Pyrimido five-membered heterocyclic compound as well as preparation method and application thereof

Abstract

Relates to a pyrimido five-membered heterocyclic compound, a preparation method and application thereof, and a pharmaceutical composition containing the pyrimido five-membered heterocyclic compound. Specifically, the invention relates to a compound shown as a formula (I) or a pharmaceutically acceptable form thereof, and the compound shows better Wee1 inhibitory activity, can be used as an efficient Wee1 inhibitor, and is used for preventing and / or treating Wee1 related diseases. # imgabs0 #

Description

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

[0001] This application claims priority to Chinese Patent Application No. 2022108377886 filed on July 15, 2022, Chinese Patent Application No. 2023103912164 filed on April 12, 2023, and Chinese Patent Application No. 2023108424468 filed on July 10, 2023. The entire contents of the aforementioned Chinese patent applications are incorporated herein by reference. Technical Field

[0002] The present invention relates to the field of medicinal chemistry, and in particular to a pyrazolopyrimidinone compound used as a Wee1 inhibitor, a preparation method thereof, a pharmaceutical composition thereof, and use thereof in preventing and / or treating Wee1 kinase-related diseases. Background Art

[0003] 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 lack control of the G1 / S checkpoint and therefore rely heavily on the G2 / M checkpoint for regulation. Wee1 kinase is a key regulator of the G2 / M checkpoint (CJ Matheson, DS Backos, P. Reigan, Trends in Pharmacological Sciences, 2016, 37:872). It is an atypical tyrosine kinase that phosphorylates Cdk1 (also known as Cdc2) at tyrosine 15 (Y15), leading to its functional inactivation. Cdk1 recruits cyclins A and B to initiate mitosis. Abrogating the G2 checkpoint with Wee1 inhibitors may selectively sensitize p53-deficient cancer cells to DNA damage while avoiding effects on surrounding normal tissues. Wee1 also regulates CDK activity during S phase, preventing the induction of DNA damage during normal S phase progression. Furthermore, Wee1 plays an active regulatory role in homologous recombination (HR) repair, an important pathway for repairing DNA double-strand breaks.

[0004] Wee1 is highly expressed in many cancers, including breast, lung, cervical, head and neck, ovarian, prostate, melanoma, leukemia, glioblastoma, medulloblastoma, and hepatocellular carcinoma. Furthermore, elevated Wee1 expression is associated with poor prognosis in multiple cancer types. Inhibiting Wee1 kinase activity and thereby removing the G2 / M checkpoint function is a potential strategy to drive tumor cells into unplanned mitosis, thereby experiencing mitotic arrest and leading to cell death. This forced entry into mitosis without completing DNA replication is highly toxic to cells and represents a novel mechanism for inducing tumor cell death. Therefore, Wee1 inhibitors hold great promise as drugs.

[0005] Some Wee1 inhibitors have been reported (for example, WO2007126122A1, WO2019173082A1, WO2018011569A1, WO2018162932A1, WO2018090939, WO2019074981, etc.), but the field is still in urgent need of new Wee1 inhibitors, especially Wee1 inhibitors with high activity and other excellent properties, to meet the unmet clinical drug needs.

[0006] Summary of the Invention

[0007] In a first aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotope-labeled compound, metabolite or prodrug thereof,

[0008] in,

[0009] Ring A is selected from phenyl, 5-10 membered heteroaromatic ring; said ring A is optionally substituted by one or more R a replace;

[0010] Every R a Each 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; the C 1-6 Alkyl, C 2-4 Alkenyl, C 2-4 Each alkynyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, oxo, -OH, -CN, -NH2, -C 1-6 Substitution of alkyl groups;

[0011] R1 Selected from halogen, -CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, -C 3-12 Cycloalkyl, 4-12 membered heterocyclic group, -O-(4-12 membered heterocyclic group), -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 heterocyclic group are each optionally substituted by one or more selected from halogen, oxo, -CN, -OR b 、-NR b R d 、-C 1- 6-alkyl, -halogenated C 1-6 Alkyl, -halogenated C 1-6 Alkoxy, -C 1-6 Alkylene-OR b 、-C 1-6 Alkylene-NR b R d 、-C 3-8 Cycloalkyl, 4-8 membered 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)2R d 、-N(R b )S(O)2R d Substituents are substituted; further, the C 3-8 Cycloalkyl, 4-8 membered heterocyclic group are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -C 1-6 Alkyl, -halogenated C 1-6 Alkyl, -NHC 1-6 Alkyl, -N(C 1-6 alkyl)2 substituent;

[0012] R b 、R d Each independently selected from H, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, 4-12 membered heterocyclic group; the C1-6 Alkyl, C 3-6 Cycloalkyl, 4-12 membered heterocyclic group are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -NH2, -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 Alkylene-N(C 1-6 alkyl)2 is substituted with a substituent;

[0013] R 2 Selected from hydrogen, halogen, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, 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, heteroaryl are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -NH2, -C 1-6 Substitution of alkyl groups;

[0014] Ring B is selected from phenyl, 5-10 membered heteroaryl; said ring B is optionally substituted by one or more substituents selected from hydrogen, halogen, -CN, -OCH3;

[0015] The E ring is selected from -C 3-12 Cycloalkyl, 4-12 membered heterocyclic group; the E ring is optionally replaced by one or more R c replace;

[0016] Every R c are independently selected from hydrogen, halogen, -OH, oxo, cyano, -C 1-6 Alkyl, -OC 1-6 alkyl;

[0017] X is selected from CH or N;

[0018] L 1 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 3-6Cycloalkylene-, -C(O)-(3-6 membered heterocyclylene)- and -C(O)-, wherein the C 1-6 Alkylene, 3-6 membered heterocyclylene and C 3-6 The cycloalkylene groups are each optionally substituted with one or more substituents selected from hydrogen, halogen, oxo, -OH, and amino;

[0019] R 3 Select from the following conditions:

[0020] i)L 1 When it 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)2C 1-6 Alkyl; the C 1-6 Each alkyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, -OH, cyano, -O-(C 1-6 alkyl) is substituted with a substituent;

[0021] ii)L 1 When it is not a single bond, R 3 Selected from halogen, amino, -OH, cyano, -halogenated C 1-6 Alkyl, -S(O)2C 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)NH2, -OC(O)NHC 1-6 Alkyl and -N(C 1-6 alkyl)C(O)C 1-6 Alkyl, wherein the C 1-6 The alkyl group is optionally substituted with one or more radicals selected from halogen, oxo, -OH, amino and -O-(C 1-6 alkyl) is substituted with a substituent.

[0022] In some embodiments of the present invention, L 1 When it is a single bond, R 3 It may also be a cyano group.

[0023] In some embodiments of the present invention, L 1 When it is a single bond, R3 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)2C 1-6 Alkyl; the C 1-6 Each alkyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, -OH, cyano, -O-(C 1-6 alkyl) is substituted with a substituent.

[0024] The pharmaceutically acceptable form of the present invention is selected from pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites or prodrugs.

[0025] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein,

[0026] Ring A is selected from phenyl, pyridyl, thienyl and pyrazolyl, and said ring A is optionally substituted by one or more R a Replace, each R a Each independently selected from H, F, Cl, -OH, -C 1-3 Alkyl and -OC 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with one or more substituents selected from F, Cl, -OH and -CN;

[0027] Preferably, the A ring is selected from phenyl, said phenyl being optionally substituted with one or more R a Replace, each R a Each independently selected from H, F, Cl, -OH, -C 1-3 Alkyl and -OC 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with one or more substituents selected from F, Cl, -OH and -CN;

[0028] More preferably, the A ring is selected from phenyl, said phenyl being optionally substituted by one or more R a Replacement, R a Selected from H, F, Cl, -CH3, -OCH3 and -CH2OH.

[0029] In some embodiments of the present invention, ring A is preferably phenyl, more preferably

[0030] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 1 Selected from C 1-6 Alkyl, 4-6 membered heterocyclic group, C 4-6 Cycloalkyl, -O-(4-6 membered heterocyclyl), -NH-(4-6 membered heterocyclyl), -C(O)N(C 1-6 alkyl) 2, the 4-6 membered heterocyclic group and C 4- 6 cycloalkyl groups are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -OCH3, -NH2, -N(C 1-3 Alkyl)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(C 1-3 Alkyl)2, -C(O)OC 1- 3 alkyl and -S(O)2C 1-3 Substitution of alkyl groups;

[0031] Preferably, R 1 pyrrolidinyl, tetrahydropyranyl, piperidinyl, tetrahydropyridinyl, morpholinyl and piperazinyl; said pyrrolidinyl, tetrahydropyranyl, piperidinyl, tetrahydropyridinyl, morpholinyl and piperazinyl are each optionally substituted by one or more selected from F, Cl, -OH, -CN, oxo, -OCH3, -NH2, -N(CH3)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(CH3)2, -C(O)OC 1-3 Alkyl and -S(O)2C 1-3 Substitution of alkyl groups;

[0032] Preferably, R 1 Selected from

[0033] In some embodiments of the present invention, R 1 It is preferably a 4-12 membered heterocyclic group, more preferably a 4-6 membered heterocyclic group, and further preferably a 6 membered heterocyclic group; the heterocyclic group is optionally substituted by one or more -C 1-6 Alkyl or -deuterated C 1-6 Alkyl substituted, preferably optionally with one or more -C1-3 Alkyl or -deuterated C 1-3 The heterocyclic group is substituted with an alkyl group, more preferably substituted with -CH3 or -CD3. 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.

