TW202404977A - Pyrimido-five-membered heterocyclic compound, and preparation method therefor and use thereof - Google Patents

Abstract

The present invention relates to a pyrimido-five-membered heterocyclic compound, a preparation method therefor and a use thereof, and a pharmaceutical composition comprising same. Specifically, the present invention relates to a compound represented by formula (I) or a pharmaceutically acceptable form thereof, which exhibits good Wee1 inhibitory activity, can serve as an efficient Wee1 inhibitor, and is used for preventing and/or treating Wee1-associated diseases.

Description

A pyrimido five-membered heterocyclic compound, its preparation method and use

This application claims the priority of Chinese patent application 2022108377886 with a filing date of 2022/7/15, Chinese patent application 2023103912164 with a filing date of 2023/4/12, and Chinese patent application 2023108424468 with a filing date of 2023/7/10 . This application cites the full text of the above-mentioned Chinese patent application.

The present invention relates to the field of medicinal chemistry, and specifically relates to a pyrazolopyrimidinone compound used as a Wee1 inhibitor, its preparation method, pharmaceutical composition and its use in preventing and/or treating Wee1 kinase-related diseases.

The cell cycle is a highly regulated and controlled process. The normal cell cycle has checkpoints at the G1/S transition, S phase and G2/M transition, allowing sufficient time for DNA damage repair. Due to TP53 mutations, many human cancer cells suffer from lack of regulation by the G1/S checkpoint and therefore rely heavily on G2/M checkpoint regulation. Wee1 kinase is a key regulator of the G2/M checkpoint (C.J.Matheson, D.S.Backos, P.Reigan, Trends in Pharmacological Sciences, 2016, 37:872). It is an atypical tyrosine kinase that operates on tyrosine 15 (Y15 ) phosphorylates Cdk1 (also known as Cdc2), resulting in its functional inactivation. Cdk1 recruits cyclins A and B to initiate mitosis. Abolishing the G2 checkpoint through Wee1 inhibitors may selectively sensitize P53-deficient cancer cells to DNA damage and avoid affecting surrounding normal tissues. Wee1 also regulates CDK activity in S phase and prevents the induction of DNA damage during normal S phase progression. In addition, Wee1 plays a positive regulatory role in homologous recombination (HR) repair, which is an important pathway for DNA double-strand break repair.

Wee1 is highly expressed in many cancers, including breast, lung, cervical, head and neck, ovarian, prostate, melanoma, leukemia, glioblastoma, medulloblastoma, and hepatocellular carcinoma. Furthermore, high expression of Wee1 is associated with poor prognosis in multiple types of cancer. Inhibiting Wee1 kinase activity and thereby removing G2/M checkpoint function is a potential strategy to drive tumor cells into unscheduled mitosis, thereby experiencing mitotic blockage and leading to cell death. This method, in which cells are forced to enter mitosis without completing DNA replication, is highly toxic to cells and represents a novel mechanism for inducing tumor cell death. Therefore, Wee1 inhibitors have good application prospects as drugs.

Some Wee1 inhibitors have been reported (for example: WO2007126122A1, WO2019173082A1, WO2018011569A1, WO2018162932A1, WO2018090939, WO2019074981, etc.), but there is still a great need for new Wee1 inhibitors in this field, especially Wee1 inhibitors with high activity and other excellent properties. e1 inhibitor , to meet clinical unmet drug needs.

Summary of the invention

In a first aspect, the present invention provides compounds of formula (I) or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, N-oxides, and isotopic labels thereof of compounds, metabolites or prodrugs, Wherein, A ring is selected from phenyl and 5-10 membered heteroaromatic rings; the A ring is optionally substituted by one or more R ^a ; each R ^a is independently selected from H, halogen, -OH, -CN, -C _1-6 alkyl, -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C _2-4 alkenyl, -C _{2 -4} alkynyl; each of the C _1-6 alkyl, C _2-4 alkenyl, and C _2-4 alkynyl groups is optionally replaced by one or more hydrogen, halogen, oxo, -OH, -CN, -NH ₂ , -C _1-6 alkyl substituent substitution; R ¹ is selected from halogen, -CN, C _1-6 alkyl, halogenated C _1-6 alkyl, -C _3-12 ring Alkyl, 4-12-membered heterocyclyl, -O-(4-12-membered heterocyclyl), -N(R ^b )-(4-12-membered heterocyclyl), -C(O)(4-12 membered heterocyclyl), -C(O)NR ^b R ^d , -N=S(O)R ^b R ^d ; the C _1-6 alkyl, C _3-12 cycloalkyl, 4-12 membered Each heterocyclyl group is optionally substituted by one or more elements selected from the group consisting of halogen, oxo, -CN, -OR ^b , -NR ^b R ^d , -C _1-6 alkyl, -halo C _1-6 alkyl. , -halogenated C _1-6 alkoxy group, -C _1-6 alkylene group -OR ^b , -C _1-6 alkylene group -NR ^b R ^d , -C _3-8 cycloalkyl group, 4-8 Member heterocyclic group, -SR ^b , -C(O)R ^b , -C(O)OR ^b , -C(O)NR ^b R ^d , -N(R ^b )C(O)R ^d , -N (R ^b )C(O)OR ^d , -S(O) ₂ R ^d , -N(R ^b )S(O) ₂ R ^d are substituted with substituents; further, the C _3-8 cycloalkane The base and the 4-8 membered heterocyclyl are each optionally substituted by one or more selected from the group consisting of halogen, oxo, -OH, -CN, -C _1-6 alkyl, -halo C _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ substituent substitution; R ^b and R ^d are each independently selected from H, -C _1-6 alkyl, -C _3-6 cycloalkyl base, 4-12 membered heterocyclyl; each of the C _1-6 alkyl, C _3-6 cycloalkyl, and 4-12 membered heterocyclyl is optionally replaced by one or more members selected from halogen, oxo base, -OH, -CN, -NH ₂ , -N(C _1-6 alkyl) ₂ , -C _1-6 alkylene -OH, -OC _1-6 alkyl, -C _1-6 alkylene -NHC _1-6 alkyl, -C _1-6 alkyl-N(C _1-6 alkyl) ₂ substituent substitution; R ² is selected from hydrogen, halogen, -C _1-6 alkyl, - C _2-6 alkenyl, -C _2-6 alkynyl, -C _3-8 cycloalkyl, 4-8 membered heterocyclyl, phenyl, 5-6 membered heteroaryl; the C _1-6 Alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, 4-8 membered heterocyclyl, phenyl, and heteroaryl are each optionally selected from one or more Substituted by halogen, oxo, -OH, -CN, -NH ₂ , -C _1-6 alkyl substituents; B ring is selected from phenyl, 5-10 membered heteroaryl; the B ring is optional is substituted by one or more substituents selected from hydrogen, halogen, -CN, -OCH ₃ ; E ring is selected from -C _3-12 cycloalkyl, 4-12 membered heterocyclyl; the E ring is optional Optionally substituted by one or more R ^c ; Each R ^c is independently selected from hydrogen, halogen, -OH, oxo, cyano, -C _1-6 alkyl, -OC _1-6 alkyl; X is selected from CH or N; L ¹ is selected from single bond, -C _1-6 alkylene-, -C _3-6 cycloalkylene-, -(3-6 membered heterocyclylene)-, -NHC (O)-S(O) ₂ -(C _1-6 alkylene)-, -C(O)C _1-6 alkylene-, -C(O)C _3-6 cycloalkylene-, -C(O)-(3-6 membered heterocyclylene)- and -C(O)-, wherein the C _1-6 alkylene group, 3-6 membered heterocyclylene group and C _3-6 membered heterocyclylene group Each cycloalkyl group is optionally substituted by one or more substituents selected from hydrogen, halogen, oxo group, -OH and amine group; R ³ is selected from the following conditions: i) When L ¹ is a single bond, R ³ Selected from cyano, -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -NHC(O)C _1-6 alkyl, -C(O) C _1-6 alkyl, -NHC(O)OC _1-6 alkyl, -S(O) ₂ C _1-6 alkyl; each of the C _1-6 alkyl groups is optionally replaced by one or more Substituted with a substituent selected from hydrogen, halogen, -OH, cyano, -O-(C _1-6 alkyl); ii) When L ¹ is not a single bond, R ³ is selected from halogen, amine, -OH, Cyano group, -halo C _1-6 alkyl, -S(O) ₂ C _1-6 alkyl, -OC _1-6 alkyl, -NHC _1-6 alkyl -N(C _1-6 alkyl ) ₂ , -NH-C(O)O(C _1-6 alkyl), -N(C _1-6 alkyl)-C(O)OC _1-6 alkyl, -OC(O)NH ₂ , -OC(O)NHC _1-6 alkyl and -N(C _1-6 alkyl)C(O)C _1-6 alkyl, wherein the C _1-6 alkyl is optionally replaced by one or more Substituted with a substituent selected from halogen, oxo, -OH, amine and -O-(C _1-6 alkyl).

In some embodiments of the present invention, when L ¹ is a single bond, R ³ can also be a cyano group.

In some embodiments of the invention, when L ¹ is a single bond, R ³ is selected from -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -NHC (O)C _1-6 alkyl, -C(O)C _1-6 alkyl, -NHC(O)OC _1-6 alkyl, -S(O) ₂ C _1-6 alkyl; the described Each C _1-6 alkyl group is optionally substituted with one or more substituents selected from hydrogen, halogen, -OH, cyano, -O-(C _1-6 alkyl).

The pharmaceutically acceptable form of the present invention is selected from pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, nitrogen oxides, isotope markers, and metabolites or prodrugs.

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, wherein A ring is selected from phenyl, pyridyl, thienyl and pyrazolyl, said A ring is optionally substituted by one or more R ^a , each R ^a is independently selected from H, F, Cl, -OH, -C _1-3 alkyl and -OC _1-3 alkyl, the C _1-3 alkyl is optionally replaced by one or more selected from F, Cl, -OH and -CN Substituted with substituents; Preferably, the A ring is selected from phenyl, and the phenyl is optionally substituted by one or more R ^a , each R ^a is independently selected from H, F, Cl, -OH, -C _1-3 alkyl and -OC _1-3 alkyl, the C _1-3 alkyl is optionally substituted by one or more substituents selected from F, Cl, -OH and -CN; more preferably Specifically, ring A is selected from phenyl, and said phenyl is optionally substituted by one or more ^{R a} selected from H, F, Cl, -CH ₃ , -OCH ₃ and -CH ₂ OH.

In some embodiments of the invention, Ring A is preferably phenyl, more preferably .

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, wherein R ¹ is selected from C _1-6 alkyl, 4-6 membered heterocyclyl, C _4-6 cycloalkyl, -O-(4-6 membered heterocyclyl), -NH-(4- 6-membered heterocyclyl), -C(O)N(C _1-6 alkyl) ₂ , each of the 4-6-membered heterocyclyl and C _4-6 cycloalkyl is optionally selected from one or more From halogen, oxo, -OH, -CN, -OCH ₃ , -NH ₂ , -N(C _1-3 alkyl) ₂ , -C _1-3 alkyl, -halo C _1-3 alkyl , -halogenated C _1-3 alkoxy, -C _1-3 alkylene -OH, -C _1-3 alkylene -N(C _1-3 alkyl) ₂ , -C(O)OC _{1 -3} alkyl and -S(O) ₂ C _1-3 alkyl substituents are substituted; Preferably, R ¹ is selected from pyrrolidinyl, tetrahydropyranyl, piperidyl, tetrahydropyridyl, morpholine base and piperazinyl; each of the pyrrolidinyl, tetrahydropyranyl, piperidinyl, tetrahydropyridyl, morpholinyl and piperazinyl groups is optionally replaced by one or more selected from F, Cl, - OH, -CN, oxo group, -OCH ₃ , -NH ₂ , -N(CH ₃ ) ₂ , -C _1-3 alkyl, -halogenated C _1-3 alkyl, -halogenated C _1-3 Alkoxy, -C _1-3 alkylene-OH, -C _1-3 alkylene-N(CH ₃ ) ₂ , -C(O)OC _1-3 alkyl and -S(O) ₂ C _1-3 alkyl substituents substituted; Preferably, R ¹ is selected from , , , and .

In some embodiments of the invention, R ¹ is preferably a 4-12 membered heterocyclyl, more preferably a 4-6 membered heterocyclyl, and further preferably a 6-membered heterocyclyl; the heterocyclyl is optionally one or more -C _1-6 alkyl or -deuterated C _1-6 alkyl substituted, preferably optionally substituted by one or more -C _1-3 alkyl or -deuterated C _1-3 alkyl, More preferably substituted by _-CH3 or _-CD3 . Wherein, the heteroatom in the heterocyclic group is selected from N, O and S, preferably N. The number of heteroatoms in the heterocyclic group is 1, 2 or 3, preferably 1 or 2.

In some embodiments of the invention, R ¹ is preferably piperidinyl, tetrahydropyridinyl or piperazinyl, which is optionally replaced by one or more -C _{1 -3} alkyl or -deuterated C _1-3 alkyl substituted; more preferably piperazinyl, which is optionally substituted by one or more -CH ₃ or -CD ₃ .

In some embodiments of the invention, ^R1 is preferably selected from and , preferably .

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, wherein R ² is selected from -CH ₃ , -CH ₂ CH ₃ , cyclopropyl, isopropyl, allyl, propargyl, -CH ₂ -cyclopropyl, -CH ₂ CF ₃ ; preferably , R ² is selected from allyl.

In some embodiments of the present invention, R ² is preferably -C _2-6 alkenyl, more preferably -C _2-4 alkenyl, and further preferably allyl.

