WO2024153250A1 - Compound as usp1 inhibitor - Google Patents

Abstract

Provided in the present invention is a class of compounds. Specifically, provided in the present invention is a compound having a structure as shown in formula (I), or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate and a solvate thereof. The compound can effectively inhibit USP1, and is used for treating or preventing diseases or conditions related to the activity or expression level of USP1.

Description

Compounds as USP1 inhibitors Technical Field

The present invention relates to the field of medicinal chemistry; specifically, the present invention relates to a class of novel compounds, a synthesis method thereof and an application thereof as a USP1 inhibitor in the preparation of drugs for treating various diseases such as tumors.

Background technique

Ubiquitination is the process of covalently attaching ubiquitin molecules to target proteins and is one of the most common post-translational modifications of proteins. Ubiquitin is a small molecule protein composed of 76 amino acids. It is widely present in all eukaryotic cells and its sequence is highly conserved. There are many types of ubiquitination, including monoubiquitination, polyubiquitination, and polyubiquitination. The types of ubiquitination chains are also diverse, such as those connected through Lys6, Lys11, Lys27, Lys33, Lys48, and Lys63. Different ubiquitination types regulate different functions. For example, proteins with polyubiquitin chains connected to Lys48 are usually degraded by the proteasome pathway, while monoubiquitination or polyubiquitin chains connected through other lysines are usually involved in cell cycle regulation, DNA damage repair, transcription, immune response, and endocytosis.

Ubiquitination is a reversible post-translational modification of proteins. Deubiquitinating enzymes can reverse ubiquitination modification by hydrolyzing peptide bonds or isopeptide bonds between ubiquitin molecules or between ubiquitin and substrate proteins. USP1 (Ubiquitin-specific protease 1) is a deubiquitinating enzyme that is mainly involved in regulating DNA damage repair processes, including translesion synthesis (TLS) and Fanconi anemia pathway (FA). PCNA plays a key role in translesion DNA replication. PCNA is monoubiquitinated by RAD6 and RAD18 in response to replication fork stalling. Monoubiquitinated PCNA allows the recruitment of TLS polymerases to bypass DNA damage for replication. Fanconi anemia is a rare autosomal recessive genetic disease. Key gene defects in the Fanconi anemia signaling pathway lead to defects in the cell's DNA interstrand cross-link repair function. Fifteen FA genes have been identified, of which eight (FANCA, FANCB, FANCC, FANCE, FANCF, FANCG, FANCL, and FANCM) encode proteins that form the FA core complex, which monoubiquitinates FANCD2 and FANCI and recruits the intrachain cross-link repair complex. USP1 interacts with UAF1 to form a complex and exerts a deubiquitination function. The USP1/UAF1 complex regulates the DNA damage repair process by deubiquitinating PCNA, FANCD2, and FANCI. Inhibiting the enzymatic activity of USP1 can significantly inhibit the growth of tumor cells with BRCA mutations or other homologous recombination repair defects. Therefore, USP1 inhibitors have broad anti-tumor prospects.

Summary of the invention

The purpose of the present invention is to provide a new type of USP1 inhibitors.

In the first aspect of the present invention, there is provided a compound having a structure as shown in the following formula (I), or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvate thereof:

In formula (I):

B is selected from Formula (Ia) or Formula (Ib):

represents the site of connection between the compound of formula (Ia) or formula (Ib) and A in the compound of formula (I); represents the site where the compound of formula (Ia) or (Ib) is connected to the benzene ring in the compound of formula (I);

Each R ¹ is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, CN; m is selected from 0, 1, or 2;

Each R ² is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _2-4 alkenyl, C _2-4 alkynyl, CN; n is selected from 0, 1, 2 _, 3, or 4;

^R3 and ^R4 are each independently selected from hydrogen, halogen, _C1-4 alkyl; or ^R3 and ^R4 together with the same carbon atom to which they are attached form a 3- to 6-membered ring structure, which optionally contains 0 or 1 heteroatom selected from N, O, S;

Each R ⁵ _is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _2-4 alkenyl, C _2-4 alkynyl, CN; p is selected from 0, 1, or 2;

each R ⁶ is independently selected from hydrogen, halogen, C _1-4 alkyl, C 1-4 haloalkyl, C _2-4 alkenyl _, C _2-4 alkynyl, C _3-6 cycloalkyl, C _3-6 cycloalkyl-C _2-4 alkynyl, 3- to 6-membered heterocyclyl, 3- to 6-membered heterocyclyl-C _2-4 alkynyl, CN, OR ^f , SR ^f , NR ^d R ^d , C(O) R ^g , C(O)OR ^f , or S(O) ₂ R ^g ; q is selected from 0, 1, 2, or 3;

A is selected from Formula (Ic), Formula (Id), Formula (Ie), Formula (If), or Formula (Ig):

represents the site at which B in the compound of formula (Ic), formula (Id), formula (Ie), formula (If), or formula (Ig) is connected to B;

represents a single bond or a double bond;

each ^Ra is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to ₈ -membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , C(O) ^NRdRd , or S(O) _2NRdRd ; the ^alkyl , cycloalkyl, 3- to 8-membered heterocyclyl, aryl, and ^heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C2-4 alkenyl, _C2-4 alkynyl ^, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl ^{, heteroaryl, CN, ORf, SRf, NRdRd, C(O)Rg , C ( O} ) ^ORf , OC(O) ^Rg or the ^{cycloalkyl and} 3- ^{to 8} - ^{membered heterocyclyl described} _in ^{Ra are optionally substituted by =} _T , wherein ^T is selected from ^O or ^{CRmRn ; Rm} and ^{Rn are} each _{independently} selected from hydrogen, ^{halogen , or} _C1-4 ^alkyl ;

Each R ^b is independently selected from hydrogen, halogen, C _1-4 alkyl; or two R ^b and the carbon atom to which they are attached together form a C _3-6 cycloalkyl; each k is independently selected from 0, 1, 2, or 3;

R ^c is selected from hydrogen, halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 haloalkenyl, C _2-4 alkynyl, CN, C (O) R ^g , C (O) OR ^f , C (O) NR ^d R ^d ; the alkyl, alkenyl or alkynyl is optionally substituted by one or more groups selected from the group consisting of halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, OR ^f , SR ^f , NR ^d R ^d , C (O) R ^g , C (O) OR ^f , OC (O) R ^g , C (O) NR ^d R ^d , NR ^d C (O) R ^g , S (O) ₂ NR ^d R ^d , or NR ^d S (O) ₂ R ^g ;

Each R ^d is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl;

Each R ^d' is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, hydroxy, C _1-4 alkoxy, or CN; each t is independently selected from 0, 1, 2, 3, or 4;

k ¹ , k ² , k ³ , and k ⁴ are each independently selected from 0, 1, 2, 3, 4 or 5;

D is selected from a chemical bond, O, NR ^e , or CR ^b R ^b ; wherein, R ^e is selected from hydrogen or C _1-4 alkyl; R ^b is as defined above;

M is selected from O or CR ^h R ⁱ ; wherein R ^h and R ⁱ are each independently selected from hydrogen, halogen, or C _1-4 alkyl; the alkyl is optionally substituted with one or more groups selected from the group consisting of halogen, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, aryl, heteroaryl, CN, OR ^f , SR ^f , NR ^d R ^d , C(O)R ^g , C(O)OR ^f , OC(O)R ^g , C(O)NR ^d R ^d , NR ^d C(O)R ^g , NR ^d C(O)NR ^d R ^d , OC(O)NR ^d R ^d , NR ^d C(O)OR ^f , OC(O)OR ^f , S(O) ₂ NR ^d R ^d , NR ^d S(O) ₂ R ^g , or NR ^d S(O) ₂ NR ^d R ^d ; or R ^h and R ⁱ together with the carbon atom to which they are attached form a 3- to -8-membered cyclic structure, which optionally contains 0, 1, or 2 heteroatoms selected from N, O, S;

^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are each independently selected from N or ^CRk ; provided that at most two of ^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are selected from N; each ^Rk is each independently selected from hydrogen, halogen, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^{Rg ,} or C(O) ^ORf ;

Each of the above R ^d is independently selected from hydrogen, C _1-4 alkyl, C _1-4 haloalkyl, C _3-6 cycloalkyl, or 3- to 6-membered heterocyclyl; each of the R ^f is independently selected from hydrogen, C _1-4 alkyl, C _1-4 haloalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, aryl, or heteroaryl; each of the R ^g is independently selected from hydrogen, C _1-4 alkyl, C _1-4 haloalkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, aryl, or heteroaryl;

wherein each of the above-mentioned alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, cyclic structure, aryl and heteroaryl groups is optionally and independently substituted with 1-3 substituents each independently selected from the group consisting of halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-8 cycloalkyl, 3- to 8-membered heterocyclic group, aryl, heteroaryl, CN, NO ₂ , OR ^f , SR ^f , NR ^d R ^d , C(O) R ^g , C(O)OR ^f , C(O)NR ^d R ^d , NR ^d C(O) R ^g , S(O) ₂ R ^g , or NR ^d S(O) ₂ R ^g , provided that the chemical structure formed is stable and meaningful; wherein R ^d , R ^f , and R ^g are as defined above;

Unless otherwise specified, the above-mentioned aryl group is an aromatic group containing 6 to 12 carbon atoms; the heteroaryl group is a 5- to 15-membered heteroaromatic group; and the cyclic structure is a saturated or unsaturated cyclic group containing or not containing heteroatoms.

In another preferred embodiment, the formula (I) is formula (IIa) or formula (IIb):

The definitions of the groups in formula (IIa) or (IIb) are as described above.

In another preferred embodiment, the formula (I) is formula (IIc) or formula (IId):

each ^Ra is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, _3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C( ^O ) ^NRdRd ; the alkyl, cycloalkyl, 3- to 8-membered heterocyclyl, aryl and heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^Rg , C(O) ^ORf , ^OC (O) ^Rg , C(O)NR ^d R ^d , NR ^d C(O) R ^g , or NR ^d S(O) ₂ R ^g ; or the cycloalkyl and 3- to 8-membered heterocyclyl described in ^Ra are optionally substituted by =T, wherein T is selected from CR ^m R ⁿ ; R ^m and R ⁿ are each independently selected from hydrogen, halogen, or C _1-4 alkyl;

R ^d , R ^f , and R ^g are as defined above.

In another preferred embodiment, the formula (I) is formula (IIe) or formula (IIf):

each ^Ra is independently selected _from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, _3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C(O) ^NRdRd ; the alkyl, cycloalkyl, 3- to 8-membered heterocyclyl, aryl and ^heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^ORf , ^NRdRd ; or the cycloalkyl and 3- to 8-membered heterocyclyl described in ^Ra are optionally substituted with =T, wherein T is selected ^{from CRmRn} ; ^Rm and ^{Rn are} independently selected from hydrogen, halogen, or _C1-4 alkyl;

R ^d , R ^f , and R ^g are as defined above.

In another preferred embodiment, each ^Ra in the formula (IIe) or formula (IIf) is independently a group selected from the following group:

In another preferred embodiment, the formula (I) is formula (IIIa) or formula (IIIb):

each ^Ra is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, _3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C( ^O ) ^NRdRd ; the alkyl, cycloalkyl, 3- to 8-membered heterocyclyl, aryl and heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^Rg , C(O) ^ORf , ^OC (O) ^Rg , C(O)NR ^d R ^d , NR ^d C(O) R ^g , or NR ^d S(O) ₂ R ^g ; or the cycloalkyl and 3- to 8-membered heterocyclyl described in ^Ra are optionally substituted by =T, wherein T is selected from CR ^m R ⁿ ; R ^m and R ⁿ are each independently selected from hydrogen, halogen, or C _1-4 alkyl;

R ^d , R ^f , and R ^g are as defined above.

In another preferred embodiment, each ^Ra in Formula (IIIa) or (IIIb) is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C(O) ^NRdRd ; the _alkyl , cycloalkyl, 3- to 8-membered heterocyclyl, aryl ^and heteroaryl in ^Ra are optionally substituted by one or more groups selected from the group consisting of halogen, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^{ORf , NRdRd} ; or the cycloalkyl and 3- to 8-membered heterocyclyl in ^Ra are optionally substituted by =T, wherein T is selected from ^CRmRn ; ^Rm and ^{R n} are each independently selected from hydrogen, halogen, or C _1-4 alkyl;

R ^d , R ^f , and R ^g are as defined above.

In another preferred embodiment, each ^Ra in the formula (IIIa) or formula (IIIb) is independently selected from the group consisting of:

In another preferred embodiment, the formula (I) is formula (IVa) or formula (IVb):

The definitions of the various groups in formula (IVa) or formula (IVb) are as described above.

In another preferred embodiment, the formula (I) is formula (V):

R ^c is selected from halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, CN, C(O)R ^g , C(O)OR ^f , C(O)NR ^d R ^d ; the alkyl, alkenyl or alkynyl is optionally substituted by one or more groups selected from the group consisting of halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, OR ^f , SR ^f , NR ^d R ^d , C(O)R ^g , C(O)OR ^f , OC(O)R ^g , C(O)NR ^d R ^d , NR ^d C(O)R ^g , or NR ^d S(O) ₂ R ^g ;

^k1 is selected from 1, 2, 3, or 4;

R ^d' , t are defined as above;

R ^d , R ^f , and R ^g are as defined above.

In another preferred embodiment, the formula (I) is formula (VIa) or formula (VIb):

The definitions of the various groups in formula (VIa) or formula (VIb) are as described above.

In another preferred embodiment, the formula (I) is formula (VIIa) or formula (VIIb):

The definitions of the various groups in formula (VIIa) or formula (VIIb) are as described above.

In another preferred embodiment, the formula (I) is formula (VIIIa) or formula (VIIIb):

^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are each independently selected from N or ^CRk ; provided that at most two of ^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are selected from N; each ^Rk is each independently selected from hydrogen, halogen, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^{Rg ,} or C(O) ^ORf ;

R ^d , R ^f , and R ^g are as defined above.

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

“*” indicates a chiral center.

The second aspect of the present invention provides a pharmaceutical composition comprising the compound described in the first aspect of the present invention, or its optical isomers, pharmaceutically acceptable salts, prodrugs, deuterated derivatives, hydrates, solvates, and pharmaceutically acceptable carriers.

The second aspect of the present invention provides a use of the compound described in the first aspect of the present invention, or its optical isomers, pharmaceutically acceptable salts, prodrugs, deuterated derivatives, hydrates, solvates, for preparing a pharmaceutical composition for treating diseases, disorders or conditions associated with USP1 activity or expression.

In another preferred embodiment, the disease, disorder or condition is selected from the following group: breast cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, cutaneous granular cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, pharyngeal cancer, pancreatic cancer, prostate cancer, glioma, ovarian cancer, endometrial cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, skin cancer, gastric cancer, mesothelioma, osteosarcoma, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, T-cell lymphoma, monocytic leukemia, eosinophilic syndrome, multiple myeloma and other solid tumors and blood tumors.

Detailed ways

After long-term and in-depth research, the inventors unexpectedly discovered a class of USP1 inhibitors with novel structures, as well as their preparation methods and applications. The compounds of the present invention can be used to treat various diseases related to the activity of the USP1. Based on the above findings, the inventors completed the present invention.

the term

Unless otherwise specified, "or" mentioned in this document has the same meaning as "and/or" (referring to "or" and "and").

Unless otherwise specified, in all compounds of the present invention, each chiral carbon atom (chiral center) may be optionally in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

As used herein, the term "alkyl" refers to a straight chain (i.e., unbranched) or branched saturated hydrocarbon group containing only carbon atoms, or a combination of straight and branched hydrocarbon groups, when used alone or as part of other substituents. When the alkyl group is preceded by a carbon number limit (e.g., C _1-10 ), it means that the alkyl group contains 1-10 carbon atoms. For example, C _1-8 alkyl refers to an alkyl group containing 1-8 carbon atoms, including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.