[0034] In some embodiments of the present invention, R 1 Preferably, the piperidinyl, tetrahydropyridinyl or piperazinyl groups are optionally substituted by one or more -C 1-3 Alkyl or -deuterated C 1-3 Alkyl is substituted; more preferably piperazinyl, which is optionally substituted with one or more -CH3 or -CD3.

[0035] In some embodiments of the present invention, R 1 Preferably selected from Preferably

[0036] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 2 is selected from -CH3, -CH2CH3, cyclopropyl, isopropyl, allyl, propargyl, -CH2-cyclopropyl, -CH2CF3; preferably, R 2 Selected from allyl.

[0037] In some embodiments of the present invention, R 2 Preferably -C 2-6 Alkenyl, more preferably -C 2-4 The alkenyl group is more preferably an allyl group.

[0038] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled, metabolite or prodrug thereof, wherein the B ring is selected from phenyl, pyridyl, pyrimidinyl, thienyl and thiazolyl, and the B ring is optionally substituted with one or more substituents selected from hydrogen, halogen and -CN;

[0039] Preferably, the B ring is selected from pyridyl and phenyl.

[0040] 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 even more preferably a 6 membered heteroaryl group. 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.

[0041] In some embodiments of the present invention, Ring B is preferably pyrimidinyl or pyridinyl, more preferably pyridinyl, and further preferably

[0042] In some embodiments of the present invention, the compound of formula (I) or its pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug, wherein the E ring is selected from a 4-6 membered heterocyclic group; the E ring is optionally replaced by one or more R c Replace, each R c Each is independently selected from hydrogen, F, Cl, -OH, oxo, -CN, -CH3, -OCH3;

[0043] Preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl.

[0044] In some embodiments of the present invention, the E ring is preferably azetidinyl or pyrrolidinyl, more preferably More preferably The E ring is optionally replaced by one or more R c Replace, each R c Each is independently selected from H, F, Cl, -OH, -CN, -CH3, -OCH3.

[0045] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein L 1 is selected from methylene, -C(O)-methylene-O- and -C(O)-; wherein the methylene is optionally substituted with one or more substituents selected from hydrogen, F, and -CH3. In some embodiments of the present invention, L 1 Preferably selected from single bond, -C 1-6 Alkylene-, -C 3-6 Cycloalkylene- and -C(O)-; the -C 1-6 Alkylene-preferably -C 1-3 Alkylene-, more preferably methylene; the -C 3-6 Cycloalkylene- is preferably cyclopropylene.

[0046] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 3 Selected from:

[0047] i)L 1 When it is a single bond, R 3Selected from -OCH3, -NHCH3, -NHC(O)CH3, -C(O)CH3 and -S(O)2CH3;

[0048] ii)L 1 When it is not a single bond, R 3 is selected from F, amino, -OH, -CN, -CHF2, -CH2F, -S(O)2CH3, -OCH3, -OCHF2, -NH-C(O)OCH3; preferably, R 3 Selected from F, -OH, -CN, -NH2-CHF2, -S(O)2CH3 and -CH3.

[0049] In some embodiments of the present invention, R 3 Preferably selected from:

[0050] i)L 1 When it is a single bond, R 3 Selected from cyano, -OC 1-6 Alkyl and -S(O)2C 1-6 Alkyl, wherein the -OC 1-6 The alkyl group is optionally substituted with one or more substituents selected from halogen; the -OC 1-6 The alkyl group is preferably -O-halogenated C 1-3 Alkyl, more preferably -OCF2; the -S(O)2C 1-6 The alkyl group is preferably -S(O)2C 1-3 Alkyl, more preferably -S(O)2CH3;

[0051] ii)L 1 -C 1-6 Alkylene-, -C 3-6 Cycloalkylene- or -C(O)- (preferably -C 1-6 Alkylene-, more preferably -C 1-3 When alkylene-), R 3 Selected from halogen, amino, -OH, cyano, C 1-6 Alkyl, -halogenated C 1-6 Alkyl, -S(O)2C 1-6 Alkyl and -OC 1-6 Alkyl; the -halogenated C 1-6 Alkyl is preferably -haloC 1-3 Alkyl, more preferably fluorinated C 1-3 Alkyl, for example, is CHF2 or CH2F; the -S(O)2C 1- 6 alkyl is preferably -S(O)2C 1-3 Alkyl, more preferably -S(O)2CH3; the -OC 1-6 The alkyl group is preferably -OC 1-3Alkyl, more preferably -OCH3.

[0052] In some embodiments of the present invention, L 1 When it is methylene, R 3 Selected from halogen, amino, -OH, cyano, C 1-6 Alkyl, -halogenated C 1-6 Alkyl, -S(O)2C 1-6 Alkyl and -OC 1-6 Alkyl; preferably halogen, amino, -OH, cyano, C 1-3 Alkyl, -halogenated C 1-3 Alkyl, -S(O)2C 1-3 Alkyl and -OC 1-3 alkyl.

[0053] In some embodiments of the present invention, the structural unit Preferably selected from More preferably

[0054] In some embodiments of the present invention, the structural unit Preferably selected from

[0055] In some embodiments of the present invention, the structural unit for Preferably

[0056] In some embodiments of the present invention, the structural unit Preferably

[0057] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled, metabolite or prodrug thereof is selected from:

[0058] Among them, R 1 As described in any one of the present invention;

[0059] The E ring is selected from a 4-6 membered heterocyclic group, and the E ring is optionally replaced by one or more R c Replace, each R c Each is independently selected from hydrogen, F, Cl, -OH, oxo, -CN, -CH3, -OCH3; preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl;

[0060] L 1As described in any one of the present invention;

[0061] R 3 As described in any one of the present invention.

[0062] In some embodiments of the present invention, in the compound of formula (I-1), ring E is preferably azetidinyl or pyrrolidinyl, more preferably More preferably The E ring is optionally replaced by one or more R c Replace, each R c Each is independently selected from H, F, Cl, -OH, -CN, -CH3, -OCH3.

[0063] In some embodiments of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled, metabolite or prodrug thereof is selected from:

[0064] in,

[0065] R a Selected from H, F, Cl, -OH, -CN, -C 1-3 Alkyl and -OC 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with one or more substituents selected from F, Cl, -OH and -CN;

[0066] R 1-1 Selected from H, -C 1-6 Alkylene-OH, -C 1-6 Alkyl, -halogenated C 1-6 Alkyl and deuterated C 1-6 alkyl;

[0067] The E ring is selected from a 4-6 membered heterocyclic group, wherein the heteroatom of the 4-6 membered heterocyclic group is N, and the number of heteroatoms is 1, 2 or 3;

[0068] The E ring is optionally replaced by one or more R c Replacement, R c Each independently selected from H, F, Cl, -OH, -CN, -C 1-6 Alkylene-OH, -C 1-6 Alkyl and -halogenated C 1-6 alkyl;

[0069] R 3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -OC 1-6 Alkyl, -NHC 1-6 Alkyl, -N(C 1-6Alkyl)2, -NHC(O)C 1- 6-alkyl, -C(O)C 1-6 Alkyl, -NHC(O)OC 1-6 Alkyl, -S(O)2C 1-6 Alkyl, -C 1-6 Alkyl S(O)2C 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-NH2, -C 1-6 Alkyl OC 1-6 Alkyl and -cyclopropyl-OH; the C 1-6 Each alkyl group is optionally substituted with one or more substituents selected from hydrogen, halogen, -OH, amino, and cyano.

[0070] In some embodiments of the present invention, the compound of formula (I) or formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R a Selected from H, F, Cl, -CH3, -CN, -OCH3 and -CH2OH.

[0071] In some embodiments of the present invention, in the compound of formula (I-2), R 1-1 Selected from -C 1-6 Alkyl and deuterated C 1-6 Alkyl; the -C 1-6 The alkyl group is preferably -C 1-3 Alkyl, more preferably -CH3; the -deuterated C 1-6 The alkyl group is preferably a deuterated C 1-3 Alkyl, more preferably -CD3.

[0072] In some embodiments of the present invention, the compound of formula (I) or formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 1-1 It is -CH3 or -CD3.

[0073] In some embodiments of the present invention, the compound of formula (I) or formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl.

[0074] In some embodiments of the present invention, the compound of formula (I) or formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R c Each is independently selected from hydrogen, F, Cl, -OH, -CN, -CH3, -OCH3 and -CH2OH.

[0075] In some embodiments of the present invention, the compound of formula (I) or formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -OC 1-3 Alkyl, -C 1-3 Alkyl-OH, -S(O)2CH3, -CH2S(O)2CH3, -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.

[0076] In some embodiments of the present invention, the compound of formula (I) or formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 3 Selected from -CN, -CH2OH, -CHF2, -CH2F and -CF3.

[0077] In some embodiments of the present invention, the compound of formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein,

[0078] R a Selected from H, F, Cl, -CH3, -CN, -OCH3 and -CH2OH;

[0079] R 1-1 For -CH3 and -CD3;

[0080] Ring E is selected from azetidinyl, pyrrolidinyl and piperidinyl;

[0081] The E ring is optionally replaced by one or more R c Replacement, R c Each independently selected from hydrogen, F, Cl, -OH, -CN, -CH3, -OCH3 and -CH2OH;

[0082] R3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -OC 1-3 Alkyl, -C 1-3 Alkyl-OH, -S(O)2CH3, -CH2S(O)2CH3, -halogenated C 1-3 Alkyl, -O-halogenated C 1-3 Alkyl, -C 1-3 Alkyl-CN and -C 1-3 Alkyl-OC 1-3 Alkane; preferably, R 3 Selected from -CN, -CH2OH, -CHF2, -CH2F and -CF3.