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or A prodrug, wherein the B ring is selected from phenyl, pyridyl, pyrimidinyl, thienyl and thiazolyl, and the B ring is optionally substituted by one or more substituents selected from hydrogen, halogen and -CN; Preferably, ring B is selected from pyridyl and phenyl.

In some embodiments of the present invention, ring B is preferably a 5-10 membered heteroaryl group, more preferably a 5-6 membered heteroaryl group, and further preferably a 6-membered heteroaryl group. Wherein, the heteroatom in the heteroaryl group is selected from N, O and S, preferably N. The number of heteroatoms in the heteroaryl group is 1 or 2, preferably 1.

In some embodiments of the present invention, B ring is preferably pyrimidinyl or pyridyl, more preferably pyridyl, further preferably .

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, wherein the E ring is selected from 4-6 membered heterocyclyl; the E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from hydrogen, F, Cl, -OH , oxo group, -CN, -CH ₃ , -OCH ₃ ; Preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl.

In some embodiments of the invention, E ring is preferably azetidinyl or pyrrolidinyl, more preferably or , further preferably ; The E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from H, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ .

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or A prodrug, wherein L ¹ is selected from methylene, -C(O)-methylene-O- and -C(O)-; wherein said methylene is optionally replaced by one or more members selected from hydrogen, F, and -CH ₃ substituents are substituted. In some embodiments of the invention, L ¹ is preferably selected from a single bond, -C _1-6 alkylene-, -C _3-6 cycloalkylene-, and -C(O)-; said -C _{1 -6} Alkylene- is preferably -C _1-3 Alkylene-, more preferably methylene; the -C _3-6 cycloalkylene- is preferably cyclopropylene.

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, wherein R ³ is selected from: i) When L ¹ is a single bond, R ³ is selected from -OCH ₃ , -NHCH ₃ , -NHC(O)CH ₃ , -C(O)CH ₃ and -S(O ) ₂ CH ₃ ; ii) When L ¹ is not a single bond, R ³ is selected from F, amine group, -OH, -CN, -CHF ₂ , -CH ₂ F, -S(O) ₂ CH ₃ , -OCH _3. -OCHF ₂ , -NH-C(O)OCH ₃ ; Preferably, R ³ is selected from F, -OH, -CN, -NH ₂ -CHF ₂ , -S(O) ₂ CH ₃ and -CH ₃ .

In some embodiments of the invention, R ³ is preferably selected from: i) When L ¹ is a single bond, R ³ is selected from cyano, -OC _1-6 alkyl and -S(O) ₂ C _1-6 alkyl base, wherein the -OC _1-6 alkyl group is optionally substituted by one or more substituents selected from halogen; the -OC _1-6 alkyl group is preferably -O-halo C _1-3 Alkyl group, more preferably -OCF ₂ ; the -S(O) ₂ C _1-6 alkyl group is preferably -S(O) ₂ C _1-3 alkyl group, more preferably -S(O) ₂ CH ₃ ; ii) L ¹ is -C _1-6 alkylene-, -C _3-6 cycloalkylene- or -C(O)- (preferably -C _1-6 alkylene-, more preferably -C _1-3 alkylene-), R ³ is selected from halogen, amine, -OH, cyano, C _1-6 alkyl, -halogenated C _1-6 alkyl, -S(O) ₂ C _{1 -6} alkyl and -OC _1-6 alkyl; the -halo C _1-6 alkyl is preferably - halo C _1-3 alkyl, more preferably fluoro C _1-3 alkyl, for example It is CHF ₂ or CH ₂ F; the -S(O) ₂ C _1-6 alkyl group is preferably -S(O) ₂ C _1-3 alkyl group, and more preferably -S(O) ₂ CH ₃ ; The -OC _1-6 alkyl group is preferably -OC _1-3 alkyl group, and more preferably -OCH ₃ .

In some embodiments of the invention, when L ¹ is methylene, R ³ is selected from halogen, amine, -OH, cyano, C _1-6 alkyl, -halo C _1-6 alkyl, - S(O) ₂ C _1-6 alkyl and -OC _1-6 alkyl; preferably halogen, amine, -OH, cyano, C _1-3 alkyl, -halo C _1-3 alkyl, -S(O) ₂ C _1-3 alkyl and -OC _1-3 alkyl.

In some embodiments of the invention, the structural units Preferably selected from and ;More preferably .

In some embodiments of the invention, the structural units Preferably selected from .

In some embodiments of the invention, the structural units for , , , , , , , , or , preferably , or .

In some embodiments of the invention, the structural units Preferably , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, said compound is selected from: Wherein, R ¹ is as described in any one of the present invention; E ring is selected from 4-6 membered heterocyclyl, and the E ring is optionally substituted by one or more R ^c , and each R ^c is independently selected. From hydrogen, F, Cl, -OH, oxo group, -CN, -CH ₃ , -OCH ₃ ; Preferably, E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl; L ¹ is as any one of the present invention ^R3 is as described in any one of the present invention.

In some embodiments of the present invention, in the compound of formula (I-1), the E ring is preferably azetidinyl or pyrrolidinyl, more preferably or , more preferably ; The E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from H, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ .

In some embodiments of the invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or Prodrug, said compound is selected from: Wherein, R ^a is selected from H, F, Cl, -OH, -CN, -C _1-3 alkyl and -OC _1-3 alkyl, the C _1-3 alkyl is optionally replaced by one or more Substituted with substituents selected from F, Cl, -OH and -CN; R ^1-1 is selected from H, -C _1-6 alkylene-OH, -C _1-6 alkyl, -halo C _1-6 Alkyl and -deuterated C _1-6 alkyl; E ring is selected from 4-6 membered heterocyclyl, the heteroatom of the 4-6 membered heterocyclyl is N, and the number of heteroatoms is 1 or 2 Or 3; the E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from H, F, Cl, -OH, -CN, -C _1-6 alkylene-OH, -C _1-6 alkyl and -halo C _1-6 alkyl; R ³ is selected from -CN, -C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-6 alkyl, - NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -NHC(O)C _1-6 alkyl, -C(O)C _1-6 alkyl, -NHC(O)OC _{1 -6} alkyl, -S(O) ₂ C _1-6 alkyl, -C _1-6 alkylS(O) ₂ C _1-6 alkyl, -halogenated C _1-6 alkyl, -O- Halogenated C _1-3 alkyl, C _1-6 alkyl-CN, C _1-6 alkyl-OH, C _1-6 alkyl-NH ₂ , -C _1-6 alkyl OC _1-6 alkyl and -cyclopropyl-OH; each of the C _1-6 alkyl groups is optionally substituted by one or more substituents selected from hydrogen, halogen, -OH, amine and cyano.

In some embodiments of the invention, the compound of Formula (I) or Formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate thereof, Isotope label, metabolite or prodrug, wherein R ^a is selected from H, F, Cl, -CH ₃ , -CN, -OCH ₃ and -CH ₂ OH.

In some embodiments of the present invention, in the compound of formula (I-2), R ^1-1 is selected from -C _1-6 alkyl and -deuterated C _1-6 alkyl; the -C _1-6 alkyl The group is preferably -C _1-3 alkyl, more preferably -CH ₃ ; the -deuterated C _1-6 alkyl group is preferably - deuterated C _1-3 alkyl, more preferably -CD ₃ .

In some embodiments of the invention, a compound of Formula (I) or Formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate thereof, Isotope label, metabolite or prodrug, wherein R ^1-1 is -CH ₃ or -CD ₃ .

In some embodiments of the invention, the compound of Formula (I) or Formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate thereof, Isotope label, metabolite or prodrug, wherein the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl.

In some embodiments of the invention, a compound of Formula (I) or Formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate thereof, Isotopic label, metabolite or prodrug, wherein R ^c is each independently selected from hydrogen, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ and -CH ₂ OH.

In some embodiments of the invention, a compound of Formula (I) or Formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate thereof, Isotope label, metabolite or prodrug, wherein R ³ is selected from -CN, -C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-3 alkyl, -C _1-3 alkyl -OH, -S(O) ₂ CH ₃ , -CH ₂ S(O) ₂ CH ₃ , -halogenated C _1-3 alkyl, -O-halogenated C _1-3 alkyl, -C _1-3 Alkyl-CN and -C _1-3 alkyl-OC _1-3 alkyl.

In some embodiments of the invention, a compound of Formula (I) or Formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate thereof, Isotope label, metabolite or prodrug, wherein R ³ is selected from -CN, -CH ₂ OH, -CHF ₂ , -CH ₂ F and -CF ₃ .

In some embodiments of the invention, the compound of formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolism substance or prodrug, wherein R ^a is selected from H, F, Cl, -CH ₃ , -CN, -OCH ₃ and -CH ₂ OH; R ^1-1 is -CH ₃ and -CD ₃ ; E ring is selected from Azetidinyl, pyrrolidinyl and piperidinyl; The E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from hydrogen, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ and -CH ₂ OH; R ³ is selected from -CN, -C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-3 alkyl, -C _1-3 alkyl-OH, -S(O) ₂ CH ₃ , -CH ₂ S(O) ₂ CH ₃ , -halogenated C _1-3 alkyl, -O-halogenated C _1-3 alkyl, -C _1-3 alkyl- CN and -C _1-3 alkyl-OC _1-3 alkyl; preferably, R ³ is selected from -CN, -CH ₂ OH, -CHF ₂ , -CH ₂ F and -CF ₃ .

In some embodiments of the invention, the compound of formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolism substance or prodrug, said compound being selected from: Among them, R ^1-1 , R ^c , R ₃ and R ^a are as described in any one of the present invention.

In some embodiments of the present invention, the compound of formula (I-3) or formula (I-4) or formula (I-5) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer thereof , polymorph, solvate, isotope label, metabolite or prodrug, wherein R ^1-1 is -CH ₃ and -CD ₃ ; R ^a is selected from H, F, Cl, -CH ₃ , -CN , -OCH ₃ and -CH ₂ OH; R ^c is each independently selected from hydrogen, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ and -CH ₂ OH; R ³ is selected from -CN, - C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-3 alkyl, -C _1-3 alkyl -OH, -S(O) ₂ CH ₃ , -CH ₂ S(O) ₂ CH ₃ , -halo C _1-3 alkyl, -O-halo C _1-3 alkyl, -C _1-3 alkyl-CN and -C _1-3 alkyl-OC _1-3 alkyl; preferred Ground, R ³ is selected from -CN, -CH ₂ OH, -CHF ₂ , -CH ₂ F and -CF ₃ .

In some embodiments of the present invention, the compound of formula (I-2) or formula (I-3) or formula (I-4) or formula (I-5) or a pharmaceutically acceptable salt, ester, stereoisomer thereof A conformer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, the compound is selected from: Among them, R ^c , R ₃ and R ^a are as described in any one of the present invention.

In some embodiments of the invention, formula (I-3-1) or formula (I-3-2) or formula (I-4-1) or formula (I-4-2) or formula (I-5 -1) A compound or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R ^a is selected from H , F, Cl, -CH ₃ , -CN, -OCH ₃ and -CH ₂ OH; R ^c is each independently selected from hydrogen, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ and -CH ₂ OH; R ³ is selected from -CN, -C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-3 alkyl, -C _1-3 alkyl -OH, -S(O) ₂ CH ₃ , -CH ₂ S(O) ₂ CH ₃ , -halo C _1-3 alkyl, -O-halo C _1-3 alkyl, -C _1-3 alkyl-CN and -C _{1- 3alkyl} -OC _1-3alkyl .

Preferably, ^R3 is selected from -CN, _-CH2OH , _-CHF2 , _-CH2F and _-CF3 .

In the present invention, when L ¹ is a single bond, R ³ is selected from cyano group, -OC _1-6 alkyl group, -NHC _1-6 alkyl group, -NHC(O)C _1-6 alkyl group, -C(O )C _1-6 alkyl, -NHC(O)OC _1-6 alkyl, -S(O) ₂ C _1-6 alkyl; each of the C _1-6 alkyl groups is optionally replaced by one or more Substituted with a substituent selected from hydrogen, halogen, -OH, cyano, -O-(C _1-6 alkyl).

In the present invention, the compound is not the following compound: .

In the present invention, the 5-10 membered heteroaromatic ring is preferably a 5-6 membered heteroaryl group, and more preferably is a pyridyl or pyrimidinyl group.

The halogen or halogen is preferably fluorine, chlorine or bromine, more preferably fluorine.

The -C _1-6 alkyl group is preferably -C _1-3 alkyl group, more preferably methyl, ethyl or isopropyl group.

The -OC _1-6 alkyl group is preferably -OC _1-3 alkyl group, and more preferably is a methoxy group.

The -NHC _1-6 alkyl group is preferably -NHC _1-3 alkyl group.

The -N(C _1-6 alkyl) ₂ is preferably -N(C _1-3 alkyl) ₂ .

The -NHC(O)C _1-6 alkyl group is preferably -NHC(O)C _1-3 alkyl group.

The -C _2-6 alkenyl group is preferably -C _2-4 alkenyl group, more preferably vinyl or propenyl group.

The -C _2-6 alkynyl group is preferably -C _2-4 alkynyl group, more preferably an ethynyl group or a propynyl group.

The -halo C _1-6 alkyl group is preferably - halo C _1-3 alkyl group, more preferably - halomethyl group or - haloethyl group.

The -C _3-12 cycloalkyl group is preferably -C _3-8 cycloalkyl group, and more preferably is -C _3-6 cycloalkyl group.

The -4-12-membered heterocyclic group is preferably a -4-8-membered heterocyclyl group, more preferably a -4-6-membered heterocyclic group, and further preferably an azetidinyl, pyrrolidinyl, piperidinyl or piperazinyl group .