As used herein, the term "alkenyl" refers to a linear or branched carbon chain group having at least one carbon-carbon double bond, either alone or as part of another substituent. Alkenyl groups may be substituted or unsubstituted. When the number of carbon atoms before the alkenyl group is limited (e.g., C _2-8 ), it means that the alkenyl group contains 2-8 carbon atoms. For example, C _2-8 alkenyl refers to an alkenyl group containing 2-8 carbon atoms, including vinyl, propenyl, 1,2-butenyl, 2,3-butenyl, butadienyl, or similar groups.

As used herein, the term "alkynyl" refers to an aliphatic hydrocarbon group having at least one carbon-carbon triple bond, either alone or as part of another substituent. The alkynyl group may be straight or branched, or a combination thereof. When the alkynyl group is preceded by a carbon number limit (e.g., _C2-8 alkynyl), the alkynyl group contains 2-8 carbon atoms. For example, the term " _C2-8 alkynyl" refers to a straight or branched alkynyl group having 2-8 carbon atoms, including ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, sec-butynyl, tert-butynyl, or the like.

As used herein, when used alone or as part of other substituents, the term "cycloalkyl" refers to a saturated or partially saturated unit ring, a bicyclic or polycyclic (condensed, bridged or spiro) ring system group. When a cycloalkyl group is preceded by a carbon atom number limit (such as C _3-10 ), it means that the cycloalkyl group contains 3-10 carbon atoms. In some preferred embodiments, the term "C _3-8 cycloalkyl" refers to a saturated or partially unsaturated monocyclic or bicyclic alkyl group with 3-8 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, or similar groups. "Spirocycloalkyl" refers to a bicyclic or polycyclic group in which a carbon atom (called a spiro atom) is shared between monocyclic rings, which may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. "Condensed cycloalkyl" refers to a full carbon bicyclic or polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. "Bridged cycloalkyl" refers to an all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. The atoms contained in the cycloalkyl are all carbon atoms. The following are some examples of cycloalkyl groups, and the present invention is not limited to the following cycloalkyl groups.

Unless otherwise stated, the following terms used in the specification and claims have the following meanings. "Aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group with a conjugated π electron system, such as phenyl and naphthyl. The aryl ring can be fused to other cyclic groups (including saturated and unsaturated rings), but cannot contain heteroatoms such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent must be on a carbon atom on the ring with a conjugated π electron system. Aryl groups can be substituted or unsubstituted. The following are some examples of aryl groups, and the present invention is not limited to the aryl groups described below.

"Heteroaryl" refers to a monocyclic or polycyclic group with aromaticity containing one to multiple heteroatoms (optionally selected from nitrogen, oxygen and sulfur), or a polycyclic group formed by condensing a heterocyclic group (containing one to multiple heteroatoms selected from nitrogen, oxygen and sulfur) with an aryl group, and the connection site is located on the aryl group. The heteroaryl group can be optionally substituted or unsubstituted. The following are some examples of heteroaryl groups, and the present invention is not limited to the following heteroaryl groups.

"Heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl. Polycyclic heterocyclyls refer to heterocyclyls including spiro, condensed rings and bridged rings. "Spirocyclic heterocyclyl" refers to a polycyclic heterocyclic group in which each ring in the system shares an atom (called a spiro atom) with other rings in the system, wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. "Condensed ring heterocyclyl" refers to a polycyclic heterocyclic group in which each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, and wherein one or more ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. "Bridged heterocyclic group" refers to a polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected. These may contain one or more double bonds, but none of the rings has a completely conjugated π-electron system, and one or more of the ring atoms are selected from nitrogen, oxygen or sulfur, and the remaining ring atoms are carbon. If there are both saturated rings and aromatic rings in the heterocyclic group (for example, a saturated ring and an aromatic ring are fused together), the point of connection to the parent must be on the saturated ring. Note: When the point of connection to the parent is on the aromatic ring, it is called a heteroaryl group, not a heterocyclic group. The following are some examples of heterocyclic groups, and the present invention is not limited to the following heterocyclic groups.

As used herein, the term "halogen" by itself or as part of another substituent refers to F, Cl, Br and I.

As used herein, the term "substituted" (with or without the "arbitrarily" modifier) refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent is a substituent described accordingly in the foregoing text, or a substituent appearing in the embodiments. Unless otherwise specified, a certain arbitrarily substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. A cyclic substituent, such as a heterocyclic group, may be connected to another ring, such as a cycloalkyl group, to form a spirobicyclic system, i.e., two rings have a common carbon atom. It will be understood by those skilled in the art that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible. The substituents include, but are not limited to, _C1-8 alkyl, _C2-8 alkenyl, _C2-8 alkynyl, _C3-8 cycloalkyl, 3- to ₁₂ -membered heterocyclic group, aryl, heteroaryl, halogen, hydroxyl, carboxyl (-COOH), _C1-8 aldehyde, _C2-10 acyl, _C2-10 ester, and amino.

For the sake of convenience and in accordance with common understanding, the term "arbitrary substitution" or "optionally substituted" only applies to sites that can be substituted by substituents, and does not include those substitutions that are chemically unfeasible.

As used herein, unless otherwise specified, the term "pharmaceutically acceptable salt" refers to a salt that is suitable for contact with the tissue of a subject (e.g., a human) without producing undue side effects. In some embodiments, a pharmaceutically acceptable salt of a compound of the present invention includes a salt of a compound of the present invention having an acidic group (e.g., potassium salt, sodium salt, magnesium salt, calcium salt) or a salt of a compound of the present invention having a basic group (e.g., sulfate, hydrochloride, phosphate, nitrate, carbonate).

use:

The present invention provides a class of compounds of formula (I), or their deuterated derivatives, their salts, isomers (enantiomers or diastereomers, if any), hydrates, pharmaceutically acceptable carriers or excipients for use in inhibiting USP1.

The compound of the present invention is useful as a USP1 inhibitor.

The present invention is a single inhibitor of USP1, which can prevent, alleviate or cure diseases by regulating the activity of USP1. The diseases include, but are not limited to, breast cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, skin granular cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, pharyngeal cancer, pancreatic cancer, prostate cancer, glioma, ovarian cancer, endometrial cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, skin cancer, gastric cancer, mesothelioma, osteosarcoma, acute myeloid leukemia, myelofibrosis, B cell lymphoma, T cell lymphoma, monocytic leukemia, eosinophilic syndrome, multiple myeloma and other solid tumors and blood tumors.

The compounds of the present invention and their deuterated derivatives, as well as pharmaceutically acceptable salts or isomers thereof (if present) or hydrates thereof and/or compositions can be formulated with pharmaceutically acceptable excipients or carriers, and the resulting compositions can be administered to mammals, such as men, women and animals, in vivo for the treatment of conditions, symptoms and diseases. The compositions can be tablets, pills, suspensions, solutions, emulsions, capsules, aerosols, sterile injections, sterile powders, etc. In some embodiments, pharmaceutically acceptable excipients include microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, mannitol, hydroxypropyl-β-cyclodextrin, β-cyclodextrin (increased), glycine, disintegrants (such as starch, cross-linked sodium carboxymethyl cellulose, complex silicates and high molecular weight polyethylene glycol), granulation binders (such as polyvinyl pyrrolidone, sucrose, gelatin and gum arabic) and lubricants (such as magnesium stearate, glycerol and talc). In a preferred embodiment, the pharmaceutical composition is a dosage form suitable for oral administration, including but not limited to tablets, solutions, suspensions, capsules, granules, and powders. The amount of the compound of the present invention or the pharmaceutical composition administered to the patient is not fixed, and is usually administered in a pharmaceutically effective amount. At the same time, the amount of the compound actually administered can be determined by the physician according to actual conditions, including the disease to be treated, the selected route of administration, the actual compound administered, the individual conditions of the patient, etc. The dosage of the compound of the present invention depends on the specific use of the treatment, the mode of administration, the patient's state, and the physician's judgment. The ratio or concentration of the compound of the present invention in the pharmaceutical composition depends on various factors, including dosage, physicochemical properties, route of administration, etc.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form new or preferred technical solutions.

Pharmaceutical compositions and methods of administration

Since the compounds of the present invention have excellent inhibitory activity against USP1, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate diseases related to USP1 activity or expression.

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 5-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The administration method of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic; (c) humectants, such as glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

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

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

The main advantages of the present invention include:

1. Provided is a compound as shown in formula I.

2. Provided is a USP1 inhibitor with a novel structure, and its preparation and use. The inhibitor can inhibit the activity of USP1 at extremely low concentrations.

3. Provides a USP1 inhibitor with good oral absorption.

4. Provided is a pharmaceutical composition for treating diseases associated with USP1 activity.

The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Some representative compounds of the present invention can be prepared by the following synthetic methods. In the following reaction formulas, the reagents and conditions of each step can be selected from conventional reagents or conditions for such preparation methods in the art. After the structure of the compound of the present invention is disclosed, the above selection can be made by those skilled in the art based on the knowledge in the art.

abbreviation

Boc＝tert-butyloxycarbonyl

CN＝cyano

m-CPBA＝3-chloroperoxybenzoic acid

DCM = dichloromethane

DIPEA or DIEA = N,N-diisopropylethylamine

DIBAL-H = Diisobutylaluminum hydride

DMF＝N,N-dimethylformamide

DMSO = dimethyl sulfoxide

DDQ = 2,3-dichloro-5,6-dicyanobenzoquinone

EtOAc or EA＝Ethyl acetate

Et＝ethyl

HATU = N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate)

LiHMDS = Lithium Bis(trimethylsilyl)amide

Me = methyl

MeOH = methanol

NMP = N-methylpyrrolidone

Ph = Phenyl

PMB = p-methoxybenzyl

Pd(PPh ₃ ) ₂ Cl ₂ = bis(triphenylphosphine)palladium dichloride

PhNTf ₂ = phenylbis(trifluoromethanesulfonyl)imide

SOCl ₂ = thionyl chloride

TIPS = triisopropylsilyl

TMS = trimethylsilyl

TBAF = Tetra-n-butylammonium fluoride

TEA = triethylamine

TFA = trifluoroacetic acid

THF = Tetrahydrofuran

TsCl = p-Toluenesulfonyl chloride

TBDPSCl = tert-butyldiphenylsilyl chloride

Example 1: Preparation of Compound 1

Compound 1-a (25 mg, 0.05 mmol) (the synthesis of common intermediate 1-a can be prepared by referring to the method in patent WO2020/132269Al) and compound 1-b (27 mg, 0.10 mmol) were dissolved in DMF (1 mL) solution, and then cesium carbonate (49 mg, 0.15 mmol) was added. The reaction mixture was heated and stirred at 90 ° C overnight. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The white solid compound 1 (3 mg, yield 10%) was obtained by separation and purification on a preparative thin layer plate (ethyl acetate: petroleum ether = 1: 1). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.38 (d, J = 1.0Hz, 1H), 7.53 (d, J = 8.3Hz, 2H), 7.42 (d, J = 8.3Hz, 2H), 5.79 (s, 2H), 4.95 -4.87(m,1H),3.85(s,3H),3.20-3.15(m,4H),1.68-1.63(m,1H),1.08-1.04(m,2H),0.87-0.84(m,2H). MS m/z 595.2[M+H] ⁺ .

Example 2: Preparation of Compound 2

3,3-Difluorocyclobutanol (200 mg, 1.85 mmol) and p-toluenesulfonyl chloride (371 mg, 1.94 mmol) were dissolved in dichloromethane (10 mL), and sodium hydrogen (148 mg, 3.70 mmol, 60%) was added under ice bath. The reaction solution was stirred for 0.5 hours. The reaction solution was quenched with saturated ammonium chloride, and the mixed solution was extracted with ethyl acetate (3×20 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:20) to obtain a colorless oily compound 2-b (399 mg, yield 82%).

Compound 1-a (24 mg, 0.05 mmol) and compound 2-b (26 mg, 0.10 mmol) were dissolved in DMF (1 mL), and cesium carbonate (49 mg, 0.15 mmol) was added. The reaction mixture was heated and stirred at 90 ° C overnight. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The mixture was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain a white solid compound 2 (11 mg, yield 41%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.50 (s, 1H), 7.60 (d, J = 8.2Hz, 2H), 7.45 (d, J = 8.2Hz, 2H), 7.06 (d, J = 14.1Hz, 1H), 6.98- 6.88(m,1H),5.80(s,2H),4.94(dd,J＝17.0,3.2Hz,1H),4.79(dd,J＝50.6,3.2Hz,1H),3.85(s,3H),1.68-1.62(m,1H),1.08-1.03(m,2H),0.89-0.81(m, 2H). MS m/z 563.2[M+H] ⁺ .

Example 3: Preparation of Compound 3

Compound 2 (5 mg, 0.008 mmol) was dissolved in a hydrogen fluoride pyridine solution (0.5 mL). The reaction solution was stirred at room temperature overnight. The reaction solution was quenched with a saturated sodium bicarbonate aqueous solution, and the mixed solution was extracted with methyl tert-butyl ether (3×5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to give a white solid compound 3 (3 mg, yield 58%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.55-8.51(m,2H),7.62-7.58(m,2H),7.49-7.43(m,2H),7.21-7.17(m,1H),6.67-6.60(m,1H),5 .80(s,2H),3.85(s,3H),1.78(t,J＝18.7Hz,3H),1.70-1.63(m,1H),1.07-1.03(m,2H),0.87-0.83(m,2H). MS m/z 583.2[M+H] ⁺ .

Example 4: Preparation of Compound 4

Compound 1-a (50 mg, 0.10 mmol), methyl 2,4-dibromobutyrate (28 mg, 0.11 mmol), and cesium carbonate (99 mg, 0.30 mmol) were dissolved in DMF (2 mL). The reaction mixture was stirred at room temperature overnight. After the reaction was completed, water was added to quench the mixture, and the mixed solution was extracted with ethyl acetate (3×10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 4-a (31 mg, yield 50%). MS m/z 591.2[M+H] ⁺ .

Compound 4-a (31 mg, 0.05 mmol) was dissolved in tetrahydrofuran (3 mL), and lithium aluminum hydride (4 mg, 0.10 mmol) was slowly added under ice bath. The reaction solution was stirred at room temperature for 0.5 hours. After the reaction was completed, water was added to quench the mixture, and the mixed solution was extracted with ethyl acetate (3×10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 4-b (29 mg, yield 98%). MS m/z 565.2 [M+H] ⁺ .

Compound 4-b (29 mg, 0.05 mmol) was dissolved in dichloromethane (3 mL), and 2,3-dichloro-5,6-dicyanobenzoquinone (23 mg, 0.10 mmol) was added. The reaction was heated and stirred at 50°C for 1 hour. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The white solid compound 4-c (11 mg, yield 38%) was obtained by separation and purification by preparative thin layer plate. MS m/z 563.2 [M+H] ⁺ .

Compound 4-c (8 mg, 0.01 mmol) and p-toluenesulfonyl chloride (3 mg, 0.01 mmol) were dissolved in dichloromethane (2 mL), and sodium hydrogen sulfide (1.1 mg, 0.03 mmol, 60%) was added under ice bath. The reaction solution was stirred for 0.5 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. Compound 4-d (9 mg, yield 88%) was obtained by separation and purification by preparative thin layer plate. MS m/z 717.2 [M+H] ⁺ .