[0083] In some embodiments of the present invention, the compound of formula (I-2) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof is selected from:

[0084] in,

[0085] R 1-1 、R c , R3, R a As described in any one of the present invention.

[0086] 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, polymorph, solvate, isotope-labeled, metabolite or prodrug thereof, wherein,

[0087] R 1-1 For -CH3 and -CD3;

[0088] R a Selected from H, F, Cl, -CH3, -CN, -OCH3 and -CH2OH;

[0089] R c Each independently selected from hydrogen, F, Cl, -OH, -CN, -CH3, -OCH3 and -CH2OH;

[0090] R 3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -OC 1-3 Alkyl, -C 1-3 Alkyl-OH, -S(O)2CH3, -CH2S(O)2CH3, -halogenated C 1-3 Alkyl, -O-halogenated C 1-3 Alkyl, -C 1-3 Alkyl-CN and -C 1-3Alkyl-OC 1-3 Alkane; preferably, R 3 Selected from -CN, -CH2OH, -CHF2, -CH2F and -CF3.

[0091] 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, tautomer, polymorph, solvate, isotopically labeled, metabolite or prodrug thereof is selected from:

[0092] Among them, R c , R3, R a As described in any one of the present invention.

[0093] In some embodiments of the present invention, the compound of formula (I-3-1) or formula (I-3-2) or formula (I-4-1) or formula (I-4-2) or formula (I-5-1) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein,

[0094] R a Selected from H, F, Cl, -CH3, -CN, -OCH3 and -CH2OH;

[0095] R c Each independently selected from hydrogen, F, Cl, -OH, -CN, -CH3, -OCH3 and -CH2OH;

[0096] R 3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -OC 1-3 Alkyl, -C 1-3 Alkyl-OH, -S(O)2CH3, -CH2S(O)2CH3, -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.

[0097] Preferably, R 3 Selected from -CN, -CH2OH, -CHF2, -CH2F and -CF3.

[0098] In the present invention, L 1 When it is a single bond, R 3 Selected from cyano, -OC 1-6 Alkyl, -NHC 1-6 Alkyl, -NHC(O)C 1-6Alkyl, -C(O)C 1- 6-alkyl, -NHC(O)OC 1-6 Alkyl, -S(O)2C 1-6 Alkyl; the C 1-6 Each alkyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, -OH, cyano, -O-(C 1-6 alkyl) is substituted with a substituent.

[0099] In the present invention, the compound is not the following compound:

[0100] In the present invention, the 5-10 membered heteroaromatic ring is preferably a 5-6 membered heteroaromatic group, more preferably a pyridyl group or a pyrimidinyl group.

[0101] The halogen or halo is preferably fluorine, chlorine or bromine, more preferably fluorine.

[0102] Said-C 1-6 The alkyl group is preferably -C 1-3 The alkyl group is more preferably a methyl group, an ethyl group or an isopropyl group.

[0103] Said-OC 1-6 The alkyl group is preferably -OC 1-3 Alkyl, more preferably methoxy.

[0104] Said-NHC 1-6 The alkyl group is preferably -NHC 1-3 alkyl.

[0105] The -N(C 1-6 Alkyl)2 is preferably -N(C 1-3 Alkyl)2.

[0106] The -NHC(O)C 1-6 The alkyl group is preferably -NHC(O)C 1-3 alkyl.

[0107] Said-C 2-6 Alkenyl is preferably -C 2-4 The alkenyl group is more preferably vinyl or propenyl.

[0108] Said-C 2-6 Alkynyl is preferably -C 2-4 Alkynyl, more preferably ethynyl or propynyl.

[0109] The -halogenated C 1-6 Alkyl is preferably -haloC 1-3 The alkyl group is more preferably a halomethyl group or a haloethyl group.

[0110] Said-C 3-12 The cycloalkyl group is preferably -C 3-8Cycloalkyl, more preferably -C 3-6 Cycloalkyl.

[0111] The 4- to 12-membered heterocyclic group is preferably a 4- to 8-membered heterocyclic group, more preferably a 4- to 6-membered heterocyclic group, and further preferably an azetidinyl group, a pyrrolidinyl group, a piperidinyl group, or a piperazinyl group.

[0112] The -halogenated C 1-6 Alkoxy is preferably -haloC 1-3 The alkoxy group is more preferably a halomethoxy group or a haloethoxy group.

[0113] Said-C 1-6 Alkylene-preferably -C 1-3 Alkylene- is more preferably -methylene, -ethylene or -propylene.

[0114] Said-C 3-6 Cycloalkylene is preferably cyclopropylene.

[0115] The -NHC(O)-S(O)2-(C 1-6 Alkylene)-preferably -NHC(O)-S(O)2-(C 1-3 alkylene)-.

[0116] The -C(O)C 1-6 Alkylene - preferably -C(O)C 1-3 Alkylene-.

[0117] The -C(O)C 1-6 The alkyl group is preferably -C(O)C 1-3 alkyl.

[0118] The -NHC(O)OC 1-6 The alkyl group is preferably -NHC(O)OC 1-3 alkyl.

[0119] -S(O)2C 1-6 The alkyl group is preferably -S(O)2C 1-3 Alkyl, more preferably -S(O)2CH3alkyl.

[0120] Said-NHC 1-6 Alkyl-N(C 1-6 Alkyl)2 is preferably -NHC 1-3 Alkyl-N(C 1-3 Alkyl)2.

[0121] The -NH-C(O)O(C 1-6 Alkyl) is preferably -NH-C(O)O(C 1-3 alkyl).

[0122] The -N(C 1-6 alkyl)-C(O)OC 1-6 The alkyl group is preferably -N(C 1-3 alkyl)-C(O)OC 1-3 alkyl.

[0123] The -OC(O)NHC 1-6 The alkyl group is preferably -OC(O)NHC 1-3 alkyl.

[0124] The -N(C 1-3 alkyl)C(O)C 1-3 The alkyl group is preferably -N(C 1-3 alkyl)C(O)C 1-3 alkyl.

[0125] The present invention provides the following compounds or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites or prodrugs thereof, wherein the compounds are selected from:

[0126] In another aspect, the present invention provides a method for preparing a compound of formula (I), comprising the following steps:

[0127] 1. Compounds IN1-1 and IN1-2 are subjected to a substitution reaction or a metal-catalyzed coupling reaction (such as Pd(PPh3)4, PdCl2(dpPf)2, etc.) in an organic solvent (such as DMF, DMSO, NMP, dioxane, dichloromethane, toluene, tetrahydrofuran, chloroform, acetonitrile, etc.) in the presence of an organic base (such as Et3N, DIPEA, N-methylmorpholine, DBU, sodium hydride, etc.) or an inorganic base (such as potassium carbonate, cesium carbonate, etc.) to obtain compound IN1-3; 2. Compound IN1-3 is first reacted with an oxidant (such as m-CPBA, potassium persulfate, etc.) to generate an intermediate sulfoxide, which is then subjected to a substitution reaction with compound IN1-4 in the presence of a base (such as Et3N, DIPEA, N-methylmorpholine, DBU, sodium hydride, etc.) to obtain a compound of formula (I).

[0128] Among them, LG 1 represents a halogen leaving group, and the remaining groups are as defined in formula (I).

[0129] In some embodiments of the present invention, LG 1 Preferably chlorine or bromine;

[0130] The starting materials for the preparation process of the present invention can be obtained from commercial sources or can be prepared according to known methods.

[0131] Those skilled in the art will appreciate that, depending on the desired product structure, the order of the reaction steps may be appropriately adjusted, and protection / deprotection reaction steps may be added or omitted.

[0132] In another aspect, the present invention provides a pharmaceutical composition comprising a compound as described herein or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0133] A further object of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, which comprises combining the compound of the present invention or a pharmaceutically acceptable form thereof, or a mixture thereof, with one or more pharmaceutically acceptable carriers.

[0134] The pharmaceutically acceptable carrier that can be used in the pharmaceutical composition of the present invention is a pharmaceutically acceptable carrier. Examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (2005).

[0135] The pharmaceutical composition can be administered in any form, as long as it prevents, alleviates, prevents or cures the symptoms of a human or animal patient. For example, it can be prepared into various suitable dosage forms according to the route of administration.

[0136] In other embodiments, the administration of the compound or pharmaceutical composition of the present invention can be combined with another treatment method. The other treatment method can be selected from, but not limited to: radiation therapy, chemotherapy, immunotherapy, or a combination thereof.

[0137] 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 preparation is in the form of a solid preparation, a semi-solid preparation, a liquid preparation, or a gaseous preparation.

[0138] A further object of the present invention is to provide an article of manufacture, for example, in the form of a kit. As used herein, an article of manufacture is intended to include, but is not limited to, a kit and a package. The article of manufacture of the present invention comprises: (a) a first container; (b) a pharmaceutical composition in the first container, wherein the composition comprises: a first therapeutic agent, the first therapeutic agent comprising: a compound of the present invention or a pharmaceutically acceptable form thereof, or a mixture thereof; (c) an optional package insert indicating that the pharmaceutical composition can be used to treat a neoplastic condition (as defined below); and (d) a second container.