The -halo C _1-6 alkoxy group is preferably - halo C _1-3 alkoxy group, and more preferably - halomethoxy or -haloethoxy group.

The -C _1-6 alkylene group- is preferably -C _1-3 alkylene group-, more preferably -methylene, -ethylene or propylene.

The -C _3-6 cycloalkylene group - is preferably - cyclopropylene group.

The -NHC(O)-S(O) ₂ -(C _1-6 alkylene)- is preferably -NHC(O)-S(O) ₂ -(C _1-3 alkylene)-.

The -C(O)C _1-6 alkylene group- is preferably -C(O)C _1-3 alkylene group-.

The -C(O)C _1-6 alkyl group is preferably -C(O)C _1-3 alkyl group.

The -NHC(O)OC _1-6 alkyl group is preferably -NHC(O)OC _1-3 alkyl group.

The -S(O) ₂ C _1-6 alkyl group is preferably -S(O) ₂ C _1-3 alkyl group, and more preferably is -S(O) ₂ CH ₃ alkyl group.

The -NHC _1-6 alkyl-N(C _1-6 alkyl) ₂ is preferably -NHC _1-3 alkyl-N(C _1-3 alkyl) ₂ .

The -NH-C(O)O(C _1-6 alkyl) is preferably -NH-C(O)O(C _1-3 alkyl).

The -N(C _1-6 alkyl)-C(O)OC _1-6 alkyl group is preferably -N(C _1-3 alkyl)-C(O)OC _1-3 alkyl group.

The -OC(O)NHC _1-6 alkyl group is preferably -OC(O)NHC _1-3 alkyl group.

The -N(C _1-3 alkyl)C(O)C _1-3 alkyl group is preferably -N(C _1-3 alkyl)C(O)C _1-3 alkyl group.

The present invention provides the following compounds or their pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotopic labels, metabolites or prodrugs, and the compounds are selected from the group consisting of: : .

In another aspect, the present invention provides a method for preparing a compound of formula (I), which method includes the following steps: 1. Compounds IN1-1 and IN1-2 are dissolved in an organic solvent (such as: DMF, DMSO, NMP, dioxane , methylene chloride, toluene, tetrahydrofuran, chloroform, acetonitrile, etc.), in organic bases (such as: Et ₃ N, DIPEA, N-methylmorpholine, DBU, sodium hydride, etc.) or inorganic bases (such as: potassium carbonate, In the presence of cesium carbonate, etc.), compound IN1-3 is obtained through a substitution reaction or a metal-catalyzed coupling reaction [such as: Pd(PPh3) ₄ , PdCl ₂ (dppf) _2, etc.]; 2. Compound IN1-3 is first treated with an oxidizing agent (such as : m-CPBA, potassium hydrogen persulfate, etc.) to generate the intermediate sulfoxide, and then react with compound IN1-4 in an alkali (such as: Et ₃ N, DIPEA, N-methylmorpholine, DBU, sodium hydride, etc.) Substitution reaction gives compounds of formula (I).

Among them, LG ¹ represents a halogen leaving group, and the remaining groups are as defined in formula (I).

In some embodiments of the present invention, LG ¹ is preferably chlorine or bromine; the starting materials for the preparation method of the present invention can be from commercial sources or can be prepared according to known methods.

Those skilled in the art will understand that according to the desired product structure, the order of the reaction steps can be appropriately adjusted, and the protection/deprotection reaction steps can be added or omitted.

In yet another aspect, the invention provides a pharmaceutical composition comprising a compound described herein or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope thereof a marker, metabolite or prodrug, and one or more pharmaceutically acceptable carriers.

A further object of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, which method includes combining the compound of the present invention or a pharmaceutically acceptable form thereof, or a mixture thereof, with one or more pharmaceutically acceptable Accepted carrier combinations.

Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention are pharmaceutically acceptable carriers, and examples of suitable pharmaceutically acceptable carriers are described in Remington’s Pharmaceutical Sciences (2005).

The pharmaceutical composition may be administered in any form as long as it achieves prevention, alleviation, prevention or cure of symptoms in human or animal patients. For example, various suitable dosage forms can be prepared according to the route of administration.

In other embodiments, administration of a compound or pharmaceutical composition of the invention may be combined with additional treatments. The additional treatment methods may be selected from, but are not limited to: radiotherapy, chemotherapy, immunotherapy, or combinations thereof.

The present invention also relates to a pharmaceutical preparation comprising a compound of the present invention or a pharmaceutically acceptable form thereof, or a mixture thereof as an active ingredient, or a pharmaceutical composition of the present invention. In some embodiments, the formulation is in the form of a solid, semi-solid, liquid or gaseous formulation.

A further object of the invention is to provide an article of manufacture, for example in the form of a kit. Article of manufacture as used herein is intended to include, but is not limited to, kits and packaging. The article of the present invention includes: (a) a first container; (b) a pharmaceutical composition located in the first container, wherein the composition includes: a first therapeutic agent, and the first therapeutic agent includes: a compound of the present invention or a pharmaceutically acceptable form thereof, or a mixture thereof; (c) optionally a package insert stating that the pharmaceutical composition may be used to treat a neoplastic condition (as defined below); and (d) a second container.

The first container is a container used to contain a pharmaceutical composition. This container can be used for preparation, storage, shipping and/or individual/bulk sales. The first container is intended to encompass bottles, jars, vials, flasks, syringes, tubes (eg for cream preparations), or any other container used for preparing, containing, storing or dispensing a pharmaceutical product.

The second container is a container for holding the first container and optional package inserts. Examples of the second container include, but are not limited to, boxes (such as cartons or plastic boxes), boxes, cartons, bags (such as paper bags or plastic bags), pouches and cheesecloth bags. The package insert may be physically adhered to the outside of the first container via ties, glue, staples, or other means of attachment, or it may be placed inside the second container without being attached to the first container. Any physical tool to which the container is attached. Alternatively, the package insert is located on the outside of the second container. When located on the outside of the second container, it is preferred that the package insert is physically attached via ties, glue, staples or other means of attachment. Alternatively, it may abut or contact the exterior of the second container without physical adhesion.

The package insert is a trademark, label, label, etc., which lists information related to the pharmaceutical composition located in the first container. The information listed is generally determined by the regulatory agency (eg, the United States Food and Drug Administration) that has jurisdiction over the region in which the product is marketed. Preferably the package insert specifically lists the indications for which the pharmaceutical composition is approved. The package insert may be made of any material from which the information contained in or on it may be read. Preferably, the package insert is a printable material (eg paper, plastic, cardboard, foil, adhesive paper or plastic, etc.) on which the required information can be formed (eg printed or applied).

In another aspect, the present invention provides the use of a compound described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention in the preparation of a medicament for preventing or treating Wee1 kinase-related diseases.

In yet another aspect, the present invention provides the use of a compound described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention in the preparation of a Wee1 inhibitor. In the described uses, the Wee1 inhibitor can be used in vivo in mammalian organisms; it can also be used in vitro, mainly for experimental purposes, for example: as a standard sample or control sample to provide comparison, or prepared according to conventional methods in this field. Complete set to provide rapid detection of the inhibitory effects of Wee1.

In yet another aspect, the present invention provides the use of a compound described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention in the preparation of a medicament for the treatment of cancer. In further embodiments, the cancers include, but are not limited to, breast, colorectal, colon, lung, and prostate cancers, as well as bile duct, bone, bladder, head and neck, kidney, liver, gastrointestinal tissue cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vulvar cancer, and leukemia [including chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL) ) and chronic myelogenous leukemia (CML)], multiple myeloma, and lymphoma.

In some embodiments, the lung cancer is lung adenocarcinoma. The colorectal cancer is colorectal adenocarcinoma.

In yet another aspect, the invention provides a method for preventing or treating Wee1 kinase-related diseases, the method comprising administering a compound as described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the invention to an individual in need thereof. things.

In another aspect, the present invention provides a compound as described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention for preventing or treating Wee1 kinase-related diseases.

In yet another aspect, the invention provides a method for preventing or treating Wee1 kinase-related diseases in combination with a compound as described herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the invention, in combination with an additional treatment method, the additional treatment method being Treatment methods include, but are not limited to: radiotherapy, chemotherapy, immunotherapy, or combinations thereof.

In some embodiments, the Wee1 kinase-associated disease is a disease that is sensitive or responsive to Wee1 kinase inhibition.

In some embodiments, the Wee1 kinase-associated disease is cancer. In further embodiments, the cancers include, but are not limited to, breast, colorectal, colon, lung, and prostate cancers, as well as bile duct, bone, bladder, head and neck, kidney, liver, gastrointestinal tissue cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vulvar cancer, and leukemia [including chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL) ) and chronic myelogenous leukemia (CML)], multiple myeloma, and lymphoma. In some embodiments, the lung cancer is lung adenocarcinoma. The colorectal cancer is colorectal adenocarcinoma.

In a further preferred embodiment, the compounds of the invention can be used in combination with chemoradiotherapy or immunotherapy to prevent or treat cancer.

Dosage regimens can be adjusted to provide the optimal desired response. For example, when administered in the form of an injection, a single injection, bolus injection, and/or continuous infusion may be administered, among others. For example, several divided doses may be administered over time, or the dosage may be proportionally reduced or increased as the exigencies of the therapeutic situation indicate. It is noted that dosage values may vary depending on the type and severity of the condition to be alleviated, and may include single or multiple doses. Generally, the dosage of treatment will vary depending on considerations such as: age, sex and general health of the patient to be treated; frequency of treatment and nature of the desired effect; extent of tissue damage; severity of symptoms duration; and other variables that can be adjusted by individual physicians. It is further understood that, for any particular individual, specific dosage regimens should be adjusted over time based on the individual needs and the professional judgment of the person administering or supervising the administration of the composition. The administration amount and administration regimen of the pharmaceutical composition can be easily determined by those of ordinary skill in the clinical field. For example, the composition or compound of the present invention can be administered in divided doses from 4 times a day to once every 3 days, and the dosage can be, for example, 0.01 to 1000 mg/time. The required dosage may be administered in one or more administrations to obtain the desired results. Pharmaceutical compositions according to the invention may also be provided in unit dosage form.

The present invention provides a class of novel highly active Wee1 inhibitors that can achieve at least one of the following technical effects: (1) high inhibitory activity against Wee1 kinase; (2) excellent physical and chemical properties (such as solubility, physical and/or chemical stability (3) Excellent pharmacokinetic properties (such as good bioavailability, suitable half-life and duration of action); (4) Excellent safety (lower toxicity and/or fewer side effects) , wider treatment range), etc.

General terms and definitions

Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technology as used herein are intended to mean technology as commonly understood in the art, including those variations or equivalent technology that would be apparent to those skilled in the art. Although the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

The terms "comprises," "comprising," "having," "containing," or "involving" and their other variations herein are inclusive or open-ended and do not exclude other unrecited elements or method steps. . Those skilled in the art will understand that the above terms such as "comprising" encompass the meaning of "consisting of."

The term "about" means within ±10% of the stated numerical value, preferably within ±5%, more preferably within ±2%.

Unless otherwise stated, concentrations are by weight and proportions (including percentages) are on a molar basis.

The term "one or more" or the similar expression "at least one" may mean, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 (species) or more (species).

Where the lower and upper limits of a numerical range are disclosed, any number falling within that range and any included range are specifically disclosed. In particular, every range of values disclosed herein (in the form "about a to b," or equivalently, "about a to b," or equivalently, "about a-b"), is to be understood to mean encompassing Every value and range within a wider range.

For example, the expression "C ₁ - ₆ " should be understood to encompass any subrange therein and every point value, such as C ₂ - ₅ , C ₃ - ₄ , C ₁ - ₂ , C ₁ - ₃ , C ₁ - ₄ , C ₁ - ₅ etc., and C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ etc. For example, the expression "C ₃ - ₁₀ " should also be understood in a similar manner, e.g. to cover any subranges and point values contained therein, such as C ₃ - ₉ , C ₆ - ₉ , C ₆ - ₈ , C ₆ - ₇ , C ₇ - ₁₀ , C ₇ - ₉ , C ₇ - ₈ , C ₈ - _9, etc. and C ₃ , C ₄ , C ₅ , C ₆ , C _{7, C 8} , C ₉ , C ₁₀ , _etc. As another example, the expression "3-10 players" should be understood to cover any sub-range and every point value therein, such as 3-4 players, 3-5 players, 3-6 players, 3-7 players, 3-8 players members, 3-9 members, 4-5 members, 4-6 members, 4-7 members, 4-8 members, 5-7 members, 5-8 members, 6-7 members, etc. and 3, 4, 5, 6 , 7, 8, 9, 10 members, etc. For example, the expression "5-10 players" should also be understood in a similar manner, for example, it can cover any sub-range and point value included therein, such as 5-6 players, 5-7 players, 5-8 players, 5- 9 members, 5-10 members, 6-7 members, 6-8 members, 6-9 members, 6-10 members, 7-8 members, etc. and 5, 6, 7, 8, 9, 10 members, etc.

The term "alkyl" when used herein alone or in combination with other groups refers to a saturated straight or branched chain hydrocarbon group. As used herein, the term "C _1-6 alkyl" refers to a saturated straight or branched chain hydrocarbon radical having 1 to 6 carbon atoms (eg, 1, 2, 3, 4, 5 or 6 carbon atoms). "C _1-6 alkyl" is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl base or n-hexyl base, etc.