Compound 4-d (9 mg, 0.01 mmol) was dissolved in tetrahydrofuran (2 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (0.2 mL, 1 M) was added dropwise. The reaction system was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and purified by preparative thin layer plate separation to obtain white solid compound 4 (4 mg, yield 56%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.52 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.02 (d, J = 0.8Hz, 1H), 7.75-7.72 (m, 2H), 7.39-7.36 (m, 2H), 5.79 (s, 2H), 4.68 (s, 1H) ),4.58(s,1H),3.85(s,3H),1.68-1.63(m,1H),1.20-1.17(m,2H),1.16-1.13(m,2H),1.08-1.04(m,2H),0.87-0.84(m,2H). MS m/z 565.1[M+H] ⁺ .

Example 5: Preparation of Compounds 6 and 7

3-(Benzyloxy)-1-cyclobutanone (3.0 g, 17.02 mmol) and triphenylphosphine (22.3 g, 85.12 mmol) were dissolved in acetonitrile (50 mL), and carbon tetrabromide (14.1 g, 42.56 mmol) was added under ice bath. The reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction system was filtered, and the filtrate was concentrated under reduced pressure. The product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a colorless oily compound 6-b (5.4 g, yield 96%).

Dissolve compound 6-b (2.1 g, 6.32 mmol) in dichloromethane (30 mL), cool to -78 ° C, and slowly drop a dichloromethane solution of boron tribromide (3.8 mL, 2.5 M). Stir the reaction at this temperature for 0.5 hours. After the reaction is completed, quench with saturated sodium bicarbonate aqueous solution, and extract the mixed solution with dichloromethane (3×20 mL). The combined organic phases are washed with saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain a dichloromethane solution containing compound 6-c, which is directly used for the next step reaction.

To the dichloromethane solution of compound 6-c, tert-butyldiphenylsilyl chloride (1.7 g, 6.32 mmol) was added, and imidazole (861 mg, 12.65 mmol) was added under ice bath. The reaction solution was stirred at room temperature for 1 hour. The mixture was quenched with saturated aqueous ammonium chloride solution, and extracted with dichloromethane (3×20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to give a colorless oily compound 6-d (2.8 g, yield 92%).

Cuprous iodide (7.9 g, 41.60 mmol) was added to tetrahydrofuran (20 mL), and a solution of methyl lithium in ether (52 mL, 1.6 M) was added dropwise under an ice bath, and the reaction mixture was stirred at this temperature for 5 minutes. A solution of compound 6-d (2.0 g, 4.16 mmol) in tetrahydrofuran (20 mL) was added dropwise. After the addition, the reaction mixture was stirred at room temperature overnight, and then iodomethane (2.9 g, 20.80 mmol) was added dropwise under an ice bath to the reaction system, and the mixture was stirred for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a colorless oily compound 6-e (940 mg, yield 64%).

Dissolve compound 6-e (610 mg, 1.74 mmol) in tetrahydrofuran (10 mL), slowly add TBAF tetrahydrofuran solution (0.9 mL, 4 M) dropwise. Stir the reaction solution at room temperature overnight. Add p-toluenesulfonyl chloride (499 mg, 2.61 mmol) and sodium hydride (209 mg, 5.22 mmol, 60%) to the reaction solution under ice bath. The reaction mixture was stirred at this temperature for 0.5 hours. After the reaction was completed, the mixture was filtered, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain a colorless oily compound 6-f (130 mg, yield 28%).

Intermediate 1-a (40 mg, 0.08 mmol) and compound 6-f (43 mg, 0.16 mmol) were dissolved in acetonitrile (1 mL), and cesium carbonate (79 mg, 0.24 mmol) was added. The reaction mixture was heated and stirred at 90 ° C overnight. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The mixture was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 6 and white solid compound 7 (4 mg, yield 8%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.53(s,1H),8.24(s,1H),7.57-7.51(m,2H),7.43-7.39(m,2H),5.79(s,2H),4.77-4.69(m,1H), 3.85(s,3H),3.11-3.04(m,2H),2.98-2.92(m,2H),1.67-1.62(m,1H),1.50(s,6H),1.07-1.04(m,2H),0.88-0.83(m,2H). MS m/z 587.3[M+H] ⁺ .

White solid compound 7 (4 mg, yield 8%). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 9.51 (s, 1H), 8.71 (s, 1H), 8.52 (s, 1H), 7.63 (s, 1H), 7.55-7.49 (m, 2H), 7.41-7.38 (m, 2H), 5.80 (s, 2H), 4.95-4.86 (m, 1H), 3.85 (s, 3H), 2.95-2.87 (m, 2H), 2.80-2.72 (m, 2H), 1.68-1.62 (m, 1H), 1.35 (s, 6H), 1.08-1.04 (m, 2H), 0.87-0.83 (m, 2H). MS m/z 587.4 [M+H] ⁺ .

Example 6: Preparation of Compounds 28 and 9

Compound 4c (25 mg, 0.04 mmol) was dissolved in dichloromethane (2 mL), and DMP oxidant (85 mg, 0.20 mmol) was added. The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the filtrate was filtered and concentrated under reduced pressure. The filtrate was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 28 (18 mg, yield 72%). MS m/z 561.2 [M+H] ⁺ .

Compound 28 (7 mg, 0.01 mmol), sodium difluorochloroacetate (5 mg, 0.03 mmol), and triphenylphosphine (8 mg, 0.03 mmol) were dissolved in DMF (1 mL). The reaction solution was heated at 90 °C overnight under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 9 (0.5 mg, yield 7%). MS m/z 595.2 [M+H] ⁺ .

Example 7: Preparation of Compounds 10A, 10B and 11

Compound 28 (6 mg, 0.01 mmol) was dissolved in methanol, and dimethyl (1-diazo-2-oxopropyl)phosphonate (4 mg, 0.02 mmol) and potassium carbonate (3 mg, 0.02 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The mixture was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 11 (4 mg, yield 67%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.52(s,1H),8.71(s,1H),8.53(s,1H),8.18(d,J＝1.2Hz,1H),7.93-7.88(m,2H),7.47-7.43(m,2H),5.80(s,2H),3.86(s,3 H),3.63(s,1H),1.70-1.64(m,1H),1.46-1.44(m,2H),1.41-1.39(m,2H),1.08-1.05(m,2H),0.87-0.85(m,2H). MS m/z 557.2[M+H] ⁺ .

Compound 11 (9 mg, 0.016 mmol) was dissolved in a hydrofluoric acid solution of pyridine (0.5 mL). The reaction solution was stirred at room temperature overnight. The reaction solution was quenched with a saturated aqueous sodium bicarbonate solution, and the mixed solution was extracted with ethyl acetate (3×5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to give white solid compound 10A (0.3 mg, yield 3%) and white solid compound 10B (1.0 mg, yield 11%).

Compound 10A: ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.16 (d, J = 1.2Hz, 1H), 7.75 -7.71(m,2H),7.41-7.39(m,2H),5.78(s,2H),4.88(dd,J＝17.4,4.2Hz,1H),4.24(dd,J＝49.8,4.2Hz,1H ),3.85(s,3H),1.67-1.63(m,1H),1.52-1.49(m,2H),1.38-1.35(m,2H),1.07-1.04(m,2H),0.87-0.85(m ,2H). MS m/z 577.1 [M+H] ⁺ .

Compound 10B: ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.05 (d, J = 1.3Hz, 1H), 7.66 -7.63(m,2H),7.44-7.41(m,2H),5.80(s,2H),5.70(br.s,1H),5.38(br.s,1H),3.85(s,3H),2.43 -2.38(m,2H),2.32-2.26(m,2H),1.67-1.63(m,1H),1.07-1.04(m,2H),0.87-0.84(m,2H). MS m/z 577.2[M+H] ⁺ .

Example 8: Preparation of Compound 12

Compound 1-a (17 mg, 0.035 mmol), compound 12-a (8 mg, 0.069 mmol) and cesium carbonate (34 mg, 0.104 mmol) were dissolved in N, N-dimethylformamide (2 mL), and the reaction mixture was stirred at 90 ° C overnight. After the reaction was completed, water was added to dilute and quench, and the mixture was extracted with ethyl acetate (3x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30: 1) to obtain a light yellow solid compound 12 (10.66 mg, yield 53%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.50 (s, 1H), 8.71 (s, 1H), 8.51 (s, 1H), 8.39 (d, J = 1.3Hz, 1H), 8.32 (d, J = 5.4Hz, 1H), 7.41 (s, 1H), 7.38–7.35 (m, 2H), 7.33–7. 27(m,3H),5.73(s,2H),3.82(s,3H),1.67–1.58(m,1H),1.07–1.02(m,2H),0.85–0.78(m,2H)ppm. MS m/z 588.2[M+H] ⁺ .

Example 9: Preparation of Compound 13

Lithium aluminum hydride (1.64 g, 43.24 mmol) was dissolved in ether (35 mL) at -5 °C, and aluminum chloride (1.92 g, 14.41 mmol) was slowly added. The reaction mixture was stirred at -5 °C for 30 minutes, and compound 13-a (1.5 g, 14.41 mmol) was slowly added dropwise. The mixed solution was stirred at -5 °C for 1 hour. After the reaction was completed, the mixture was quenched with saturated sodium sulfate solution, and the obtained mixture was filtered and washed with ether. The filtrate was concentrated under reduced pressure to obtain an ether solution of compound 13-b, which was directly used in the next step.

The ether solution of compound 13-b (theoretical yield: 1.1 g, 14.41 mmol) was dissolved in dichloromethane (10 mL), and the mixture was cooled to -20 ° C. Triethylamine (2.19 g, 21.69 mmol) and p-toluenesulfonyl chloride (3.31 g, 17.35 mmol) were added in sequence, and the reaction solution was stirred for 1 hour and slowly warmed to room temperature. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain a colorless oil compound 13-c (1.6 g, yield 48%). MS m/z 253.1 [M + Na] ⁺ .

Compound 1-a (22 mg, 0.045 mmol), compound 13-c (31 mg, 0.134 mmol) and cesium carbonate (44 mg, 0.134 mmol) were dissolved in N, N-dimethylformamide (2 mL) and stirred at room temperature for 2 hours. After the reaction was completed, water was added to dilute, and the mixed solution was extracted with ethyl acetate (3 x 5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 25: 1) to obtain white solid compound 13 (19.59 mg, yield 80%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.52 (s, 1H), 7.97 (s, 1H), 7.61 (d, J = 8.3Hz, 2H), 7.43 (d, J = 8.3Hz, 2H), 5.79 (s, 2H), 4.92 (d, J = 12.8 Hz,2H),4.83(dd,J＝17.0,3.7Hz,1H),4.54(dd,J＝50.1,3.7Hz,1H),3.85(s,3H),1.69–1.63(m,1H),1.08–1.04(m,2H),0.88–0.83(m,2H)ppm. MS m/z 551.2[M+H] ⁺ .

Example 10: Preparation of Compound 15

Compound 2 (20 mg, 0.036 mmol), 1 drop of acetic acid and palladium carbon (10%, 10 mg) were added to methanol (3 mL) at room temperature, and the reaction mixture was stirred for 15 minutes under a hydrogen atmosphere at room temperature. After the reaction was completed, the mixed solution was filtered and washed with diatomaceous earth, and the filtrate was concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 25: 1) to obtain a colorless oily crude product 15-a (15 mg, yield 75%). MS m/z 567.2 [M+H] ⁺ .

The crude product 15-a (15 mg, 0.026 mmol) was dissolved in toluene (1.5 mL), and 2,3-dichloro-5,6-dicyanobenzoquinone (5 mg) was added and stirred at 60°C for 1 hour. After the reaction was completed, the mixed solution was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 25: 1) to obtain a white solid compound 15 (1.79 mg, yield 12%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.52(s,1H),7.91(s,1H),7.59(d,J＝8.2Hz,2H),7.41(d,J＝8.2Hz,2H),5.79(s,2H),5.07–4.94(m ,1H),4.68(d,J＝7.0Hz,2H),3.85(s,3H),1.85(d,J＝17.5Hz,3H),1.70–1.60(m,1H),1.09–1.03(m,2H),0.90–0.82(m,2H)ppm. MS m/z 565.2[M+H] ⁺ .

Example 11: Preparation of Compound 16

Iodine (310 mg, 1.22 mmol) was dissolved in acetonitrile (4 mL) at room temperature, triphenylphosphine (321 mg, 1.22 mmol) was added and stirred for 2 hours. Triethylamine (124 mg, 1.22 mmol) and compound 16-a (100 mg, 1.02 mmol) were added to the reaction in sequence, and the mixture was refluxed and stirred for 3 hours. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain a light yellow solid product 16-b (109 mg, yield 51%). MS m/z 209.0 [M + H] ⁺ .

Compound 1-a (95 mg, 0.193 mmol), compound 16-b (96 mg, 0.463 mmol), methanesulfonic acid (2-dicyclohexylphosphine-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-methylamino-1,1'-biphenyl-2-yl) palladium (II) (17 mg, 0.019 mmol), 2-dicyclohexylphosphine-2',6'-diisopropyloxy-1,1'-biphenyl (18 mg, 0.039 mmol) and cesium carbonate (189 mg, 0.579 mmol) were dissolved in 1,4-dioxane (3 mL), and the system was replaced with nitrogen. The reaction mixture was stirred at 100 ° C overnight. After the reaction was completed, water was added to dilute. The mixture was extracted with ethyl acetate (3×10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol=25:1) to give a white solid product 16 (20 mg, yield 18%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.49 (d, J = 1.3Hz, 1H), 7.57 (d, J = 8.3Hz, 2H), 7.39 (d, J = 8.3Hz, 2H), 5.80 (s, 2H), 5.60 ( t,J＝1.5Hz,1H),3.84(s,3H),3.04–2.97(m,2H),2.46–2.43(m,2H),1.67–1.61(m,1H),1.08–1.02(m,2H),0.88–0.82(m,2H)ppm. MS m/z 573.1[M+H] ⁺ .

Example 12: Preparation of Compounds 29 and 22

Triethyl 2-phosphonopropyl ester (2.0 g, 8.51 mmol) was dissolved in tetrahydrofuran (20 mL), and sodium hydride (295 mg, 7.38 mmol, 60%) was slowly added under an ice bath nitrogen atmosphere. The reaction solution was stirred for 1 hour at this temperature, and then 3-(benzyloxy)-1-cyclobutanone (1.0 g, 5.67 mmol) was added. The reaction solution was stirred at room temperature overnight. After the reaction was completed, saturated ammonium chloride aqueous solution was quenched, and the mixed solution was extracted with ethyl acetate (3×20 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a colorless oily compound 29-a (1.1 g, yield 76%).

Compound 29-a (580 g, 2.23 mmol) was dissolved in dichloromethane (10 mL), cooled to -70 °C, and boron tribromide (559 mg, 3.34 mmol) was slowly added dropwise, and the reaction solution was stirred for 0.5 hours. The reaction solution was quenched with saturated sodium bicarbonate aqueous solution, and the mixed solution was extracted with dichloromethane (3×10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered, and the filtrate containing compound 29-b was directly used for the next step reaction.

To the dichloromethane solution containing compound 29-b (379 mg, 2.23 mmol), p-toluenesulfonyl chloride (468 mg, 2.45 mmol) was added, and sodium hydride (178 mg, 4.46 mmol, 60%) was slowly added under an ice bath, and the reaction solution was stirred for 0.5 hours. After the reaction was completed, saturated ammonium chloride aqueous solution was quenched, and the mixed solution was extracted with dichloromethane (3×20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a colorless oil compound 29-c (534 mg, yield 74%).

Compound 1-a (26 mg, 0.05 mmol) and 29-c (26 mg, 0.08 mmol) were dissolved in DMF (1 mL), and cesium carbonate (49 mg, 0.15 mmol) was added. The reaction mixture was heated and stirred at 90°C overnight. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The compound 29 (20 mg, yield 59%) was obtained by separation and purification on a preparative thin layer plate (ethyl acetate: petroleum ether = 1:1). MS m/z 645.3 [M+H] ⁺ .

