Wee1 protein degradation agent and use thereof

Abstract

Provided in the present invention are a Wee1 protein degradation agent and the use thereof. The Wee1 protein degradation agent of the present invention is a compound of following formula I, a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, an isotopically labeled compound or a pharmaceutically acceptable salt thereof, or a mixture of same, wherein each group in the formula is as described herein. The compound of the present invention can be used for treating diseases by means of reducing the level of Wee1 protein. Q-L-U (I)

Description

Wee1 protein degrader and its application Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a Wee1 protein degrader and its application. Background Technology

[0002] The cell cycle is the process of cell growth and division, mainly consisting of four phases: G1, S, G2, and M. In the G1 phase, the cell actively metabolizes and synthesizes necessary substances; in the S phase, DNA replication occurs; in the G2 phase, growth and DNA inspection take place; and in the M phase, mitosis produces two daughter cells. Checkpoints are established throughout the process to ensure accurate replication of genetic information. Each cell cycle checkpoint is a system of multiple factors working through complex mechanisms. For example, the so-called G2-M checkpoint, before the cell completes DNA replication and enters mitosis, is composed of a very complex system that uses complex mechanisms, including phosphorylation, to check for DNA damage or defects after DNA synthesis, thus determining whether the cell will proceed to the next phase of chromosome separation (M phase). An important kinase in this cell cycle checkpoint is Cdk1, which forms a complex with Cyclin-B1 (Nurse, P., 1990, Nature 344, 503-508). The activation and inactivation of Cdk1 play a crucial role in the cell's transition from the G2 phase to mitosis (M phase) and the subsequent completion of mitosis. The activity of Cdk1 is regulated through multiple mechanisms, including binding to cyclin A or cyclin B, as well as phosphorylation and dephosphorylation.

[0003] Wee1 is a tyrosine kinase that inhibits Cdk1 activity by phosphorylating tyrosine 15 (Y15) on Cdk1 (McGowan, CH, et al., 1993, The EMBO journal 12, 75-85; Parker, LL, et al., 1992, Science 257, 1955-1957). Therefore, Wee1 is a key inhibitory regulator of Cdk1 activity, playing a crucial role at the G2-M phase checkpoint (O'Connell, et al., 1997, The EMBO journal 16, 545-554). Loss or inactivation of Wee1 leads to premature mitosis, resulting in mitotic failure and cell death (Stumpff, J., et al., 2004, Curr Biol 14, 2143-2148). Some tumor cells have functional defects or loss of the G1-S phase cell cycle checkpoint, heavily relying on the G2-M phase checkpoint to ensure cell growth and division (Sancar, A. et al., 2004, Annual review of biochemistry 73, 39-85). In these cancer cells that have lost the ability to use the G1 phase checkpoint, further loss of the G2-M phase checkpoint due to factors such as p53 deficiency leading to reduced Wee1 expression or activity makes these tumor cells highly sensitive to DNA damage (Wang, Y. et al., 2004, Cancer biology & therapy 3, 305-313).

[0004] Currently, several Wee1 kinase inhibitors, such as Zn-c3, Debio0123, SC0191, and SY4835, are in the clinical research stage. Although these Wee1 kinase inhibitors have shown certain clinical efficacy, they still have problems such as large clinical toxic side effects and poor patient tolerance.

[0005] Protein degrading agents represent a novel therapeutic modality that utilizes the cell's natural protein degradation mechanism (ubiquitin-proteasome system) to selectively target and eliminate proteins associated with disease pathogenesis. In recent years, due to their unique mechanism of action and broad application prospects, they have occupied an important position in the field of new drug development, providing new directions for drug discovery. Compared with traditional small molecule inhibitors, protein degrading agents offer several advantages. They can target proteins that traditional small molecule inhibitors struggle to bind to, including "undruggable" targets that lack active sites or have inaccessible active sites. Protein degrading agents function through a catalytic mechanism; once the degradation of the target protein is induced, the effect can persist even as drug concentrations decrease. Theoretically, their catalytic properties allow for therapeutic effects at lower doses, potentially helping to reduce drug side effects and improve safety. Protein degrading agents may offer a wider therapeutic window because they do not require sustained occupation of the target protein but rather achieve therapeutic effects by inducing its degradation. Because protein degrading agents eliminate the entire protein rather than merely inhibiting its activity, they can reduce drug resistance issues caused by target protein mutations or over-regulation. For targets that are structural proteins involved in disease processes, protein degraders can disrupt their structure and function by degrading these proteins, which is often difficult to achieve with traditional small molecule inhibitors.

[0006] Wee1 protein degradation can effectively eliminate Wee1 kinase activity, thereby blocking downstream signal transduction. Weel degraders represent an emerging strategy for cancer treatment and show broad application prospects. In recent years, various Weel degraders have been reported and disclosed, such as WO2020069105, CN113402520, WO2023083194, WO2024098692, and WO2024146502. Summary of the Invention

[0007] The present invention provides compounds with structures as shown in Formula I (including Formulas Ia, Ib, IIa, IIb, IIIa and IIIb), which can act as Weel degrading agents and reduce Wee1 protein levels.

[0008] The present invention also provides a pharmaceutical composition comprising an effective amount of a compound of formula I (including formulas Ia, Ib, IIa, IIb, IIIa and IIIb) for the treatment of cancer.

[0009] In one specific embodiment, the pharmaceutical composition may also contain one or more pharmaceutically acceptable carriers or diluents for the treatment of cancer.

[0010] In one specific embodiment, the pharmaceutical composition may also contain at least one known anticancer drug or a pharmaceutically acceptable salt of said anticancer drug for the treatment of cancer.

[0011] This invention also relates to methods for preparing novel compounds of structural formula I (including formulas Ia, Ib, IIa, IIb, IIIa and IIIb). Attached Figure Description

[0012] Figure 1 shows the protein levels of WEE1 and downstream pCDK1 (Y15) in the LoVo cell line after treatment with the exemplary compounds of the present invention. Detailed Implementation

[0013] It should be understood that the features of the various embodiments described herein can be combined arbitrarily to form the technical solutions described herein; the definitions of each group herein apply to any embodiment described herein, for example, the definitions of alkyl substituents herein apply to any embodiment described herein, unless the alkyl substituents have been clearly defined in the embodiment.

[0014] The term "hydrogen (H)" as used in this article includes its isotopes deuterium (D) and tritium (T).

[0015] As used in this article, "alkyl" refers to an alkyl group itself or a straight-chain or branched group with up to ten (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) carbon atoms. Useful alkyl groups include straight-chain or branched C atoms. 1-10 Alkyl, preferably C 1- 6-alkyl. In some embodiments, the alkyl group is C6. 1-4 Alkyl group. Typical C 1-10 Alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, 3-pentyl, hexyl, and octyl, which can be optionally substituted.

[0016] The term "alkoxy" as used in this article refers to the alkoxy group that is reacted with the aforementioned C 1-10 Alkyl, preferably C 1-6 Alkyl or C 1-4 Alkyl-substituted oxygen groups, such as methoxy, ethoxy, etc. The alkyl group in the alkoxy group may optionally be substituted. Substituents in the alkoxy group include, but are not limited to, halogens, amino groups, and carboxyl groups (including their ester groups), wherein the amino group includes alkylamino and dialkylamino groups.

[0017] The term "amino" as used herein can be represented as -NR′R″, where R′ and R″ are each independently hydrogen, and optionally substituted C. 1-10 Alkyl, optionally substituted cycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; or R′ and R″ together with the N to which they are attached form an optionally substituted 4- to 7-membered cycloamino group, said cycloamino group optionally containing one or more (e.g., 2, 3) additional heteroatoms selected from O, N, and S. Preferred amino groups include NH2, and at least one of R′ and R″ is C. 1-6 Alkyl groups.

[0018] The term "oxygenation" as used in this article refers to =O.

[0019] As used herein, "aryl" refers to a monocyclic, bicyclic, or tricyclic aromatic group containing 6 to 14 carbon atoms, either as a component of another group. An aryl group may be substituted by one or more of the substituents described herein.

[0020] Useful aryl groups include C 6-14 Aryl, preferably C 6-10 Aryl. Typical C 6-14 Aryl groups include phenyl, naphthyl, phenanthryl, anthraceneyl, indyl, azulel, biphenyl, biphenylene, and fluoroyl.

[0021] Useful halogens or halogen groups include fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0022] An acyl group is a functional group containing a carbonyl group (carbon-oxygen double bond). Exemplary acyl groups can be represented as RC(O)-, where R can be, for example, an alkyl group as described herein. Useful acyl groups include C... 1-6 An acyl group, or a C1-3 acyl group, such as an acetyl group. The acyl group may optionally be substituted with a group selected from halogens, amino groups, and aryl groups, wherein the amino and aryl groups may optionally be substituted. When substituted with a halogen, the number of halogen substituents may range from 1 to 5. Examples of substituted acyl groups include chloroacetyl and pentafluorobenzoyl. When substituted with an amino group, the amino group may be substituted with one or two substituents as described herein.

[0023] In this document, sulfonyl group refers to RS(=O)2-, where R can be, for example, an alkyl group as described herein. In this document, C... 1-4 sulfonyl group refers to R being C 1-4 Alkyl sulfonyl group.

[0024] As used herein, "heteroaryl" refers to a group containing 5-14, preferably 5-10, ring atoms, and having 6, 10, or 14 π electrons shared in the ring system. The ring atoms in a heteroaryl group are carbon atoms and 1-3 heteroatoms selected from oxygen, nitrogen, and sulfur. A heteroaryl group may be substituted by one or more of the substituents described herein.

[0025] As used herein, "heterocyclic group" refers to a saturated or partially saturated 3-7 membered monocyclic group, 7-10 membered bicyclic group, 11-14 membered tricyclic group, helical ring group, or bridged ring group, consisting of a carbon atom and 1-4 heteroatoms selected from O, N, and S, wherein the nitrogen and sulfur heteroatoms can be arbitrarily oxidized, and the nitrogen can be arbitrarily quaternized. Heterocyclic groups also include fused heterocycles formed by the fusion of any heterocycle defined above in the aforementioned bicyclic system with a benzene ring. If the resulting compound is stable, then the carbon or nitrogen atom of the heterocycle can be substituted. The heterocyclic group can be substituted by one or more substituents as described herein.