As used herein alone or in combination with other groups, the term "alkenyl" refers to a straight or branched hydrocarbon group containing one or more double bonds. Illustrative examples of alkenyl groups include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl, hexenyl, and the like. Illustrative examples of C2-6 alkenyl groups with more than one double bond include butadienyl, pentadienyl, hexadienyl and hexatrienyl and branched forms thereof. The position of the unsaturated bond (double bond) can be anywhere in the carbon chain.

When used herein alone or in combination with other groups, the term "alkynyl" refers to an unsaturated straight or branched chain alkynyl group, such as ethynyl, 1-propynyl, propargyl, butynyl, and the like.

The term "alkylene" when used herein alone or in combination with other groups refers to a saturated straight or branched chain divalent hydrocarbon radical. As used herein, the term "C _1-6 alkylene" refers to a saturated straight or branched chain divalent hydrocarbon radical having 1 to 6 carbon atoms. "C _1-6 alkylene group" includes, but is not limited to, methylene, ethylene, propylene or butylene, for example.

When used herein alone or in combination with other groups, the term "alkenylene" refers to a linear or branched divalent aliphatic hydrocarbon group having one or more carbon-carbon double bonds to which two Groups (or fragments) can be connected to the same carbon atom or to different carbon atoms. For example, the term "C _2-6 alkenylene" as used herein refers to an alkenylene group having 2 to 6 carbon atoms (e.g. , , , wait).

When used herein alone or in combination with other groups, the term "alkynylene" refers to a linear or branched divalent aliphatic hydrocarbon group having one or more carbon-carbon triple bonds to which two Groups (or fragments) are connected to different carbon atoms. For example, the term "C _2-6 alkynylene" as used herein refers to an alkynylene group having 2 to 6 carbon atoms (e.g. , wait).

When used herein alone or in combination with other groups, the term "alicyclic" or "aliphatic hydrocarbon ring" refers to aliphatic hydrocarbons that are monocyclic or polycyclic (including fused rings, bridged rings, and spiro rings, such as bicyclic rings) Ring system, including cycloalkane ring, cycloalkene ring, cycloalkyne ring, etc.

When used herein alone or in combination with other groups, the terms "cycloalkyl", "carbocyclic" or "cycloalkylene" refer to saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) non-cyclic Aromatic hydrocarbon group. Common cycloalkyl groups include (but are not limited to) monocyclic cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclobutene, cycloalkyl Pentene, cyclohexene, etc.; or bicyclic cycloalkyl, including fused ring, bridged ring or spiro ring, such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl base, bicyclo[5.2.0]nonyl, decalinyl, etc. For example, "C _3-12 cycloalkyl" refers to a cycloalkyl group having 3 to 12 ring carbon atoms (eg, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12).

When used herein alone or in combination with other groups, the term "heterocycloalkyl" or "heterocyclyl" refers to a saturated or partially saturated, monocyclic or polycyclic ring (such as a bicyclic ring, for example: a bridged ring, a bridged ring or spirocyclic) non-aromatic group, the ring atoms of which are composed of carbon atoms and at least one (for example, 1, 2, 3 or 4) heteroatoms selected from nitrogen, oxygen and sulfur. If the valence bond requirements are met, the heterocycloalkyl group can be connected to the rest of the molecule through any ring atom. For example, "3-8 membered heterocycloalkyl" refers to a heterocycloalkyl group having 3 to 8 ring atoms. Common heterocycloalkyl groups include (but are not limited to) oxiranyl, oxocyclobutanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, homopiperazinyl , cyclotenine, etc.

When used herein alone or in combination with other groups, the term "heteroaryl" or "heteroaryl ring" refers to an aromatic ring having a conjugated pi electron system in which one or more (e.g., 1, 2, or 3 ) ring atoms are heteroatoms selected from N, O, P and S, and the remaining ring atoms are C. Heteroaryl or heteroaromatic rings can be characterized by the number of ring atoms. For example, a 5-12 membered heteroaryl group may contain 5-12 (eg, 5, 6, 7, 8, 9, 10, 11 or 12) ring atoms, especially 5, 6, 9, 10 ring atoms. . Examples of heteroaryl groups are, for example, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, isoxazolyl, isothiazolyl, oxadiazole base, triazolyl, thiadiazolyl, etc.; this term also covers the situation where the heteroaryl or heteroaromatic ring can optionally be further fused to the aryl or heteroaryl ring to form a fused ring.

When used herein alone or in combination with other groups, the term "haloalkyl" refers to an alkyl group as described above, in which one or more hydrogen atoms are replaced by halogen. For example, the term "C _1-6 haloalkyl" refers to a C _1-6 alkyl group optionally substituted with one or more (eg, 1-3) halogens. Those skilled in the art will understand that when there is more than one halogen substituent, the halogens may be the same or different and may be located on the same or different C atoms. Examples of haloalkyl groups are, for example, -CH ₂ F, -CHF ₂ , -CF ₃ , -CCl ₃ , -C ₂ F ₅ , -C ₂ Cl ₅ , -CH ₂ CF ₃ , -CH ₂ Cl or -CH ₂ CH ₂ CF ₃ etc.

The term "alkoxy" when used herein alone or in combination with other groups means an alkyl group as described above with an oxygen atom attached to the parent molecular moiety. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, n-butoxy, tert-butoxy, pentyloxy, and the like.

The term "halogen" when used herein alone or in combination with other groups refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). When used herein alone or in combination with other groups, the term "hydroxy" refers to -OH.

When used herein alone or in combination with other groups, the term "cyano" refers to -CN.

When used herein alone or in combination with other groups, the term "nitro" refers to _-NO2 .

As used herein alone or in combination with other groups, the term "amine group" refers to _-NH2 .

When used herein alone or in combination with other groups, the term "oxo" refers to =O.

The term "each independently" used herein means that at least two groups (or fragments) present in the structure with the same or similar value ranges can have the same or different meanings under specific circumstances. For example, the substituent X and the substituent Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl or aryl. When the substituent X is hydrogen, the substituent Y can be either hydrogen, halogen, or Hydroxy, cyano, alkyl or aryl; similarly, when the substituent Y is hydrogen, the substituent X can be either hydrogen or halogen, hydroxyl, cyano, alkyl or aryl

The term "substitution" and its other variants herein refer to the replacement of one or more (such as 1, 2, 3 or 4) atoms or groups of atoms (such as hydrogen atoms) on the designated atoms with other equivalents , provided that the normal valency of the specified atom or group of atoms in the current situation is not exceeded and that a stable compound can be formed. If an atom or group of atoms is described as "optionally substituted," it may be either substituted or unsubstituted. Unless otherwise stated, the attachment location of a substituent herein may be from any suitable position on the substituent. When the bond in a substituent is shown as a chemical bond passing through two atoms connected to each other in the ring system, it means that the substituent can be connected to any ring-forming atom in the ring system.

Solid lines may be used in this article ( ), solid wedge ( ) or virtual wedge ( ) depicts the carbon-carbon bonds of the compounds of the invention. The use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom are included (eg, a specific enantiomer, a racemic mixture, etc.). The use of solid or imaginary wedges to depict bonds to asymmetric carbon atoms is intended to demonstrate that the stereoisomers shown exist. When present in a racemic mixture, solid and imaginary wedges are used to define relative stereochemistry rather than absolute stereochemistry. Unless otherwise specified, the compounds of the present invention may be present as stereoisomers [which include cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, It exists in the form of geometric isomers, rotamers, conformational isomers, atropisomers and mixtures thereof]. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (eg, racemic mixtures and pairs of diastereoisomers).

The invention also encompasses all possible crystalline forms or polymorphs of the compounds of the invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

It will also be understood that certain compounds of the present invention may exist in free form for therapeutic use, or, where appropriate, as pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites or prodrugs that can be directly administered to a patient in need thereof. Or indirectly provide the compound of the present invention or its metabolite. Therefore, when reference is made herein to "a compound of the invention", it is also intended to encompass the various derivative forms of the compound described above.

Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof. Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. For reviews of suitable salts, see, for example, Remington's Pharmaceutical Sciences , Mack Publishing Company, Easton, Pa., (2005); and Handbook of Pharmaceutical Salts : Properties, Selection , and Use ), Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

As used herein, the term "ester" means an ester derived from the compounds described herein, which includes physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the free acid or alcohol form of the compound of the invention) . The compounds of the present invention may themselves be esters.

The compounds of the invention may exist in the form of solvates, preferably hydrates, wherein the compounds of the invention comprise as structural elements of the crystal lattice of the compounds a polar solvent, in particular such as water, methanol or ethanol. The amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming nitrogen oxides because nitrogen requires available lone pairs of electrons to oxidize to oxides. One skilled in the art will recognize nitrogen-containing heterocycles capable of forming nitrogen oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming nitrogen oxides. Synthetic methods for the preparation of nitrogen oxides of heterocyclic and tertiary amines are well known to those skilled in the art and include the use of peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (mCPBA), hydrogen peroxide, alkyl peroxybenzoic acid, etc. Hydrogen oxides such as tert-butyl hydroperoxide, sodium perborate and dioxirane such as dimethyldioxirane oxidize heterocycles and tertiary amines. These methods for the preparation of nitrogen oxides have been extensively described and reviewed in the literature, see for example: TL Gilchrist, Comprehensive Organic Synthesis , vol. 7, pp 748-750 (AR Katritzky and AJ Boulton, Eds., Academic Press) ; and GWH Cheeseman and ESG Werstiuk, Advances in Heterocyclic Chemistry , vol. 22, pp 390-392 (AR Katritzky and AJ Boulton, Eds., Academic Press).

Also included within the scope of the invention are metabolites of the compounds of the invention, ie substances formed in the body upon administration of the compounds of the invention. The metabolites of a compound can be identified by techniques well known in the art, and their activity can be characterized by experimental methods. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Accordingly, the present invention includes metabolites of a compound of the invention, including compounds prepared by contacting a compound of the invention with a mammal for a time sufficient to produce a metabolite thereof.

The invention further includes within its scope prodrugs of the compounds of the invention, which are certain derivatives of the compounds of the invention which may themselves have little or no pharmacological activity when administered into or onto the body. can be converted by, for example, hydrolytic cleavage to the compounds of the invention having the desired activity. Typically such prodrugs will be functional derivatives of the compound that are readily converted in vivo to the desired therapeutically active compound. Additional information on the use of prodrugs can be found in “Pro-drugs as Novel Delivery Systems,” Volume 14, ACS Symposium Series (T. Higuchi and V. Stella). Prodrugs of the invention can be prepared, for example, by using certain moieties known to those skilled in the art as "pro-moiety [eg as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)]" Prepared by substituting appropriate functional groups present in the compounds of the invention.

The invention also encompasses compounds of the invention containing protecting groups. In any process for preparing the compounds of the invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any relevant molecules, thereby forming chemically protected forms of the compounds of the invention. This can be accomplished by conventional protecting groups, such as those described in T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 2006, which references are incorporated herein by reference. The protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

The present invention also encompasses methods of preparing the compounds described herein. It will be appreciated that the compounds of the present invention may be synthesized using the methods described below as well as synthetic methods known in the art of synthetic organic chemistry or variations thereof known to those skilled in the art. Preferred methods include, but are not limited to, those described below. The reaction can be carried out in a solvent or solvent mixture suitable for the reagents and materials used and suitable for achieving transformation.

The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating one or more symptoms of a target disorder or condition.

As used herein, the term "effective amount" (e.g., "therapeutically effective amount" or "prophylactically effective amount") refers to an amount of an active ingredient that, when administered, will achieve the desired effect, such as one that alleviates the condition being treated. or symptoms or prevention of the occurrence of a condition or its symptoms.

Unless otherwise indicated, the term "treat" or "treat" as used herein means reversing, alleviating, inhibiting the progression of one or more symptoms of a disorder or condition to which such term applies, or preventing such a disorder. or a condition or one or more symptoms of such a disease or condition.

"Individual" as used herein includes humans or non-human animals. Exemplary human subjects include human subjects (referred to as patients) suffering from a disease, such as a disease described herein, or normal subjects. In the present invention, "non-human animals" include all vertebrates, such as non-mammals (such as birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals and/or domesticated animals (such as sheep, dogs, cats, cows, pigs, etc.). Detailed implementation

The invention includes all combinations of the specific embodiments described. Further embodiments and the full scope of applicability of the invention will become apparent from the detailed description provided below. It is to be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are provided by way of illustration only, as various changes and modifications within the spirit and scope of the invention will be made hereunder in detail. The description will become apparent to those familiar with the art. All publications, patents, and patent applications cited herein, including citations, are hereby incorporated by reference in their entirety for all purposes.

Experimental methods without specifying specific conditions in the following examples should be selected according to conventional methods and conditions, or according to product specifications. Example

The following examples and test examples illustrate the present invention in detail, but they do not limit the scope of the present invention, and changes can be made without departing from the scope of the present invention.

Mass spectrometry (MS) was measured using an Agilent (ESI) mass spectrometer, manufacturer: Agilent, model: Agilent 6120B.

Preparative high performance liquid chromatography (HPLC) used Shimadzu LC-8A preparative liquid chromatography (YMC, ODS, 250×20 mm chromatography column).

Thin layer chromatography was used for purification using GF 254 (0.4 ~ 0.5 nm) silica plates produced in Yantai.

The reaction is monitored by thin layer chromatography (TLC) or liquid chromatography mass spectrometry (LC-MS). The developing solvent systems used include but are not limited to: dichloromethane and methanol systems, n-hexane and ethyl acetate systems. As well as the petroleum ether and ethyl acetate system, the volume ratio of the solvent is adjusted according to the polarity of the compound, or triethylamine, etc. is added for adjustment.