[0139] The first container is a container for holding a pharmaceutical composition. This container can be used for preparation, storage, transportation and / or individual / bulk sales. The first container is intended to encompass bottles, jars, vials, flasks, syringes, tubes (e.g., for cream products), or any other container for preparing, holding, storing, or dispensing pharmaceutical products.

[0140] The second container is a container for accommodating the first container and optional package insert. Examples of the second container include, but are not limited to, boxes (e.g., paper or plastic boxes), boxes, cartons, bags (e.g., paper or plastic bags), pouches, and sacks. The package insert can be physically adhered to the outside of the first container via a cable tie, glue, staples, or other adhesion methods, or it can be placed inside the second container without any physical tool for adhering to the first container. Alternatively, the package insert is located outside the second container. When located outside the second container, it is preferred that the package insert is physically adhered via a cable tie, glue, staples, or other adhesion methods. Alternatively, it can abut or contact the outside of the second container without physical adhesion.

[0141] The package insert is a trademark, label, or indicia that lists information about the pharmaceutical composition within the first container. The information listed is typically determined by the regulatory agency (e.g., the U.S. Food and Drug Administration) that governs the region in which the product is to be sold. Preferably, the package insert specifically lists the indications for which the pharmaceutical composition is approved. The package insert can be made of any material from which the information contained therein or thereon can be read. Preferably, the package insert is a printable material (e.g., paper, plastic, cardboard, foil, adhesive paper, or plastic, etc.) onto which the desired information can be formed (e.g., printed or applied).

[0142] In another aspect, the present invention provides 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 a Wee1 kinase-related disease.

[0143] 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 Wee1 inhibitor. In such use, the Wee1 inhibitor can be used in vivo in mammals or in vitro, primarily for experimental purposes, for example, as a standard or control sample for comparison, or prepared into a kit according to conventional methods in the art for rapid detection of Wee1 inhibitory effects.

[0144] 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 use in the preparation of a medicament for treating cancer. In a further embodiment, the cancer includes but is not limited to breast cancer, colorectal cancer, colon cancer, lung cancer and prostate cancer, as well as bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, gastrointestinal tissue cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vulvar cancer, as well as leukemia (including chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL) and chronic myeloid leukemia (CML)), multiple myeloma and lymphoma.

[0145] In some embodiments, the lung cancer is lung adenocarcinoma, and the colorectal cancer is colorectal adenocarcinoma.

[0146] In another aspect, the present invention provides a method for preventing or treating a Wee1 kinase-related disease, comprising administering to an individual in need thereof a compound as described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention.

[0147] 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 use in preventing or treating Wee1 kinase-related diseases.

[0148] In another aspect, the present invention provides a method for preventing or treating a Wee1 kinase-related disease in combination with a compound as described herein or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention, wherein the additional treatment method includes but is not limited to: radiotherapy, chemotherapy, immunotherapy, or a combination thereof.

[0149] In some embodiments, the Wee1 kinase-associated disease is a disease that is sensitive or responsive to Wee1 kinase inhibition.

[0150] In some embodiments, the Wee1 kinase-related disease is cancer. In further embodiments, the cancer includes but is not limited to breast cancer, colorectal cancer, colon cancer, lung cancer and prostate cancer, as well as bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, gastrointestinal tissue cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vulvar cancer, as well as leukemia (including chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL) and chronic myeloid leukemia (CML)), multiple myeloma and lymphoma. In some embodiments, the lung cancer is lung adenocarcinoma. The colorectal cancer is colorectal adenocarcinoma.

[0151] In a further preferred embodiment, the compounds of the present invention can be used in combination with chemoradiotherapy or immunotherapy to prevent or treat cancer.

[0152] The dosage regimen can be adjusted to provide the optimal desired response. For example, when administered as an injectable, a single bolus, bolus, and / or continuous infusion can be administered, among others. For example, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the therapeutic situation. It should be noted that dosage values ​​can vary depending on the type and severity of the condition to be alleviated and can include single or multiple doses. Generally, the dosage for treatment varies, depending on considerations such as: the age, sex, and general health of the patient to be treated; the frequency of treatment and the nature of the desired effect; the extent of tissue damage; the duration of symptoms; and other variables that can be adjusted by the individual physician. It will be further understood that for any particular individual, the specific dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the composition. The dosage and administration regimen of the pharmaceutical composition can be readily determined by one 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, with the dosage amount being, for example, 0.01 to 1000 mg / dose. The required dose may be administered in one or more doses to achieve the desired result.The pharmaceutical composition according to the present invention may also be provided in unit dosage form.

[0153] The present invention provides a novel class of highly active Wee1 inhibitors that can achieve at least one of the following technical effects: (1) high inhibitory activity against Wee1 kinase; (2) excellent physicochemical properties (e.g., solubility, physical and / or chemical stability); (3) excellent pharmacokinetic properties (e.g., good bioavailability, suitable half-life and duration of action); (4) excellent safety (lower toxicity and / or fewer side effects, wider therapeutic window), etc.

[0154] General Terms and Definitions

[0155] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including variations of technology or substitutions of equivalent technology that would be apparent to those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

[0156] The terms "include," "comprising," "having," "containing," or "involving," and their variations herein, are inclusive or open-ended and do not exclude other unrecited elements or method steps. Those skilled in the art will appreciate that the above terms, such as "comprising," encompass the meaning of "consisting of."

[0157] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0158] Unless otherwise indicated, concentrations are by weight and ratios (including percentages) are by molar amounts.

[0159] 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 or more.

[0160] When the lower and upper limits of a numerical range are disclosed, any value and any included range falling within the range are specifically disclosed. In particular, each range of values ​​disclosed herein (in the form "about a to b," or equivalently, "approximately a to b," or equivalently, "about a b") should be understood to represent each value and range encompassed within the broader range.

[0161] For example, the statement "C 1-6 " should be understood to include any sub-ranges therein and each point value, such as C 2-5 、C 3-4 、C 1-2 、C 1-3 、C 1-4 、C 1-5 etc., as well as C1, C2, C3, C4, C5, C6, etc. For example, the expression “C 3-10 ” should also be understood in a similar manner, for example, any sub-ranges and point values ​​contained therein may be included, for example, 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 C3, C4, C5, C6, C7, C8, C9, C 10etc. For another example, the expression "3-10 yuan" should be understood to include any sub-ranges and point values ​​therein, such as 3-4 yuan, 3-5 yuan, 3-6 yuan, 3-7 yuan, 3-8 yuan, 3-9 yuan, 4-5 yuan, 4-6 yuan, 4-7 yuan, 4-8 yuan, 5-7 yuan, 5-8 yuan, 6-7 yuan, etc., as well as 3, 4, 5, 6, 7, 8, 9, 10 yuan, etc. For another example, the expression "5-10 yuan" should also be understood in a similar manner, such as it can include any sub-ranges and point values ​​contained therein, such as 5-6 yuan, 5-7 yuan, 5-8 yuan, 5-9 yuan, 5-10 yuan, 6-7 yuan, 6-8 yuan, 6-9 yuan, 6-10 yuan, 7-8 yuan, etc., as well as 5, 6, 7, 8, 9, 10 yuan, etc.

[0162] As used herein, the term "alkyl" refers to a saturated straight or branched chain hydrocarbon group, alone or in combination with other groups. 1-6 "Alkyl" refers to a saturated straight or branched chain hydrocarbon group having 1 to 6 carbon atoms (e.g., 1, 2, 3, 4, 5 or 6 carbon atoms). 1- The "alkyl" is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or n-hexyl.

[0163] When used alone or in combination with other groups herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon group containing one or more double bonds. Illustrative examples of alkenyl include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl, and hexenyl. Illustrative examples of C2-6 alkenyl groups with one or more double bonds include butadienyl, pentadienyl, hexadienyl, and hexatrienyl, as well as branched forms thereof. The position of the unsaturated bond (double bond) can be at any position of the carbon chain.

[0164]

[0046] The term "alkynyl" as used herein alone or in combination with other groups refers to unsaturated straight or branched chain alkynyl groups, for example, ethynyl, 1-propynyl, propargyl, butynyl, and the like.

[0165] As used herein, the term "alkylene" refers to a saturated, linear or branched, divalent hydrocarbon group. 1-6 "Alkylene" refers to a saturated straight or branched divalent hydrocarbon group having 1 to 6 carbon atoms. 1-6 Examples of "alkylene" include, but are not limited to, methylene, ethylene, propylene, or butylene.

[0166] When used herein alone or in combination with other groups, the term "alkenylene" refers to a straight or branched divalent aliphatic hydrocarbon group having one or more carbon-carbon double bonds, and the two groups (or fragments) connected thereto may be connected to the same carbon atom or to different carbon atoms. For example, the term "C 2-6 "Alkenylene" refers to an alkenylene group having 2 to 6 carbon atoms (e.g. wait).

[0167] As used herein, the term "alkynylene" refers to a straight or branched divalent aliphatic hydrocarbon group having one or more carbon-carbon triple bonds, wherein the two groups (or fragments) connected thereto are each connected to a different carbon atom. For example, as used herein, the term "C 2-6 "Alkyne" refers to an alkynyl group having 2 to 6 carbon atoms (e.g. wait).

[0168] When used herein alone or in combination with other groups, the term "alicyclic" or "aliphatic hydrocarbon ring" refers to a monocyclic or polycyclic (including fused rings, bridged rings and spiro rings, such as bicyclic) aliphatic hydrocarbon ring system, including cycloalkane rings, cycloalkene rings and cycloalkyne rings.