[0026] In this document, unless otherwise stated, when substituted, the alkyl, alkoxy, amino, acyl, aryl, or heteroaryl groups described in any embodiment herein may be substituted by one or more (e.g., 1, 2, 3, or 4) substituents selected from the following groups: halogen, hydroxyl, carboxyl, amino, nitro, cyano, C 1-6 Acylamino, C 1-6 Alkoxy, C 1-6 Alkyl, C 1-6 Acyl group, C 6-10 Aryl, C 3-8 Cycloalkyl, heteroaryl, and carbonyl groups, etc. The substituents themselves may also be optionally substituted. More preferably, the substituents include, but are not limited to, halogens, hydroxyl groups, cyano groups, amino groups, and C-groups. 1-6 Alkoxy, C 1-6 Alkyl and C 1-6 Acyl group.

[0027] It should be understood that in the various embodiments described herein, when the substituent is a heterocyclic group, aryl group, or heteroaryl group, the number of such heterocyclic group, aryl group, or heteroaryl group is usually one.

[0028] The compounds described in this invention, or their pharmaceutically acceptable salts, can effectively degrade Wee1 protein, exhibiting excellent antitumor proliferative activity and oral absorption.

[0029] Specifically, the present invention provides compounds, stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, as shown in Formula I: QLU (I) Where: L is a linking group; U is an E3 ubiquitin ligase-binding ligand; Q is a Wee1 protease-binding ligand having the structure shown in Formula Q-1 or Q-2: Where: A is N or CR a Ring B is an optionally substituted carbocyclic group, an optionally substituted heterocyclic group, an optionally substituted aryl group, or an optionally substituted heteroaryl group; Ring W is absent, an optionally substituted carbocyclic group, or an optionally substituted heterocyclic group; R1 is hydrogen, an optionally substituted C 1-8 Alkyl, optionally substituted C 2-8alkenyl, optionally substituted C 3-8 Cycloalkyl, optionally substituted aryl, optionally substituted heterocyclic, or optionally substituted heteroaryl; R2-R8 and R a Each can be hydrogen, halogen, optionally substituted amino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkoxy, optionally substituted alkenyl, optionally substituted alkynyl, nitro, cyano, hydroxyl, mercapto, acyloxy, azide, carboxyl, ethylenedioxy, hydroxyacylamino, or optionally substituted alkylthio; *1 indicates the position where Q and L are connected.

[0030] The compound of formula I of this invention may be a compound represented by formulas Ia and Ib below, or a stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt thereof, or a mixture thereof: Among them, A, ring B, ring W, R1-R8, L and U are as defined in Equation I.

[0031] In one or more embodiments of the compound of formula I, A is N.

[0032] In one or more embodiments of the compound of formula I, ring B is optionally substituted with C. 3-8 Carbocyclic group, optionally substituted 4-10 membered heterocyclic group, optionally substituted C 6-14 Aryl or optionally substituted 5-10 heteroaryl groups. Exemplary C3-8 carbocyclic groups are C... 3-8 cycloalkyl or C 3-8 Cycloalkenyl, preferably C 3-6 Cycloalkenyl or C 3-6 Cycloalkyl groups. Exemplary 4-10 membered heterocyclic groups can be 4-10 membered heterocyclic groups containing nitrogen and / or oxygen and / or sulfur, preferably 4, 5, 6, or 7 membered heterocyclic groups containing nitrogen and / or oxygen, such as oxobutyryl, azabutyryl, pyrrolidinyl, piperazine, and piperidinyl, etc. Exemplary C 6-14 The aryl group can be phenyl or naphthyl. Exemplary 5-10-membered heteroaryl groups can be nitrogen- and / or oxygen- and / or sulfur-containing 5-10-membered heteroaryl groups, preferably 5 or 6-membered nitrogen- and / or oxygen-containing heteroaryl groups. In some embodiments, ring B is an optionally substituted phenyl or an optionally substituted 5-6-membered heteroaryl group, preferably an optionally substituted phenyl group. Preferably, ring B is optionally surrounded by 1, 2, 3, or 4 groups selected from halogens, hydroxyl groups, cyano groups, C... 1-4 Alkyl, Halogenated C 1-4 Alkyl, C 1-4 Substitution of alkoxy groups and NR'R”, wherein R' and R” are each independently H or C. 1-4 Alkyl group. In some embodiments, the ring B is unsubstituted, or is replaced by one or two carbon atoms. 1-4 Alkyl substitution; in some embodiments, ring B is unsubstituted, or substituted with one C. 1-3Alkyl substitution. In some embodiments, ring B is optionally replaced by one carbon atom. 1-3 Alkyl-substituted phenyl groups.

[0033] In one or more embodiments of the compound of formula I, ring W is optionally substituted C 3-8 Carbocyclic group or optionally substituted 5-12 membered heterocyclic group. Exemplary C 3-8 The carbonyl group is C 3-8 cycloalkyl or C 3-8 Cycloalkenyl, preferably C 3-6 Cycloalkenyl or C 3-6 Cycloalkyl. Exemplary 5-12 membered heterocyclic groups can be 5-12 membered nitrogen- and / or oxygen- and / or sulfur-containing heterocyclic groups, preferably 5, 6, or 7 membered nitrogen- and / or oxygen-containing heterocyclic groups or 9-12 membered nitrogen-containing spirocyclic groups, such as pyrrolidinyl, piperazine, and piperidinyl. In some embodiments, ring W is an optionally substituted 5-12 membered heterocyclic group, preferably a 5-12 membered nitrogen-containing heterocyclic group, more preferably a 5-6 membered nitrogen-containing heterocyclic group or a 9-12 membered nitrogen-containing spirocyclic group. Preferably, ring W is optionally surrounded by 1, 2, 3, 4, or 5 groups selected from halogens, hydroxyl groups, cyano groups, C... 1-4 Alkyl, Halogenated C 1-4 Alkyl, C 1-4 Substitution of alkoxy groups and NR'R”, wherein R' and R” are each independently H or C. 1-4 Alkyl group. In some embodiments, ring W is not substituted. In some embodiments, ring W is substituted with a cyano group at its C-position connected to ring B.

[0034] In one or more embodiments of the compound of formula I, R1 is hydrogen, optionally substituted C. 1-8 Alkyl, optionally substituted C 2-8 alkenyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 6-14 Aryl, optionally substituted 4-10 membered heterocyclic groups, or optionally substituted 5-10 membered heteroaryl groups. Exemplary 4-10 membered heterocyclic groups can be 4-10 membered heterocyclic groups containing nitrogen and / or oxygen and / or sulfur, preferably 4, 5, 6, or 7 membered heterocyclic groups containing nitrogen and / or oxygen, such as oxobutyryl, azabutyryl, pyrrolidinyl, piperazine, and piperidinyl, etc. Exemplary C 6-14 The aryl group can be phenyl or naphthyl. Exemplary 5-10-membered heteroaryl groups can be nitrogen- and / or oxygen- and / or sulfur-containing 5-10-membered heteroaryl groups, preferably 5 or 6-membered nitrogen- and / or oxygen-containing heteroaryl groups. In some embodiments, R1 is an optionally substituted C 6-14 The aryl group or optionally substituted 5-10 heteroaryl group, preferably an optionally substituted phenyl group. Preferably, R1 is optionally surrounded by 1, 2, 3, 4 or 5 groups selected from halogen, hydroxyl, cyano, C 1-4 Alkyl, Halogenated C 1-4Alkyl, C 1-4 Substitution of alkoxy groups and NR'R”, wherein R' and R” are each independently H or C. 1-4 Alkyl group. In some embodiments, R1 is a phenyl group optionally substituted with 1 to 3 halogens. In some embodiments, R1 is a phenyl group substituted with halogens, especially chlorine, at positions 2 and 6.

[0035] In one or more embodiments of the compound of formula I, R2-R8 and R a When substituted, the substituents can be independently selected from halogens, hydroxyl groups, cyano groups, and NR'R". The number of substituents can be 1, 2, 3, 4, or 5, wherein R' and R" are each independently H or C. 1-4 Alkyl groups. In some embodiments, R2-R8 and R a Each can be independently hydrogen, halogen, or C1-3 alkyl.

[0036] In one or more embodiments of the compound of formula I, L has the following structure: Wherein: L1 and L2 are each independently bonded, optionally substituted alkylene groups or -(CR) L1 R L2 ) n C(O)-; Ring Z1 和 Each ring Z2 is independent and absent, or is an optionally substituted heterocyclic group or an optionally substituted heteroaryl group; R L1 and R L2 Each can be independently H, halogen, or C. 1-3 Alkyl, or R L1 and R L2 Together with the C atom to which it is attached, it forms a carbonyl or heterocyclic group; n is 0, 1, 2 or 3; *1 and *2 represent the attachment positions with L, Q and U, respectively.

[0037] In one or more embodiments of the compound of formula I, L1 and L2 are each independently an optionally substituted C 1- 4. Alkylene. In some embodiments, when L1 and L2 are substituted, the substituents can be 1 to 5, selected from halogens, hydroxyl groups, cyano groups, and NR'R", wherein R' and R" are each independently H or C. 1-4 Alkyl group. In some embodiments, L1 and L2 are each independently an unsubstituted C12. 1-3 Alkylene.

[0038] In some implementation schemes, R L1 and R L2 Each can be H or C independently. 1-3 Alkyl group. In some embodiments, R L1 and R L2 All are H. In some implementations, R L1 and RL2 Together with the attached C atom, it forms a 3-6 membered carbon cyclic group or a 4-6 membered heterocyclic group, preferably a 3-6 membered cycloalkyl group or a 4-6 membered nitrogen-containing heterocyclic group, such as aziridine, pyrrolidinyl, piperidinyl and piperazine.

[0039] In one or more embodiments of the compound of formula I, ring Z1 and ring Z2 are each independently an optionally substituted 5-12-membered N-containing heterocyclic group or an optionally substituted 5-6-membered heteroaryl group.