Compound 29 (20 mg, 0.03 mmol) was dissolved in tetrahydrofuran (2 mL), and lithium aluminum hydride (3 mg, 0.06 mmol) was slowly added under ice bath. The mixture was stirred at room temperature for 1 hour. After the reaction was completed, tetrahydrofuran (10 mL) was added to dilute the mixture, and sodium sulfate decahydrate was quenched. The mixture was stirred at room temperature for 15 minutes, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude compound 22-a (10 mg, yield 53%) was directly used for the next step reaction. MS m/z 605.4 [M+H] ⁺ .

Compound 22-a (10 mg, 0.02 mmol) was dissolved in toluene (2 mL), and 2,3-dichloro-5,6-dicyanobenzoquinone (7 mg, 0.03 mmol) was added. The reaction solution was heated and stirred at 40°C for 0.5 hours. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The compound 22 (3 mg, yield 30%) was obtained by separation and purification on a preparative thin layer plate (ethyl acetate: petroleum ether = 1:1). ¹ H NMR (500 MHz, DMSO-d ₆ )δ9.51(s,1H),8.71(s,1H),8.52(s,1H),8.24(d,J＝0.8Hz,1H),7.58-7.52(m,2H),7.44-7.39(m,2H),5.79(s,2H),4.79-4.72(m,1H),4.53(t,J＝5. 5Hz,1H),3.85( d,J＝1.9Hz,3H),3.80-3.71(m,2H),3.21-3.16(m,1H),3.13-3.08(m,1H),3.03-2.96(m,2H),1.68-1.62(m,1H),1.51(s,3H),1.08-1.04(m,2H),0 .88-0.83(m,2H). MS m/z 603.2[M+H] ⁺ .

Example 13: Preparation of Compound 23

Compound 22 (17 mg, 0.03 mmol) was dissolved in dichloromethane (2 mL), thionyl chloride (18 mg, 0.15 mmol) was added dropwise, and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the obtained crude compound 23-a (17 mg, yield 97%) was directly used for the next step reaction.

Compound 23-a (17 mg, 0.03 mmol) was dissolved in acetonitrile (2 mL), and dimethylamine tetrahydrofuran solution (0.2 mL, 2 M) was added dropwise. The reaction solution was heated and stirred at 50°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 23 (3 mg, yield 17%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.52(s,1H),8.71(s,1H),8.53(s,1H),8.32(s,1H),7.58-7.53(m,2H),7.45-7.40(m,2H),5.79(s,2H),4.86-4.80(m,1H),3 .85(s,3H),3.54-3.46(m,2H),3.26-3.21(m,2H),3.16-3.10(m,2H),2.66(s,6H),1.72-1.62(m,4H),1.08-1.04(m,2H),0.89-0.85(m,2H). MS m/z 630.3[M+H] ⁺ .

Example 14: Preparation of Compound 24

Sodium iodide (1.39 g, 9.25 mmol) was dissolved in acetic acid (6 mL), and compound 24-a (600 mg, 8.81 mmol) was added. The reaction mixture was stirred at 70 ° C overnight. After the reaction was cooled to room temperature, ether (10 mL) and water (10 mL) were added in sequence. After the organic phase and the aqueous phase were separated, the aqueous phase was extracted with ether (2 x 15 mL). The combined organic phase was washed with 2M sodium hydroxide solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain yellow solid compound 24-b (860 mg, yield 50%). ¹ H NMR (500 MHz, CDCl ₃ ) δ7.83 (d, J = 15.1 Hz, 1H), 7.15 (d, J = 15.1 Hz, 1H), 2.25 (s, 3H) ppm.

Compound 1-a (35 mg, 0.071 mmol), compound 24-b (56 mg, 0.284 mmol) and cesium carbonate (69 mg, 0.213 mmol) were dissolved in N, N-dimethylformamide (2 mL) and stirred overnight at 90 ° C. After the reaction was completed, it was cooled to room temperature, diluted with water, and the mixed solution was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 35: 1) to obtain white solid compound 24 (2.9 mg, yield 7%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.52 (s, 1H), 8.71 (s, 1H), 8.67 (d, J = 1.1Hz, 1H), 8.53 (s, 1H), 7.68 (d, J = 14.4Hz, 1H), 7.64 (d, J = 8.3Hz, 2H), 7.48 (d, J = 8.3Hz, 2H), 6.89 (d, J = 14.4Hz, 1H), 5.82 (s, 2H), 3.85 (s, 3H), 2.25 (s, 3H), 1.69–1.62 (m, 1H), 1.09–1.02 (m, 2H), 0.89–0.83 (m, 2H)ppm. MS m/z 561.2[M+H] ⁺ .

Example 15: Preparation of Compounds 25A and 25B

Sodium iodide (3.0 g, 20.0 mmol) was dissolved in acetic acid (10 mL), and compound 25-a (2.0 mL, 19.7 mmol) was added. The reaction mixture was stirred at 70 ° C overnight. After the reaction was cooled to room temperature, ether (20 mL) and water (20 mL) were added in sequence. After the organic phase and the aqueous phase were separated, the aqueous phase was extracted with ether (2x 15 mL). The combined organic phase was washed with 2M sodium hydroxide solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ether = 15:1) to obtain a colorless oil compound 25-b (4.45 g, yield 100%).

Compound 1-a (30 mg, 0.061 mmol), compound 25-b (55 mg, 0.244 mmol) and cesium carbonate (60 mg, 0.183 mmol) were dissolved in N, N-dimethylformamide (2 mL) and stirred overnight at 90 ° C. After the reaction was cooled to room temperature, it was diluted with water and the mixed solution was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (petroleum ether: ether = 1: 3) to give white solid compound 25A (23 mg, yield 64%) and white solid compound 25B (3 mg, yield 8%). 25A: ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.72–8.70 (m, 2H), 8.53 (s, 1H), 7.67 (d, J = 14.2Hz, 1H), 7.61 (d, J = 8.3Hz, 2H), 7.48 (d, J = 8.3Hz, 2H), 6.69 (d . MS m/z 591.2 [M+H] ⁺ .

25B: ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.52 (s, 1H), 8.24 (d, J = 1.2Hz, 1H), 7.62 ( d,J＝8.3Hz,2H),7.43(d,J＝8.4Hz,2H),7.27(d,J＝9.3Hz,1H),6.03(d,J＝9.3Hz,1H),5.77(s, 2H), 4.01 (q, J = 7.1Hz, 2H), 3.85 (s, 3H), 1.70–1.61 (m, 1H), 1.09–1.04 (m, 5H), 0.88–0.83 (m, 2H)ppm. MS m/z 591.2[M+H] ⁺ .

Example 16: Preparation of Compound 30

Compound 30-a (125 mg, 0.991 mmol) (compound 30-a synthesis reference: Angew Chem. Int. Ed. doi: 10.1002/anie.202014308.) was dissolved in methanol (4 mL), and sodium borohydride (75 mg, 1.980 mmol) was added in batches. The reaction mixture was stirred at room temperature for 30 minutes. After the reaction was completed, water was added to quench the mixture, and the mixture was concentrated under reduced pressure. The obtained crude product was extracted with ether (3x 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product 30-b containing compound was obtained and used directly in the next reaction.

The crude compound 30-b was dissolved in dichloromethane (6 mL), and triethylamine (200 mg, 1.980 mmol) and p-toluenesulfonyl chloride (283 mg, 1.49 mmol) were added in sequence at 0°C. The mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain a yellow oily compound 30-c (15 mg, yield 5%).

Compound 1-a (2 mg, 0.004 mmol), compound 30-c (12 mg, 0.040 mmol) and cesium carbonate (4 mg, 0.012 mmol) were dissolved in N, N-dimethylformamide (1.5 mL) and stirred at 90 ° C for 2 hours. After the reaction was cooled to room temperature, it was diluted with water and the mixed solution was extracted with ethyl acetate (3 x 5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 30: 1) to obtain white solid compound 30 (0.50 mg, yield 20%). MS m/z 605.2 [M + H] ⁺ .

Example 17: Preparation of Compound 31

Compound 32 (13 mg, 0.024 mmol), sodium difluorochloroacetate (15 mg, 0.095 mmol) and triphenylphosphine (8 mg, 0.029 mmol) were dissolved in N, N-dimethylformamide (2 mL) and stirred at 100 ° C for 4 hours. After the reaction was cooled to room temperature, it was diluted with water and the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (petroleum ether: methyl tert-butyl ether = 1:5) to give a white solid compound 31 (5 mg, yield 36%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.72 (s, 1H), 8.56 (d, J = 1.0Hz, 1H), 8.53 (s, 1H), 7.58 (d, J = 8.3Hz, 2H), 7.46 (d, J = 8.4Hz, 2H), 7.09 (d, J = 13.9Hz, 1H ), 6.76(dd,J＝13.7,11.1Hz,1H),5.79(s,2H),5.70–5.61(m,1H),3.86(s,3H),1.69–1.63(m,1H),1.09–1.04(m,2H),0.89–0.85(m,2H)ppm. MS m/z 581.2[M+H] ⁺ .

Example 18: Preparation of Compound 32

Compound 25A (64 mg, 0.108 mmol) was dissolved in dichloromethane (4 mL), and a hexane solution of diisobutylaluminum hydride (DIBAL-H, 1.0 M, 0.8 mL) was slowly added dropwise at -78 °C. The reaction solution was stirred at -78 °C for 5 minutes. After the reaction was completed, a saturated sodium tartrate solution was added to quench the reaction. After warming to room temperature, the mixture was diluted with dichloromethane and stirred for 1 hour, and then extracted with dichloromethane (3 x 15 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product containing compound 32-a was directly used in the next step. MS m/z 551.2 [M + H] ⁺ .

The crude product containing compound 32-a (theoretical yield: 60 mg, 0.108 mmol) was dissolved in dichloromethane (3 mL), and Dess-Martin periodinane (DMP, 55 mg, 0.130 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction mixture was filtered and washed with diatomaceous earth. The filtrate was concentrated under reduced pressure and purified by preparative thin layer chromatography (petroleum ether: methyl tert-butyl ether = 1:4) to obtain a white solid compound 32 (20 mg, two-step yield 34%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.61 (d, J = 7.6Hz, 1H), 9.52 (s, 1H), 8.73 (d, J = 1.1Hz, 1H), 8.72 (s, 1H), 8.54 (s, 1H), 8.01 (d, J = 14.1Hz, 1H), 7.67 (d, J = 8.3Hz, 2H ),7.50(d,J＝8.3Hz,2H),6.81(dd,J＝14.1,7.6Hz,1H),5.82(s,2H),3.86(s,3H),1.70–1.63(m,1H),1.10–1.04(m,2H),0.90–0.84(m,2H)ppm. MS m/z 547.2[M+H] ⁺ .

Example 19: Preparation of Compound 34

Compound 34-a (110 mg, 0.22 mmol) (the synthesis of intermediate 34-a can be prepared by referring to the method in patent WO2020/132269Al) was dissolved in N, N-dimethylformamide (5 mL), and potassium carbonate (89 mg, 0.65 mmol) and N-phenylbis(trifluoromethanesulfonyl)imide (386 mg, 1.08 mmol) were added in sequence. After addition, the mixture was stirred at room temperature for one hour. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: ethyl acetate = 10: 1) to give a white solid compound 34-b (100 mg, yield 72%).

Compound 34-b (50 mg, 0.08 mmol), tert-butyl 3-ethynyl-1-azetidinecarboxylate (21 mg, 0.12 mmol), cuprous iodide (2 mg, 0.01 mmol), bistriphenylphosphine palladium dichloride (6 mg, 0.008 mmol), Pyridine (12 mg, 0.16 mmol) was dissolved in N, N-dimethylformamide (0.5 mL), and the reaction mixture was placed in a sealed tube and heated to 65 ° C under a nitrogen atmosphere and stirred for 3 hours. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain a yellow solid compound 34-c (40 mg, yield 76%).

Compound 34-c (40 mg, 0.06 mmol) was dissolved in methanol (1 mL), and a hydrochloric acid dioxane solution (4.0 M, 0.5 mL) was added. The reaction mixture was stirred at 30 ° C for 2 hours. After the reaction was completed by TLC monitoring, the reaction system was cooled to room temperature and concentrated under reduced pressure. The resulting mixture was dissolved in a small amount of methanol, neutralized with ammonia water, and then concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to give a white solid compound 34 (20 mg, yield 59%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 7.74-7.69 (m, 2H), 7.48 (d, J = 8.6Hz, 2H), 7.28 (s, 1H), 5.80 (s, 2H), 3.85 (s, 3H), 3. 65-3.60(m,1H),3.60-3.55(m,2H),3.46(br.,3H),1.68-1.60(m,1H),1.08-1.02(m,2H),0.87-0.83(m,2H). MS m/z 572.3[M+H] ⁺ .

Example 20: Preparation of Compound 35

Compound 34 (10 mg, 0.02 mmol) and paraformaldehyde (3 mg, 0.10 mmol) were dissolved in methanol (0.5 mL), and then acetic acid (1 drop) was added dropwise. The reaction was stirred at room temperature for half an hour, and sodium cyanoborohydride (11 mg, 0.05 mmol) was added. The reaction mixture was stirred at room temperature for 12 hours. After the reaction was completed by TLC monitoring, the reaction mixture was filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain white solid compound 35 (2.60 mg, yield 25%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.52(s,1H),7.73-7.69(m,2H),7.51-7.47(m,2H),7.27(s,1H),5.81(s,2H),3.85(s,3H),3.41(t ,J＝7.2Hz,2H),2.86(t,J＝6.7Hz,2H),2.08(s,3H),2.02-1.95(m,1H),1.68-1.61(m,1H),1.08-1.03(m,2H),0.87-0.83(m,2H). MS m/z 586.3[M+H] ⁺ .

Example 21: Preparation of Compound 36

Compound 24 (25 mg, 0.045 mmol) was dissolved in tetrahydrofuran (3 mL), and methyl lithium ether solution (1.6 M, 0.03 mL) was slowly added dropwise at -78 ° C. The reaction system was stirred at -78 ° C for 1 hour. After the reaction was completed, saturated ammonium chloride solution was added dropwise to quench. The mixed solution was extracted with ethyl acetate (3 x 10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 30: 1) to give white solid compound 36 (3.82 mg, yield 15%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.52 (s, 1H), 8.31 (d, J = 1.1Hz, 1H), 7.60 (d, J = 8.3Hz, 2H), 7.44 (d, J = 8.4Hz, 2H), 6.86 (d, J = 14.0Hz, 1H ), 6.37(d,J＝14.0Hz,1H),5.79(s,2H),4.85(s,1H),3.85(s,3H),1.69–1.62(m,1H),1.22(s,6H),1.08–1.02(m,2H),0.91–0.83(m,2H)ppm. MS m/z 577.2[M+H] ⁺ .

Example 22: Preparation of Compound 37

Compound 1-a (50 mg, 0.101 mmol), iodobenzene (41 mg, 0.203 mmol), cuprous iodide (2 mg, 0.010 mmol), 8-hydroxyquinoline (2 mg, 0.010 mmol) and potassium carbonate (42 mg, 0.305 mmol) were dissolved in dimethyl sulfoxide (2.5 mL). The reaction mixture was stirred overnight at 100 ° C under a nitrogen atmosphere. After the reaction was cooled to room temperature, it was diluted with water. The mixed solution was extracted with ethyl acetate (3×10 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (petroleum ether: ether = 1:4) to obtain a white solid product 37 (2.4 mg, yield 4%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.48(s,1H),8.71(s,1H),8.48(s,1H),8.20(s,1H),7.50–7.46(m,3H),7.40–7.36(m,2H),7.28(d,J＝8.3Hz,2H),7.22(d,J＝ 8.2Hz,2H),5.68(s,2H),3.80(s,3H),1.67–1.58(m,1H),1.07–1.02(m,2H),0.83–0.78(m,2H)ppm. MS m/z 569.3[M+H] ⁺ .