Column chromatography generally uses Qingdao Ocean 200 ~ 300 mesh silica gel as the stationary phase. The eluent system includes but is not limited to dichloromethane and methanol system and n-hexane and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine can also be added for adjustment.

Unless otherwise specified in the examples, the reaction temperature is room temperature (20°C to 30°C).

Unless otherwise specified, the reagents used in the examples were purchased from Acros Organics, Aldrich Chemical Company, Nanjing Yaoshi Technology, or Shanghai Shuya Pharmaceutical Technology. Example 1 : Preparation of compound Preparation example 1 : 2-allyl-1-(6-(3-(hydroxymethyl)azetidin-1-yl)pyridin-2-yl)-6-(4 -(4-methylpiperazin-1-yl)phenyl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Preparation of Compound A1) Step 1: Preparation of 2-bromo-6-(3-((tert-butyldiphenylsilyl)oxy)methyl)azetidin-1-yl)pyridine (compound A1-2 )

Add A1-1 (3.60 g, 11.08 mmol), 2,6-dibromopyridine (2.62 g, 11.08 mmol), and potassium carbonate (3.06 g, 22.16 mmol) to dioxane (35 mL), stir and heat React to 80°C for 4 hrs. After TLC confirmed that the reaction was complete, water was added to the reaction solution, extracted with ethyl acetate, the combined organic phases were washed with brine, dried over saturated sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound A1-2 (1.4 g , yield 26.2%). Step 2: 2-allyl-1-(6-(3-((tert-butyldiphenylsilyl)oxy)methyl)azetidin-1-yl)pyridin-2-yl) Preparation of -6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (compound A1-4 )

Compound A1-3 (300 mg, 1.35 mmol), compound A1-2 (779 mg, 1.61 mmol), copper iodide (257 mg, 1.35 mmol), potassium carbonate (279 mg, 2.02 mmol) and N,N' -Dimethylethylenediamine (131 mg, 1.49 mmol) was added to dioxane (5 ml), and heated to 95°C under nitrogen protection for 12 hours. LCMS monitoring showed that the raw materials reacted completely. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A1-4 (400 mg, 47.6%). Step 3: 2-allyl-1-(6-(3-(tert-butyldiphenylsilyl)oxy)methyl)azetidin-1-yl)pyridin-2-yl)- 6-(4-(4-methylpiperazin-1-yl)phenyl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (compound A1-6 ) Preparation

Compound A1-4 (200 mg, 0.32 mmol) was dissolved in toluene (2 mL), m-chloroperoxybenzoic acid (69 mg, 0.32 mmol, 80%) was added at 25°C, reacted at this temperature for 1 hour, and then Compound N,N-diisopropylethylamine (215.80 mg, 1.67 mmol) and compound A1-5 (67 mg, 0.35 mmol) were added, and the reaction was continued for 16 hours. LCMS showed that the reaction was complete. Water was added to the reaction solution and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride, dried and filtered over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified with a silica gel column to obtain compound A1-6 (100 mg, 40%). Step 4: 2-allyl-1-(6-(3-(hydroxymethyl)azetidin-1-yl)pyridin-2-yl)-6-(4-(4-methylpiperidine) Azin-1-yl)phenyl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (preparation of compound A1 )

Compound A1-6 (100 mg, 0.13 mmol) was dissolved in methanol (3 mL), amine fluoride (14.5 mg, 0.39 mmol) was added and the reaction was carried out for 2 hours. LCMS showed that the reaction was complete. The reaction solution was filtered and concentrated under reduced pressure. The crude The product was purified by preparative high-performance liquid chromatography to obtain compound A1 (10 mg, 14.5%).

LCMS (ESI) m/z: 528.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, CD ₃ OD) δ ppm 8.79 (s, 1H), 7.70 (t, J = 7.6 Hz, 1H), 7.59 ( br d, J = 8.8 Hz, 2H), 7.05 (d, J = 7.6 Hz, 1H), 6.97 (d, J = 9.2 Hz, 2H), 6.34 (d, J = 8.0 Hz, 1H), 5.70 - 5.82 (m, 1H), 5.09 (dd, J = 10.2, 1.2 Hz, 1H), 5.01 (dd, J = 17.2, 1.2 Hz, 1H), 4.76 (br d, J = 6.4 Hz, 2H), 4.11 (t , J = 8.4 Hz, 2H), 3.82 (dd, J = 8.4, 5.2 Hz, 2H), 3.77 (d, J = 6.4 Hz, 2H), 3.18 - 3.24 (m, 4H), 2.63 - 2.69 (m, 4H), 2.51 – 2.41 (m, 1H), 2.38 (s, 3H). Preparation Example 2 : Preparation of Compound A9S Step One: Preparation of Compound A9S-2

Add A9S-1 (0.36 g, 4.13 mmol), 2,6-dibromopyridine (1 g, 4.13 mmol), and potassium carbonate (1.14 g, 8.26 mmol) to dimethylsulfoxide (8 mL), and stir Heat to 80°C and react for 2 hours. After TLC confirmed that the reaction was complete, water was added to the reaction solution, extracted with ethyl acetate, the combined organic phases were washed with brine, dried over saturated sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound A9S-2 (0.6 g , yield 60%). Step 2: Preparation of compound A9S-3

Compound A9S-2 (600 mg, 2.47 mmol), compound A1-3 (548 mg, 2.47 mmol), copper iodide (48 mg, 0.25 mmol), and potassium carbonate (279 mg, 4.94 mmol) were added to dioxane ring (5 ml), and heated to 95°C under nitrogen protection for 12 hours. LCMS monitoring showed that the raw materials reacted completely. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography to obtain compound A9S-3 (300 mg, yield 32%). Step 3: Preparation of Compound A9S

Compound A9S-3 (45 mg, 0.2 mmol) was dissolved in toluene (2 mL), m-CPBA (43 mg, 0.2 mmol, 80%) was added at 25°C, reacted at this temperature for 1 hour, and then compound N was added , N-diisopropylethylamine (168 mg, 1.3 mmol) and compound A1-5 (100 mg, 0.26 mmol), continue to react for 16 hours. LCMS showed that the reaction was complete. Water was added to the reaction solution and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride, dried and filtered over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by reverse HPLC to obtain compound A9S (10 mg, Yield 10%).

LCMS (ESI) m/z: 528.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.09 (s, 1H), 8.79 (s, 1H), 8.30 (s, 1H), 7.74 (d, J = 8.2 Hz, 1H), 7.60 (s, 2H), 7.04 (d, J = 7.6 Hz, 1H), 6.96 – 6.84 (m, 2H), 6.46 (d, J = 8.2 Hz, 1H ), 5.68 (ddt, J = 16.4, 10.2, 5.8 Hz, 1H), 5.04 (dt, J = 10.4, 1.4 Hz, 1H), 4.92 (dt, J = 17.2, 1.6 Hz, 1H), 4.59 (s, 2H), 4.27 (t, J = 7.0 Hz, 1H), 3.93 – 3.75 (m, 2H), 3.68 (td, J = 5.8, 5.2, 3.2 Hz, 2H), 3.09 (t, J = 5.0 Hz, 4H ), 2.45 (t, J = 5.0 Hz, 4H), 2.34 – 2.13 (m, 5H).