[0169] As used herein, the term "cycloalkyl", "carbocycle" or "cycloalkylene" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) non-aromatic hydrocarbon group. Common cycloalkyl groups include, but are not limited to, monocyclic cycloalkyls such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclobutene, cyclopentene, cyclohexene, etc.; or bicyclic cycloalkyls including fused rings, bridged rings or spiro rings such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]nonyl, decahydronaphthyl, etc. For example, "C 3-12 "Cycloalkyl" refers to a cycloalkyl group having 3 to 12 ring carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12).

[0170] As used herein, the term "heterocycloalkyl" or "heterocyclyl," when used alone or in combination with other groups, refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic, e.g., fused, bridged, or spirocyclic) non-aromatic group whose ring atoms consist of carbon atoms and at least one (e.g., 1, 2, 3, or 4) heteroatom selected from nitrogen, oxygen, and sulfur. A heterocycloalkyl group may be attached to the remainder of the molecule via any one of the ring atoms if valence requirements are met. For example, a "3- to 8-membered heterocycloalkyl" group refers to a heterocycloalkyl group having 3 to 8 ring atoms. Common heterocycloalkyl groups include, but are not limited to, oxiranyl, oxocyclobutane, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, homopiperazinyl, sulfolane, and the like.

[0171] As used herein, the term "heteroaryl" or "heteroaromatic ring," alone or in combination with other groups, refers to an aromatic ring having a conjugated π-electron system in which one or more (e.g., 1, 2, or 3) of the ring atoms are heteroatoms selected from N, O, P, and S, and the remaining ring atoms are C. A heteroaryl group or heteroaromatic ring can be characterized by the number of ring atoms. For example, a 5-12 membered heteroaryl group can contain 5-12 (e.g., 5, 6, 7, 8, 9, 10, 11, or 12) ring atoms, particularly 5, 6, 9, or 10 ring atoms. Examples of heteroaryl groups include, for example, thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyridinyl, pyrazinyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, and the like; the term also encompasses heteroaryl groups or heteroaromatic rings that are optionally further fused to an aryl or heteroaryl ring to form a fused ring.

[0172] The term "haloalkyl" as used herein, alone or in combination with other groups, refers to an alkyl group as described above in which one or more hydrogen atoms are replaced by a halogen. For example, the term "C 1-6 "Haloalkyl" refers to a C 1-6 Alkyl. It will be understood by those skilled in the art that when there are 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 include -CH2F, -CHF2, -CF3, -CCl3, -C2F5, -C2Cl5, -CH2CF3, -CH2Cl or -CH2CH2CF3.

[0173] The term "alkoxy" as 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, pentoxy, and the like.

[0174] The term "halogen" as used herein alone or in combination with other groups refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). The term "hydroxy" as used herein alone or in combination with other groups refers to -OH.

[0175]

[0046] The term "cyano," as used herein, alone or in combination with other groups, refers to -CN.

[0176]

[0046] The term "nitro," as used herein, alone or in combination with other groups, refers to -NO2.

[0177] The term "amino," as used herein, alone or in combination with other groups, refers to -NH2.

[0178] The term "oxo," as used herein alone or in combination with other groups, refers to =0.

[0179] As used herein, the term "each independently" means that at least two groups (or fragments) present in a structure with the same or similar value ranges may have the same or different meanings under specific circumstances. For example, if substituent X and substituent Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y may be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X may be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl.

[0180] The term "substituted" and its other variant forms in this article refer to one or more (such as 1, 2, 3 or 4) atoms or atomic groups (such as hydrogen atoms) on the specified atom being replaced by other equivalents, provided that the normal valence of the specified atom or atomic group in the current situation is not exceeded and a stable compound can be formed. If an atom or atomic group is described as "optionally substituted by...", it can be substituted or unsubstituted. Unless otherwise indicated, the attachment site of a substituent herein can be from any suitable position of the substituent. When the connecting bond in a substituent is shown as a chemical bond between two atoms connected to each other in a ring system, it means that the substituent can be connected to any ring-forming atom in the ring system.

[0181] In this article, solid lines can be used Solid wedge or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention may exist as stereoisomers, which include cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, 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 (e.g., racemic mixtures and diastereomeric pairs).

[0182] The present invention also encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0183] It should also be understood that certain compounds of the present invention may be used therapeutically in free form or, where appropriate, in the form of 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, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0184] 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 a review 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 present invention are known to those skilled in the art.

[0185] As used herein, the term "ester" refers to esters derived from compounds described herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the invention in the free acid or alcohol form). The compounds of the invention may themselves also be esters.

[0186] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0187] Those skilled in the art will appreciate that, since nitrogen requires an available lone pair of electrons to be oxidized to oxides, not all nitrogen-containing heterocycles are capable of forming nitrogen oxides. Those skilled in the art will recognize nitrogen-containing heterocycles that are 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 preparing nitrogen oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidizing heterocycles and tertiary amines with peroxyacids such as Peracetic Acid and Metachloroperbenzoic Acid (mCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxirane such as dimethyldioxirane. These methods for preparing 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 ESGWerstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392 (AR Katritzky and AJ Boulton, Eds., Academic Press).

[0188] Also included within the scope of the present invention are metabolites of the compounds of the present invention, i.e., substances formed in vivo upon administration of the compounds of the present invention. Metabolites of the compounds can be identified using techniques known in the art, and their activity can be characterized by assays. Such products can be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Thus, the present invention includes metabolites of the compounds of the present invention, including compounds produced by methods that contact the compounds of the present invention with a mammal for a period of time sufficient to produce their metabolites.

[0189] The present invention further includes within its scope prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity and can be converted into compounds of the present invention having the desired activity by, for example, hydrolytic cleavage when administered to the body or thereon. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compounds in vivo. Further 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 present invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)).

[0190] The present invention also encompasses compounds of the present invention that contain protecting groups. In any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in TW Greene & PGM Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 2006, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0191] The present invention also encompasses methods for preparing the compounds described herein. It should be understood that the compounds of the present invention can 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 that is appropriate for the reagents and materials used and suitable for the transformation to be achieved.

[0192] 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.

[0193] As used herein, the term "effective amount" (e.g., "therapeutically effective amount" or "prophylactically effective amount") refers to an amount of active ingredient that, after administration, will achieve the desired effect to some extent, such as alleviating one or more symptoms of the condition being treated or preventing the appearance of the condition or its symptoms.

[0194] As used herein, unless otherwise indicated, the terms "treat," ...

[0195] As used herein, "subject" includes humans and non-human animals. Exemplary human subjects include human subjects suffering from diseases (e.g., the diseases described herein) (referred to as patients) or normal individuals. "Non-human animals" herein include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.). DETAILED DESCRIPTION

[0196] The present invention includes all combinations of the specific embodiments described. Further embodiments of the present invention and the full scope of its applicability will become apparent from the detailed description provided below. However, it should be understood that although the detailed description and specific examples indicate preferred embodiments of the present invention, these descriptions and examples are provided by way of illustration only, as various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art from this detailed description. All publications, patents, and patent applications cited herein, including citations, are incorporated herein by reference in their entirety for all purposes.

[0197] In the following examples, the experimental methods without specific conditions were carried out according to conventional methods and conditions, or selected according to the product instructions.

[0198] Example

[0199] The present invention will be described in detail below with reference to Examples and Test Examples. However, these examples are not intended to limit the scope of the present invention, and variations are possible without departing from the scope of the present invention.

[0200] Mass spectrometry (MS) was measured using an Agilent (ESI) mass spectrometer, manufactured by Agilent, model: Agilent 6120B.

[0201] Preparative high performance liquid chromatography (HPLC) was performed using a Shimadzu LC-8A preparative liquid chromatograph (YMC, ODS, 250×20 mm column).

[0202] Thin layer chromatography purification was performed using GF 254 (0.4-0.5 nm) silica gel plates produced in Yantai.

[0203] 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, and petroleum ether and ethyl acetate systems. The volume ratio of the solvents is adjusted according to the polarity of the compounds or by adding triethylamine.

[0204] Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as the stationary phase. Eluent systems include, but are not limited to, dichloromethane and methanol systems and n-hexane and ethyl acetate systems. The volume ratio of the solvents is adjusted according to the polarity of the compound, and a small amount of triethylamine can also be added for adjustment.

[0205] Unless otherwise specified in the examples, the reaction temperature is room temperature (20°C to 30°C).

[0206] 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.

[0207] Example 1: Preparation of compounds

[0208] 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)

[0209] Step 1: Preparation of 2-bromo-6-(3-((tert-butyldiphenylsilyl)oxy)methyl)azetidin-1-yl)pyridine (Compound A1-2)

[0210] 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) were added to dioxane (35 mL) and heated to 80°C with stirring for 4 hr. After TLC confirmed the reaction was complete, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with brine, dried over saturated sodium sulfate, and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography to afford compound A1-2 (1.4 g, 26.2% yield).

[0211] Step 2: Preparation of 2-allyl-1-(6-(3-((tert-butyldiphenylsilyl)oxy)methyl)azetidin-1-yl)pyridin-2-yl)-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound A1-4)

[0212] Compound A1-3 (300 mg, 1.35 mmol), compound A1-2 (779 mg, 1.61 mmol), cuprous iodide (257 mg, 1.35 mmol), potassium carbonate (279 mg, 2.02 mmol), and N,N′-dimethylethylenediamine (131 mg, 1.49 mmol) were added to dioxane (5 ml) and heated to 95° C. under nitrogen for 12 hours. LCMS monitoring showed that the starting 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. The crude product was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound A1-4 (400 mg, 47.6%).

