, wherein: R _{0 is} selected from methyl; R _{1 is} selected from halogen or C _1-4 alkyl, and the alkyl is optionally further substituted with 1-3 substituents selected from D or halogen; R _{3 is} selected from C _1-6 alkyl, C _3-6 carbocyclic group, C _4-8 heterocyclic group, -C _1-2 alkylene-C _3-8 carbocyclic group or -C _1-2 alkylene Alkyl-C _4-8 heterocyclic group, the heterocyclic group may contain 1 to 3 heteroatoms selected from N or O, and the alkyl group, carbocyclic group, heterocyclic group, and alkylene group may be any Optionally further substituted by one or more substituents selected from -OH, halogen, cyano, =OC _1-4 alkoxy or C _1-4 alkyl; and the alkyl in the substituents is any Optionally further substituted by one or more substituents selected from -OH, carboxyl, cyano or halogen; n is selected from 0, 1 or 2; p is selected from 1. The compound according to any one of claims 1 to 5, or its stereoisomer, solvate, prodrug, metabolite, deuterated product, pharmaceutically acceptable salt or co-crystal, wherein the compound is selected from:

or

An intermediate for preparing the compound of any one of claims 1-6, wherein the intermediate is selected from:

A pharmaceutical composition, the pharmaceutical composition comprising: (1) The compound according to any one of claims 1 to 6 or its stereoisomers, solvates, prodrugs, metabolites, deuterated products, pharmaceutically acceptable salts or co-crystals; (2) Optional one or more other active ingredients; and (3) A pharmaceutically acceptable carrier and/or excipient. The compound of any one of claims 1-6 or its stereoisomers, solvates, prodrugs, metabolites, deuterated substances, pharmaceutically acceptable salts or co-crystals or the pharmaceutical combination of claim 8 In the preparation of drugs for the treatment of cancer. The compound of any one of claims 1-6 or its stereoisomers, solvates, prodrugs, metabolites, deuterated substances, pharmaceutically acceptable salts or co-crystals or the pharmaceutical combination of claim 8 In the preparation of DNA-PK inhibitors.