Example 23: Preparation of Compound 38

Compound 1-a (50 mg, 0.101 mmol), potassium hydroxide (40 mg, 0.711 mmol), potassium carbonate (35 mg, 0.254 mmol) and tetrabutylammonium bromide (3 mg, 0.010 mmol) were dissolved in 1,2-dichloroethane (3 mL), and the reaction mixture was stirred at 50 ° C overnight. After the reaction was completed, water was added to dilute. The mixture was extracted with dichloromethane (3×15 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain a yellow oily crude product 38-a. MS m/z 555.0 [M+H] ⁺ .

The crude product 38-a (theoretical yield: 56 mg, 0.101 mmol) and potassium hydroxide (40 mg, 0.711 mmol) were dissolved in methanol (3 mL), and the reaction mixture was stirred at reflux temperature for 1.5 hours. After the reaction was completed, it was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30: 1) to obtain a white solid product 38 (48 mg, two-step yield 91%). ¹ H NMR(500MHz,DMSO-d ₆ )δ9.51(s,1H),8.71(s,1H),8.52(s,1H),8.44(d,J＝1.0Hz,1H),7.58(d,J＝8.3Hz,2H),7.45(d,J＝8.3Hz,2H),7.00(dd,J＝15.5,8.7Hz,1H),5.79(s,2H),5.74(dd,J＝15.4,1.4Hz,1H),5.12(dd,J＝8.7,1.4Hz,1H),3.85(s,3H),1.69–1.61(m,1H),1.08–1.02(m,2H),0.88–0.83(m,2H)ppm。 MS m/z 519.2 [M+H] ⁺ .

Example 24: Preparation of Compound 39

Compound 34-b (35 mg, 0.05 mmol), tetrahydrofuran solution of propyne (0.2 mL, 1 M), cuprous iodide (2 mg, 0.10 mmol), bistriphenylphosphine palladium dichloride (4 mg, 0.005 mmol), pyridine (12 mg, 0.15 mmol) were dissolved in N, N-dimethylformamide (0.5 mL), and the reaction mixture was placed in a sealed tube and heated to 65 ° C under a nitrogen atmosphere and stirred for 3 hours. TLC monitored the completion of the reaction, and after the reaction mixture was cooled to room temperature, it was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to obtain yellow solid compound 39 (4.13 mg, yield 14%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.52(s,1H),8.71(s,1H),8.53(s,1H),7.70(d,J＝8.2Hz,2H),7.47(d,J＝8.2Hz,2H),7.24(s,1H),5.80(s,2H),3.85(s,3H), 2.05(s,3H),1.65(br,1H),1.10-1.02(m,2H),0.92-0.75(m,2H). MS m/z 531.1[M+H] ⁺ .

Example 25: Preparation of Compound 40

Compound 34-b (20 mg, 0.03 mmol), 4-ethynyl-2-fluoropyridine (6 mg, 0.05 mmol), cuprous iodide (1 mg, 0.06 mmol), bistriphenylphosphine palladium dichloride (2 mg, 0.003 mmol), pyridine (7 mg, 0.09 mmol) were dissolved in N, N-dimethylformamide (0.5 mL), and the reaction mixture was placed in a sealed tube and heated to 65 ° C under a nitrogen atmosphere and stirred for 3 hours. After the reaction was completed by TLC monitoring, the reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to give a white solid compound 40 (4.03 mg, yield 21%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.69 (s, 1H), 8.53 (s, 1H), 8.31 (d, J = 5.1Hz, 1H), 7.82-7.78 (m, 2H), 7.59 (s, 1H), 7.55-7.49 (m, 2H), 7.39-7.37 (m,1H),7.31(br,1H),5.82(s,2H),3.82(s,3H),1.61(m,1H),1.04-0.99(m,2H),0.82-0.77(m,2H). MS m/z 612.1[M+H] ⁺ .

Example 26: Preparation of Compound 41

Compound 1-a (30 mg, 0.061 mmol), compound 41-a (40 mg, 0.152 mmol) and cesium carbonate (60 mg, 0.183 mmol) were dissolved in N, N-dimethylformamide (2 mL) and stirred at room temperature for 3.5 hours. After the reaction was completed, water was added to dilute the mixture, and the mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 30: 1) to give a white solid compound 41 (2.85 mg, yield 8%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.51(s,1H),7.99(d,J＝1.0Hz,1H),7.56(d,J＝8.3Hz,2H),7.41(d,J＝8.3Hz,2H),5.77(s,2H),3.85 (s,3H),3.63(s,3H),1.69–1.61(m,1H),1.09–1.01(m,4H),0.89–0.80(m,4H)ppm. MS m/z 591.2[M+H] ⁺ .

Example 27: Preparation of Compound 42

Compound 38 (36 mg, 0.069 mmol), 4-iodopyridine (43 mg, 0.208 mmol), palladium acetate (2 mg, 0.007 mmol) and tri(o-methylphenyl)phosphine (4 mg, 0.014 mmol) were dissolved in N,N-dimethylformamide (2 mL), and triethylamine (21 mg, 0.208 mmol) was added. The reaction solution was stirred at 100 ° C overnight. After the reaction was completed, it was cooled to room temperature, diluted with water to quench, and the mixed solution was extracted with ethyl acetate (3x 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 30: 1) to give a white solid compound 42 (3.39 mg, yield 8%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.56–8.53 (m, 3H), 8.53 (s, 1H), 7.78 (d, J = 14.4Hz, 1H), 7.67 (d, J = 8.3Hz, 2H), 7.50–7.44 (m, 4H), 7 .23(d,J＝14.4Hz,1H),5.80(s,2H),3.83(s,3H),1.71–1.61(m,1H),1.07–1.01(m,2H),0.85–0.81(m,2H)ppm. MS m/z 596.2[M+H] ⁺ .

Example 28: Preparation of Compound 43

Compound 29-b (209 mg, 1.23 mmol) was dissolved in DMF (3 mL), and tert-butyldiphenylsilyl chloride (375 mg, 1.35 mmol) and imidazole (251 mg, 3.69 mmol) were added respectively. The reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a colorless oil compound 43-a (290 mg, yield 58%).

Compound 43-a (290 mg, 0.71 mmol) was dissolved in tetrahydrofuran (5 mL), and lithium aluminum hydride (54 mg, 1.42 mmol) was slowly added under ice bath. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was diluted with tetrahydrofuran (20 mL), quenched with sodium sulfate decahydrate, and stirred at room temperature for 15 minutes, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude compound 43-b (243 mg, yield 96%) was directly used for the next step reaction.

Compound 43-b (243 mg, 0.66 mmol) was dissolved in tetrahydrofuran (5 mL) solution, and sodium hydride (79 mg, 1.98 mmol, 60%) was slowly added under ice bath. The reaction mixture was stirred for 0.5 hours, and then iodomethane (469 mg, 3.30 mmol) was added dropwise, and the reaction solution was heated and stirred at 50 ° C overnight. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure, and separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a colorless oil compound 43-c (133 mg, yield 53%).

Compound 43-c (81 mg, 0.21 mmol) was dissolved in tetrahydrofuran (3 mL), and tetrabutylammonium fluoride tetrahydrofuran solution (0.1 mL, 4 M) was added dropwise. The reaction solution was stirred at room temperature for 1 hour. Then, p-toluenesulfonyl chloride (60 mg, 0.32 mmol) and sodium hydride (17 mg, 0.42 mmol, 60%) were added to the reaction solution under an ice bath. The reaction mixture was stirred for 0.5 hours under an ice bath. After the reaction was completed, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the colorless oil compound 43-d (44 mg, yield 70%) was obtained by separation and purification by silica gel column chromatography.

Compound 1-a (21 mg, 0.04 mmol) and compound 43-d (38 mg, 0.13 mmol) were dissolved in acetonitrile (1 mL), and cesium carbonate (28 mg, 0.09 mmol) was added. The reaction mixture was heated and stirred at 90 ° C overnight. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The mixture was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 43 (5 mg, yield 19%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.28 (d, J = 1.0Hz, 1H), 7.55 (d, J = 8.3Hz, 2H), 7.41 (d, J = 8.3Hz, 2H), 5.79 (s, 2H), 4.82-4 .74(m,1H),3.85(s,3H),3.69(s,2H),3.23-3.11(m,5H),3.07-3.01(m,2H),1.68-1.62(m,1H),1.50(s,3H),1.08-1.04(m,2H),0.87-0.84(m,2H). MS m/z 617.2 [M+H] ⁺ .

Example 29: Preparation of Compound 44

Compound 34-b (20 mg, 0.03 mmol), 4-ethynyl-2-fluoropyridine (6 mg, 0.05 mmol), cuprous iodide (1 mg, 0.06 mmol), bistriphenylphosphine palladium dichloride (2 mg, 0.003 mmol), pyridine (7 mg, 0.09 mmol) were dissolved in N, N-dimethylformamide (0.5 mL), and the reaction mixture was placed in a sealed tube and heated to 65 ° C under a nitrogen atmosphere and stirred for 3 hours. TLC monitored the completion of the reaction, and after the reaction mixture was cooled to room temperature, it was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to give a white solid compound 44 (3.92 mg, yield 22%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.53(s,1H),7.73(d,J＝8.6Hz,2H),7.48(d,J＝8.6Hz,2H),7.33(s,1H),5.81(s,2H),4.75-4.69(m ,2H),4.50-4.44(m,2H),4.16-4.10(m,1H),3.85(s,3H),1.64(m,1H),1.08-1.02(m,2H),0.87-0.83(m,2H). MS m/z 573.1[M+H] ⁺ .

Example 30: Preparation of Compound 45

Compound 34 (5 mg, 0.01 mmol), acetic acid (1 mg, 0.02 mmol), N,N-diisopropylethylamine (4 mg, 0.03 mmol) were dissolved in acetonitrile (0.5 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11 mg, 0.03 mmol) was added. After the addition, the system was reacted at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated and purified by preparative silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain a white solid compound 45 (2.67 mg, yield 50%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 7.73 (d, J = 8.5Hz, 2H), 7.47 (d, J = 8.5Hz, 2H), 7.34 (s, 1H), 5.80 (s, 2H), 4.35 (t, J = 8.2Hz ,1H),4.09(t,J=8.4Hz,1H),4.04-4.00(m,1H),3.85(s,3H),3.75-3.67(m,2H),1.71(s,3H),1.67-1.62(m,1H),1.08-1.03(m,2H),0.87-0.83(m,2H ). MS m/z 614.2 [M+H] ⁺ .

Example 31: Preparation of Compound 46

Compound 34-b (4 mg, 0.006 mmol), 3-ethynylpyridine (1.93 mg, 0.018 mmol), cuprous iodide (1 mg, 0.006 mmol), bistriphenylphosphine palladium dichloride (2 mg, 0.003 mmol), pyridine (2 mg, 0.03 mmol) were dissolved in N, N-dimethylformamide (0.5 mL), and the reaction mixture was placed in a sealed tube and heated to 65 ° C under a nitrogen atmosphere and stirred for 3 hours. TLC monitored the completion of the reaction. After the reaction mixture was cooled to room temperature, it was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1, 2% ammonia water) to give a white solid compound 46 (2 mg, yield 54%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.69(s,1H),8.66-8.63(m,2H),8.53(s,1H),7.91-7.87(m,1H),7.83-7.79(m,2H),7.54-7.49(m,3H),7.49-7 .44(m,1H),5.82(s,2H),3.81(s,3H),1.65-1.57(m,1H),1.04-0.99(m,2H),0.88-0.82(m,2H). MS m/z 594.1[M+H] ⁺ .

Example 32: Preparation of Compound 47

Compound 34-b (100 mg, 0.156 mmol), trimethylethynylsilane (46 mg, 0.468 mmol), cuprous iodide (3 mg, 0.016 mmol), bistriphenylphosphine palladium dichloride (11 mg, 0.016 mmol), pyridine (36 mg, 0.468 mmol) were dissolved in N, N-dimethylformamide (0.5 mL), the reaction mixture was placed in a sealed tube, heated to 65 ° C under nitrogen atmosphere and stirred for 3 hours. TLC monitored the completion of the reaction. After the reaction mixture was cooled to room temperature, it was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 30: 1) to obtain compound 47-a (80 mg, yield 87%).

Compound 47-a (20 mg, 0.034 mmol), 3-iodothiophene (14 mg, 0.068 mmol), bistriphenylphosphine palladium dichloride (2 mg, 0.003 mmol), cesium carbonate (33 mg, 0.10 mmol) were dissolved in N, N-dimethylformamide (0.5 mL), the reaction mixture was placed in a sealed tube, heated to 80 ° C under a nitrogen atmosphere and stirred for 0.5 hours. TLC monitored the completion of the reaction. After the reaction mixture was cooled to room temperature, it was filtered and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to obtain compound 47 (5.72 mg, yield 28%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.70(s,1H),8.53(s,1H),7.93(dd,J＝2.9,1.2Hz,1H),7.80-7.75(m,2H),7.68-7.64(m,1H),7.53-7.49(m,2H) ,7.42(s,1H),7.15(dd,J=5.0,1.2Hz,1H),5.81(s,2H),3.82(s,3H),1.65-1.59(m,1H),1.05-0.99(m,2H),0.90-0.82(m,2H). MS m/z 599.0[M+H] ⁺ .

Example 33: Preparation of Compound 48

Compound 47-a (15 mg, 0.03 mmol) and compound 48-a (17 mg, 0.06 mmol) (synthesis of 48-a according to the method in the article DOI: 10.1039/d0sc02213f) were dissolved in tetrahydrofuran (1.0 mL), and triethylamine (9 mg, 0.09 mmol), cuprous chloride (2 mg, 0.02 mmol), copper acetylacetonate (5 mg, 0.02 mmol) and cesium fluoride (23 mg, 0.15 mmol) were added, and the mixture was irradiated with a blue light lamp under a nitrogen atmosphere and stirred at room temperature overnight. The reaction system was filtered, and the organic phase was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (ethyl acetate 100%) to obtain white solid compound 48 (2.5 mg, yield 16%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.52(s,1H),7.75-7.66(m,2H),7.52-7.43(m,2H),7.30(s,1H),5.80(s,2H),3.85(s,3H),3.28- 3.21(m,1H),3.03-2.89(m,2H),2.63-2.52(m,2H),1.68-1.61(m,1H),1.09-1.00(m,2H),0.88-0.80(m,2H). MS m/z 607.1[M+H] ⁺ .

Example 34: Preparation of Compound 49

Compound 1-a (92 mg, 0.19 mmol), (2-bromoethynyl)triisopropylsilane (247 mg, 0.95 mmol), 2-(2-pyridine)-benzimidazole (7 mg, 0.04 mmol), cuprous iodide (4 mg, 0.02 mmol), cesium carbonate (124 mg, 0.38 mmol) were dissolved in dioxane (2 mL), and the reaction mixture was heated and stirred at 100 ° C overnight under a nitrogen atmosphere. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:1) to obtain a white solid compound 49-a (52 mg, yield 41%). MS m/z 673.3 [M+H] ⁺ .

Compound 49-a (30 mg, 0.04 mmol) was dissolved in DMF (2 mL), and cesium fluoride (33 mg, 0.22 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 49 (14 mg, yield 61%). MS m/z 517.1 [M+H] ⁺ .