The following compounds were synthesized with reference to the method of Preparation Example 1 or Preparation Example 2. compound structure LCMS([M+H] ⁺ and/or NMR A3 m/z: 530.3 ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.65 (s, 1 H) 7.60 - 7.72 (m, 3 H) 6.86 - 7.07 (m, 3 H) 6.31 (d, J =8.0 Hz, 1 H) 5.62 - 5.79 (m, 1 H) 5.10 (d, J =10.4 Hz, 1 H) 4.96 - 5.04 (m, 1 H) 4.77 (brd, J =6.4 Hz, 2 H) 4.69 (d, J =5.2 Hz, 1 H) 4.58 (d, J =5.2 Hz, 1 H) 4.17 (t, J =8.0 Hz, 2 H) 3.94 (dd, J =8.4, 5.4 Hz, 2 H) 2.97 - 3.19 (m , 3 H) 3.37 - 3.73 (m, 6 H) 2.90 (s, 3 H). A4 m/z: 576.2 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.80 (d, J = 0.6 Hz, 1H), 7.82 (s, 1H), 7.60 (s, 2H), 7.17 (d, J = 7.8 Hz, 1H), 6.91 (d, J = 9.0 Hz, 2H), 6.42 (d, J = 8.0 Hz, 1H), 5.73– 5.61 (m, 1H), 5.03 (dd, J = 10.2, 1.2 Hz, 1H), 4.93 (dd, J = 17.2, 1.4 Hz, 1H), 4.63 (s, 2H), 4.50 – 4.37 (m, 1H), 4.29 (t, J = 8.6 Hz, 2H), 4.19 (dd, J = 9.0, 5.2 Hz, 2H), 3.10 (s, 4H), 3.06 (s, 3H), 2.69 – 2.52 (m, 4H), 2.24 (s, 3H). A5 m/z: 541.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.09 (s, 1H), 8.79 (s, 1H), 7.76 (s, 1H), 7.60 (s, 2H), 7.49 (s, 1H), 7.09 (d, J = 7.6 Hz, 1H), 7.03 (s, 1H), 6.91 (d, J = 9.0 Hz, 2H), 6.33 (d, J = 8.2 Hz, 1H), 5.72 – 5.64 ( m, 1H), 4.98 (dd, J = 13.6, 1.2 Hz, 2H), 4.63 (s, 2H), 4.09 (t, J = 8.2 Hz, 2H), 4.04 – 3.92 (m, 2H), 3.46 – 3.31 (m, 1H), 3.18 – 3.00 (m, 4H), 2.48 – 2.41 (m, 4H), 2.23 (s, 3H). A9R m/z: 528.2 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.09 (s, 1H), 8.79 (s, 1H), 7.75 (s, 1H), 7.59 (s, 2H), 7.04 (d, J = 7.8 Hz, 1H), 6.90 (d, J = 9.0 Hz, 2H), 6.46 (d, J = 8.2 Hz, 1H), 5.72 – 5.63 (m, 1H), 5.04 (dd, J = 10.2, 1.2 Hz, 1H), 4.94 (d, J = 5.2 Hz, 1H), 4.93 – 4.88 (m, 1H), 4.59 (s, 2H), 4.33 – 4.20 (m, 1H), 3.92 – 3.74 (m, 2H) , 3.72 – 3.59 (m, 2H), 3.14 – 3.02 (m, 4H), 2.48 – 2.42 (m, 4H), 2.32 – 2.24 (m, 1H), 2.22 (s, 3H), 2.21 – 2.13 (m, 1H). A10S m/z: 542.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.07 (s, 1H), 8.79 (s, 1H), 7.72 (t, J = 7.8 Hz, 1H), 7.60 (s, 2H) , 6.98 (d, J = 7.6 Hz, 1H), 6.89 (d, J = 9.1 Hz, 2H), 6.46 (d, J = 8.4 Hz, 1H), 5.69 (dq, J = 10.4, 5.6 Hz, 1H) , 5.04 (dd, J = 10.4, 1.2 Hz, 1H), 4.91 (dd, J = 17.2, 1.2 Hz, 1H), 4.79 (t, J = 5.6 Hz, 1H), 4.64 (s, 2H), 3.99 ( s, 1H), 3.59 – 3.51 (m, 1H), 3.47 (t, J = 8.2 Hz, 1H), 3.27 (dd, J = 17.2, 8.6 Hz, 2H), 3.10 (s, 4H), 2.49 (s , 4H), 2.24 (s, 3H), 2.01 (d, J = 9.0 Hz, 2H), 1.95 – 1.83 (m, 2H). A11S m/z: 542.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.06 (s, 1H), 8.79 (s, 1H), 7.71 (s, 1H), 7.61 (s, 2H), 6.97 (d, J = 7.6 Hz, 1H), 6.90 (d, J = 9.2 Hz, 2H), 6.37 (d, J = 8.2 Hz, 1H), 5.73 – 5.64 (m, 1H), 5.03 (d, J = 9.2 Hz, 1H), 4.91 (d, J = 17.2 Hz, 1H), 4.71 – 4.62 (m, 2H), 3.55 – 3.36 (m, 5H), 3.22 – 3.04 (m, 5H), 2.70 – 2.64 (m, 1H) , 2.47 – 2.36 (m, 3H), 2.35 – 2.31 (m, 1H), 2.26 (s, 3H), 2.20 – 1.94 (m, 1H), 1.80 – 1.67 (m, 1H). A11R m/z: 542.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.06 (s, 1H), 8.79 (s, 1H), 7.71 – 7.61 (m, 3H), 6.97 (d, J = 7.6 Hz, 1H), 6.89 (d, J = 9.2 Hz, 2H), 6.37 (d, J = 8.4 Hz, 1H), 5.72 – 5.64 (m, 1H), 5.03 (dd, J = 10.2, 1.2 Hz, 1H), 4.91 (dd, J = 17.2, 1.2 Hz, 1H), 4.71 – 4.64 (m, 2H), 3.55 – 3.36 (m, 5H), 3.17 – 3.03 (m, 5H), 2.69 – 2.63 (m, 1H), 2.46 – 2.34 (m, 3H), 2.35 – 2.31 (m, 1H), 2.22 (s, 3H), 2.18 – 1.91 (m, 1H), 1.77 – 1.68 (m, 1H). A13 m/z: 556.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.11 (s, 1H), 8.79 (s, 1H), 8.25 (s, 1H), 7.79 (s, 1H), 7.59 (s, 2H), 7.01 (d, J = 7.6 Hz, 1H), 6.94 – 6.80 (m, 2H), 6.59 (d, J = 8.2 Hz, 1H), 5.74 – 5.56 (m, 1H), 5.18 (s, 1H ), 5.04 (dq, J = 10.2, 1.2 Hz, 1H), 4.92 (dd, J = 17.2, 1.6 Hz, 1H), 4.49 (s, 1H), 4.13 (dd, J = 8.6, 6.8 Hz, 1H) , 3.90 – 3.70 (m, 2H), 3.08 (t, J = 5.0 Hz, 4H), 2.46 (d, J = 5.0 Hz, 4H), 2.38 – 2.24 (m, 1H), 2.22 (s, 3H), 2.11 (dq, J = 19.4, 7.8 Hz, 1H), 1.15 (s, 3H), 1.04 (s, 3H). A14 m/z: 523.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.12 (s, 1H), 8.80 (s, 1H), 7.82 (s, 1H), 7.60 (s, 2H), 7.17 (d, J = 7.7 Hz, 1H), 6.91 (d, J = 9.0 Hz, 2H), 6.40 (d, J = 8.0 Hz, 1H), 5.68 (dq, J = 10.4, 5.8 Hz, 1H), 4.98 (ddd, J = 18.4, 13.6, 1.2 Hz, 2H), 4.61 (d, J = 5.1 Hz, 2H), 4.27 (t, J = 8.4 Hz, 2H), 4.15 (dd, J = 8.0, 5.6 Hz, 2H), 3.88 (ddd, J = 8.6, 7.2, 4.2 Hz, 1H), 3.30 (s, 3H), 3.12 (s, 4H), 2.57 – 2.52 (m, 4H). A15 m/z: 537.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.12 (s, 1H), 8.80 (s, 1H), 7.82 (t, J = 7.6 Hz, 1H), 7.60 (s, 2H) , 7.18 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 9.2 Hz, 2H), 6.40 (d, J = 8.2 Hz, 1H), 5.72 – 5.64 (m, 1H), 5.04 (dd, J = 10.2, 1.2 Hz, 1H), 4.93 (dd, J = 17.2, 1.2Hz, 1H), 4.60 (d, J = 5.2 Hz, 2H), 4.30 (d, J = 8.2 Hz, 2H), 3.97 ( d, J = 8.2 Hz, 2H), 3.12 – 3.08 (m, 4H), 2.52 (s, 3H), 2.38 – 2.21 (m, 4H), 1.66 (s, 3H). A16 m/z: 531.3 A17 m/z: 542.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.08 (s, 1H), 8.79 (s, 1H), 7.72 (s, 1H), 7.59 (s, 2H), 7.05 (d, J = 7.6 Hz, 1H), 6.90 (d, J = 9.0 Hz, 2H), 6.28 (d, J = 8.2 Hz, 1H), 5.81 – 5.62 (m, 1H), 5.03 (d, J = 10.2 Hz, 1H), 4.91 (dd, J = 15.2, 9.8 Hz, 1H), 4.63 (s, 2H), 3.82 (d, J = 7.8 Hz, 2H), 3.55 (d, J = 7.8 Hz, 2H), 3.41 ( d, J = 5.4 Hz, 2H), 3.29 (s, 1H), 3.10 – 3.07 (m, 4H), 2.47 – 2.44 (m, 4H), 2.22 (s, 3H), 1.24 (s, 3H). A18 m/z: 546.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.10 (s, 1H), 8.80 (s, 1H), 7.77 (s, 1H), 7.61 (s, 2H), 7.11 (d, J = 7.4 Hz, 1H), 6.92 (d, J = 9.2 Hz, 2H), 6.38 (d, J = 8.2 Hz, 1H), 6.12 (s, 1H), 5.77 – 5.60 (m, 1H), 5.09 – 4.99 (m, 1H), 4.92 (dd, J = 17.2, 1.2 Hz, 1H), 4.62 (d, J = 5.2 Hz, 2H), 4.56 (s, 1H), 4.44 (s, 1H), 4.02 (d , J = 8.8 Hz, 2H), 3.81 (dd, J = 8.8, 2.6 Hz, 2H), 3.18 – 3.10 (m, 4H), 2.63 – 2.58 (m, 4H), 2.33 – 2.28 (m, 3H). A19 m/z: 546.2 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.12 (s, 1H), 8.80 (s, 1H), 7.79 (t, J = 7.6 Hz, 1H), 7.68 – 7.53 (m, 2H), 7.14 (d, J = 7.6 Hz, 1H), 6.92 (d, J = 9.0 Hz, 2H), 6.41 (d, J = 8.2 Hz, 1H), 5.69 (dq, J = 10.4, 5.8 Hz, 1H), 5.33 (s, 1H), 5.04 (dd, J = 10.2, 1.2 Hz, 1H), 4.92 (dd, J = 17.2, 1.3 Hz, 1H), 4.62 (d, J = 4.6 Hz, 2H), 4.14 (dd, J = 17.6, 10.2 Hz, 2H), 4.01 (dd, J = 20.8, 9.8 Hz, 2H), 3.70 (d, J = 21.0 Hz, 2H), 3.16 (s, 4H), 2.73 – 2.59 (m, 4H), 2.35 (d, J = 9.2 Hz, 3H). A20 m/z: 542.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.09 (s, 1H), 8.79 (s, 1H), 7.73 (s, 1H), 7.60 (s, 2H), 7.04 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 9.2 Hz, 2H), 6.28 (d, J = 8.2 Hz, 1H), 5.72 – 5.65 (m, 1H), 5.03 (dd, J = 10.2, 1.2 Hz, 1H), 4.92 (dd, J = 17.2, 1.2 Hz, 1H), 4.74 (d, J = 4.8 Hz, 1H), 4.65 – 4.63 (m, 2H), 3.97 – 3.94 (m, 2H), 3.84 – 3.80 (m, 1H), 3.75 – 3.71 (m, 2H), 3.13 – 3.12 (m, 4H), 2.69 – 2.52 (m, 4H), 2.33 – 2.29 (m, 4H), 1.03 (d, J = 6.2 Hz, 3H). A23 m/z: 542.3 A24 m/z: 558.3 A25 m/z: 546.3 A26 m/z: 558.3 A27 m/z: 553.3 A33 m/z: 560.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.09 (s, 1H), 8.79 (s, 1H), 7.75 (s, 1H), 7.60 (s, 2H), 7.09 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 9.2 Hz, 2H), 6.33 (d, J = 8.2 Hz, 1H), 5.72 – 5.65 (m, 1H), 5.03 (dd, J = 10.2, 1.2 Hz, 2H), 4.92 (dd, J = 17.2, 1.2 Hz, 1H), 4.67 – 4.62 (m, 3H), 4.55 (s, 1H), 3.81 – 3.73 (m, 4H), 3.60 (s, 2H) , 3.12 – 3.05 (m, 4H), 2.50 – 2.48 (m, 4H), 2.22 (s, 3H). A35 m/z: 590.3 ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.09 (s, 1H), 8.79 (s, 1H), 7.76 (s, 1H), 7.60 (s, 2H), 7.10 (d, J = 7.6 Hz, 1H), 6.97 – 6.80 (m, 2H), 6.33 (d, J = 8.1 Hz, 1H), 5.72 – 5.64 (m, 1H), 5.09 – 4.85 (m, 2H), 4.62 (d , J = 5.8 Hz, 2H), 4.17 (t, J = 8.2 Hz, 2H), 3.83 (dd, J = 8.2, 6.0 Hz, 2H), 3.56 (d, J = 7.4 Hz, 2H), 3.26 (s , 1H), 3.10 (t, J = 4.9 Hz, 4H), 2.99 (s, 3H), 2.47 (d, J = 4.4 Hz, 4H), 2.24 (s, 3H) A41 m/z: 542.3 A42 m/z: 530.3 A50 m/z: 549.3 A51 m/z: 570.2, ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.05 (s, 1H), 8.79 (s, 1H), 7.76 – 7.51 (m, 3H), 6.92 (dd, J = 24.8, 8.4 Hz, 3H), 6.38 (d, J = 8.3 Hz, 1H), 5.69 (dq, J = 10.8, 5.8 Hz, 1H), 5.03 (dd, J = 10.3, 1.2 Hz, 1H), 4.92 (dd, J = 17.2, 1.3 Hz, 1H), 4.65 (s, 2H), 4.37 (s, 1H), 3.66 – 3.42 (m, 2H), 3.26 (dd, J = 17.8, 7.9 Hz, 2H), 3.10 – 3.05 (m, 4H), 2.47 – 2.43 (m, 4H), 2.29 (dd, J = 17.9, 10.0 Hz, 1H), 2.22 (s, 3H), 1.97 – 1.82 (m, 2H), 1.14 (d, J = 6.9 Hz, 6H). A52 m/z: 553.2 ¹ H NMR (400 MHz, CD ₃ OD) δ 8.76 (s, 1H), 7.75 (t, J = 8.0 Hz, 1H), 7.57 (d, J = 8.6 Hz, 2H), 7.17 ( d, J = 7.6 Hz, 1H), 7.01 – 6.89 (m, 2H), 6.41 (d, J = 8.0 Hz, 1H), 5.73 – 5.64 (m, 1H), 5.07 (dd, J = 10.2, 1.2 Hz , 1H), 4.98 (dd, J = 17.2, 1.2 Hz, 1H), 4.72 (d, J = 6.0 Hz, 2H), 4.27 (d, J = 8.4 Hz, 2H), 4.13 (d, J = 8.4 Hz , 2H), 3.91 (s, 2H), 3.22 – 3.14 (m, 4H), 2.74 – 2.64 (m, 4H), 2.41 (s, 3H). Preparation Example 3 : Reference Compound 1 and Reference Compound 2 were prepared according to the methods of Example 131 and Example 19 in patent WO2023072301 . , Example 2 : Wee1 kinase activity inhibition experiment

In the Wee1 kinase activity inhibition experiment, ADP-Glo luminescent kit was used to study the effects of different compounds on Wee1 kinase activity. The Wee1 enzyme was purchased from Carna bioscience, and Poly [Lys Tyr (4:1)] was used as the enzyme reaction substrate. After incubating the compound (maximum concentration 1000 nM, 3-fold gradient dilution) with Wee1 (2 nM) for 30 minutes, add the reaction substrate and ATP and react at room temperature for 1 hour, add ADP-Glo stop reagent, and react in the dark at room temperature for 40 minutes. Add ADP-Glo kinase detection reagent, incubate at room temperature for 1 hour in the dark, detect the Luminescence value, and calculate the half inhibitory concentration of the compound on kinase activity. Table 1. Results of Wee1 kinase activity inhibition assay compound IC ₅₀ (nM) compound IC ₅₀ (nM) Compound A1 +++ Compound A18 +++ Compound A3 +++ Compound A19 +++ Compound A4 ++++ Compound A20 +++ Compound A5 +++ Compound A23 ++++ Compound A9S ++++ Compound A24 +++ Compound A9R +++ Compound A25 ++++ Compound A10S ++++ Compound A26 ++++ Compound A11R ++++ Compound A27 ++++ Compound A11S ++++ Compound A33 +++ Compound A14 +++ Compound A35 +++ Compound A15 +++ Compound A41 +++ Compound A16 ++++ Compound A42 +++ Compound A17 +++ Compound A50 ++++ ++++：IC ₅₀ ＜1 nM; +++：1 nM＜IC ₅₀ ＜10 nM

The results show that the compound of the present application has good Wee1 enzyme inhibitory activity. Example 3 : Cell proliferation inhibition experiment one

In A427 cells, the effect of each compound on cell proliferation was detected. Specifically, after incubating the serially diluted compounds for 120 hours, the CellTiter-Glo® chemiluminescent cell viability assay (CTG method) was used to evaluate and calculate the half inhibitory concentration (IC ₅₀ ) of the compounds on several cell lines. Table 2. Inhibitory results of compounds on different cell proliferation activities compound IC ₅₀ (nM) compound IC ₅₀ (nM) Compound A1 +++ Compound A18 +++ Compound A4 +++ Compound A19 +++ Compound A5 +++ Compound A23 ++++ Compound A9S ++++ Compound A24 ++++ Compound A10S ++++ Compound A25 ++++ Compound A14 +++ Compound A26 ++++ Compound A15 +++ Compound A27 +++ Compound A16 ++++ Compound A33 +++ Compound A17 +++ Compound A50 ++++ Reference compound 1 ++ Reference compound 2 ++ ++++：IC ₅₀ ＜200 nM; +++：200 nM＜IC ₅₀ ＜400 nM; ++：IC ₅₀ ＞400 nM

The results show that the compound of the present application has good cell proliferation inhibitory activity. Example 4 : Cell Proliferation Inhibition Experiment 2

In HT29, SNU5, SNU16, and A427 cells, the effect of each compound on cell proliferation was detected. Specifically, after incubating the serially diluted compounds for 120 hours, the CellTiter-Glo® chemiluminescent cell viability assay (CTG method) was used to evaluate and calculate the half inhibitory concentration (IC ₅₀ ) of the compounds on several cell lines. Table 3. Inhibitory results of compounds on different cell proliferation activities compound IC ₅₀ (μM) HT29 SNU5 SNU16 A427 AZD1775 303.06 125.08 913.81 208.9 Compound A9S 183.96 94.71 410.29 182.2

The results show that compared with AZD1775, the compound of the present application has better cell proliferation inhibitory activity in HT29 and SNU16 cells. Example 5 : Balb/c mouse pharmacokinetic (PK) study

Compounds A9S and AZD1775 were administered to female and male Balb/c mice via intravenous and intragastric administration respectively to study the pharmacokinetic properties. The intravenous dosage is 1 mg/kg, the drug solvent is 5% DMSO+5% Solutol+90% Saline; the intragastric dosage is 10 mg/kg, the drug solvent is 0.5% MC (sodium methylcellulose) . Blood was collected at different time points after intravenous and intragastric administration. The blood was anticoagulated with EDTA.K2 and stored at -80°C.