[0040] In some embodiments, when ring Z1 and ring Z2 are substituted, each is independently replaced by 1, 2, 3, 4, or 5 groups selected from halogen, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Substitution of alkoxy groups and NR'R”, wherein R' and R” are each independently H or C. 1-4 alkyl.

[0041] In one or more embodiments of the compound of formula I, L is a group that: Wherein, Z1 and Z2 are each independently an optionally substituted 5-12 member nitrogen-containing heterocyclic group or an optionally substituted 5 member nitrogen-containing heteroaryl group; m and n are each independently 0, 1, or 2; *l and *2 represent the connection positions with L and Q and U, respectively. In some embodiments, when both Z1 and Z2 are present, one is an optionally substituted 5-12 member nitrogen-containing heterocyclic group, and the other is an optionally substituted 5 member nitrogen-containing heteroaryl group. In a preferred embodiment, L is a group that: Among them, Y1 and Y2 are each independently CR L3 Or N, R L3 H, halogen, hydroxyl, cyano, C 1-3 Alkyl or C 1-3 Alkoxy group; m and n are each 0, 1 or 2 independently; p1, p2, p3 and p4 are each 0 or 1 independently; *1 and *2 indicate the connection positions with L, Q and U respectively.

[0042] In one or more embodiments of the compound of formula I, U is an E3 ubiquitin ligase ligand capable of binding to CRBN. In some embodiments, the E3 ubiquitin ligase binding ligand comprises at least the following groups: In the formula, X is N or CR d2 ;R d2 H, halogen, hydroxyl, cyano, C 1-3 Alkyl or C 1-3 Alkoxy group; *3 indicates the position where this group is attached to the rest of the compound of formula I.

[0043] In one or more embodiments, U is a group that includes: Wherein, ring D is absent, or is an optionally substituted aryl group, an optionally substituted heterocyclic group, or an optionally substituted heteroaryl group; V is a bond, -NR d1 -、-C(O)NR d1 -or-NR d1 C(O)-; X is N or CR d2 ;R d1 For H or C 1-3 Alkyl; R d2 H, halogen, hydroxyl, cyano, C 1-3 Alkyl or C 1-3 Alkoxy group; *2 indicates the connection position of the U group and L.

[0044] In one or more embodiments, ring D is: an optionally substituted 6-14-membered aryl group, such as a phenyl group; or an optionally substituted 4-10-membered heterocyclic group, such as an optionally substituted 4-10-membered nitrogen-containing heterocyclic group; or an optionally substituted 5-10-membered heteroaryl group, preferably an optionally substituted 5-10-membered nitrogen-containing heteroaryl group. Preferably, ring D is optionally surrounded by 1, 2, or 3 groups selected from halogens, oxo (=O), C... 1-3 Alkyl, C 3-6 cycloalkyl and C 1-3 Substitution of alkoxy groups.

[0045] In some implementations, X is CR d2 In some implementations, R d2 For H or C 1-3 alkyl.

[0046] In one or more embodiments, U is selected from the following groups: Among them, d1, d2, d3, and d4 are each independently N or CR. d3 R d3 H, halogen, C 1-3 Alkyl, C 3-6 cycloalkyl or C 1-3 alkoxy group; X is CH or N; T is CH2 or C=O; R d4 For H or C 1-3 Alkyl group; *2 indicates the connection position of the U group and L.

[0047] In some implementation schemes, CR d3 It is H, halogen, or C1-3 alkoxy.

[0048] In some embodiments, the ring containing d1, d2, d3, and d4 is a benzene ring, and the benzene ring is optionally surrounded by one, two, or three elements selected from halogens and C. 1-3 The alkoxy group is substituted, preferably the benzene ring is unsubstituted, or is substituted with one halogen or one carbon atom. 1-3 Alkyl-substituted.

[0049] In some implementations, U is: In the formula, X represents N; d1, d2, and d4 represent CH, and d3 represents CR. d3 R d3 H, halogen or C 1-3 Alkyl group.

[0050] In some implementations, U is: In the formula, d3 and d4 are CH; one of d1 and d2 is CH and the other is CR. d3 Among them, CR d3 It is a halogen.

[0051] The preferred compounds of Formula I of the present invention are represented by compounds of Formula II (including Formulas IIa and IIb) or their stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds or pharmaceutically acceptable salts, or mixtures thereof: Among them, A, ring B, ring W, R1-R8, L1, L2, Z1, Z2 and ring D are as described in any of the above embodiments.

[0052] The preferred compounds of Formula I of the present invention are represented by compounds of Formula III (including Formulas IIIa and IIIb) or their stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds or pharmaceutically acceptable salts, or mixtures thereof: Wherein, L1, L2, Z1, Z2, ring D, and X are as described in any of the preceding embodiments; R9 and R 10 Each is independently selected from halogens and C 1-3 Alkyl; R 11 and R 12 Each C is independently selected from hydrogen, halogens, and optionally substituted C. 1-3 Alkyl; T is N or CR 13 R 13 It is hydrogen, cyano, hydroxyl, C 1-3 Alkyl or C 1-3 Alkyl group.

[0053] In some embodiments of formulas IIIa and IIIb, R9 and R 10 Each is independently selected from F, Cl, and methyl. In some embodiments, R9 and R 10 Both are Cl.

[0054] In some implementations of formulas IIIa and IIIb, R 11 and R 12 Each is independently selected from hydrogen, halogen, and C. 1-3 Alkyl, Halogenated C 1-3 alkyl and hydroxy substituted C 1-3Alkyl group. In a preferred embodiment, R 11 and R 12 Each is independently selected from hydrogen, F, Cl, and methyl. In some embodiments, R 11 and R 12 Each can be either hydrogen or methyl.

[0055] In some implementations of formulas IIIa and IIIb, R 13 It can be hydrogen, cyano, hydroxy, methyl, or methoxy. In some embodiments, R 13 It is either hydrogen or cyano.

[0056] In some embodiments of formulas IIIa and IIIb, L1 and L2 are each independently of the optionally substituted C. 1-4 Alkylene. In some embodiments, when L1 and L2 are substituted, the substituents can be 1 to 5, selected from halogens, hydroxyl groups, cyano groups, and NR'R", wherein R' and R" are each independently H or C. 1-4 Alkyl group. In some embodiments, L1 and L2 are each independently an unsubstituted C12. 1-3 Alkylene.

[0057] In some embodiments of formulas IIIa and IIIb, ring Z1 and ring Z2 are each independently a optionally substituted 5-12-membered N-containing heterocyclic group or an optionally substituted 5-6-membered heteroaryl group. In some embodiments, when ring Z1 and ring Z2 are substituted, they are each independently replaced by 1, 2, 3, 4, or 5 groups selected from halogens, hydroxyl groups, cyano groups, C... 1-4 Alkyl, C 1-4 Substitution of alkoxy groups and NR'R”, wherein R' and R” are each independently H or C. 1-4 alkyl.

[0058] In some embodiments of formulas IIIa and IIIb, the groups composed of L1, Z1, Z2, and L2 are selected from the following structures: Wherein, *1 and *2 represent the connection positions of the above groups with N and X in the structure of compound IIIa or IIIb, respectively. Y1 and Y2 are each independently CR L3 Or N, R L3 H, halogen, hydroxyl, cyano, C 1-3 Alkyl or C 1-3 Alkoxy group; m and n are each independently 0, 1 or 2; p1, p2, p3 and p4 are each independently 0 or 1; *1 and *2 indicate the connection positions with L, Q and U respectively.

[0059] In some implementations of formulas IIIa and IIIb, X is N or CR. d2 ;R d2 H, halogen, hydroxyl, cyano, C 1-3Alkyl or C 1-3 Alkyl group.

[0060] It should be understood that, although the above text refers to A, ring B, ring W, and R1-R in equation I (including equations Ia, Ib, IIa, IIb, IIIa, and IIIb), 13 T, L1, L2, Z1, Z2 and ring D are described separately, but the described features can be combined arbitrarily to form a range of different Formula I (including Formulas Ia, Ib, IIa, IIb, IIIa and IIIb) compounds of the present invention.

[0061] Preferred compound examples of Formula I include, but are not limited to, the following compounds, or their stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof:

[0062] Some of the compounds of this invention may exist as stereoisomers, including optical isomers. This invention includes all stereoisomers and racemic mixtures of such stereoisomers, as well as individual enantiomers that can be isolated according to methods well known to those skilled in the art.

[0063] Examples of medicinal salts include inorganic and organic acid salts, such as hydrochloride, hydrobromide, phosphate, sulfate, citrate, lactate, tartrate, maleate, fumarate, mandelate, and oxalate; as well as inorganic and organic base salts formed with bases such as sodium hydroxyl, tris(hydroxymethyl)aminomethane (TRIS, tromethamine), and N-methylglucosamine.

[0064] Examples of prodrugs of the compounds of the present invention include simple esters of compounds containing carboxylic acids (e.g., by reacting with C according to methods known in the art). 1-4 Esters obtained by alcohol condensation; esters of compounds containing hydroxyl groups (e.g., obtained by reacting with C according to methods known in the art). 1-4 Carboxylic acid, C 3-6 esters obtained by condensation of diacids or their anhydrides, such as succinic anhydride and fumaric anhydride; imines of compounds containing amino groups (e.g., obtained by condensation with C according to methods known in the art). 1-4 Imines obtained by condensation of aldehydes or ketones; urethanes of compounds containing amino groups, such as those esters described by Leu et al. (J.Med.Chem.42:3623-3628(1999)) and Greenwald et al. (J.Med.Chem.42:3657-3667(1999)); aldol acetals or ketal acetals of compounds containing alcohols (e.g., those acetals obtained by condensation with chloromethyl methyl ether or chloromethyl ethyl ether according to methods known in the art).

