Aminopyrimidine compounds or salts thereof as proteolysis agents, and methods of making and medical uses thereof

Abstract

The application discloses an aminopyrimidine compound as a protein degradation agent, which is a compound shown in formula (I) or a pharmaceutically acceptable salt, stereoisomer or solvate thereof. The application also discloses a preparation method and medical application of the compound. The compound has FLT3 and CHK1 degradation activity, has proliferation inhibition activity on various tumor cell strains, is effective on various AML mutations such as internal tandem duplication mutations of a near-membrane domain and D835 point mutations of an activation loop in a kinase domain, can overcome drug resistance caused by point mutations in clinic, and can be applied to preparation of an antitumor drug.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and more particularly to an aminopyrimidine compound or its salt as a protein degrading agent, a method for preparing the compound, and its pharmaceutical uses. Background Technology

[0002] Fms-like tyrosine kinase 3 (FLT3) is a class III tyrosine kinase receptor that activates normal hematopoiesis and cell proliferation processes in primitive hematopoietic stem cells and progenitor cells, playing a crucial role in the expansion of pluripotent progenitor cells in the bone marrow. FLT3 gene mutations are found in nearly 30% of AML patients, with internal tandem repeats (ITDs) occurring in the juxtamembrane domain accounting for approximately 20%–25% of AML patients, and point mutations (TKDs) occurring in the tyrosine kinase domain accounting for approximately 7%–10%. These AML patients often have a poor prognosis. FLT3 consists of an extracellular ligand-binding domain containing five immunoglobulin-like domains, a transmembrane domain, a juxtamembrane domain (JM), and a highly conserved intracellular kinase domain. Under normal conditions, FLT3 ligand (FL) binds to the FLT3 extracellular domain, causing FLT3 dimerization and phosphorylation, promoting cell survival, proliferation, and differentiation by activating multiple downstream signaling pathways (PI3K / Akt, RAS / MAPK, and JAK / STAT5). Both FLT3-ITD and FLT3-TKD mutations can lead to ligand-independent dimerization and autophosphorylation, thereby continuously activating the FLT3 signaling pathway and ultimately causing abnormal proliferation of malignant cells.

[0003] Currently, three FLT3 inhibitors have been approved for marketing by the FDA / Japan, and multiple candidate drugs are in clinical trials for AML. Novartis' multi-kinase inhibitor midostaurin was approved by the FDA in April 2017 for the treatment of adult AML patients with FLT3 mutations. Astellas' gilteritinib was approved by the FDA in 2018 for the treatment of adult AML patients with relapsed / refractory disease. Daiichi Sankyo's quizartinib was also approved in Japan in 2019 for the treatment of relapsed / refractory (R / R) AML patients. FLT3 inhibitor treatment has significantly improved the survival and prognosis of AML patients. Despite significant progress in developing more potent and specific next-generation FLT3 inhibitors, the emergence of resistance remains a major challenge. Most patients develop resistance and unexpected safety issues after initial remission. Short response periods and high relapse rates due to resistance remain major problems in the treatment of AML patients.