Example 35: Preparation of Compound 50

Compound 49-a (22 mg, 0.03 mmol) was dissolved in tetrahydrofuran (2 mL), tetrabutylammonium fluoride (1 M, 0.10 mL) was added dropwise, and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the obtained crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 50 (1 mg, yield 7%). ¹ H NMR (500MHz, CD ₃ OD) δ9.40 (s, 1H), 8.64 (s, 1H), 8.41 (s, 1H), 7.93 (s, 1H), 7.66-7.63 (m, 2H), 7.55-7.51 (m, 2H), 6.86 (dd, J＝76.6, 3.9Hz, 1H), 6.48 (dd,J=29.9,3.9Hz,1H),5.81(s,2H),3.92(s,3H),1.70-1.65(m,1H),1.18-1.14(m,2H),0.92-0.88(m,2H). MS m/z 537.1[M+H] ⁺ .

Example 35: Preparation of Compound 51

Compound 49 (6 mg, 0.01 mmol) was dissolved in tetrahydrofuran (1 mL), and acetone (1 mg, 0.02 mmol) and sodium amide (1 mg, 0.03 mmol) were added. The reaction solution was stirred at room temperature for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 51 (0.7 mg, yield 10%). ¹ H NMR (500MHz, CD ₃ OD) δ9.40(s,1H),8.64(s,1H),8.42(s,1H),8.00-7.96(m,3H),7.53-7.49(m,2H),5.81(s,2H),3.91(s,3H),1.68-1.65(m,1H), 1.48(s,6H),1.17-1.14(m,2H),0.92-0.89(m,2H). MS m/z 575.1[M+H] ⁺ .

Example 36: Preparation of Compound 52

Anhydrous chromium chloride (1.66 g, 13.50 mmol) was dissolved in tetrahydrofuran (20 mL). A solution of tert-butyl 3-formylazetidine-1-carboxylate (500 mg, 2.70 mmol) and iodoform (2.13 g, 5.40 mmol) in tetrahydrofuran (10 mL) was added dropwise under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the colorless oily compound 52-a (226 mg, yield 27%) was obtained by separation and purification by silica gel column chromatography.

Compound 1-a (50 mg, 0.10 mmol), compound 52-a (63 mg, 0.20 mmol), 2-(2-pyridine)-benzimidazole (4 mg, 0.02 mmol), cuprous iodide (2 mg, 0.01 mmol), cesium carbonate (65 mg, 0.20 mmol) were dissolved in dioxane (2 mL), and the reaction mixture was heated and stirred at 100 ° C overnight under a nitrogen atmosphere. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain white solid compound 52-b (10 mg, yield 15%). MS m/z 674.3 [M+H] ⁺ .

Compound 52-b (10 mg, 0.01 mmol) was dissolved in dichloromethane (2 mL), and a solution of hydrogen chloride in dioxane (4 M, 0.2 mL) was added dropwise under ice bath. The reaction solution was stirred at room temperature for 1 hour. The pH of the reaction solution was adjusted to 8-10 with aqueous ammonia, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate to obtain white solid compound 52 (7 mg, yield 82%). MS m/z 574.1 [M+H] ⁺ .

Example 37: Preparation of Compound 53

Compound 52 (6 mg, 0.01 mmol) was dissolved in methanol (1 mL), and paraformaldehyde (1 mg, 0.03 mmol), anhydrous zinc chloride (3 mg, 0.02 mmol), and sodium cyanoborohydride (2 mg, 0.03 mmol) were added. The reaction mixture was stirred at room temperature overnight, and the reaction solution was quenched with water. The mixed solution was extracted with ethyl acetate (3×5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate to obtain white solid compound 53 (3 mg, yield 49%). ¹ H NMR (500 MHz, DMSO-d ₆ )δ9.52(s,1H),8.72(s,1H),8.53(s,1H),8.34(s,1H),7.63-7.57(m,2H),7.46-7.41(m,2H),6.85(d,J＝13.9Hz,1H),6.40(dd,J＝13.9,8.9Hz,1H),5. 79(s,2H),3.86(s,3H),3.59-3.53(m,2H),3.27-3.20(m,1H),3.14-3.08(m,2H),2.32(s,3H),1.69-1.63(m,1H),1.09-1.05(m,2H),0.89-0.84( m,2H). MS m/z 588.2 [M+H] ⁺ .

Example 38: Preparation of Compound 54

Compound 1-a (35 mg, 0.07 mmol), compound 54-a (47 mg, 0.213 mmol), 2-dicyclohexylphosphine-2',6'-diisopropyl-1,1'-biphenyl (6 mg, 0.014 mmol), (2-dicyclohexylphosphine-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-methylamino-1,1'-biphenyl-2-yl) palladium (II) methanesulfonate (6 mg, 0.007 mmol), cesium carbonate (69 mg, 0.213 mmol) were dissolved in 1,4-dioxane solution (1 mL), and the reaction mixture was placed in a sealed tube and heated to 100 ° C under nitrogen atmosphere and stirred for 3 hours. TLC monitored the completion of the reaction. After the reaction mixture was cooled to room temperature, it was filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol=25:1) to give compound 54 (31 mg, yield 75%). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ9.52 (s, 1H), 8.71 (s, 1H), 8.53 (s, 1H), 8.29 (d, J = 1.0 Hz, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.37 (d, J = 8.3 Hz, 2H), 5.93 (s, 1H), 5.77 (s, 2H), 3.84 (s, 3H), 1.69-1.61 (m, 2H), 1.10 (d, J = 6.0 Hz, 1H), 1.07-1.02 (m, 3H), 0.94 (d, J = 6.0 Hz, 1H), 0.88-0.82 (m, 3H). The solvent peak contains one hydrogen. MS m/z 587.2 [M+H] ⁺ .

Example 39: Preparation of Compounds 55 and 56

2-Fluoropyridine-4-carboxaldehyde (100 mg, 0.80 mmol) was dissolved in methanol (3 mL). Dimethyl (1-diazo-2-oxopropyl)phosphonate (230 mg, 1.20 mmol) was added dropwise, followed by potassium carbonate (21 mg, 1.60 mmol). The reaction mixture was stirred at room temperature for 3 hours. The reaction solution was quenched with water, and the mixed solution was extracted with ethyl acetate (3×10 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:20) to give white solid compound 55-a (23 mg, yield 22%).

Compound 1-a (50 mg, 0.10 mmol), compound 55-a (23 mg, 0.19 mmol), and cesium carbonate (65 mg, 0.20 mmol) were dissolved in acetonitrile (2 mL). The reaction mixture was heated and stirred at 90 ° C overnight. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (methanol: dichloromethane = 1:20) to obtain compound 55 (11 mg, yield 21%) and compound 56 (3 mg, yield 6%).

Compound 55: ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.55-8.54 (m, 1H), 8.53 (s, 1H), 8.21 (d, J＝5.2Hz,1H),7.89(d,J＝14.4Hz,1H),7.69-7.65(m,2H),7.50-7.46(m,2H),7.45-7.43(m,1H),7.32(s ,1H),7.29(d,J＝14.4Hz,1H),5.80(s,2H),3.83(s,3H),1.68-1.63(m,1H),1.06-1.03(m,2H),0.86- 0.82(m,2H). MS m/z 614.0 [M+H] ⁺ .

Compound 56: ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 8.71 (s, 1H), 8.52 (s, 1H), 8.01 (d, J = 5.2Hz, 1H), 7.92 -7.91(m,1H),7.60-7.56(m,2H),7.37(m,2H),7.27(d,J＝8.8Hz,1H),6.74(d,J＝8.8Hz,1H),6.68- 6.65(m,1H),6.54(s,1H),5.74(s,2H),3.84(s,3H),1.67-1.63(m,1H),1.07-1.04(m,2H),0.87-0.84( m,2H). MS m/z 614.0 [M+H] ⁺ .

Example 40: Preparation of Compound 57

Compound 1-a (50 mg, 0.101 mmol) and compound 3-ethynylpyridine (21 mg, 0.203 mmol) were dissolved in acetonitrile (1 mL), and cesium carbonate (99 mg, 0.304 mmol) was added, and the mixture was heated to 80°C for 14 hours. After the reaction was completed, water was added to the reaction solution and then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by thin layer chromatography on silica gel plate (dichloromethane: methanol = 25: 1) to obtain compound 57 (3.42 mg, yield 5.6%). ¹ H NMR (500 MHz, DMSO-d ₆ )δ9.50(s,1H),8.71(s,1H),8.51(s,1H),8.38(dd,J＝4.7,1.5Hz,1H),8.08(d,J＝2.0Hz,1H),7.88(s,1H),7.65(d,J＝8.3Hz,2H),7.34(d,J＝8.3Hz,2H), 7.22-7.19(m,1H),7.17(d,J＝8.8Hz,1H),7.10(d,J＝8.0Hz,1H),6.75(d,J＝8.8Hz,1H),5.73(s,2H),3.83(s,3H),1.67-1.61(m,1H),1.07-1.01(m,2 H),0.87-0.81(m,3H). MS m/z 596.1 [M+H] ⁺ .

Example 41: Preparation of Compounds 58 and 63

Compound 1-a (50 mg, 0.10 mmol), 2-ethynylpyridine (21 mg, 0.19 mmol), and cesium carbonate (65 mg, 0.20 mmol) were dissolved in acetonitrile (2 mL). The reaction mixture was heated and stirred at 90 °C overnight. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (methanol: dichloromethane = 1:20) to obtain compound 63 (2 mg, yield 3%). ¹ H NMR (500 MHz, DMSO-d ₆ )δ9.52(s,1H),8.71(s,1H),8.63(s,1H),8.53(s,1H),8.51-8.49(m,1H),7.86(d,J＝14.0Hz,1H),7.82-7.78(m,1H),7.64(d,J＝8.2Hz,2H),7.48(d, J＝8.2Hz,2H),7.38(d,J＝7.8Hz,1H),7.33(d,J＝14.0Hz,1H),7.30-7.27(m,1H),5.82(s,2H),3.84(s,3H),1.68-1.64(m,1H),1.06-1.02(m,2H),0.87 -0.82(m,2H). MS m/z 596.0 [M+H] ⁺ .

Compound 58 (9 mg, yield 15%). ¹ H NMR (500 MHz, DMSO-d ₆ )δ9.50(s,1H),8.71(s,1H),8.51(s,1H),8.39-8.37(m,1H),7.87-7.86(m,1H),7.66(dd,J＝8.1,1.7Hz,3H),7.35(d,J＝8.3Hz,2H),7.22-7.19(m,1H) ,7.08(d,J＝9.1Hz,1H),7.03(d,J＝7.9Hz,1H),6.70(d,J＝9.1Hz,1H),5.73(s,2H),3.83(s,3H),1.67-1.61(m,1H),1.07-1.03(m,2H),0.86-0.82(m, 2H). MS m/z 596.0 [M+H] ⁺ .

Example 42: Preparation of Compounds 59 and 60

Compound 1-a (50 mg, 0.101 mmol) and compound 5-ethynylpyrimidine (21 mg, 0.203 mmol) were dissolved in acetonitrile (1 mL), and cesium carbonate (99 mg, 0.304 mmol) was added, and the mixture was heated to 80°C for 14 hours. After the reaction was completed, water was added to the reaction solution and then extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by thin layer chromatography on silica gel plates (dichloromethane: methanol = 25:1) to obtain compound 59 (2 mg, yield 3.3%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51 (s, 1H), 9.10 (s, 1H), 8.95 (s, 2H), 8.71 (s, 1H), 8.54-8.51 (m, 2H), 7.82 (d, J = 14.5Hz, 1H), 7.69 (d, J = 8.2Hz, 2H), 7.47 (d ,J＝8.2Hz,2H),7.24(d,J＝14.5Hz,1H),5.80(s,2H),3.83(s,3H),1.68-1.63(m,1H),1.07-1.02(m,2H),0.88-0.81(m,2H). MS m/z 597.0[M+H] ⁺ .

Compound 60 (5.86 mg, yield 9.7%): ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.50 (s, 1H), 8.98 (s, 1H), 8.71 (s, 1H), 8.51 (s, 1H),8.23(s,2H),7.97(d,J＝0.9Hz,1H),7.64(d,J＝8.3Hz,2H),7.36(d,J＝8.3Hz,2H),7.25(d, J＝8.8Hz,1H),6.73(d,J＝8.8Hz,1H),5.74(s,2H),3.85(s,3H),1.67-1.61(m,1H),1.08-1.02(m,2H ),0.87-0.81(m,2H). MS m/z 597.0 [M+H] ⁺ .

Example 43: Preparation of Compounds 61 and 62

Anhydrous chromium chloride (864 mg, 7.03 mmol) was dissolved in tetrahydrofuran (10 mL). A solution of 1-tert-butyloxycarbonylpiperidine-4-carboxaldehyde (500 mg, 2.34 mmol) and iodoform (1.85 g, 4.69 mmol) in tetrahydrofuran (10 mL) was added dropwise under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the colorless oily compound 61-a (210 mg, yield 26%) was obtained by separation and purification by silica gel column chromatography.

Compound 1-a (100 mg, 0.203 mmol), compound 61-a (137 mg, 0.41 mmol), 2-(2-pyridine)-benzimidazole (8 mg, 0.041 mmol), cuprous iodide (4 mg, 0.02 mmol), cesium carbonate (132 mg, 0.41 mmol) were dissolved in dioxane (2 mL), and the reaction mixture was heated and stirred at 100 ° C overnight under a nitrogen atmosphere. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography to obtain white solid compound 61-b (22 mg, yield 15%). MS m/z 702.3 [M+H] ⁺ .

Compound 61-b (22 mg, 0.03 mmol) was dissolved in dichloromethane (2 mL), and a solution of hydrogen chloride in dioxane (4 M, 0.2 mL) was added dropwise under ice bath. The reaction solution was stirred at room temperature for 1 hour. The pH of the reaction solution was adjusted to 8-10 with aqueous ammonia, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate to obtain white solid compound 62 (10 mg, yield 53%). MS m/z 602.2 [M+H] ⁺ .

Compound 62 (10 mg, 0.016 mmol) was dissolved in methanol (1 mL), paraformaldehyde (1.5 mg, 0.048 mmol) was added, and acetic acid (1 drop) was added. After stirring at room temperature for 30 minutes, sodium cyanoborohydride (2 mg, 0.032 mmol) was added. The reaction mixture was stirred at room temperature overnight, the reaction solution was quenched with water, and the mixed solution was extracted with ethyl acetate (3×5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate to obtain white solid compound 61 (1.89 mg, yield 18%). ¹ H NMR (500 MHz, DMSO-d ₆ )δ9.51(s,1H),8.72(s,1H),8.52(s,1H),8.28(s,1H),7.61(d,J＝8.3Hz,2H),7.44(d,J＝8.3Hz,2H),6.77(d,J＝14.0Hz,1H),6.17(dd,J＝14.1,7.4Hz,1H ),5.78(s,2H),3.86(s,3H),2.81-2.71(m,2H),2.18(s,3H),2.13-2.04(m,2H),2.02-1.85(m,3H),1.69-1.59(m,3H),1.10-1.03(m,2H),0.90-0 .78(m,2H). MS m/z 616.0 [M+H] ⁺ .

Example 44: Preparation of Compound 64

Compound 64-a (100 mg, 0.55 mmol) was dissolved in toluene (2.0 mL), and azobisisobutyronitrile (25 mg, 0.15 mmol) and tributyltin (1 mL, 3.7 mmol) were added. The mixture was stirred at 90 degrees for 2 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain a yellow oil 64-b (250.0 mg, yield 97%).

Compound 64-b (250 mg, 0.53 mmol) was dissolved in dichloromethane (10 mL), and iodine (147 mg, 0.58 mmol) was added and stirred at room temperature for 1 hour. The reaction solution was washed with potassium fluoride aqueous solution and sodium thiosulfate aqueous solution, extracted with dichloromethane, and the organic phase was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain a light yellow solid compound 64-c (150.0 mg, yield 91%). MS m/z 334.1 [M + Na] +.