[0065] The compounds of the present invention can be prepared using methods known to those skilled in the art or new methods of the present invention. Specifically, the compounds of the present invention having formula I (including formulas II and III) can be prepared as shown in the reaction examples in reaction scheme 1. 3-Amino-4-chlorobenzoic acid, acrylic acid and urea react under acetic acid conditions to give 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)yl)benzoic acid, 1-(2-methyl-4-nitrophenyl)piperazine and 9-formyl-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester react under triethylamine and sodium triacetoxyborohydride conditions to give 9-((4-(2-methyl-4-nitrophenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester, 9-((4-(2-methyl-4-nitrophenyl)piperazine ... The reaction of 9-((4-(4-amino-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester under Pd / C and H2 conditions yields 9-((4-(4-amino-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester, 9-(((4-(4-amino-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester, and 6-(2,6-dichlorophenyl)-2-(methylsulfonyl)-8,9-dihydroimidazo[1,2] The reaction of 9-((4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5,5]undecane-3-carboxylic acid tert-butyl ester, 9-((4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[ The reaction of tert-butyl undecane-3-carboxylic acid with 4M ethyl hydrochloride solution yields 2,2-((4-((3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-3-methylphenyl)amino)-6-(2,6-dichlorophenyl)-8,9-dihydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-5(6H)-one, 2,2-((4- ...5.5]undecane-3-carboxylic acid tert-butyl ester and 4M ethyl hydrochloride solution.5] Undecane-9-yl)methyl)piperazin-1-yl)-3-methylphenyl)amino)-6-(2,6-dichlorophenyl)-8,9-dihydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-5(6H)-one and 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid in (7-azobenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate and N,N- The reaction under diisopropylethylamine conditions yields 1-(2-chloro-5-(9-(4-(((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5,5]undecane-3-carbonyl)phenyl)dihydropyrimidin-2,4(1H,3H)-dione. Reaction scheme 1.

[0066] Other related compounds can be prepared using a similar method. For example, by replacing 1-(2-methyl-4-nitrophenyl)piperidine with 4-(2-methyl-4-nitrophenyl)piperidine, the target compound 1-(2-chloro-5-(9-(4-(((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidin-2,4(1H,3H)-dione can be prepared. Replacing 1-(2-methyl-4-nitrophenyl)piperazine with 1-(4-nitrophenyl)piperazine yields the target compound 1-(2-chloro-5-(9-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)phenyl)piperazine-1-yl)methyl)-3-azaspiro[5,5]undecane-3-carbonyl)phenyl)dihydropyrimidin-2,4(1H,3H)-dione. Replacing 1-(2-methyl-4-nitrophenyl)piperidine-4-onitrile with 4-(4-nitrophenyl)piperidine-4-onitrile yields the target compound 1-((3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)phenyl)piperidine-4-onitrile.

[0067] An important aspect of this invention is the discovery that compounds of formula I (including formulas II and III) are Wee1 protein degraders. Therefore, compounds of formula I (including formulas II and III), or their stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, can be used to treat diseases, such as cancer, by reducing Wee1 protein levels.

[0068] The present invention also includes a treatment method involving the administration of an effective amount of a compound of formula I (including formulas II and III) or a pharmaceutically acceptable salt or prodrug thereof to an animal. The treatment method is used to treat diseases such as cancer by lowering Wee1 protein levels. Diseases that can be treated or prevented by the methods or pharmaceutical compositions of the present invention include, but are not limited to, liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer (such as small cell lung cancer), Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, gastric cancer, colon cancer, malignant pancreatic islet tumor, malignant carcinoid cancer, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, piloblastic leukemia, rhabdomyosarcoma, Kaposi's sarcoma, urogenital tumors, thyroid cancer, esophageal cancer, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, idiopathic thrombocythemia, adrenocortical carcinoma, skin cancer, and prostate cancer.

[0069] Therefore, the present invention provides a method for treating or preventing diseases or conditions by reducing Weel protein levels, the method comprising administering to a desired subject an effective amount of a compound of formula I (including formulas II and III) or its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, or a pharmaceutical composition containing an effective amount of a compound of formula I (including formulas II and III) or its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof. In this invention, the subject includes mammals, more specifically humans.

[0070] In implementing the treatment method of the present invention, an effective amount of a pharmaceutical preparation is administered to a patient with one or more of these symptoms. The pharmaceutical preparation contains an effective therapeutic concentration of a compound of formula I (including formulas II and III), formulated for oral, intravenous, topical, or external administration, for the treatment of cancer and other diseases. The dosage is the amount of medicine that effectively improves or eliminates one or more symptoms. For the treatment of a specific disease, an effective amount is an amount sufficient to improve or alleviate, in some way, the symptoms associated with the disease. Such a dosage may be administered as a single dose or may be administered according to an effective treatment regimen. The dosage may cure the disease, but administration is generally intended to improve the symptoms of the disease. Repeated administration is generally required to achieve the desired symptom improvement.

[0071] In another embodiment, a pharmaceutical composition is provided comprising a compound of formula I (including formulas II and III) containing a Wee1 protein degrader, or a stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt thereof, or a mixture thereof.

[0072] Another embodiment of the present invention relates to a pharmaceutical composition that can effectively treat cancer, comprising a Wee1 protein degrader of formula I (including formulas II and III), or a stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt thereof, or a mixture thereof, in combination with at least one known anticancer drug or a pharmaceutically acceptable salt thereof. In particular, it is used in combination with other anticancer drugs related to DNA damage and repair mechanisms, including PARP inhibitors olaparib, niraparib, rucaparib, talazoparib, senaparib, and saruparib; HDAC inhibitors vorinostat, romidesin, palbistat, and belistat; and so on. It is also used in combination with other anticancer drugs related to cell division checkpoints, including Chk1 / 2 inhibitors, CDK4 / 6 inhibitors such as palbociclib, ATM / ATR inhibitors, and so on. Other anticancer drugs that can be used in combination therapy include, but are not limited to, alkylating agents such as busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, and carboplatin; topoisomerase I inhibitors such as camptothecin, irinotecan, and topotecan; topoisomerase II inhibitors such as doxorubicin, epirubicin, aclarubicin, mitoxantrone, methyl hydroxyrosine, and mentholtoporp; RNA / DNA inhibitors. Metabolites such as 5-azacytidine, gemcitabine, 5-fluorouracil, and methotrexate; DNA antimetabolites such as 5-fluoro-2′-deoxyuridine, fludarabine, nelarabine, cytarabine, pralatrexate, pemetrexed, hydroxyurea, and thioguanine; antimitotic agents such as colchicine, vincristine, vinorelbine, paclitaxel, ixaprilone, cabazitaxel, and docetaxel; antibodies such as monoclonal antibodies, panitumumab, nizotocin, nivolumab, pembrolizumab, and rabeclomethasone. Mocizumab, Bevacizumab, Pertuzumab, Trastuzumab, Cetuximab, Obinutuzumab, Ofamumab, Rituximab, Alemtuzumab, Tiimumab, Tosimomab, Bentuximab, Daremumab, Erotozumab, T-DM1, Ofatumumab, Dinutuximab, Blinatumomab, Ipilimumab, Avastin, Herceptin, and Rituximab; kinase inhibitors such as Imatinib, Genomicon. Fertrinib, Erlotinib, Ostinib, Afatinib, Ceritinib, Alectinib, Crizotinib, Erlotinib, Lapatinib, Sorafenib, Regorafenib, Vemurafenib, Dabrafenib, Aflibercept, Sunitinib, Nilotinib, Dasatinib, Bosutinib, Pramipinib, Ibrutinib, Cabozantinib, Lenvatinib, Vandetanib, Trametinib, Carbitinib, Axitinib, Tessiromoxetine, Idelalisib, Pazopanib, Tetracycline, and Everolimus.Other known anticancer drugs that can be used in combination therapy include tamoxifen, letrozole, fulvestrant, mitoxantridine, octreotide, retinoid, arsenic, zoledronic acid, bortezomib, carfilzomib, Ixazomib, vemodega, sondega, denosumab, thalidomide, lenalidomide, venetoclax, Aldesleukin (recombinant human interleukin-2), and Sipueucel-T (prostate cancer treatment vaccine).

[0073] In carrying out the method of the present invention, the compound of the present invention may be administered together with at least one known anticancer drug as a single pharmaceutical composition. Alternatively, the compound of the present invention may be administered separately from at least one known anticancer drug. In one embodiment, the compound of the present invention and at least one known anticancer drug are administered approximately simultaneously, i.e., all drugs are administered simultaneously or sequentially, as long as the compound simultaneously reaches therapeutic concentrations in the blood. In another embodiment, the compound of the present invention and at least one known anticancer drug are administered according to their respective dosage regimens, as long as the compound reaches therapeutic concentrations in the blood.

[0074] Another embodiment of the invention is a biocoupler, consisting of the said compound, that effectively inhibits tumors as a kinase inhibitor. This tumor-inhibiting biocoupler comprises the said compound with at least one known therapeutically active antibody, such as Herceptin or Rituxan, or a growth factor, such as EGF or FGF, or a cytokine, such as interleukin-2 or 4, or any molecule capable of binding to the cell surface. The antibody, along with other molecules, can deliver the compound to its target site, making it an effective anticancer drug. This biocoupler can also enhance the anticancer effects of therapeutically active antibodies, such as Herceptin or Rituxan.

[0075] Another embodiment of the present invention relates to a pharmaceutical composition that can effectively inhibit tumors, comprising a Wee1 protein degrader of formula I (including formulas II and III), or a stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt thereof, or a mixture thereof, for combination therapy with radiotherapy. In this embodiment, the compound of the present invention and radiotherapy may be administered at the same time or at different times.

[0076] Another embodiment of the invention relates to a pharmaceutical composition effective for postoperative treatment of cancer, comprising a Wee1 protein degrader of formula I (including formulas II and III), or a stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt thereof, or a mixture thereof. The invention also relates to a treatment method involving surgical removal of a tumor followed by treatment of cancer in a mammal with the pharmaceutical composition of the invention.

[0077] The pharmaceutical compositions of the present invention comprise pharmaceutical formulations in which the contents of all the compounds of the present invention effectively achieve their intended objectives. Although individual needs vary, those skilled in the art can determine the optimal dosage of each component of the pharmaceutical formulation. Generally, the compounds, or their available salicies, are administered orally to mammals daily at a dosage of about 0.0025 to 50 mg / kg body weight. However, it is preferable to administer orally at a dosage of about 0.01 to 10 mg / kg. If a known anticancer drug is also administered, its dosage should effectively achieve its intended purpose. The optimal dosages of these known anticancer drugs are well known to those skilled in the art.