Blood or plasma samples are processed to precipitate proteins and then analyzed by LC-MS/MS. WinNonlin 6.3 software was used to calculate pharmacokinetic parameters using the non-compartmental model. The results are shown in Table 4 below. Table 4. Pharmacokinetic parameters of compounds in the blood of male and female Balb/c mice Compound to be tested AZD1775 Compound A9S Dosing method intravenous administration Administration by gavage intravenous administration Administration by gavage Mouse sex male female male female male female male female Cl_obs mL/min/kg 124 107 / / 42.9 37.5 / / T _1/2 h 0.248 0.259 0.672 0.667 1.29 1.10 1.08 0.854 C ₀ ng/mL 501 503 / / 675 606 / / C _max ng/mL / / 1124 474 / / 1156 1095 AUC _last h*ng/mL 131 153 817 621 370 425 2505 2646 V _{ss_obs} L/kg 1.82 1.75 / / 2.99 2.64 / / F % / / 61.2 40.0 / / 64.7 58.7 Note: "/" means does not exist

Conclusion: The compound A9S of the present invention reaches a higher exposure level in the systemic circulation of female and male mice at an injection dose of 1 mg/kg and an oral dose of 10 mg/Kg, and its comprehensive properties are significantly better than AZD1775, showing Excellent pharmacokinetic properties. Example 6 : Biochemical hERG inhibition test 1. Experimental materials

Stable expression of hERG channel (Cat.K1236) HEK 293 (human embryonic kidney cells, purchased from Invitrogen) culture of stably transfected cell lines: cells in a medium containing 85% DMEM, 10% dialyzed fetal calf serum, 0.1 mM NEAA, 25 mM HEPES , 100 U/mL penicillin-streptomycin, 5 μg/mL Blasticidin and 400 μg/mL Geneticin. The cells were cultured in a cell culture flask at 25 cm ² , 5% CO ₂ , and 37°C. Use TrypLE™Express to subculture cells approximately three times per week to maintain a confluency of approximately 40% to 80%. Before the experiment, cells were induced with doxycycline (1 μg/mL) for 48 hours. On the day of the experiment, the induced cells were resuspended and seeded on a 3.5 cm cell culture dish containing a coverslip (5 × 10 ⁵ cells/3.5 cm cell culture dish) before use, and cultured in a medium without Blasticidin and Geneticin.

Note: The number of subcultures of cell lines used for safety evaluation tests is less than 60. 2. Preparation of working solutions of test compounds

Compounds were dissolved in DMSO and formulated at 30 mM, 10 mM, 3.33 mM, 1.11 mM, and 0.37 mM. Extracellular fluid was then added, with the final concentrations of test compounds being 30 µM, 10 µM, 3.33 µM, 1.11 µM and 0.37 µM, and the final concentration of DMSO being 0.1%. 3. Experimental plan

1) Take a coverslip with a large number of single HEK 293 hERG cells growing evenly on the surface, place it in the continuous recording tank on an inverted microscope, perfuse extracellular fluid (approximately 1 ml per minute) and continue recording, waiting for the current to stabilize. 2) Use standard whole-cell recording mode to record HERG channel currents of single cells. First, the membrane voltage was clamped at -80 mV, and the cells were given a +20 mV voltage stimulus for 5 s to activate the hERG potassium channel, and then repolarized to -50 mV for 5 s to generate an outward tail current, and the current was continuously perfused. Stable, at this time the peak value of the tail current is the control current value. 3) Then perfuse the extracellular fluid containing the drug to be tested and continue recording until the inhibitory effect of the drug on hERG current reaches a stable state. At this time, the peak value of the tail current is the current value after adding the drug. 4) Perfuse the cells with extracellular solution again until the hERG current returns to or is close to the level before adding the drug. Then you can continue to perfuse and test other concentrations or drugs. One or more compounds or drug concentrations can be tested on each cell. 5) Use Dofetilide (TRC) as a positive control in the experiment to ensure that the cells used react normally. 4. Data acceptance criteria

The following criteria were used to determine the acceptability of information. 1) Initial seal resistance＞1 GΩ; 2) Leakage current is less than 50% of the controlled peak tail current at any time; 3) Peak tail amplitude＞250 pA; 4) Thin film resistance Rm＞500 MΩ; 5) Access resistance ( Ra)<15 MΩ; 6) The apparent drop in peak current is <2.5%/min. 5. Experimental results Table 5. hERG inhibition test results Compound number IC ₅₀ (μM) Compound A9S >30 AZD1775 13.31 Reference compound 1 5.40 Reference compound 2 4.59

Conclusion: The test results show that compared with AZD1775, reference compound 1, and reference compound 2, the compound of the present invention has a very weak inhibitory effect on hERG potassium channels and has better drug potential. Example 7 : Biochemical CYP enzyme (cytochrome P450) inhibition test 1. Test system:

P450-Glo™ CYP1A2 Screening System, (Promega); P450-Glo™ CYP2D6 Screening System, (Promega); P450-Glo™ CYP3A4 Screening System, (Promega). 2. Test instruments:

BMG PHERAstar FS Luminescent. 3. Test method: Carry out the test according to the kit instructions, the steps are as follows: 3.1. Inhibition of CYP1A2:

Test group: Add different concentrations of the compounds to be tested into the microwell culture plate, add Luciferin-ME (100 μM), K ₃ PO ₄ (100 mM) and CYP1A2 (0.01 pmol/μL) to each well, and place in the chamber Pre-incubate at room temperature for 10 min, then add the NADPH regeneration system, react at room temperature for 30 min, finally add an equal volume of detection buffer, incubate at room temperature for 20 min, and then perform chemiluminescence detection.

Negative control group: The experimental method is the same as that of the test group, except that the compound to be tested is not added.

Blank control group: The experimental method is the same as that of the test group, except that the compound to be tested is not added, and CYP1A2 Membrance (0.01 pmol/μL) is used instead of CYP1A2. 3.2. Inhibition of CYP2D6:

Test group: Add different concentrations of test compounds to microwell culture plates, add Luciferin-ME EGE (3 μM), K ₃ PO ₄ (100 mM) and CYP2D6 (5 nM) to each well, at room temperature Pre-incubate for 10 min at room temperature, then add NADPH regeneration system, react at 37°C for 30 min, finally add an equal volume of detection buffer, incubate at room temperature for 20 min, and then perform chemiluminescence detection.

Negative control group: The experimental method is the same as that of the test group, except that the compound to be tested is not added.

Blank control group: The experimental method is the same as that of the test group, except that the compound to be tested is not added, and CYP2D6 Membrance (5 nM) is used instead of CYP2D6. 3.3. Inhibition of CYP3A4:

Test group: Add different concentrations of test compounds to microwell culture plates, add Luciferin-IPA (3 μM), K ₃ PO ₄ (100 mM) and CYP3A4 (2 nM) to each well at room temperature. Pre-incubate for 10 min, then add NADPH regeneration system, react at room temperature for 30 min, finally add an equal volume of detection buffer, incubate at room temperature for 20 min, and then perform chemiluminescence detection.

Negative control group: The experimental method is the same as that of the test group, except that the compound to be tested is not added.

Blank control group: The experimental method is the same as that of the test group, except that the compound to be tested is not added, and CYP3A4 Membrance (2 nM) is used instead of CYP3A4. 4. Data processing:

Percent inhibition rate = [1-(Chemiluminescence signal value of the test compound concentration group-Chemiluminescence signal value of the blank control group)/(Chemiluminescence signal value of the negative control group-Chemiluminescence signal value of the blank control group)]× 100%.

When the percentage inhibition rate is between 30-80%, estimate the half inhibitory concentration (IC ₅₀ ) or range of the compound on CYP enzyme according to the following formula: IC ₅₀ =X×(1-percent inhibition rate)/percent inhibition rate, where X is the test concentration of the compound. 5. Test results:

The inhibition of three CYP enzymes by the compounds of the present invention was determined according to the above method, and the results are shown in Table 6 below. Table 6. CYP enzyme inhibition test results Example number IC ₅₀ (μM) CYP1A2 CYP2D6 CYP3A4 Compound A9S >10 >10 >10 Reference compound 1 >10 >10 6.35 Reference compound 2 >10 >10 2.57 6. Conclusion:

The above results show that the compound of the present invention has no obvious inhibitory effect on the three major CYP enzyme subtypes, indicating that its potential drug interaction possibility is relatively low and is superior to reference compound 1 and reference compound 2.

The above embodiments do not limit the solution of the present application in any way. Various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications, journal articles, books, and any other disclosures, is incorporated by reference in its entirety.

Claims (17)