[0213] Step 3: Preparation of 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)

[0214] Compound A1-4 (200 mg, 0.32 mmol) was dissolved in toluene (2 mL), and m-chloroperbenzoic acid (69 mg, 0.32 mmol, 80%) was added at 25°C. The reaction was allowed to proceed at this temperature for 1 hour. Subsequently, 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 the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound A1-6 (100 mg, 40%).

[0215] Step 4: 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)

[0216] [1] Compound A1-6 (100 mg, 0.13 mmol) was dissolved in methanol (3 mL), and ammonium fluoride (14.5 mg, 0.39 mmol) was added and reacted for 2 hours. LCMS showed that the reaction was complete. The reaction solution was filtered and concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography to obtain compound A1 (10 mg, 14.5%).

[0217] LCMS (ESI) m / z: 528.1 [M+H] + ;

[0218] 1 H NMR (400MHz, CD3OD) δppm 8.79 (s, 1H), 7.70 (t, J=7.6Hz, 1H), 7.59 (br d, J=8.8Hz, 2H), 7.05 (d, J=7.6Hz, 1H), 6.97 (d, J=9.2Hz, 2H), 6.34 (d, J=8.0Hz, 1H), 5 .70-5.82 (m, 1H), 5.09 (dd, J=10.2, 1.2Hz, 1H), 5.01 (dd, J=17.2, 1.2Hz, 1H), 4.76 (br d, J=6.4Hz, 2H), 4.11 (t, J=8.4Hz, 2H), 3.82 (dd, J=8.4, 5.2Hz, 2H), 3.77 (d, J=6 .4Hz, 2H), 3.18-3.24(m, 4H), 2.63-2.69(m, 4H), 2.51-2.41(m, 1H), 2.38(s, 3H).

[0219] Preparation Example 2: Preparation of Compound A9S

[0220] Step 1: Preparation of compound A9S-2

[0221] 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) were added to dimethyl sulfoxide (8 mL) and heated to 80°C with stirring for 2 hours. After TLC confirmed the completion of the reaction, water was added to the reaction solution, and the mixture was 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, 60% yield).

[0222] Step 2: Preparation of compound A9S-3

[0223] Compound A9S-2 (600 mg, 2.47 mmol), compound A1-3 (548 mg, 2.47 mmol), cuprous iodide (48 mg, 0.25 mmol), and potassium carbonate (279 mg, 4.94 mmol) were added to dioxane (5 ml) and heated to 95°C under nitrogen for 12 hours. LCMS monitoring indicated complete reaction. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography to afford compound A9S-3 (300 mg, 32% yield).

[0224] Step 3: Preparation of compound A9S

[0225] Compound A9S-3 (45 mg, 0.2 mmol) was dissolved in toluene (2 mL), and m-CPBA (43 mg, 0.2 mmol, 80%) was added at 25°C. The mixture was reacted at this temperature for 1 hour. Subsequently, N,N-diisopropylethylamine (168 mg, 1.3 mmol) and compound A1-5 (100 mg, 0.26 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 the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse HPLC to obtain compound A9S (10 mg, 10% yield).

[0226] LCMS (ESI) m / z: 528.2 [M+H] + ;

[0227] 1 H NMR (400MHz, DMSO-d6) δ10.09 (s, 1H), 8.79 (s, 1H), 8.30 (s, 1H), 7.74 (d, J=8.2Hz, 1H), 7.60 (s, 2H), 7. 04 (d, J=7.6Hz, 1H), 6.96-6.84 (m, 2H), 6.46 (d, J=8.2Hz, 1H), 5.68 (ddt, J=16.4, 10.2, 5.8Hz, 1H), 5.04 (dt, J=10.4, 1.4Hz, 1H), 4.92 (dt, J=17.2, 1.6Hz, 1H), 4.59 (s, 2H), 4.27 (t, J=7.0Hz, 1H), 3.93-3.75 (m , 2H), 3.68 (td, J=5.8, 5.2, 3.2Hz, 2H), 3.09 (t, J=5.0Hz, 4H), 2.45 (t, J=5.0Hz, 4H), 2.34-2.13 (m, 5H).

[0228] The following compounds were synthesized by referring to the method of Preparation Example 1 or Preparation Example 2:

[0229] Preparation Example 3: Reference compound 1 and reference compound 2 were prepared according to the method of Example 131 and Example 19 in patent WO2023072301.

[0230] Example 2: Wee1 enzyme activity inhibition experiment

[0231] In the Wee1 enzyme inhibition assay, the effects of various compounds on Wee1 activity were investigated using the ADP-Glo ​​luminescent kit. Wee1 enzyme was purchased from Carna bioscience, and Poly(Lys Tyr(4:1)) was used as the substrate. Compounds (maximum concentration 1000 nM, in a 3-fold serial dilution) were incubated with Wee1 (2 nM) for 30 minutes. The reaction substrate and ATP were then added and incubated at room temperature for 1 hour. ADP-Glo ​​stop reagent was then added, and the reaction was incubated in the dark for 40 minutes at room temperature. ADP-Glo ​​kinase detection reagent was then added, and the reaction was incubated in the dark for 1 hour at room temperature. Luminescence was measured, and the half-maximal inhibitory concentration (50%) of the compound on enzyme activity was calculated.

[0232] Table 1. Results of Wee1 enzyme activity inhibition test

[0233] ++++:IC 50 <1nM;+++: 1nM <IC 50 <10 nM; the results show that the compound of the present application has good Wee1 enzyme inhibitory activity.

[0234] Example 3: Cell proliferation inhibition experiment 1

[0235] The effects of each compound on cell proliferation were tested in A427 cells. Specifically, after incubation of the compound with gradient dilution for 120 hours, The chemiluminescent cell viability assay (CTG method) was used to evaluate and calculate the half-maximal inhibitory concentration (IC50) of the compounds on these cell lines.

[0236] Table 2. Inhibitory effects of compounds on cell proliferation

[0237] ++++:IC 50 <200nM;+++:200nM<IC 50 <400nM;++>400nM

[0238] The results show that the compound of the present application has good cell proliferation inhibitory activity.

[0239] Example 4: Cell proliferation inhibition experiment 2

[0240] The effects of each compound on cell proliferation were tested in HT29, SNU5, SNU16, and A427 cells. Specifically, after incubation of the compound with gradient dilution for 120 hours, The chemiluminescent cell viability assay (CTG method) was used to evaluate and calculate the half-maximal inhibitory concentration (IC50) of the compounds on these cell lines.

[0241] Table 3. Inhibitory effects of compounds on proliferation of different cells

[0242] The results showed that the compound of the present application had better cell proliferation inhibitory activity in HT29 and SNU16 cells compared with AZD1775.

[0243] Example 5: Pharmacokinetic (PK) study in Balb / c mice

[0244] The pharmacokinetic properties of compounds A9S and AZD1775 were evaluated in female and male Balb / c mice by intravenous and oral administration. The intravenous dose was 1 mg / kg in a solvent consisting of 5% DMSO, 5% Solutol, and 90% Saline; the oral dose was 10 mg / kg in a solvent consisting of 0.5% sodium methylcellulose (MC). Blood was collected at various time points after intravenous and oral administration, anticoagulated with EDTA-K2, and stored at -80°C.

[0245] Blood or plasma samples were treated with protein precipitation and then analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using WinNonlin 6.3 software and a non-compartmental model. The results are shown in Table 4 below.

[0246] Table 4. Pharmacokinetic parameters of the compounds in the blood of male and female Balb / c mice Note: “ / ” means it does not exist

[0247] Conclusion: Compound A9S of the present invention achieved higher exposure 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. Its comprehensive properties were significantly better than those of AZD1775, and it exhibited excellent pharmacokinetic properties.

[0248] Example 6: Biochemical hERG inhibition assay

[0249] 1. Experimental Materials

[0250] Culture of stably transfected HEK 293 cells (human embryonic kidney cells, purchased from Invitrogen) expressing hERG channels (Cat. K1236): cells were cultured in a medium containing 85% DMEM, 10% dialyzed fetal bovine serum, 0.1 mM NEAA, 25 mM HEPES, 100 U / mL penicillin-streptomycin, 5 μg / mL Blasticidin, and 400 μg / mL Geneticin. 2 , 5% CO2, 37℃ in cell culture flasks. TM Express cells were passaged approximately three times per week and maintained at approximately 40% to 80% confluency. 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 onto 3.5 cm cell culture dishes (5 × 10 5 cells / 3.5 cm cell culture dish) and cultured in a medium without blasticidin and geneticin.

[0251] Note: The cell line used for safety evaluation tests has a passage number of less than 60.

[0252] 2. Preparation of Working Solutions of Test Compounds

[0253] The compounds were dissolved in DMSO at 30 mM, 10 mM, 3.33 mM, 1.11 mM and 0.37 mM, and then added to the extracellular fluid to give final concentrations of 30 μM, 10 μM, 3.33 μM, 1.11 μM and 0.37 μM, respectively. The final concentration of DMSO was 0.1%.