[0078] A single oral dose may comprise about 0.01 to 50 mg, preferably about 0.1 to 10 mg, of the compound of the present invention. A single dose may be administered once or multiple times daily as one or more tablets, each tablet containing about 0.1 to 50 mg, preferably about 0.25 to 10 mg, of the compound of the present invention or a solvate thereof.

[0079] In topical formulations, the concentration of the compounds of the present invention can be from about 0.01 to 100 mg per gram of carrier.

[0080] The compounds of the present invention can be administered as unprocessed pharmaceutical products. They can also be administered as part of a suitable pharmaceutical formulation containing a pharmaceutically acceptable carrier (including excipients and adjuvants). These pharmaceutically acceptable carriers facilitate the processing of the compounds into pharmaceutically acceptable formulations. Preferred pharmaceutical formulations, particularly those for oral administration and preferred routes of administration such as tablets, lozenges, and capsules, as well as solutions suitable for injection or oral administration, contain about 0.01% to 99%, preferably from about 0.25% to 75%, of the active compound and excipients.

[0081] The scope of this invention also includes non-toxic, pharmaceutically acceptable salts of the compounds of this invention. Acid addition salts are formed by mixing a solution of a non-toxic, pharmaceutically acceptable acid with a solution of the compound of this invention. Examples of the acids include hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, etc. Base addition salts are formed by mixing a solution of a non-toxic, pharmaceutically acceptable base with a solution of the compound of this invention. Examples of the bases include sodium hydroxide, potassium hydroxide, hydrocholine, sodium carbonate, tris(hydroxymethyl)aminomethane, N-methylglucosamine, etc.

[0082] The pharmaceutical formulations of this invention can be administered to any mammal, provided they achieve the therapeutic effects of the compounds of this invention. Humans and veterinary animals are most important among these mammals, although this invention is not intended to be so limited.

[0083] The pharmaceutical formulation of this invention can be administered via any route to achieve its intended purpose. For example, it can be administered via parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial, nasal, or topical routes. Alternatively or concurrently, it can be administered orally. The dosage of the drug will be determined based on the patient's age, health and weight, the type of concurrent treatment, the frequency of treatment, and the desired therapeutic benefit.

[0084] The pharmaceutical formulations of the present invention can be manufactured using known methods. For example, they can be manufactured by conventional mixing, granulation, tableting, dissolving, or freeze-drying processes. When manufacturing oral formulations, solid excipients and active compounds can be combined, and the mixture can be selectively ground. If desired or necessary, appropriate excipients can be added, and the granular mixture can be processed to obtain tablets or tablet cores.

[0085] Suitable excipients, especially fillers, include sugars such as lactose or sucrose, mannitol or sorbitol; cellulose preparations and / or calcium phosphates, such as tricalcium phosphate or dicalcium phosphate; and binders, such as starch pastes including corn starch, wheat starch, rice starch, potato starch, gelatin, astragalus gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone. If desired, disintegrants, such as the starches mentioned above, as well as carboxymethyl starch, croscarmellose, agar, or alginate or its salts, such as sodium alginate, may be added. Adjuvants, especially flow conditioners and lubricants, include silica, talc, stearic acid or its salts, such as magnesium stearate or calcium stearate, and / or polyethylene glycol. If desired, a suitable coating that resists gastric juices can be provided to the tablet core. For this purpose, a concentrated sugar solution can be applied. This solution may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solution, and suitable organic solvents or solvent mixtures. To prepare a gastric juice-resistant coating, a suitable cellulose solution, such as cellulose acetate phthalate or hydroxypropyl methylcellulose phthalate, can be used. Dyes or pigments may be added to the coating of the tablet or tablet core, for example, for identification or to characterize the dosage of the active ingredient.

[0086] Other orally edible pharmaceutical formulations include compressible capsules made of gelatin, and sealed soft capsules made of gelatin and plasticizers such as glycerin or sorbitol. The compressible capsule may contain an active compound in particulate form, mixed with fillers such as lactose; binders such as starch; and / or lubricants such as talc or magnesium stearate, and stabilizers. In soft capsules, the active compound is preferably dissolved or suspended in a suitable liquid such as oils or liquid paraffin, in which stabilizers may be added.

[0087] Suitable formulations for parenteral administration include aqueous solutions of the active compound, such as solutions of water-soluble salts and alkaline solutions. Additionally, oily injectable suspensions of the appropriate active compound can be administered. Suitable lipophilic solvents or carriers include oils such as sesame oils, synthetic fatty acid esters such as ethyl oleate or triglycerides or polyethylene glycol 400, or hydrogenated castor oil, or cyclodextrin. Aqueous injectable suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, and / or dextran. Suspension stabilizers may also be included.

[0088] According to one aspect of the invention, the compounds of the invention are formulated for external and parenteral use and are used to treat skin cancer.

[0089] The topical formulations of this invention can be formulated into oils, creams, emulsions, ointments, etc., using a preferred suitable carrier. Suitable carriers include plant or mineral oils, white mineral oil (white paraffin), branched-chain fatty acids or oils, animal fats, and high molecular weight alcohols (greater than C10). 12 Preferred carriers are those in which the active ingredient can dissolve. Emulsifiers, stabilizers, moisturizers, and antioxidants may also be included, as well as agents that impart color or fragrance if desired. Furthermore, these topical formulations may contain transdermal penetration enhancers. Examples of such enhancers can be found in U.S. Patent Nos. 3,989,816 and 4,444,762.

[0090] Creams are preferably formulated with a mixture of mineral oil, self-emulsifying beeswax, and water, mixed with an active ingredient dissolved in a small amount of oil, such as almond oil. A typical example of a cream includes approximately 40 parts water, 20 parts beeswax, 40 parts mineral oil, and 1 part almond oil.

[0091] Ointments can be formulated by mixing a plant oil containing active ingredients, such as almond oil, with warm paraffin wax, and then allowing the mixture to cool. A typical example of an ointment consists of approximately 30% by weight almond oil and 70% by weight white paraffin wax.

[0092] This invention also relates to the preparation of medicaments using the compounds of this invention to treat clinical conditions that are effective against the degradation of Wee1 kinase. These medicaments may include the aforementioned pharmaceutical compositions.