[0004] CHK1 is a key regulator in the cell cycle, participating in cellular responses to DNA damage or other types of stress. When DNA is damaged, the DNA damage response (DDR) is activated to repair the damaged DNA. Upstream ATR activates CHK1 by phosphorylating S345 and S317. Activated CHK1 further phosphorylates downstream substrates, thereby inhibiting cyclin-dependent kinase (CDK) activity, leading to cell cycle arrest, preventing cells from entering mitosis, and providing time for DNA repair. Studies have shown that CHK1 is associated with the survival and drug resistance of FLT3-ITD AML cells. Increased levels of reactive oxygen species (ROS) in FLT3-ITD AML cells lead to increased DNA damage, which is detrimental to cell survival. Cancer cells can avoid this adverse condition by enhancing the DNA damage response (DDR). CHK1, as a key factor in DDR activity, has been shown to be related to the survival of FLT3-ITD AML cells. Research has shown that high expression of CHK1 in transgenic mouse models reduces oncogene-induced toxic DNA replication stress, thereby promoting tumorigenesis. Yuan et al. found that ectopic expression of CHK1 enhances drug resistance in FLT3-ITD cells and maintains H3 histone phosphorylation during DNA damage response. These results suggest that CHK1 may be associated with tumor cell survival and drug resistance. Targeting CHK1 can effectively inhibit the proliferation of FLT3-ITD AML cells. Wang et al. reported that all-trans retinoic acid (ATRA) induces fatal mitotic catastrophe in FLT3-ITD AML cells by degrading CHK1 kinase, thereby preventing DNA damage repair. High CHK1 expression may be associated with poor prognosis in AML patients. In AML patients, high CHK1 expression is associated with shorter overall, event-free, and relapse-free survival and earlier relapse. Zhang et al. used shRNA to reduce CHK1 expression and found that it significantly inhibited the proliferation of FLT3-ITD-positive MV4-11 cells and induced apoptosis.

[0005] Protein hydrolysis-targeting chimeras (PROTACs) are heterobifunctional small molecules that link a target protein (POI) ligand and an E3 ubiquitin ligase ligand via a linker. They can degrade the target protein via the ubiquitin-proteasome system (UPS) by ubiquitination of the POI onto the E3 ubiquitin ligase, followed by proteasome degradation. Compared to traditional small molecule inhibitors (SMIs), PROTACs offer several advantages as potential therapeutic agents. PROTACs can target undrugible targets and mutant proteins, improving target selectivity, prolonging drug action time, and simultaneously attenuating both kinase and non-kinase activities of the protein. Therefore, PROTACs have become an emerging technology for developing unique targeted anticancer drugs.

[0006] Therefore, it is necessary to provide bifunctional compounds that bind to both the target proteins FLT3, CHK1, FLT3 / CHK1, and E3 ubiquitin ligase, thereby targeting and ubiquitinizing FLT3, CHK1, and FLT3 / CHK1, inducing proteasomal degradation of FLT3, CHK1, and FLT3 / CHK1, for the treatment of related diseases or conditions. More advantageously, such bifunctional compounds possess good FLT3 / CHK1 specificity, improved physical and chemical properties, and / or improved pharmacokinetic properties. Moreover, by developing FLT3, CHK1, and FLT3 / CHK1 degraders, the problems of drug resistance mutations in existing FLT3 small molecule inhibitors are overcome. Summary of the Invention

[0007] The purpose of this invention is to provide a class of aminopyrimidine derivatives with FLT3 and CHK1 protein degradation activity and anti-tumor effects. These compounds have excellent biological activity and enhance the molecular diversity and novelty of the compounds. The function of these compounds is to recruit targeted proteins to E3 ubiquitin ligases for degradation.

[0008] This invention also provides an aminopyrimidine derivative tautomer, stereoisomer, optical isomer, salt, and preparation method as a protein degrading agent, as well as its application as an Fms-like tyrosine kinase 3 (FLT3), cell cycle checkpoint kinase 1 (CHK1), or dual-targeting FLT3 / CHK1 protein degrading agent in antitumor, anti-inflammatory, and anti-autoimmune disease drugs.

[0009] The technical solution of the present invention is as follows:

[0010] In a first aspect, the present invention relates to a compound having the structure shown in general formula (I):

[0011]

[0012] Or its optical isomer or its pharmaceutically acceptable salt, stereoisomer, or solvate.