A compound represented by formula (I) or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, Wherein, A ring is selected from phenyl and 5-10 membered heteroaromatic rings; the A ring is optionally substituted by one or more R ^a ; each R ^a is independently selected from H, halogen, -OH, -CN, -C _1-6 alkyl, -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -C _2-4 alkenyl, -C _{2 -4} alkynyl; each of the C _1-6 alkyl, C _2-4 alkenyl, and C _2-4 alkynyl groups is optionally replaced by one or more hydrogen, halogen, oxo, -OH, -CN, -NH ₂ , -C _1-6 alkyl substituent substitution; R ¹ is selected from halogen, -CN, C _1-6 alkyl, halogenated C _1-6 alkyl, -C _3-12 ring Alkyl, 4-12-membered heterocyclyl, -O-(4-12-membered heterocyclyl), -N(R ^b )-(4-12-membered heterocyclyl), -C(O)(4-12 membered heterocyclyl), -C(O)NR ^b R ^d , -N=S(O)R ^b R ^d ; the C _1-6 alkyl, C _3-12 cycloalkyl, 4-12 membered Each heterocyclyl group is optionally substituted by one or more elements selected from the group consisting of halogen, oxo, -CN, -OR ^b , -NR ^b R ^d , -C _1-6 alkyl, -halo C _1-6 alkyl. , -halogenated C _1-6 alkoxy group, -C _1-6 alkylene group -OR ^b , -C _1-6 alkylene group -NR ^b R ^d , -C _3-8 cycloalkyl group, 4-8 Member heterocyclic group, -SR ^b , -C(O)R ^b , -C(O)OR ^b , -C(O)NR ^b R ^d , -N(R ^b )C(O)R ^d , -N (R ^b )C(O)OR ^d , -S(O) ₂ R ^d , -N(R ^b )S(O) ₂ R ^d are substituted with substituents; further, the C _3-8 cycloalkane The base and the 4-8 membered heterocyclyl are each optionally substituted by one or more selected from the group consisting of halogen, oxo, -OH, -CN, -C _1-6 alkyl, -halo C _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ substituent substitution; R ^b and R ^d are each independently selected from H, -C _1-6 alkyl, -C _3-6 cycloalkyl base, 4-12 membered heterocyclyl; each of the C _1-6 alkyl, C _3-6 cycloalkyl, and 4-12 membered heterocyclyl is optionally replaced by one or more members selected from halogen, oxo base, -OH, -CN, -NH ₂ , -N(C _1-6 alkyl) ₂ , -C _1-6 alkylene -OH, -OC _1-6 alkyl, -C _1-6 alkylene -NHC _1-6 alkyl, -C _1-6 alkyl-N(C _1-6 alkyl) ₂ substituent substitution; R ² is selected from hydrogen, halogen, -C _1-6 alkyl, - C _2-6 alkenyl, -C _2-6 alkynyl, -C _3-8 cycloalkyl, 4-8 membered heterocyclyl, phenyl, 5-6 membered heteroaryl; the C _1-6 Alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, 4-8 membered heterocyclyl, phenyl, and heteroaryl are each optionally selected from one or more Substituted by halogen, oxo, -OH, -CN, -NH ₂ , -C _1-6 alkyl substituents; B ring is selected from phenyl, 5-10 membered heteroaryl; the B ring is optional is substituted by one or more substituents selected from hydrogen, halogen, -CN, -OCH ₃ ; E ring is selected from -C _3-12 cycloalkyl, 4-12 membered heterocyclyl; the E ring is optional Optionally substituted by one or more R ^c ; Each R ^c is independently selected from hydrogen, halogen, -OH, oxo, cyano, -C _1-6 alkyl, -OC _1-6 alkyl; X is selected from CH or N; L ¹ is selected from single bond, -C _1-6 alkylene-, -C _3-6 cycloalkylene-, -(3-6 membered heterocyclylene)-, -NHC (O)-S(O) ₂ -(C _1-6 alkylene)-, -C(O)C _1-6 alkylene-, -C(O)C _3-6 cycloalkylene-, -C(O)-(3-6 membered heterocyclylene)- and -C(O)-, wherein the C _1-6 alkylene group, 3-6 membered heterocyclylene group and C _3-6 membered heterocyclylene group Each cycloalkyl group is optionally substituted by one or more substituents selected from hydrogen, halogen, oxo group, -OH and amine group; R ³ is selected from the following conditions: i) When L ¹ is a single bond, R ³ Selected from cyano, -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -NHC(O)C _1-6 alkyl, -C(O) C _1-6 alkyl, -NHC(O)OC _1-6 alkyl, -S(O) ₂ C _1-6 alkyl; each of the C _1-6 alkyl groups is optionally replaced by one or more Substituted with a substituent selected from hydrogen, halogen, -OH, cyano, -O-(C _1-6 alkyl); ii) When L ¹ is not a single bond, R ³ is selected from halogen, amine, -OH, Cyano group, -halo C _1-6 alkyl, -S(O) ₂ C _1-6 alkyl, -OC _1-6 alkyl, -NHC _1-6 alkyl -N(C _1-6 alkyl ) ₂ , -NH-C(O)O(C _1-6 alkyl), -N(C _1-6 alkyl)-C(O)OC _1-6 alkyl, -OC(O)NH ₂ , -OC(O)NHC _1-6 alkyl and -N(C _1-6 alkyl)C(O)C _1-6 alkyl, wherein the C _1-6 alkyl is optionally replaced by one or more Substituted with a substituent selected from halogen, oxo, -OH, amine and -O-(C _1-6 alkyl). The compound according to claim 1 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, wherein, A The ring is selected from phenyl, pyridyl, thienyl and pyrazolyl, and the A ring is optionally substituted by one or more R ^a , each R ^a is independently selected from H, F, Cl, -OH , -C _1-3 alkyl and -OC _1-3 alkyl, the C _1-3 alkyl is optionally substituted by one or more substituents selected from F, Cl, -OH and -CN; preferably Ground, A ring is selected from phenyl, described phenyl is optionally substituted by one or more R ^a , each R ^a is independently selected from H, F, Cl, -OH, -C _1-3 alkane base and -OC _1-3 alkyl, the C _1-3 alkyl is optionally substituted by one or more substituents selected from F, Cl, -OH and -CN; More preferably, the A ring is selected from Phenyl, said phenyl is optionally substituted by one or more R ^a , R ^a is selected from H, F, Cl, -CH ₃ , -OCH ₃ and -CH ₂ OH. The compound according to claim 1 or 2 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or prodrug, wherein R ¹ is selected from C _1-6 alkyl, 4-6 membered heterocyclyl, C _4-6 cycloalkyl, -O-(4-6 membered heterocyclyl), -NH-(4-6 membered heterocyclyl) base), -C(O)N(C _1-6 alkyl) ₂ , each of the 4-6 membered heterocyclyl and C _4-6 cycloalkyl is optionally replaced by one or more members selected from halogen, oxygen Substitute, -OH, -CN, -OCH ₃ , -NH ₂ , -N(C _1-3 alkyl) ₂ , -C _1-3 alkyl, -halo C _1-3 alkyl, -halo C _1-3 alkoxy, -C _1-3 alkylene-OH, -C _1-3 alkylene-N(C _1-3 alkyl) ₂ , -C(O)OC _1-3 alkyl and -S(O) ₂ C _1-3 alkyl substituent substitution; Preferably, R ¹ is selected from pyrrolidinyl, tetrahydropyranyl, piperidyl, tetrahydropyridyl, morpholinyl and piperazine base; each of the pyrrolidinyl, tetrahydropyranyl, piperidinyl, tetrahydropyridyl, morpholinyl and piperazinyl groups is optionally replaced by one or more selected from F, Cl, -OH, -CN , oxo group, -OCH ₃ , -NH ₂ , -N(CH ₃ ) ₂ , -C _1-3 alkyl, -halogenated C _1-3 alkyl, -halogenated C _1-3 alkoxy, -C _1-3 alkylene-OH, -C _1-3 alkylene-N(CH ₃ ) ₂ , -C(O)OC _1-3 alkyl and -S(O) ₂ C _1-3 alkyl Substituted with substituents of the radical; Preferably, R ¹ is selected from , , , and . The compound according to any one of claims 1-3 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or Prodrug, wherein R ² is selected from -CH ₃ , -CH ₂ CH ₃ , cyclopropyl, isopropyl, allyl, propargyl, -CH ₂ -cyclopropyl, -CH ₂ CF ₃ ; preferably , R ² is selected from allyl. The compound according to any one of claims 1-4 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or A prodrug, wherein the B ring is selected from phenyl, pyridyl, pyrimidinyl, thienyl and thiazolyl, and the B ring is optionally substituted by one or more substituents selected from hydrogen, halogen and -CN; Preferably, ring B is selected from pyridyl and phenyl. The compound according to any one of claims 1-5 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or Prodrug, wherein the E ring is selected from 4-6 membered heterocyclyl; the E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from hydrogen, F, Cl, -OH , oxo group, -CN, -CH ₃ , -OCH ₃ ; Preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl. The compound according to any one of claims 1-6 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or A prodrug, wherein L ¹ is selected from methylene, -C(O)-methylene-O- and -C(O)-; wherein said methylene is optionally replaced by one or more members selected from hydrogen, F, and -CH ₃ substituents are substituted. The compound according to any one of claims 1-7 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or precursor medicine, wherein R ³ is selected from: i) When L ¹ is a single bond, R ³ is selected from -OCH ₃ , -NHCH ₃ , -NHC(O)CH ₃ , -C(O)CH ₃ and -S(O) ₂ CH ₃ ; ii) When L ¹ is not a single bond, R ³ is selected from F, amine group, -OH, -CN, -CHF ₂ , -CH ₂ F, -S(O) ₂ CH ₃ , -OCH ₃ , -OCHF ₂ , -NH-C(O)OCH ₃ ; Preferably, R ³ is selected from F, -OH, -CN, -NH ₂ -CHF ₂ , -S(O) ₂ CH ₃ and -CH ₃ . The compound according to any one of claims 1-8 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or Prodrug, said compound is selected from: Wherein, R ¹ is as described in claim 3; E ring is selected from 4-6 membered heterocyclyl, and the E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from hydrogen , F, Cl, -OH, oxo group, -CN, -CH ₃ , -OCH ₃ ; Preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl; L ¹ is as described in claim 7; R ³ is as stated in claim 8. The compound according to claim 1 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, said compound Selected from: Wherein, the E ring is selected from a 4-6 membered heterocyclyl group, the heteroatom of the 4-6 membered heterocyclyl group is N, and the number of heteroatoms is 1, 2 or 3; the E ring is optionally Substituted by one or more R ^c , each R ^c is independently selected from H, F, Cl, -OH, -CN, -C _1-6 alkylene-OH, -C _1-6 alkyl and -halo C _1-6 alkyl; R ³ is selected from -CN, -C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) ₂ , -NHC(O)C _1-6 alkyl, -C(O)C _1-6 alkyl, -NHC(O)OC _1-6 alkyl, -S(O) ₂ C _1-6 alkyl, -C _1-6 alkylS(O) ₂ C _1-6 alkyl, -halo C _1-6 alkyl, -O-halo C _1-3 alkyl, C _{1 -6} alkyl-CN, C _1-6 alkyl-OH, C _1-6 alkyl-NH ₂ , -C _1-6 alkyl OC _1-6 alkyl and -cyclopropyl-OH; the described Each C _1-6 alkyl group is optionally substituted with one or more substituents selected from hydrogen, halogen, -OH, amine and cyano. The compound according to claim 10 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopic label, metabolite or prodrug, wherein, R ^a is selected from H, F, Cl, -CH ₃ , -CN, -OCH ₃ and -CH ₂ OH; R ^1-1 is -CH ₃ or -CD ₃ ; E ring is selected from azetidinyl, pyrrolidinyl and pipera Aldyl; the E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from hydrogen, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ and -CH ₂ OH ; R ³ is selected from -CN, -C(O)NH ₂ , -OC(O)NH ₂ , -OC _1-3 alkyl, -C _1-3 alkyl -OH, -S(O) ₂ CH ₃ , -CH ₂ S(O) ₂ CH ₃ , -halo C _1-3 alkyl, -O-halo C _1-3 alkyl, -C _1-3 alkyl-CN and -C _1-3 alkyl -OC _1-3 alkyl; preferably, R ³ is selected from -CN, -CH ₂ OH, -CHF ₂ , -CH ₂ F and -CF ₃ . The compound according to any one of claims 1-11 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or precursor medicine, which meets one or more of the following conditions: 1) L ¹ is a single bond, R ³ is selected from -OC _1-6 alkyl, -NHC _1-6 alkyl, -N(C _1-6 alkyl) _2. -NHC(O)C _1-6 alkyl, -C(O)C _1-6 alkyl, -NHC(O)OC _1-6 alkyl, -S(O) ₂ C _1-6 alkyl ; Each of the C _1-6 alkyl groups is optionally substituted by one or more substituents selected from hydrogen, halogen, -OH, cyano, -O-(C _1-6 alkyl); 2) A The ring is phenyl, preferably ; 3) R ¹ is a 4-12-membered heterocyclyl, preferably a 4-6-membered heterocyclyl, more preferably a 6-membered heterocyclyl; the heterocyclyl is optionally replaced by one or more -C _{1- 6} alkyl or -deuterated C _1-6 alkyl substituted, preferably optionally substituted by one or more -C _1-3 alkyl or -deuterated C _1-3 alkyl, more preferably -CH ₃ or - CD ₃ substitution; 4) R ² is -C _2-6 alkenyl, preferably -C _2-4 alkenyl, more preferably allyl; 5) Ring B is 5-10 membered heteroaryl, preferably 5 -6-membered heteroaryl, more preferably 6-membered heteroaryl, further preferably pyridyl, particularly preferably ; 6) E ring is azetidinyl or pyrrolidinyl, preferably or , more preferably ; The E ring is optionally substituted by one or more R ^c , each R ^c is independently selected from H, F, Cl, -OH, -CN, -CH ₃ , -OCH ₃ ; 7) L ¹ is selected from single bond, -C _1-6 alkylene-, -C _3-6 cycloalkylene- and -C(O)-; the -C _1-6 alkylene- is preferably -C _{1 -3} alkylene-, more preferably methylene; the -C _3-6 cycloalkylene- is preferably cyclopropane; 8) R ³ is selected from: i) When L ¹ is a single bond, R ³ Selected from cyano, -OC _1-6 alkyl and -S(O) ₂ C _1-6 alkyl, wherein the -OC _1-6 alkyl is optionally substituted by one or more halogens base substitution; the -OC _1-6 alkyl group is preferably -O-halogenated C _1-3 alkyl group, more preferably -OCF ₂ ; the -S(O) ₂ C _1-6 alkyl group is preferably is -S(O) ₂ C _1-3 alkyl, more preferably -S(O) ₂ CH ₃ ; ii) L ¹ is -C _1-6 alkylene-, -C _3-6 cycloalkylene - or -C(O)-, preferably -C _1-6 alkylene-, more preferably -C _1-3 alkylene-, R ³ is selected from halogen, amino, -OH, cyano, C _{1 -6} alkyl, -halogenated C _1-6 alkyl, -S(O) ₂ C _1-6 alkyl and -OC _1-6 alkyl; the -halogenated C _1-6 alkyl is preferably -Halo C _1-3 alkyl, more preferably fluorinated C _1-3 alkyl, such as CHF ₂ or CH ₂ F; the -S(O) ₂ C _1-6 alkyl is preferably -S (O) ₂ C _1-3 alkyl, more preferably -S(O) ₂ CH ₃ ; the -OC _1-6 alkyl is preferably -OC _1-3 alkyl, more preferably -OCH ₃ ; 9) In the compound of formula (I-2), R ^1-1 is selected from -C _1-6 alkyl and -deuterated C _1-6 alkyl; the -C _1-6 alkyl is preferably -C _{1- 3} alkyl group, more preferably -CH ₃ ; the -deuterated C _1-6 alkyl group is preferably - deuterated C _1-3 alkyl group, more preferably - CD ₃ . The compound according to claim 12 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, which satisfies the following One or more of the conditions: 1) L ¹ is methylene, R ³ is selected from halogen, amine, -OH, cyano, C _1-6 alkyl, -halogenated C _1-6 alkyl, -S (O) ₂ C _1-6 alkyl and -OC _1-6 alkyl; preferably halogen, amine, -OH, cyano, C _1-3 alkyl, -halo C _1-3 alkyl, - S(O) ₂ C _1-3 alkyl and -OC _1-3 alkyl; 2) R ¹ is piperidinyl, tetrahydropyridinyl or piperazinyl, and the piperidinyl, tetrahydropyridinyl or piperazinyl Azinyl is optionally substituted by one or more -C _1-3 alkyl or -deuterated C _1-3 alkyl; preferably piperazinyl, said piperazinyl is optionally substituted by one or more -CH ₃ or -CD ₃ is substituted; preferably, R ¹ is selected from and , preferably ; 3) Structural unit Selected from and , preferably ; 4) Structural unit Selected from ; 5) Structural unit for , , , , , , , , or , preferably , , or ; Preferably, structural unit for , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or . The compound according to any one of claims 1-13 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or precursor Medicine, said compound is selected from: Wherein: R ^1-1 , R ^c , R ₃ and R ^a are as described in any one of claims 1-13; Preferably, the compound is selected from: Among them: R ^c , R ₃ and R ^a are as described in any one of claim items 1-13. The compound according to claim 1 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, said compound Selected from: . A pharmaceutical composition comprising the compound described in any one of claims 1-15 or its pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, and isotopes a marker, metabolite or prodrug, and one or more pharmaceutically acceptable carriers. A compound according to any one of claims 1-15 or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitrogen oxide, isotope Use of markers, metabolites or prodrugs or pharmaceutical compositions according to claim 16 in the preparation of drugs for the prevention and/or treatment of cancer or the preparation of Wee1 inhibitors; preferably, the cancer is selected from the following One or more of the following cancers: breast, colorectal, colon, lung, prostate, bile duct, bone, bladder, head and neck, kidney, liver, gastrointestinal tissue, esophagus, ovary, pancreas cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vulvar cancer, leukemia, multiple myeloma and lymphoma.