[0093] The following examples are illustrative and not intended to limit the methods and formulations of the present invention. Other appropriate modifications and improvements to various conditions and parameters that will be apparent to those skilled in the art and that are commonly encountered in clinical treatment are all within the spirit and scope of the present invention. General Description of Examples: All reagents used were commercially available, and solvents were dried and purified according to standard methods. Mass spectrometry samples were analyzed using a single quadrupole mass spectrometer (Shimadzu 2020) with electrospray ionization. Recordings were performed using a Varian 400MHz NMR spectrometer. 1¹H NMR spectra, chemical shifts are recorded in ppm starting from the low field with TMS as the internal standard (0.00 ppm), and coupling constant J values ​​are in Hz. Example 1: 1-(2-chloro-5-(9-(4-(((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5,5]undecane-3-carbonyl)phenyl)dihydropyrimidin-2,4(1H,3H)-dione a) Preparation of 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)yl)benzoic acid: 3-amino-4-chlorobenzoic acid (1.0 g, 5.8 mmol, 1.0 eq) was placed in a solution of acrylic acid (1.68 g, 23.3 mmol, 4.0 eq), stirred at 100 °C for 3 hours, and cooled to room temperature. Acetic acid (AcOH, 5 mL) was added to the mixture. After stirring at 100 °C for 10 minutes, urea (2.1 g, 34.8 mmol, 6.3 eq) was added to the mixture, and stirred overnight at 120 °C. The reaction mixture was poured into a mixture of concentrated hydrochloric acid (5 mL) and ice water (5 mL), mixed thoroughly, and the suspension was allowed to stand overnight at 0 °C. The filter cake was washed with diethyl ether (3 mL × 3) to give a yellow solid product (230 mg, 9% yield). LC-MS: m / z, 268.85 [M+H] + 266.75 [MH] - b) Preparation of 9-((4-(2-methyl-4-nitrophenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester: 9-formyl-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (198.4 mg, 0.70 mmol, 1.3 eq) was added to a solution of 1-(2-methyl-4-nitrophenyl)piperazine (201.0 mg, 0.54 mmol, 1.0 eq) in DCM (5 mL) at room temperature and under nitrogen. The mixture was stirred overnight at room temperature, and sodium triacetoxyborohydride (NaBH(OAc)3, 228.9 mg, 1.08 mmol, 2.0 eq) was added to the reaction mixture. The mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure, and the residue was purified by preparative TLC (DCM / MeOH = 15 / 1) to give a yellow solid product (57 mg, 22% yield). LC-MS: 487.15 [M+H] +c) Preparation of tert-butyl 9-((4-(4-amino-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid: Pd / C (20 mg, 10%) was added to an ethyl acetate (EA, 5 mL) solution of 9-((4-(2-methyl-4-nitrophenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid (57 mg, 0.12 mmol, 1.0 eq) at room temperature. The mixture was stirred overnight at room temperature under H2. The mixture was filtered, and the filtrate was concentrated under reduced pressure and purified by preparative TLC (PE / EA = 1 / 1) to give a white solid product (22 mg, 41% yield). LC-MS: m / z, 457.3 [M+H] + Preparation of tert-butyl 9-((4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid: tert-butyl 9-(((4-(4-amino-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid (22.0 mg, 0.05 mmol, 1.0 mmol) 6-(2,6-dichlorophenyl)-2-(methanesulfonyl)-8,9-dihydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-5(6H)-one (28.5 mg, 0.072 mmol, 1.5 eq) and trifluoroacetic acid (TFA, 2 drops) were added to a solution of acetonitrile (3 mL). The mixture was stirred at room temperature for 0.5 h under nitrogen protection. Water (10 mL) was added to the mixture, and the mixture was extracted with EA (10 mL × 3). The organic layer was dried over sodium sulfate and concentrated. The residue was purified by pre-TLC (PE / EA = 2 / 1) to give a white solid product (25 mg, 66% yield). LC-MS: m / z, 788.25 [M+H] + 786.20 [MH] -Preparation of 2,2-((4-(((3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-3-methylphenyl)amino)-6-(2,6-dichlorophenyl)-8,9-dihydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-5(6H)-one: At room temperature, 4 M ethyl acetate solution (4 mL) of 4 M ethyl acetate solution was added to a solution of 2 mL of EA containing 25 mg, 0.032 mmol, 1.0 eq of 9-((4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (EA). The mixture was stirred overnight at room temperature. The reaction mixture was concentrated to give a yellow solid product (35 mg, crude product). LC-MS: m / z, 688.25 [M+H] + 244.20[0.5M+H] + 686.20 [MH] - f) Preparation of 1-(2-chloro-5-(9-(4-(((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperazin-1-yl)methyl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)dihydropyrimidin-2,4(1H,3H)-dione: To 22-((4-(4-((3-azaspiro[5.5]undecane-9-yl)methyl)piperazin-1-yl)-3-methylphenyl)amino)-6-(2,6-dichlorophenyl)-8,9- In a solution of dihydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-5(6H)-one (35 mg, crude) and 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (25 mg, 0.061 mmol, 1.2 eq) in DMF (4 mL), (7-azobenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (HATU, 29.1 mg, 0.77 mmol, 1.5 eq) and N,N-diisopropylethylamine (DIEA, 46 mg, 0.36 mmol, 7.0 eq) were added, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative TLC (DCM / MeOH = 10 / 1) and preparative HPLC (HCOOH, 0.1%, 0%–60%) to give the target compound as a yellow solid (6.6 mg, 22% yield). LCMS: m / z 940.10 [M+H] + . 1H NMR (DMSO-d6): δ10.51 (s, 1H), 10.29 (s, 1H), 8.66 (s, 1H), 8.38 (s, 1H), 7.67 (s, 1H), 7.65 (s, 1H), 7.64-7.61 (m, 2H), 7.56-7.51 (m, 2H), 7.38 (dd, J=8.2, 2.0Hz, 1H), 6.99 (d, J=8.8Hz, 1H), 4.22-4.13 (m, 2H), 3.81 (t, J=8.8Hz, 2 H), 3.76-3.70(m, 1H), 3.66-3.50(m, 4H), 3.45-3.40(m, 3H), 3.33-3.25(m, 3H), 2.86-2.77(m, 4H), 2.78-2.70(m, 2H), 2.23(s, 3H), 2.20-2.13(m, 2H), 1.76-1.65(m, 2H), 1.60-1.40(m, 5H), 1.36-1.20(m, 2H), 1.17-0.96(m, 4H). Example 2: 1-(3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-4-nitrile) a) Preparation of 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)yl)benzoic acid: 3-amino-4-chlorobenzoic acid (1.0 g, 5.8 mmol, 1.0 eq) was placed in a solution of acrylic acid (1.68 g, 23.3 mmol, 4.0 eq), stirred at 100 °C for 3 hours, and cooled to room temperature. Acetic acid (AcOH, 5 mL) was added to the mixture. After stirring at 100 °C for 10 minutes, urea (2.1 g, 34.8 mmol, 6.3 eq) was added to the mixture, and stirred overnight at 120 °C. The reaction mixture was poured into a mixture of concentrated hydrochloric acid (5 mL) and ice water (5 mL), mixed thoroughly, and the suspension was allowed to stand overnight at 0 °C. The filter cake was washed with diethyl ether (3 mL × 3) to give a yellow solid product (230 mg, 9% yield). LC-MS: m / z, 268.85 [M+H] + 266.75 [MH] -b) Preparation of 9-((4-cyano-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-1-yl)methyl)-3-azaspiro[5,5]undecane-3-carboxylic acid tert-butyl ester: At room temperature and under nitrogen, 9-(4-((6-(2,6-dichlorophenyl)) 9-Formyl-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (30 mg, 0.107 mmol, 1.3 eq) was added to a solution of 5 mL of DCM containing 45.0 mg (45.0 mg, 0.082 mmol, 1.0 eq) of 5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-4-onitrile (45.0 mg, 0.082 mmol, 1.0 eq). After stirring overnight at room temperature, sodium triacetoxyborohydride (NaBH(OAc)3, 34.7 mg, 0.164 mmol, 2.0 eq) was added to the reaction mixture. The mixture was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure, and the residue was purified by preparative TLC (DCM / MeOH = 15 / 1) to give a yellow solid product (57 mg, 85% yield). LC-MS: 812.30 [M+H] + 407.10[0.5M+H] + 810.20 [MH] - c) Preparation of 1-((3-azaspiro[5.5]undecane-9-yl)methyl)-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidine-4-nitrile: at room temperature, to 9-((4-cyano-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidine-4-nitrile A solution of tert-butyl chlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-1-yl)methyl)-3-azaspiro[5,5]undecane-3-carboxylic acid (tert-butyl ester) (57 mg, 0.12 mmol, 1.0 eq) in EA (2 mL) was mixed with 4 M ethyl acetate hydrochloride solution (4 mL). The mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure to give a colorless oily crude product (50 mg, crude product). LC-MS: m / z, 712.20 [M+H] + 356.70 [0.5M+H] + 710.15 [MH] -d) Preparation of 1-((3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-4-nitrile: To 1-((3-azaspiro[5.5]undecane-9-yl)methyl)-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimido[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-4-nitrile Pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-4-onitrile (50 mg, crude) and 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (22.6 mg, 0.084 mmol, 1.2 eq) were added to dry DMF (4 mL), along with (7-azobenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (HATU, 41 mg, 0.108 mmol, 1.5 eq) and N,N-diisopropylethylamine (DIEA, 65 mg, 0.504 mmol, 7.0 eq) at room temperature under nitrogen. The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative TLC (DCM / MeOH = 10 / 1) and preparative HPLC (HCOOH, 0.1%, 0%–60%) to give a white solid 1-((3-(4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoyl)-3-azaspiro[5.5]undecane-9-yl)methyl)-4-(4-((6-(2,6-dichlorophenyl)-5-oxo-5,6,8,9-tetrahydroimidazo[1,2-a]pyrimidino[5,4-e]pyrimidin-2-yl)amino)-2-methylphenyl)piperidin-4-onitrile (13.5 mg, 20% yield). LCMS: m / z 964.15 [M+H] + 4782.80[0.5M+H] + 962.00 [MH] - . 1H NMR (DMSO-d6): δ10.51 (s, 1H), 10.45 (s, 1H), 8.71 (s, 1H), 8.32 (s, 1H), 7.72-7.65 (m, 3H), 7.63 (d, J=8.2Hz, 1H), 7. 57-7.52 (m, 2H), 7.38 (dd, J=8.2, 2.0Hz, 1H), 7.33 (d, J=8.8Hz, 1H), 4.20 (t, J=10.3Hz, 2H), 3.83 (t, J=8.7Hz, 2H), 3 0.80-3.70 (m, 2H), 3.68-3.50 (m, 4H), 2.94 (d, J = 11.1 Hz, 2H), 2.77-2.70 (m, 2H), 2.56 (s, 3H), 2.35-2.25 (m, 4H), 2.24-2.17 (m, 2H), 1.96-1.86 (m, 2H), 1.69 (d, J = 10.8 Hz, 2H), 1.61-1.40 (m, 5H), 1.38-1.21 (m, 2H), 1.18-0.97 (m, 4H). Examples 3-36: Examples 3-36 were prepared using a synthetic method similar to that of Example 1. The compounds in each example are as follows: Example 37: The inhibitory effect of the compound of the present invention on Wee1 enzyme activity was demonstrated using ADP-Glo ​​Kinase reagent (Promega, #V9103). The reaction buffer consisted of 50 mM HEPES, 1 mM EGTA, 10 mM MgCl2, 0.01% Brij 35, and 2 mM DTT (pH 7.5). First, the stock solution of the compound to be tested was serially diluted with DMSO at a ratio of 1:3 to 10 concentrations. Then, 0.05 μL of the diluted compound was added to a 384-well plate (Greiner, #784075) using an Echo 665 instrument, followed by 2.5 μL of Wee1 enzyme (Cama, #05-177) solution. The plate was centrifuged at 1000 rpm for 1 minute. After incubation at 25°C for 10 minutes, 2.5 μL of an ATP / substrate mixture was added to initiate the reaction, which was then incubated at 25°C for 60 minutes. Subsequently, 4 μL of ADP-Glo ​​reagent was added and incubated at 25°C for 40 minutes, followed by 8 μL of kinase detection reagent, and incubation continued at 25°C for another 40 minutes. Finally, the chemiluminescence value was detected using a BMG Pherastar FSX instrument. The inhibition rate % was calculated as: (Chemiluminescence value of wells without compound - Chemiluminescence value of wells containing test compound) / (Chemiluminescence value of wells without compound - Chemiluminescence value of negative control wells) × 100. A nonlinear regression equation was then used: Y = Bottom + (Top - Bottom) / (1 + 10^(LogIC)). 50 The IC was calculated using the fitted curve (-X)*HillSlope) and the software XLfit 5.5.0. 50 The inhibitory effect of the compound of this application on Wee1 enzyme activity was determined by the above experimental methods, and the measured IC50 value was... 50 The values ​​are shown in Table 1 below. Where +++ indicates IC 50 ≤10nM. Table 1 The results showed that the compounds of this invention had a good inhibitory effect on Wee1 kinase activity. Example 38: Degradation of Wee1 protein by the compounds of this invention. Resuscitated human colon cancer cells (LoVo) were passaged to a good growth state. When the confluence reached approximately 90%, they were digested with trypsin and seeded at a certain density in 12-well plates (Coming, #3513) and cultured overnight at 37°C in a 5% CO2 cell culture incubator. Different concentrations of the test compound were added to the 12-well cell plates and incubated for 24 hours and 4 hours respectively in a cell culture incubator. The culture medium was aspirated, and the cells in the plates were washed with pre-warmed PBS. Residual PBS was aspirated, and the plates were frozen at -80°C for later use. Cell lysis buffer (Thermo Fisher, #FNN0011) was added to the cell plates, and after thorough grinding, lysis was performed on ice for 30 minutes, followed by centrifugation at 12000 rpm at 4°C for 20 minutes. The supernatant was transferred to pre-chilled EP tubes for aliquoting. The protein concentration in the lysis buffer was determined using the BCA method, and the protein concentration of all samples was adjusted to be uniform using the lysis buffer. Add an appropriate amount of sample to the loading buffer and incubate in a 100°C metal bath for 10 minutes. After the sample cools to room temperature, centrifuge at 12,000 rpm for 5 minutes. Perform electrophoresis using a 4-12% SDS-PAGE precast gel (ThermoFisher, #WG1403BOX), loading 10 μg of sample per well. Electrophore at 120V for 10 minutes, then switch to 150V for 50 minutes, followed by wet transfer at 300mA for 1 hour. Block with blocking buffer (Bioss, #C1066) for 1 hour. Incubate the membrane with WEE1 antibody (CST, #4936) or phosphorylated cdc2 (Tyr15)(10A11) antibody (CST, #4539) overnight at 4°C, then wash three times with TBS-T for 5 minutes each time. Incubate the membrane with HRP-conjugated secondary antibody at room temperature for 1 hour, then wash three times with TBS-T for 5 minutes each time. After ECL staining, imaging was performed using a chemiluminescence imaging system (Bio-Rad). The compounds in Examples 1 and 2 of this application, at nanomolar concentrations, showed significant degradation effects on Wee1 protein over both 4 and 24 hours, and effectively inhibited its downstream target pCDK1 (Y15). This effect increased with increasing concentration, exhibiting a clear concentration-dependent effect (Figure 1). Example 39: Inhibitory effect of the compounds of this invention on LoVo cell growth. Newly revived LoVo cells were passaged to the third generation, exhibiting good growth and approximately 90% confluence, and were then used for experiments. LoVo cells were digested with trypsin, centrifuged at 800 rpm for 5 min, the supernatant was discarded, and the cells were resuspended in fresh culture medium and counted. Cells were seeded at a density of 6000 cells per well into 96-well cell culture plates and incubated overnight at 37°C in a 5% CO2 incubator.The stock solution of the test compound was serially diluted with DMSO at ratios of 1:3 and 1:10 to eight concentrations (the last concentration being a DMSO negative control): 10 μM, 3.3 μM, 1 μM, 0.33 μM, 0.1 μM, 0.033 μM, 0.01 μM, and 0 μM (final DMSO concentration was 1‰). 5 μL of each concentration was added to 120 μL of medium (25-fold dilution) and vortexed. Cells cultured overnight were removed from the medium, and 195 μL of fresh medium was added to each well, followed by 5 μL of the diluted test compound containing the corresponding concentration. The culture plate was then incubated at 37°C in a 5% CO2 incubator for 3 days. After removing the stock solution, 90 μL of fresh serum-free 1640 medium was added to each well, followed by 10 μL of CCK-8 assay reagent. The plate was incubated for another 2 hours, and the absorbance (OD) values ​​at 450 / 650 nm were read using a multi-reader. Data were analyzed using Graph Pad Prism 5.0 software. The inhibitory activity of the compound on cell proliferation was plotted on a coordinate system of cell viability and compound concentration. IC. 50 The value was fitted with an S-shaped dose-response curve equation, which is: Y = 100 / (1 + 10^(LogC - LogIC)) 50 Where C is the compound concentration. The compound of this application has an inhibitory effect on the growth of LoVo cells. The IC50 was determined using the above experimental methods. 50 The values ​​are shown in Table 2 below. Where +++ indicates IC 50 ≤100nM, ++ means 100nM <IC 50 ≤500 nM. Table 2: IC50 of the compounds in the examples inhibiting the proliferation activity of LoVo cells. 50 Example 40: Inhibitory Effect of the Compound of the Present Invention on HCC1569 Cell Growth The cytotoxicity of the compound of the present invention against human breast cancer cells HCC1569 was detected using a luminescent cell viability assay. HCC1569 cells were seeded into 96-well cell culture plates at an appropriate seeding density. The culture plates were placed in a humidified incubator at 37°C and 5% CO2 overnight. The next day, after removing the stock solution, 95 μL of fresh culture medium was added to each well of the cell culture plate. The test compound was serially diluted 3-fold and 10-fold using 100% DMSO, resulting in 8 concentration points. In another plate, 245 μL of culture medium and 5 μL of the serially diluted test compound were mixed (50-fold dilution). 5 μL of the diluted test compound (20-fold dilution) was added to each well of the 96-well cell culture plate containing 95 μL of fresh culture medium. The cell culture plates were then returned to the 37°C and 5% CO2 incubator for 7 days. On day 4, culture medium was replenished, while the concentration of the test compound remained unchanged. On day 7, 100 μL of Cell Counting-Lite 2.0 reagent (Vazyme, DD1101-02) was added to each well. The cell plate was then shaken at 350 rpm for 2 minutes and incubated at room temperature for 30 minutes. The chemiluminescence signal was read using a microplate reader. The inhibitory activity curve of the compound on cell proliferation was plotted on the coordinates of cell viability and compound concentration. Cell viability % = (RLU) 化合 物 -RLU 背景 ) / (RLU DM s o -RLU 背景 )×100. IC 50 The value is fitted by the S-type dose-response curve equation: Y = 100 / (1 + 10^(logC - logIC)) 50 C represents the compound concentration. The compound described in this application has an inhibitory effect on the growth of HCC1569 cells. The IC50 was determined using the above experimental methods. 50 The values ​​are shown in Table 3 below. ++ indicates 100 nM. <IC 50 ≤500 nM. Table 3: IC50 of the compounds in the examples inhibiting the proliferation activity of HCC1569 cells. 50 Example 41: Pharmacokinetic Study of the Compound of the Invention in Mice After a Single Oral Administration The compound of the invention was administered to CD-1 (ICR) mice by gavage at a dose of 10 mg / kg. Plasma samples were collected at eight time points: 0.250, 0.500, 1.00, 2.00, 4.00, 6.00, 8.00, and 24.0 hours after administration. The concentration of the compound was determined by LC-MS / MS. The pharmacokinetic parameters of the compound in mice after oral administration are summarized in Table 4. Table 4: Pharmacokinetic Parameters of the Compound of the Example in Mice After Oral Administration Note: t 1 / 2： Elimination half-life; C max Maximum plasma drug concentration; AUC 0-t Area under the plasma drug concentration-time curve from time 0 to the last measurable concentration; AUC 0-inf The area under the plasma drug concentration-time curve from time 0 to infinity. Results show that the compounds of this invention have good oral absorption and high exposure levels in mice.