Compound 64-c (60 mg, 0.19 mmol) and compound 1-a (30 mg, 0.06 mmol) were dissolved in DMSO (2.0 mL), and cuprous oxide (3 mg, 0.02 mmol) and N ₁ , N ₂ -di(furan-2-ylmethyl)oxyaldehyde amide (5 mg, 0.02 mmol) were added, and stirred overnight at 100 degrees under a nitrogen atmosphere. The reaction system was cooled to room temperature, quenched with water, extracted with ethyl acetate, and the organic phase was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer chromatography (ethyl acetate 100%) to obtain yellow solid compound 64-d (20 mg, yield 49%). MS m/z 676.3[M+H] ⁺ .

Compound 64-d (20 mg, 0.03 mmol) was dissolved in dichloromethane (2.0 mL), trifluoroacetic acid (1.0 mL) was added at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, dissolved in methanol (5.0 mL), ammonia water (1.0 mL) was added, stirred for 5 minutes, and concentrated under reduced pressure again. The crude product was separated and purified by preparative thin layer chromatography (dichloromethane: methanol = 10: 1) to obtain white solid compound 64 (10 mg, yield 59%). MS m/z 575.9 [M + H] ⁺ . ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.53(s,1H),8.27(s,1H),7.66-7.61(m,2H),7.45-7.41(m,2H),6.90(d,J＝14.1Hz,1H),6.32(d, J＝14.1Hz,1H),5.79(s,2H),3.85(s,3H),1.69-1.62(m,1H),1.16(s,6H),1.08-1.03(m,2H),0.88-0.84(m,2H).

Example 45: Preparation of Compound 65

Compound 36 (5 mg, 0.01 mmol) was placed in a sealed tube, tetrahydrofuran (1.0 mL) was added, sodium hydrogen sulfide (2 mg, 0.05 mmol) was added at 0 degrees, and the mixture was slowly heated to room temperature and stirred for 30 minutes. Then iodomethane (10 mg, 0.07 mmol) was added, and the reaction solution was heated to 70 degrees Celsius and stirred for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (100% methyl tert-butyl ether) to obtain white solid compound 65 (1.8 mg, yield 35%). MS m/z 590.9 [M+H] ⁺ .

Example 46: Preparation of Compound 66

Compound 4-c (18 mg, 0.03 mmol) was dissolved in 1,2-dichloroethane, and diethylaminosulfur trifluoride (19 mg, 0.12 mmol) was added dropwise. The reaction solution was stirred at room temperature for 1 hour, the reaction solution was quenched with water, and the mixed solution was extracted with dichloromethane (3×5 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:1) to obtain white solid compound 66 (8 mg, yield 44%). ¹ H NMR (500MHz, DMSO-d ₆ ) δ9.51(s,1H),8.71(s,1H),8.53(s,1H),8.29(d,J＝1.2Hz,1H),7.59-7.53(m,2H),7.42-7.37(m,2H),5.79(s,2H),3.85(s,3 H),2.64-2.54(m,2H),2.48-2.39(m,2H),1.94-1.87(m,1H),1.69-1.62(m,2H),1.08-1.04(m,2H),0.87-0.83(m,2H). MS m/z 565.8[M+H] ⁺ .

Example 47: Preparation of Compound 67

Chromium chloride (323 mg, 2.63 mmol) was placed in a 25 mL single-mouth bottle, and tetrahydrofuran (2 mL) was added. A mixed solution of compound 67-a (50 mg, 0.44 mmol) and iodoform (345 mg, 0.88 mmol) in tetrahydrofuran (3 mL) was added dropwise at 0°C. After the addition, the reaction solution was stirred at 0°C for 3 hours. Water was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was concentrated under reduced pressure. The crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain a yellow oil 67-b (30.0 mg, yield 29%).

Compound 67-b (30 mg, 0.13 mmol) and compound 1-a (20 mg, 0.04 mmol) were dissolved in DMSO (2.0 mL), and cuprous oxide (3 mg, 0.02 mmol) and N ₁ , N ₂ -di(furan-2-ylmethyl)oxyaldehyde amide (5 mg, 0.02 mmol) were added, and stirred overnight at 100 degrees Celsius under a nitrogen atmosphere. The mixture was cooled to room temperature, quenched with water, extracted with ethyl acetate, and the organic phase was concentrated under reduced pressure. The crude product was separated and purified by preparative thin-layer chromatography (dichloromethane: methanol = 20:1) to obtain white solid compound 67 (1.01 mg, yield 4%). MS m/z 603.0 [M+H] ⁺ .

Example 48: Preparation of Compound 68

Compound 24-b (125 mg, 0.64 mmol) was dissolved in methanol (3 mL), and sodium borohydride (36 mg, 0.96 mmol) was slowly added under ice bath. The reaction solution was stirred at room temperature for 1 hour. The reaction was concentrated under reduced pressure, and the crude product was separated and purified by preparative thin layer plate (ethyl acetate: petroleum ether = 1:5) to obtain compound 68-a (102 mg, yield 81%).

Compound 68-a (15 mg, 0.07 mmol), compound 1-a (18 mg, 0.04 mmol), N, N'-di(furan-2-ylmethyl)oxyaldehyde amide (2 mg, 0.01 mmol), cuprous oxide (1 mg, 0.01 mmol), cesium carbonate (39 mg, 0.12 mmol) were dissolved in N, N-dimethylformamide (1 mL), and the reaction mixture was heated and stirred at 110 ° C for 3 hours under a nitrogen atmosphere. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by preparative thin layer plate (ethyl acetate: dichloromethane = 1:1) to obtain white solid compound 68 (5 mg, yield 24%). ¹ H NMR (500 MHz, DMSO-d) δ9.51 (s, 1H), 8.72 (s, 1H), 8.53 (s, 1H), 8.34 (d, J＝1.0Hz,1H),7.62-7.58(m,2H),7.45-7.42(m,2H),6.87(dd,J＝14.0,1.4Hz,1H),6.29(dd,J＝14.0,5.1Hz,1H),5.80(s,2H),5.01(d,J＝4.4Hz,1H),4.32- 4.27(m,1H),3.85(s,3H),1.68-1.63(m,1H),1.17(d,J=6.5Hz,3H),1.07-1.04(m,2H),0.88-0.85(m,2H). MS m/z 563.0[M+H] ⁺ .

Example 49: Inhibition of USP1 enzyme activity by compounds

Method 1: Dilute the control compound ML323 to 1mM in DMSO, and then dilute it in 4-fold gradient with DMSO for a total of 10 concentrations. Dilute the test compound in 3-fold gradient with DMSO for a total of ten concentrations. Prepare 1× detection buffer, and use the detection buffer to prepare 2× USP1&UAF1 working solution and 2× substrate working solution. Use Echo to transfer 120nL/well of the test compound to a 384-well plate, with 1% DMSO as a solvent control and 10μM ML323 as a positive control. Add 6μL of 2× USP1&UAF1 solution to each well, shake for 30 seconds, and incubate at 25℃ for 15min. Add 6μL of 2× substrate solution to each well to start the reaction and seal the detection plate. Incubate at 25℃ for 60min. Use Envision 2105 to read the fluorescence value. The following formula was used for calculation: inhibition rate (%) = (DMSO well control reading - compound well reading) / (DMSO well control reading - positive control reading) × 100%. The IC ₅₀ value of each compound was analyzed by nonlinear regression method of Graphpad Prism 8 software, and the formula was: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC ₅₀ -X)*HillSlope)). Y is the inhibition rate, and X is the Log value of the concentration of the compound. The enzyme inhibition activity data of some representative compounds are shown in Table 1.

Method 2: Test compounds were diluted in DMSO gradient. Prepare 1× detection buffer (modified Tris buffer), and use the detection buffer to prepare USP1&UAF1 enzyme working solution and substrate (Ubiquitin Rhodamine 110Protein, CF(Ub-Rho)) working solution. Use Echo to transfer test compounds to a 384-well plate, with a final DMSO concentration of 1%. Add 10μL of USP1 enzyme solution to each well and incubate for 1h at room temperature. Add 10μL of substrate solution to each well to start the reaction, centrifuge for 30 seconds, and shake for 30 seconds. Use SpectraMax Paradigm to read the fluorescence value continuously for 30 minutes. Calculate according to the following formula: Inhibition rate (%) = (DMSO well control reading - compound well reading) / (DMSO well control reading - solvent control reading) × 100%. The _IC50 value of each compound was analyzed by nonlinear regression method of XL-Fit software, and the formula was: Y = Bottom + (Top-Bottom) / (1 + 10^ (( _LogIC50 -X) * HillSlope)). Y is the inhibition rate, and X is the Log value of the concentration of the compound. The enzyme inhibition activity data of some representative compounds are shown in Table 1.

Table 1: Inhibitory activity of compounds against USP1 enzyme

Example 50: Inhibition of MDA-MB-468 cell proliferation by compounds

Select MDA-MB-468 cells with good growth status and digest them with trypsin. Add fresh culture medium, mix thoroughly, and centrifuge at 1000rpm for 5 minutes. Inoculate in 96-well plates at a seeding density of 1500 cells per well and culture in a 37℃ incubator overnight. On the second day, remove the culture plate, dilute the compound in a five-fold gradient, treat with the drug, and then culture in a 37℃ incubator for 168 hours.

After the cell culture plate was taken out and equilibrated to room temperature, 100 μL of CellTiter Glo detection reagent was added to each well, shaken for 2 minutes in the dark, incubated for 10 minutes, and the chemiluminescence value was detected using Enspire. Calculated according to the following formula: Inhibition rate (%) = (1-(compound RLU-blank control RLU)/(DMSO control RLU-blank control RLU)) × 100%. Among them, the blank control wells were only added with normal culture medium without cells, and the DMSO wells were added with cells, no compounds, and contained 0.5% DMSO. XLFit was used to draw the efficacy inhibition rate curve and calculate the IC ₅₀ value. The cell inhibition activity data of some representative compounds are shown in Table 2.

Table 2: Inhibitory activity of compounds on MDA-MB-468 cell proliferation

Example 51: Pharmacokinetics in rats

Method 1: Instrument: XEVO TQ-S LC-MS instrument produced by Waters. All the measured data were collected and processed by Masslynx V4.1 software, and the data were calculated and processed by Microsoft Excel. WinNonLin 8.0 software was used to calculate the pharmacokinetic parameters by statistical moment method. The main kinetic parameters included T _max , T _1/2 , C _max , AUC _0-24h , etc. Chromatographic column: ACQUITY UPLC BEH C18 (2.1mm×50mm, 1.7μm); column temperature 40℃; mobile phase A was water (0.1% formic acid), mobile phase B was acetonitrile, the flow rate was 0.350 ml/min, gradient elution was used, and the elution gradient was 0.50min: 10% B; 1.50min: 90% B; 2.50min: 90% B; 2.51min: 10% B; 3.50min: stop. Injection volume: 1 μL.

Animals: 3 SD male rats, weighing 200-220g, were purchased and raised in the laboratory of the Experimental Animal Center for 2 days before use. They were fasted for 12 hours before administration and within 4 hours after administration, and had free access to water during the experiment. Blood samples were collected at the prescribed time after oral gavage.

Solvent: 0.4% ethanol + 0.4% Tween 80 + 99.2% (0.5% methylcellulose M450). Preparation of oral administration solution: accurately weigh the compound, add it to the solvent, ultrasonicate for 5 minutes at room temperature to completely dissolve the drug, and prepare a 0.3 mg/ml drug solution.

Drug samples: Generally, multiple samples with similar structures (molecular weight difference of more than 2 units) are taken, accurately weighed, and administered together (cassette PK). This allows multiple compounds to be screened simultaneously and their oral absorption rates to be compared. Single administration is also used to study the pharmacokinetics of drug samples in rats.

Blood was collected from the eye socket at 0.25, 0.5, 1, 2, 4, 8, 10 and 24 hours after oral administration. 50 μL of plasma sample was added with 200 μL of acetonitrile (containing internal standard verapamil 2 ng/mL), vortexed for 3 min, centrifuged at 20000 rcf, 4°C for 10 min, and the supernatant was taken for LC-MS/MS analysis.

The compound is accurately weighed and prepared into different concentrations, and quantitative analysis is performed on the mass spectrometer to establish a standard curve, and then the concentration of the compound in the above plasma is tested to obtain the concentration of the compound at different time points. All the measured data are collected and processed by relevant software, and the pharmacokinetic parameters are calculated using the statistical moment method (mainly including kinetic parameters T _max , T _1/2 , C _max , AUC _0-24h , etc.). The kinetic parameters of some representative compounds are shown in Table 3.

Method 2: Instrument: SCIEX Triple Quad 6500+ triple quadrupole LC-MS, operating software: Analyst 1.7.2 (Applied Biosystems, Inc., USA); ExionLC liquid phase system; use Microsoft Excel to calculate and process data. WinNolin 8.2 software was used to calculate pharmacokinetic parameters using the statistical moment method. Mainly including kinetic parameters Tmax, T1/2, Cmax, AUC0-24h, etc. Chromatographic column: Synergi 4μm Fusion-RP Luna C18 2mm*50mm, 4μm; column temperature 40℃; mobile phase A is water (0.1% formic acid), mobile phase B is acetonitrile, flow rate is 0.8mL/min, gradient elution is adopted, elution gradient is 0.10min: 15%B; 1.6min: 95%B; 1.90min: 95%B; 1.91min: 15%B; 2.20min: 15%B. Injection volume: 1μL.

Animals: 3 SD male rats, weighing 200-220g, were purchased and raised in the laboratory of the Experimental Animal Center for 3 days before use. They were fasted for 12 hours before administration and within 4 hours after administration, and had free access to water during the experiment. Blood samples were collected at the specified time after gavage.

Solvent: 0.4% ethanol + 0.4% Tween 80 + 99.2% (0.5% methylcellulose M450). Preparation of oral administration solution: accurately weigh the compound, add it to the solvent, vortex and sonicate until the compound is in a uniform suspension state, and prepare a drug solution of corresponding concentration.

Drug samples: Generally, multiple samples with similar structures (molecular weight difference of more than 2 units) are taken, accurately weighed, and administered together (cassette PK). This allows multiple compounds to be screened simultaneously and their oral absorption rates to be compared. Single administration is also used to study the pharmacokinetics of drug samples in rats.

Blood was collected from the jugular vein at 0.25, 0.5, 1, 2, 4, 8, 10 and 24 hours after oral administration. 20 μL of plasma sample (blank sample and internal standard blank sample plus 20 μL of blank plasma) was transferred to a 1.5 mL centrifuge tube, and 200 μL of internal standard (50% methanol acetonitrile solution (concentration 100 ng/mL)) solution was added (Double blank sample was added with 200 μL of 50% methanol acetonitrile solution). After vortexing the sample for 5 minutes, centrifuge it at 14000 rpm and 4°C for 5 minutes, transfer 80 μL and add it to 80 μL of water, mix well, and analyze by LC-MS/MS.

The compound is accurately weighed and prepared into different concentrations, and quantitative analysis is performed on the mass spectrometer to establish a standard curve, and then the concentration of the compound in the above plasma is tested to obtain the concentration of the compound at different time points. All the measured data are collected and processed by relevant software, and the pharmacokinetic parameters are calculated using the statistical moment method (mainly including kinetic parameters T _max , T _1/2 , C _max , AUC _0-24h , etc.). The pharmacokinetic data of some representative compounds are shown in Table 3.

Table 3 Pharmacokinetic parameters in rats

* Note: The test method for compounds 53 and 59 is method 2, and the test method for the remaining compounds is method 1.