[0254] 3. Experimental Plan

[0255] 1) Take a coverslip with a large number of single HEK 293 hERG cells uniformly grown on its surface and place it in a continuous recording chamber on an inverted microscope. Perfuse the cells with extracellular solution (approximately 1 ml per minute) and continuously record until the current stabilizes. 2) Use standard whole-cell recording mode to record the hERG channel current of a single cell. First, clamp the membrane voltage at -80mV. Stimulate the cells with a voltage of +20mV for 5 seconds to activate the hERG potassium channel. Then repolarize to -50mV for 5 seconds to generate an outward tail current. Continue perfusing until the current stabilizes. The peak tail current at this point is the control current value. 3) Then, perfuse the cells with extracellular solution containing the drug to be tested and continue recording until the drug's inhibitory effect on the hERG current reaches a steady state. The peak tail current at this point is the current value after drug addition. 4) Perfuse the cells again with extracellular solution until the hERG current returns to or approaches the level before drug addition. Then, continue perfusing to test other concentrations or drugs. One or more compounds or drug concentrations can be tested on each cell. 5) Dofetilide (TRC) was used as a positive control in the experiment to ensure that the cells used reacted normally.

[0256] 4. Data acceptance criteria

[0257] The following criteria were used to determine the acceptability of data.

[0258] 1) Initial sealing resistance > 1GΩ;

[0259] 2) The leakage current is less than 50% of the controlled peak tail current at any time;

[0260] 3) peak tail amplitude >250 pA;

[0261] 4) Sheet resistance Rm>500MΩ;

[0262] 5) Access resistance (Ra) < 15MΩ;

[0263] 6) The apparent decrease in peak current is less than 2.5% / min.

[0264] 5. Experimental Results

[0265] Table 5. hERG inhibition test results

[0266] 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 drugability.

[0267] Example 7: Biochemical CYP enzyme (cytochrome P450) inhibition test

[0268] 1. Test system:

[0269] P450-Glo TM CYP1A2 Screening System, (Promega);

[0270] P450-Glo TM CYP2D6 Screening System, (Promega);

[0271] P450-Glo TM CYP3A4 Screening System, (Promega).

[0272] 2. Test instruments:

[0273] BMG PHERAstar FS Luminescent.

[0274] 3. Test method:

[0275] The test was carried out according to the kit instructions, the steps are as follows:

[0276] 3.1. Inhibition of CYP1A2:

[0277] Test group: Different concentrations of the test compound were added to the microplate, and Luciferin-ME (100 μM), K3PO4 (100 mM), and CYP1A2 (0.01 pmol / μL) were added to each well. The cells were pre-incubated at room temperature for 10 min, and then the NADPH regeneration system was added. The reaction was carried out at room temperature for 30 min. Finally, an equal volume of detection buffer was added, and the cells were incubated at room temperature for 20 min. Then, chemiluminescence detection was performed.

[0278] Negative control group: The experimental method is the same as the test group, except that no test compound is added.

[0279] Blank control group: The experimental method is the same as the test group, except that the test compound is not added and CYP1A2 Membrance (0.01 pmol / μL) is used instead of CYP1A2.

[0280] 3.2. Inhibition of CYP2D6:

[0281] Test group: Different concentrations of the test compound were added to a microplate. Luciferin-ME EGE (3 μM), K3PO4 (100 mM), and CYP2D6 (5 nM) were added to each well and pre-incubated at room temperature for 10 minutes. Subsequently, an NADPH regeneration system was added and the reaction was carried out at 37°C for 30 minutes. Finally, an equal volume of detection buffer was added and incubated at room temperature for 20 minutes. Then, chemiluminescence detection was performed.

[0282] Negative control group: The experimental method is the same as the test group, except that no test compound is added.

[0283] Blank control group: The experimental method is the same as the test group, except that the test compound is not added and CYP2D6 Membrance (5nM) is used instead of CYP2D6.

[0284] 3.3. Inhibition of CYP3A4:

[0285] Test group: Different concentrations of the test compound were added to the microplate, and Luciferin-IPA (3 μM), K3PO4 (100 mM), and CYP3A4 (2 nM) were added to each well. The cells were pre-incubated at room temperature for 10 min, and then the NADPH regeneration system was added. The cells were reacted at room temperature for 30 min. Finally, an equal volume of detection buffer was added, and the cells were incubated at room temperature for 20 min. Then, chemiluminescence detection was performed.

[0286] Negative control group: The experimental method is the same as the test group, except that no test compound is added.

[0287] Blank control group: The experimental method is the same as the test group, except that the test compound is not added and CYP3A4 Membrance (2nM) is used instead of CYP3A4.

[0288] 4. Data processing:

[0289] Percent inhibition rate = (1-(chemiluminescent signal value of the test compound concentration group-chemiluminescent signal value of the blank control group) / (chemiluminescent signal value of the negative control group-chemiluminescent signal value of the blank control group)) × 100%.

[0290] When the percentage inhibition rate is between 30% and 80%, the half-maximal inhibitory concentration (IC 50 ) or range: IC 50 =X×(1-percent inhibition rate) / percent inhibition rate, where X is the test concentration of the compound.

[0291] 5. Test results:

[0292] The inhibition of the compounds of the present invention on three CYP enzymes was determined according to the above method. The results are shown in Table 5 below.

[0293] Table 5. CYP enzyme inhibition test results

[0294] 6. Conclusion

[0295] 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, which is better than reference compound 1 and reference compound 2.

[0296] The above embodiments do not limit the solutions of the present application in any way. In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art based on 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

1. A compound represented by formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, in, Ring A is selected from phenyl, 5-10 membered heteroaromatic ring; said ring A is optionally substituted by one or more R a replace; Every R a Each 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; the C 1-6 Alkyl, C 2-4 Alkenyl, C 2-4 Each alkynyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, oxo, -OH, -CN, -NH2, -C 1-6 Substitution of alkyl groups; R 1 Selected from halogen, -CN, C 1-6 Alkyl, halogenated C 1-6 Alkyl, -C 3-12 Cycloalkyl, 4-12 membered heterocyclic group, -O-(4-12 membered heterocyclic group), -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 dd ; The C 1-6 Alkyl, C 3-12 Cycloalkyl, 4-12 membered heterocyclic group are each optionally substituted by one or more selected from halogen, oxo, -CN, -OR b 、-NR b R d 、-C 1- 6-alkyl, -halogenated C 1-6 Alkyl, -halogenated C 1-6 Alkoxy, -C 1-6 Alkylene-OR b 、-C 1-6 Alkylene-NR b R d 、-C 3-8 Cycloalkyl, 4-8 membered 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)2R d 、-N(R b )S(O)2R d Substituents are substituted; further, the C 3-8 Cycloalkyl, 4-8 membered heterocyclic group are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -C 1-6 Alkyl, -halogenated C 1-6 Alkyl, -NHC 1-6 Alkyl, -N(C 1-6 alkyl)2 substituent; R b 、R d Each independently selected from H, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, 4-12 membered heterocyclic group; the C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-12 membered heterocyclic group are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -NH2, -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 Alkylene-N(C 1-6 alkyl)2 is substituted with a substituent; R 2 Selected from hydrogen, halogen, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, 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, heteroaryl are each optionally substituted by one or more selected from halogen, oxo, -OH, -CN, -NH2, -C 1-6 Substitution of alkyl groups; Ring B is selected from phenyl, 5-10 membered heteroaryl; said ring B is optionally substituted by one or more substituents selected from hydrogen, halogen, -CN, -OCH3; The E ring is selected from -C 3-12 Cycloalkyl, 4-12 membered heterocyclic group; the E ring is optionally replaced by one or more R c replace; Every R c are 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 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, 3-6 membered heterocyclylene and C 3-6 The cycloalkylene groups are each optionally substituted with one or more substituents selected from hydrogen, halogen, oxo, -OH, and amino; R 3 Select from the following conditions: i)L 1 When it 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)2C 1-6 Alkyl; the C 1-6 Each alkyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, -OH, cyano, -O-(C 1-6 alkyl) is substituted with a substituent; ii)L 1 When it is not a single bond, R 3 Selected from halogen, amino, -OH, cyano, -halogenated C 1-6 Alkyl, -S(O)2C 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)NH2, -OC(O)NHC 1-6 Alkyl and -N(C 1-6 alkyl)C(O)C 1-6 Alkyl, wherein the C 1-6 The alkyl group is optionally substituted with one or more radicals selected from halogen, oxo, -OH, amino and -O-(C 1-6 alkyl) is substituted with a substituent.