[0094] While the invention has been fully described, those skilled in the art will understand that the same practices can be carried out under broad and equivalent conditions, formulations, and other parameters without affecting the scope of the invention or any embodiments thereof. All patents, patent applications, and publications cited herein are incorporated herein by reference in their entirety.

Claims

1. A compound of formula I, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof: QLU(I) In the formula: L is a linking group; U is an E3 ubiquitin ligase-binding ligand; Q is the Wee1 protease-binding ligand, having the structure shown in formula Q-1 or Q-2: in: A is N or CR a ; Ring B can be an optionally substituted carbocyclic group, an optionally substituted heterocyclic group, an optionally substituted aryl group, or an optionally substituted heteroaryl group; Ring W can be absent, optionally substituted carbocyclic, or optionally substituted heterocyclic. R1is hydrogen, optionally substituted C 1-8 alkyl, optionally substituted C 2-8 alkenyl, optionally substituted C 3-8 cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; R2-R8and R a each independently is hydrogen, halogen, optionally substituted amino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkoxy, optionally substituted alkenyl, optionally substituted alkynyl, nitro, cyano, hydroxy, thiol, acyloxy, azido, carboxy, ethylenedioxy, hydroxyamido, or optionally substituted alkylthio; *1 indicates the position where Q and L are connected.

2. The compound as claimed in claim 1, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, characterized in that, Formulas Q-1 and Q-2 have one or more of the following characteristics: (1) A is N; (2) Ring B is optionally substituted phenyl or optionally substituted 5-6 membered heteroaryl, preferably optionally substituted phenyl; preferably, Ring B is optionally substituted with 1, 2, 3, or 4 substituents selected from halo, hydroxy, cyano, C 1-4 alkyl, haloC 1-4 alkyl, C 1-4 alkoxy, and NR’R”, wherein R’ and R” are each independently H or C 1-4 alkyl; (3) Ring W is an optionally substituted 5-12 membered heterocyclic group, preferably a 5-12 membered N-containing heterocyclic group, more preferably a 5-6 membered nitrogen-containing heterocyclic group or a 9-12 membered nitrogen-containing spirocyclic group; preferably, ring W is optionally surrounded by 1, 2, 3, 4 or 5 groups selected from halogen, hydroxyl, cyano, C 1-4 Alkyl, Halogenated C 1-4 Alkyl, C 1-4 Substitution of alkoxy groups and NR'R”, wherein R' and R” are each independently H or C. 1-4 alkyl; (4) R1is optionally substituted aryl or optionally substituted heteroaryl, preferably optionally substituted phenyl; preferably, R1is optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of halogen, hydroxy, cyano, C 1-4 alkyl, haloC 1-4 alkyl, C 1-4 alkoxy, and NR'R" wherein each of R' and R" is independently H or C 1-4 alkyl; (5) R2-R8and R a each optionally substituted with 1-5 substituents selected from the group consisting of halogen, hydroxyl, cyano, and NR'R", wherein R' and R" are each independently H or C 1-4 alkyl; preferably, R2-R8and R a each independently hydrogen, halogen, or C 1-3 alkyl.

3. The compound, stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt, or mixture thereof, as described in claim 1 or 2, characterized in that, L has the following structure: in: L1and L2are each independently a bond, optionally substituted alkylene, or -(CR L1 R L2 ) n C(O)-; Ring Z1 and ring Z2 are either independent or either a heterocyclic group or a heteroaryl group with optional substitution. R L1 and R L2 each independently is H, halogen or C 1-3 alkyl, or R L1 and R L2 and the C atom to which they are attached together form a carbocyclic or heterocyclic group; n is 0, 1, 2, or 3; *1 and *2 represent the connection positions with L, Q, and U, respectively.