Example 52: Pharmacokinetics in mice

Instrument: SCIEX Triple Quad 6500+ triple quadrupole liquid chromatography-mass spectrometer, operating software: Analyst 1.7.2 (Applied Biosystems, Inc., USA); ExionLC liquid phase system; data were calculated and processed using Microsoft Excel. Pharmacokinetic parameters were calculated using WinNolin 8.2 software and statistical moment method. Mainly including kinetic parameters Tmax, T1/2, Cmax, AUC0-24h, etc. Chromatographic column: Synergi 4μm Fusion-RP Luna C18 2mm*50mm, 4μm; column temperature 40℃; mobile phase A is water (0.1% formic acid), mobile phase B is acetonitrile, flow rate is 0.8mL/min, gradient elution is adopted, elution gradient is 0.10min: 15%B; 1.6min: 95%B; 1.90min: 95%B; 1.91min: 15%B; 2.20min: 15%B. Injection volume: 1μL.

Animals: 3 ICR male mice, weighing 25-30g, were purchased and raised in the laboratory of the Experimental Animal Center for 3 days before use. They were fasted for 12 hours before administration and within 4 hours after administration, and had free access to water during the experiment. Blood samples were collected at the scheduled time after gavage.

Solvent: 0.4% ethanol + 0.4% Tween 80 + 99.2% (0.5% methylcellulose M450). Preparation of oral administration solution: accurately weigh the compound, add it to the solvent, vortex and sonicate until the compound is in a uniform suspension state, and prepare a drug solution of corresponding concentration.

Drug samples: Generally, multiple samples with similar structures (molecular weight difference of more than 2 units) are taken, accurately weighed, and administered together (cassette PK). This allows multiple compounds to be screened simultaneously and their oral absorption rates to be compared. Single administration is also used to study the pharmacokinetics of drug samples in mice.

Blood was collected from the cheek at 0.25, 0.5, 1, 2, 4, 6, 8, 10 and 24 hours after oral administration. 20 μL plasma sample (blank sample and internal standard blank sample plus 20 μL blank plasma) was transferred to a 1.5 mL centrifuge tube, and 200 μL internal standard (50% methanol acetonitrile solution (concentration 100 ng/mL)) solution was added (Double blank sample was added with 200 μL 50% methanol acetonitrile solution). After vortexing the sample for 5 minutes, centrifuge at 6000g and 4°C for 3 minutes, 80 μL was transferred and added to 80 μL of water, mixed evenly, and analyzed by LC-MS/MS.

The compound is accurately weighed and prepared into different concentrations, and quantitative analysis is performed on the mass spectrometer to establish a standard curve, and then the concentration of the compound in the above plasma is tested to obtain the concentration of the compound at different time points. All the measured data are collected and processed by relevant software, and the pharmacokinetic parameters are calculated using the statistical moment method (mainly including kinetic parameters T _max , T _1/2 , C _max , AUC _0-24h , etc.). The pharmacokinetic data of some representative compounds are shown in Table 4.

Table 4 Pharmacokinetic parameters in mice

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (17)

A compound having a structure shown in the following formula (I), or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvate thereof: In formula (I): B is selected from Formula (Ia) or Formula (Ib): represents the site of connection between the compound of formula (Ia) or formula (Ib) and A in the compound of formula (I); represents the site where the compound of formula (Ia) or (Ib) is connected to the benzene ring in the compound of formula (I); Each R ¹ is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, CN; m is selected from 0, 1, or 2; Each R ² is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _2-4 alkenyl, C _2-4 alkynyl, CN; n is selected from 0, 1, 2, 3, or 4; ^R3 and ^R4 are each independently selected from hydrogen, halogen, _C1-4 alkyl; or ^R3 and ^R4 together with the same carbon atom to which they are attached form a 3- to 6-membered ring structure, which optionally contains 0 or 1 heteroatom selected from N, O, S; Each R ⁵ is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _2-4 alkenyl, C _2-4 alkynyl, CN; p is selected from 0, 1, or 2; each R ⁶ is independently selected from hydrogen, halogen, C _1-4 alkyl, C 1-4 haloalkyl, C _2-4 alkenyl _, C _2-4 alkynyl, C _3-6 cycloalkyl, C _3-6 cycloalkyl-C _2-4 alkynyl, 3- to 6-membered heterocyclyl, 3- to 6-membered heterocyclyl-C _2-4 alkynyl, CN, OR ^f , SR ^f , NR ^d R ^d , C(O) R ^g , C(O)OR ^f , or S(O) ₂ R ^g ; q is selected from 0, 1, 2, or 3; A is selected from Formula (Ic), Formula (Id), Formula (Ie), Formula (If), or Formula (Ig): represents the site at which B in the compound of formula (Ic), formula (Id), formula (Ie), formula (If), or formula (Ig) is connected to B; represents a single bond or a double bond; each ^Ra is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to ₈ -membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , C(O) ^NRdRd , or S(O) _2NRdRd ; the ^alkyl , cycloalkyl, 3- to 8-membered heterocyclyl, aryl, and ^heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C2-4 alkenyl, _C2-4 alkynyl ^, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl ^{, heteroaryl, CN, ORf, SRf, NRdRd, C(O)Rg , C ( O} ) ^ORf , OC(O) ^Rg or the ^{cycloalkyl and} 3- ^{to 8} - ^{membered heterocyclyl described} _in ^{Ra are optionally substituted by =} _T , wherein ^T is selected from ^O or ^{CRmRn ; Rm} and ^{Rn are} each _{independently} selected from hydrogen, ^{halogen , or} _C1-4 ^alkyl ; Each R ^b is independently selected from hydrogen, halogen, C _1-4 alkyl; or two R ^b and the carbon atom to which they are attached together form a C _3-6 cycloalkyl; each k is independently selected from 0, 1, 2, or 3; R ^c is selected from hydrogen, halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 haloalkenyl, C _2-4 alkynyl, CN, C (O) R ^g , C (O) OR ^f , C (O) NR ^d R ^d ; the alkyl, alkenyl or alkynyl is optionally substituted by one or more groups selected from the group consisting of halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, OR ^f , SR ^f , NR ^d R ^d , C (O) R ^g , C (O) OR ^f , OC (O) R ^g , C (O) NR ^d R ^d , NR ^d C (O) R ^g , S (O) ₂ NR ^d R ^d , or NR ^d S (O) ₂ R ^g ; Each R ^d is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl; Each R ^d' is independently selected from hydrogen, halogen, C _1-4 alkyl, C _1-4 haloalkyl, hydroxy, C _1-4 alkoxy, or CN; each t is independently selected from 0, 1, 2, 3, or 4; k ¹ , k ² , k ³ , and k ⁴ are each independently selected from 0, 1, 2, 3, 4 or 5; D is selected from a chemical bond, O, NR ^e , or CR ^b R ^b ; wherein, R ^e is selected from hydrogen or C _1-4 alkyl; R ^b is as defined above; M is selected from O or CR ^h R ⁱ ; wherein R ^h and R ⁱ are each independently selected from hydrogen, halogen, or C _1-4 alkyl; the alkyl is optionally substituted with one or more groups selected from the group consisting of halogen, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, aryl, heteroaryl, CN, OR ^f , SR ^f , NR ^d R ^d , C(O)R ^g , C(O)OR ^f , OC(O)R ^g , C(O)NR ^d R ^d , NR ^d C(O)R ^g , NR ^d C(O)NR ^d R ^d , OC(O)NR ^d R ^d , NR ^d C(O)OR ^f , OC(O)OR ^f , S(O) ₂ NR ^d R ^d , NR ^d S(O) ₂ R ^g , or NR ^d S(O) ₂ NR ^d R ^d ; or R ^h and R ⁱ together with the carbon atom to which they are attached form a 3- to -8-membered cyclic structure, which optionally contains 0, 1, or 2 heteroatoms selected from N, O, S; ^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are each independently selected from N or ^CRk ; provided that at most two of ^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are selected from N; each ^Rk is each independently selected from hydrogen, halogen, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^{Rg ,} or C(O) ^ORf ; Each of the above R ^d is independently selected from hydrogen, C _1-4 alkyl, C _1-4 haloalkyl, C _3-6 cycloalkyl, or 3- to 6-membered heterocyclyl; each of the R ^f is independently selected from hydrogen, C _1-4 alkyl, C _1-4 haloalkyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, aryl, or heteroaryl; each of the R ^g is independently selected from hydrogen, C _1-4 alkyl, C _1-4 haloalkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, aryl, or heteroaryl; wherein each of the above-mentioned alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic group, cyclic structure, aryl and heteroaryl groups is optionally and independently substituted with 1-3 substituents each independently selected from the group consisting of halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-8 cycloalkyl, 3- to 8-membered heterocyclic group, aryl, heteroaryl, CN, NO ₂ , OR ^f , SR ^f , NR ^d R ^d , C(O) R ^g , C(O)OR ^f , C(O)NR ^d R ^d , NR ^d C(O) R ^g , S(O) ₂ R ^g , or NR ^d S(O) ₂ R ^g , provided that the chemical structure formed is stable and meaningful; wherein R ^d , R ^f , and R ^g are as defined above; Unless otherwise specified, the above-mentioned aryl group is an aromatic group containing 6 to 12 carbon atoms; the heteroaryl group is a 5- to 15-membered heteroaromatic group; and the cyclic structure is a saturated or unsaturated cyclic group containing or not containing heteroatoms. The compound according to claim 1, characterized in that formula (I) is formula (IIa) or formula (IIb): The definitions of the groups in formula (IIa) or (IIb) are as described in claim 1. The compound according to claim 2, characterized in that formula (I) is formula (IIc) or formula (IId): each ^Ra is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, _3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C( ^O ) ^NRdRd ; the alkyl, cycloalkyl, 3- to 8-membered heterocyclyl, aryl and heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^Rg , C(O) ^ORf , ^OC (O) ^Rg , C(O)NR ^d R ^d , NR ^d C(O) R ^g , or NR ^d S(O) ₂ R ^g ; or the cycloalkyl and 3- to 8-membered heterocyclyl described in ^Ra are optionally substituted by =T, wherein T is selected from CR ^m R ⁿ ; R ^m and R ⁿ are each independently selected from hydrogen, halogen, or C _1-4 alkyl; R ^d , R ^f , and R ^g are as defined in claim 1 . The compound according to claim 3, characterized in that formula (I) is formula (IIe) or formula (IIf): Each ^{Ra is} independently selected from hydrogen, halogen, _C1-4 alkyl, _C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2- R a is optionally substituted with _halogen , C _3-6 cycloalkyl, 3- to 8-membered heterocyclyl, ^aryl , heteroaryl, CN, OR ^f , NR ^d R ^d ; or the cycloalkyl and 3- to 8-membered heterocyclyl in R ^a are optionally substituted with ＝T, wherein T is selected from CR ^{m R n ; R m and R n} are each independently selected ^from hydrogen, halogen _, or C _1-4 alkyl; R ^d , R ^f , and R ^g are as defined in claim 1 . The compound according to claim 4, characterized in that each ^Ra in formula (IIe) or formula (IIf) is independently a group selected from the following group: The compound according to claim 1, characterized in that formula (I) is formula (IIIa) or formula (IIIb): each ^Ra is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 haloalkyl, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, _3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C( ^O ) ^NRdRd ; the alkyl, cycloalkyl, 3- to 8-membered heterocyclyl, aryl and heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^Rg , C(O) ^ORf , ^OC (O) ^Rg , C(O)NR ^d R ^d , NR ^d C(O) R ^g , or NR ^d S(O) ₂ R ^g ; or the cycloalkyl and 3- to 8-membered heterocyclyl described in ^Ra are optionally substituted by =T, wherein T is selected from CR ^m R ⁿ ; R ^m and R ⁿ are each independently selected from hydrogen, halogen, or C _1-4 alkyl; R ^d , R ^f , and R ^g are as defined in claim 1 . The compound of claim 6, characterized in that each ^Ra in formula (IIIa) or formula (IIIb) is independently selected from hydrogen, halogen, _C1-4 alkyl, C2-4 _haloalkyl , _C2-4 alkenyl, _C2-4 haloalkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, C(O) ^Rg , C(O) ^ORf , or C(O ^{) NRdRd} ; the alkyl, cycloalkyl, 3- to 8-membered heterocyclyl, aryl and heteroaryl described in ^Ra are optionally substituted with one or more groups selected from the group consisting of halogen, _C3-6 cycloalkyl, 3- to 8-membered heterocyclyl, aryl, heteroaryl, CN, ^ORf , ^NRdRd ; or the cycloalkyl and 3- to 8-membered ^heterocyclyl in ^Ra are optionally substituted with =T, wherein T is selected from ^CRmRn ; ^Rm and ^{R n} are each independently selected from hydrogen, halogen, or C _1-4 alkyl; R ^d , R ^f , and R ^g are as defined in claim 1 . [Corrected 29.03.2024 in accordance with Rule 26]
The compound according to claim 7, characterized in that each ^Ra in formula (IIIa) or formula (IIIb) is independently selected from the following groups: The compound according to claim 1, characterized in that formula (I) is formula (IVa) or formula (IVb): The definitions of the various groups in formula (IVa) or (IVb) are as described in claim 1. The compound according to any one of claims 1 and 4, characterized in that formula (I) is formula (V): R ^c is selected from halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, CN, C(O)R ^g , C(O)OR ^f , C(O)NR ^d R ^d ; the alkyl, alkenyl or alkynyl is optionally substituted by one or more groups selected from the group consisting of halogen, C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, OR ^f , SR ^f , NR ^d R ^d , C(O)R ^g , C(O)OR ^f , OC(O)R ^g , C(O)NR ^d R ^d , NR ^d C(O)R ^g , or NR ^d S(O) ₂ R ^g ; ^k1 is selected from 1, 2, 3, or 4; R ^d' , t are defined as in claim 1; R ^d , R ^f , and R ^g are as defined in claim 1 . The compound according to claim 1, characterized in that formula (I) is formula (VIa) or formula (VIb): The definitions of the various groups in formula (VIa) or (VIb) are as described in claim 1. The compound according to claim 1, characterized in that formula (I) is formula (VIIa) or formula (VIIb): The definitions of the various groups in formula (VIIa) or (VIIb) are as described in claim 1. The compound according to claim 1, characterized in that formula (I) is formula (VIIIa) or formula (VIIIb): ^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are each independently selected from N or ^CRk ; provided that at most two of ^X1 , ^X2 , ^X3 , ^X4 , and ^X5 are selected from N; each ^Rk is each independently selected from hydrogen, halogen, _C2-4 alkenyl, _C2-4 alkynyl, _C3-6 cycloalkyl, 3- to 6-membered heterocyclyl, CN, ^ORf , ^SRf , ^NRdRd , C(O) ^{Rg ,} or C(O) ^ORf ; R ^d , R ^f , and R ^g are as defined in claim 1 . The compound according to claim 1, or its optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, solvate, selected from one of the following groups: “*” indicates a chiral center. A pharmaceutical composition, characterized in that it comprises the compound according to any one of claims 1 to 14, or its optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, solvate, and a pharmaceutically acceptable carrier. A use of a compound according to any one of claims 1 to 14, or an optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, or a solvate thereof, characterized in that it is used to prepare a pharmaceutical composition for treating a disease, disorder, or condition associated with USP1 activity or expression. The use according to claim 16, characterized in that the disease, disorder or condition is selected from the group consisting of breast cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, cutaneous granular cell tumor, melanoma, liver cancer, rectal cancer, bladder cancer, pharyngeal cancer, pancreatic cancer, prostate cancer, glioma, ovarian cancer, endometrial cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, skin cancer, gastric cancer, mesothelioma, osteosarcoma, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, T-cell lymphoma, monocytic leukemia, eosinophilic syndrome, multiple myeloma and other solid tumors and hematological tumors.