2. The compound according to claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein: Ring A is selected from phenyl, pyridyl, thienyl and pyrazolyl, and said ring A is optionally substituted by one or more R a Replace, each R a Each independently selected from H, F, Cl, -OH, -C 1-3 Alkyl and -OC 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with one or more substituents selected from F, Cl, -OH and -CN; Preferably, the A ring is selected from phenyl, said phenyl being optionally substituted with one or more R a Replace, each R a Each independently selected from H, F, Cl, -OH, -C 1-3 Alkyl and -OC 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with one or more substituents selected from F, Cl, -OH and -CN; More preferably, the A ring is selected from phenyl, said phenyl being optionally substituted by one or more R a Replacement, R a Selected from H, F, Cl, -CH3, -OCH3 and -CH2OH.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein R 1 Selected from C 1-6 Alkyl, 4-6 membered heterocyclic group, C 4-6 Cycloalkyl, -O-(4-6 membered heterocyclyl), -NH-(4-6 membered heterocyclyl), -C(O)N(C 1-6 alkyl) 2, the 4-6 membered heterocyclic group and C 4-6 Each cycloalkyl group is optionally substituted by one or more radicals selected from halogen, oxo, -OH, -CN, -OCH3, -NH2, -N(C 1-3 Alkyl)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(C 1-3 Alkyl)2, -C(O)OC 1-3 Alkyl and -S(O)2C 1-3 Substitution of alkyl groups; Preferably, R 1 pyrrolidinyl, tetrahydropyranyl, piperidinyl, tetrahydropyridinyl, morpholinyl and piperazinyl; said pyrrolidinyl, tetrahydropyranyl, piperidinyl, tetrahydropyridinyl, morpholinyl and piperazinyl are each optionally substituted by one or more selected from F, Cl, -OH, -CN, oxo, -OCH3, -NH2, -N(CH3)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(CH3)2, -C(O)OC 1-3 Alkyl and -S(O)2C 1-3 Substitution of alkyl groups; Preferably, R 1 Selected from 4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, wherein R 2 is selected from -CH3, -CH2CH3, cyclopropyl, isopropyl, allyl, propargyl, -CH2-cyclopropyl, -CH2CF3; preferably, R 2 Selected from allyl.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled, metabolite, or prodrug thereof, wherein the B ring is selected from phenyl, pyridyl, pyrimidinyl, thienyl, and thiazolyl, and the B ring is optionally substituted with one or more substituents selected from hydrogen, halogen, and -CN; Preferably, the B ring is selected from pyridyl and phenyl.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled, metabolite or prodrug thereof, wherein the E ring is selected from a 4-6 membered heterocyclic group; the E ring is optionally replaced by one or more R c Replace, each R c Each is independently selected from hydrogen, F, Cl, -OH, oxo, -CN, -CH3, -OCH3; Preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl.

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein L 1 is selected from methylene, -C(O)-methylene-O-, and -C(O)-; wherein the methylene is optionally substituted with one or more substituents selected from hydrogen, F, and -CH3.

8. The compound according to claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled compound, metabolite or prodrug thereof, wherein R 3 Selected from: i)L 1 When it is a single bond, R 3 Selected from -OCH3, -NHCH3, -NHC(O)CH3, -C(O)CH3 and -S(O)2CH3; ii) L 1 When it is not a single bond, R 3 is selected from F, amino, -OH, -CN, -CHF2, -CH2F, -S(O)2CH3, -OCH3, -OCHF2, -NH-C(O)OCH3; preferably, R 3 Selected from F, -OH, -CN, -NH2-CHF2, -S(O)2CH3 and -CH3.

9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound is selected from: in, R 1 As claimed in claim 3; The E ring is selected from a 4-6 membered heterocyclic group, and the E ring is optionally replaced by one or more R c Replace, each R c Each is independently selected from hydrogen, F, Cl, -OH, oxo, -CN, -CH3, -OCH3; preferably, the E ring is selected from azetidinyl, pyrrolidinyl and piperidinyl; L 1 As claimed in claim 7; R 3 As described in claim 8.

10. The compound according to claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound is selected from: in, The E ring is selected from a 4-6 membered heterocyclic group, wherein the heteroatom of the 4-6 membered heterocyclic group is N, and the number of heteroatoms is 1, 2 or 3; The E ring is optionally replaced by one or more R c Replacement, R c Each independently selected from H, F, Cl, -OH, -CN, -C 1-6 Alkylene-OH, -C 1-6 Alkyl and -halogenated C 1-6 alkyl; R 3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -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)2C 1-6 Alkyl, -C 1-6 Alkyl S(O)2C 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-NH2, -C 1-6 Alkyl OC 1-6 Alkyl and -cyclopropyl-OH; the C 1-6 Each alkyl group is optionally substituted with one or more substituents selected from hydrogen, halogen, -OH, amino, and cyano.

11. The compound according to claim 10, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, in, R a Selected from H, F, Cl, -CH3, -CN, -OCH3 and -CH2OH; R 1-1 is -CH3 or -CD3; Ring E is selected from azetidinyl, pyrrolidinyl and piperidinyl; The E ring is optionally replaced by one or more R c Replacement, R c Each independently selected from hydrogen, F, Cl, -OH, -CN, -CH3, -OCH3 and -CH2OH; R 3 Selected from -CN, -C(O)NH2, -OC(O)NH2, -OC 1-3 Alkyl, -C 1-3 Alkyl-OH, -S(O)2CH3, -CH2S(O)2CH3, -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 Selected from -CN, -CH2OH, -CHF2, -CH2F and -CF3.

12. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, which satisfies one or more of the following conditions: 1)L 1 is a single bond, R 3 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)2C 1-6 Alkyl; the C 1-6 Each alkyl group is optionally substituted by one or more radicals selected from hydrogen, halogen, -OH, cyano, -O-(C 1-6 alkyl) is substituted with a substituent; 2) Ring A is phenyl, preferably 3) R 1 is a 4-12 membered heterocyclic group, preferably a 4-6 membered heterocyclic group, more preferably a 6 membered heterocyclic group; the heterocyclic group is optionally substituted by one or more -C 1-6 Alkyl or -deuterated C 1-6 Alkyl substituted, preferably optionally with one or more -C 1-3 Alkyl or -deuterated C 1-3 Alkyl substituted, more preferably substituted by -CH3 or -CD3; 4) R 2 -C 2-6 Alkenyl, preferably -C 2-4 alkenyl, more preferably allyl; 5) Ring B is a 5-10 membered heteroaryl group, preferably a 5-6 membered heteroaryl group, more preferably a 6 membered heteroaryl group, further preferably a pyridyl group, and particularly preferably 6) Ring E is azetidinyl or pyrrolidinyl, preferably More preferably The E ring is optionally replaced by one or more R c Replace, each R c Each independently selected from H, F, Cl, -OH, -CN, -CH3, -OCH3; 7)L 1 Selected from single bond, -C 1-6 Alkylene-, -C 3-6 Cycloalkylene- and -C(O)-; the -C 1-6 Alkylene-preferably -C 1-3 Alkylene-, more preferably methylene; the -C 3-6 Cycloalkylene - preferably cyclopropylene; 8)R 3 Selected from: i)L 1 When it is a single bond, R 3 Selected from cyano, -OC 1-6 Alkyl and -S(O)2C 1-6 Alkyl, wherein the -OC 1-6 The alkyl group is optionally substituted with one or more substituents selected from halogen; the -OC 1-6 The alkyl group is preferably -O-halogenated C 1-3 Alkyl, more preferably -OCF2; the -S(O)2C 1-6 The alkyl group is preferably -S(O)2C 1-3 Alkyl, more preferably -S(O)2CH3; ii)L 1 -C 1-6 Alkylene-, -C 3-6 Cycloalkylene- or -C(O)-, preferably -C 1-6 Alkylene-, more preferably -C 1-3 When alkylene-, R 3 Selected from halogen, amino, -OH, cyano, C 1-6 Alkyl, -halogenated C 1-6 Alkyl, -S(O)2C 1-6 Alkyl and -OC 1-6 Alkyl; the -halogenated C 1-6 Alkyl is preferably -haloC 1-3 Alkyl, more preferably fluorinated C 1-3 Alkyl, for example, is CHF2 or CH2F; the -S(O)2C 1-6 The alkyl group is preferably -S(O)2C 1-3 Alkyl, more preferably -S(O)2CH3; the -OC 1-6 The alkyl group is preferably -OC 1-3 Alkyl, more preferably -OCH3; 9) In the compound of formula (I-2), R 1-1 Selected from -C 1-6 Alkyl and deuterated C 1-6 Alkyl; the -C 1-6 The alkyl group is preferably -C 1-3 Alkyl, more preferably -CH3; the -deuterated C 1-6 The alkyl group is preferably a deuterated C 1-3 Alkyl, more preferably -CD3.

13. The compound according to claim 12, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, which satisfies one or more of the following conditions: 1)L 1 is methylene, R 3 Selected from halogen, amino, -OH, cyano, C 1-6 Alkyl, -halogenated C 1-6 Alkyl, -S(O)2C 1-6 Alkyl and -OC 1-6 Alkyl; preferably halogen, amino, -OH, cyano, C 1-3 Alkyl, -halogenated C 1-3 Alkyl, -S(O)2C 1-3 Alkyl and -OC 1-3 alkyl; 2) R 1 is piperidinyl, tetrahydropyridinyl or piperazinyl, said piperidinyl, tetrahydropyridinyl or piperazinyl being optionally substituted by one or more -C 1-3 Alkyl or -deuterated C 1-3 Alkyl substituted; preferably piperazinyl, the piperazinyl optionally substituted by one or more -CH3 or -CD3; preferably, R 1 Selected from Preferably 3) Structural unit Selected from Preferably 4) Structural unit Selected from 5) Structural unit for Preferably Preferably, the structural unit for 14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound is selected from: in: R 1-1 、R c , R3, R a As claimed in any one of claims 1 to 13; Preferably, the compound is selected from: in: R c , R3, R a As described in any one of claims 1 to 13.

15. The compound according to claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound is selected from:

16. A pharmaceutical composition comprising the compound of any one of claims 1 to 15 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

17. Use of a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug thereof, or a pharmaceutical composition according to claim 16, in the preparation of a medicament for preventing and / or treating cancer or in the preparation of a Wee1 inhibitor; preferably, the cancer is selected from one or more of the following cancers: breast cancer, colorectal cancer, colon cancer, lung cancer, prostate cancer, bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, gastrointestinal tissue cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vulvar cancer, leukemia, multiple myeloma and lymphoma.

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