4. The compound as claimed in claim 3, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, characterized in that, The L has one or more of the following characteristics: (1) L1and L2are each independently C1-5alkylene optionally substituted with 1-5 substituents selected from substituted halogen, hydroxyl, cyano, and NR'R" wherein R' and R" are each independently H or C1-3alkyl; or 1-4 (2) L1and L2are each independently C1-5alkylene optionally substituted with 1-5 substituents selected from substituted halogen, hydroxyl, cyano, and NR'R" wherein R' and R" are each independently H or C1-3alkyl; or 1-4 (3) L1and L2are each independently C1-5alkylene optionally substituted with 1 (2) R L1 and R L2 each independently H or C 1-3 alkyl; (3) each of ring Z1and ring Z2is independently a 5-12 membered N-containing heterocyclyl optionally substituted with 1-5 substituents selected from the group consisting of halogen, hydroxyl, cyano, C 1-4 alkyl, C 1-4 alkoxy, and NR’R”, wherein each of R’ and R” is independently H or C 1-4 alkyl.

5. The compound of claim 3, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, characterized in that, L represents the following groups: Wherein, Z1 and Z2 are each independently a substituted 5-12 member nitrogen-containing heterocyclic group or an substituted 5 member nitrogen-containing heteroaryl group; m and n are each independently 0, 1 or 2; *1 and *2 represent the connection positions with L, Q and U respectively; Preferably, when both Z1 and Z2 are present, one of them is an optionally substituted 5-12 member nitrogen-containing heterocyclic group, and the other is an optionally substituted 5 member nitrogen-containing heteroaryl group; Preferably, L is a group that includes: wherein Y1and Y2are each independently CR L3 or N, R L3 is H, halogen, hydroxyl, cyano, C 1-3 alkyl or C 1-3 alkoxy; m and n are each independently 0, 1 or 2; p1, p2, p3and p4are each independently 0 or 1 ; *1and *2represent the positions of attachment to L to Q and U, respectively.

6. The compound, stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt, or a mixture thereof, as described in any one of claims 1 to 5, characterized in that, The E3 ubiquitin ligase binding ligand includes at least the following groups: In the formula, X is N or CR d2 ;R d2 H, halogen, hydroxyl, cyano, C 1-3 Alkyl or C 1-3 Alkoxy group; *3 indicates the position where the remaining part of the compound of formula I is attached; Preferably, U is a group consisting of: Wherein, ring D is absent, or is an optionally substituted aryl group, an optionally substituted heterocyclic group, or an optionally substituted heteroaryl group; V is a bond, -NR d1 -、-C(O)NR d1 -or-NR d1 C(O)-; X is N or CR d2 ;R d1 For H or C 1-3 Alkyl; R d2 H, halogen, hydroxyl, cyano, C 1-3 Alkyl or C 1-3 Alkoxy group; *2 indicates the connection position of the U group and L; Preferably, ring D is: an optionally substituted 6-14-membered aryl group, an optionally substituted 4-10-membered heterocyclic group, or an optionally substituted 5-10-membered heteroaryl group; preferably, ring D is optionally surrounded by 1, 2, or 3 groups selected from halogen, oxo, C 1-3 Alkyl, C 3-6 cycloalkyl and C 1-3 Substitution of alkoxy groups; Preferably, U is selected from the following groups: Among them, d1, d2, d3, and d4 are each independently N or CR. d3 R d3 H, halogen, C 1-3 Alkyl, C 3-6 cycloalkyl or C 1-3 alkoxy group; X is CH or N; T is CH2 or C=O; R d4 For H or C 1-3 Alkyl group; *2 indicates the connection position of the U group and L.

7. The compound of claim 1, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, characterized in that, The compound has the structure shown in formula IIa or IIb: Wherein, A, ring B, ring W, R1-R8, L1, L2, Z1, Z2 and ring D are as described in any one of claims 1 to 7.

8. The compound of claim 1, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, characterized in that, The compound has the structure shown in formula IIIa or IIIb: Wherein, L1, L2, Z1, Z2, ring D and X are as described in any one of claims 1 to 6; R9 and R 10 Each is independently selected from halogens and C 1-3 alkyl; R 11 and R 12 Each C is independently selected from hydrogen, halogens, and optionally substituted C. 1-3 alkyl; T is N or CR 13 ; R 13 It is hydrogen, cyano, hydroxyl, C 1-3 Alkyl or C 1-3 Alkyl group.

9. The compound of claim 1, its stereoisomers, tautomers, N-oxides, hydrates, solvates, isotopically labeled compounds, or pharmaceutically acceptable salts, or mixtures thereof, characterized in that, The compound is selected from:

10. Use of any compound, stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound, or pharmaceutically acceptable salt, or mixture thereof, according to any one of claims 1 to 9, in the preparation of a medicament for the treatment or prevention of a disease or condition by reducing Wee1 protein levels; Preferably, the disease or condition is cancer, preferably selected from liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, colon cancer, malignant pancreatic islet tumor, malignant carcinoid cancer, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, piloblastic leukemia, rhabdomyosarcoma, Kaposi's sarcoma, urogenital tumors, thyroid cancer, esophageal cancer, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, idiopathic thrombocytosis, adrenocortical carcinoma, skin cancer, and prostate cancer; Preferably, the drug further comprises at least one anticancer drug, or a pharmaceutically acceptable salt of the anticancer drug; preferably, the anticancer drug is selected from one or more of the following: olaparib, niraparib, rucaparib, talazoparib, senaparib, saruparib, vorinostat, romidesin, palbistat, belistat, palbociclib, busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, carboplatin, camptothecin, irinotecan, topotecan, Doxorubicin, Epirubicin, Aclarubicin, Mitoxantrone, Methylhydroxyrosine, Mintopopeptide, 5-azacytidine, Gemcitabine, 5-Fluorouracil, Methotrexate, 5-Fluoro-2′-Deoxyuridine, Fludarabine, Nerapine, Cytarabine, Pralatrexate, Pemetrexed, Hydroxyurea, Thioguanine, Colchicine, Vincristine, Vinorelbine, Paclitaxel, Ixaspirone, Cabazitaxel, Docetaxel, Monoclonal Antibody, Panitumumab, Netsotuzumab, Nivolumab, Pembrolizumab, Ramucirumab, Bevacizumab, Pertuzumab, Trastuzumab, Cetuximab, Obintocinumab Ofatumumab, Rituximab, Alemumab, Tiimumab, Tosimomab, Bentuximab, Daremumab, Erlotinib, T-DM1, Ofatumumab, Dinutuximab, Blinatumomab, Iprimumab, Avastin, Herceptin, Rituxan, Imatinib, Gefitinib, Erlotinib, Ostinib, Afatinib, Celitinib, Alectinib, Crizotinib, Erlotinib, Lapatinib, Sorafenib, Regorafenib, Vemurafenib, Dabrafenib, Aflibercept, Sunitinib, Nilotinib, Dasatinib, Bosutinib Pralatinib, ibrutinib, cabozantinib, lenvatinib, vandetanib, trametinib, carbitinib, axitinib, tesiromoximib, Idelalisib, pazopanib, tesiromoximib, everolimus, tamoxifen, letrozole, fulvestrant, mitoroguanidine, octreotide, retinic acid, arsenic trioxide, zoledronic acid, bortezomib, carfilzomib, Ixazomib, vemodega, sonidega, denosumab, thalidomide, lenalidomide, venetoclax, Aldesleukin (recombinant human interleukin-2), and Sipueucel-T (prostate cancer treatment vaccine); Preferably, the drug is used in combination with radiotherapy.

11. A pharmaceutical composition comprising the compound of any one of claims 1 to 9, or a stereoisomer, tautomer, N-oxide, hydrate, solvate, isotopically labeled compound or pharmaceutically acceptable salt thereof, or a mixture thereof and a pharmaceutically acceptable carrier; Preferably, the pharmaceutical composition further contains at least one anticancer drug, or a pharmaceutically acceptable salt of the anticancer drug; Preferably, the at least one anticancer drug is selected from the group consisting of: olaparib, niraparib, rucaparib, talazoparib, senaparib, saruparib, vorinostat, romidesin, palbistat, belitista, palbociclib, busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, carboplatin, camptothecin, irinotecan, topotecan, doxorubicin, epirubicin, aclarubicin, mitoxantrone, methyl hydroxyrosine, and methyltoprolol. Prescriptions, 5-azacytidine, gemcitabine, 5-fluorouracil, methotrexate, 5-fluoro-2′-deoxyuridine, fludarabine, nerabine, cytarabine, pralatrexate, pemetrexed, hydroxyurea, thioguanine, colchicine, vincristine, vinorelbine, paclitaxel, ixapril, cabazitaxel, docetaxel, monoclonal antibodies, panitumumab, nezotuzumab, nivolumab, pembrolizumab, ramucirumab, bevacizumab, pertuzumab, trastuzumab, cetuximab, oxaliplatin, ofumab, rituximab, alemtuzumab, teimozide, tosimomuzumab Bentuximab, Daremumab, Erlotinib, T-DM1, Ofatumumab, Dinutuximab, Blinatumomab, Iprimumab, Avastin, Herceptin, Rituxan, Imatinib, Gefitinib, Erlotinib, Ostinib, Afatinib, Celitinib, Alectinib, Crizotinib, Erlotinib, Lapatinib, Sorafenib, Regorafenib, Vemurafenib, Dabrafenib, Aflibercept, Sunitinib, Nilotinib, Dasatinib, Bosutinib, Pralatinib, Ibrutinib, Cabozantinib, Lenvatinib, Vandetanib Trametinib, Carbitinib, Axitinib, Tessiromoxetine, Idelalisib, Pazopanib, Testifloxacin, Everolimus, Vorinostat, Romidhizine, Pabistat, Belistat, Tamoxifen, Letrozole, Fulvestrant, Mitoguanidine, Octreotide, Retinoic acid, Arsenic trioxide, Zoledronic acid, Bortezomib, Carfilzomib, Ixazomib, Vemodil, Sonicil, Denosumab, Salidone, Lenalidomide, Venetoclax, Aldesleukin (recombinant human interleukin-2), and Sipueucel-T (prostate cancer treatment vaccine).

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