[0013] in:

[0014] R1 is selected from C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 cycloalkyl, (C 0-3 Alkyl)C 3-10 Heterocyclic alkyl groups, wherein the heteroatom is at least one of nitrogen, oxygen, and sulfur, and the alkyl, cycloalkyl, and heterocyclic alkyl moieties may be further independently bound by 1, 2, 3, or 4 R atoms. a Substitution; and multiple R groups exist on the same group. a At that time, multiple R a They are independent of each other and can be the same or different;

[0015] R2 is selected from -C 1-6 Alkyl-NH-, -C 0-3 Alkyl-C 3-6 cycloalkyl-NH-,-C 0-3 Alkyl-C 3-8 Heterocyclic alkyl-, -C 0-3 Alkyl-C 3-8 Heterocyclic alkyl groups -NH-, -C 7-18 Spirocycloalkyl groups -NH-, - spirocycloalkyl groups containing heteroatoms (e.g., C 6-18 Spirocycloalkyl)-, wherein the heteroatom is at least one of nitrogen, oxygen, and sulfur, and the alkyl, cycloalkyl, heterocycloalkyl, and spirocycloalkyl moieties may each be independently further separated by 1, 2, 3, or 4 R- atoms. b Substitution; and multiple R groups exist on the same group. b At that time, multiple R b They are independent of each other and can be the same or different;

[0016] R a R b Each group is independently selected from hydrogen, halogen, hydroxyl, amino, cyano, and C. 1-3 Alkoxy, C 1-5 Alkyl, wherein C 1-5 Alkyl groups may be further substituted with halogens, hydroxyl groups, or cyano groups;

[0017] Each of the q R3 groups is independently selected from hydrogen, halogen, hydroxyl, C. 1-3 Alkoxy, C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 Cycloalkyl, cyano, nitro, amino, wherein the alkyl, cycloalkyl, and alkoxy moieties may be further independently converted by one or more halogens, hydroxyl groups, amino groups, cyano groups, or alkoxy groups (e.g., C14, 22, 32, 43, 54 ... 1-3 alkoxy), C 1-5 Alkyl substitution;

[0018] q can be 0, 1, 2, 3 or 4.

[0019] The linker is either nonexistent or -(CH2). m - a hydrocarbon chain, wherein any one or more -CH2- units in the hydrocarbon chain can be independently selected from -CH2=CH2-, -C≡C-, -O-, -CO-, -NR L The linker may be replaced by a unit of -, -SO-, -SO2- and / or a 3-12 membered cyclic group, and any optional -CH2- unit or substitution group on the linker may be further replaced by one or more independently selected from deuterium, halogen, cyano, hydroxyl, amino, C 1-6 Alkyl, C1-6 Haloalkyl, C 1-6 Hydroxyalkyl groups, 4-8 membered heterocyclic hydrocarbon groups containing oxygen, nitrogen, or sulfur atoms, and C 3-8 Cycloalkyl substituents, wherein R L For H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl; and wherein, when two cyclic groups in the linker are adjacent, they optionally form a fused ring, spiro ring, or bridged ring structure together; the cyclic groups include cycloalkyl and heterocyclic alkyl containing oxygen, nitrogen, or sulfur atoms.

[0020] m can be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

[0021] Degron is the E3 ubiquitin ligase-binding moiety; the Degron moiety is an E3 ubiquitin ligase moiety selected from the following structures:

[0022]

[0023] V is CR c Or N, where R c C is hydrogen or optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups. 1-6 Alkyl; preferably, V is N, CH, C(C) 1-6 Alkyl); more preferably, V is N or CH;

[0024] W represents a single bond, NH, or NR. d -CO-NH-, O, R d , where R d For unreplaced C 1-6 Alkylene or C-aryl groups optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups 1-6 Alkylene; more preferably, W is -CO-NH- or a single bond;

[0025] U represents CO and CR. e R f ;R e R f They can be the same or different; more preferably, U is CO or CH2.

[0026] X1 and X2 are independent CRs. e Or N;

[0027] R e R f Each is independently selected from hydrogen, halogen, and C. 1-3 Alkoxy, C1-6 alkyl;

[0028] Each of the p R4 groups is independently a hydrogen, halogen, hydroxyl, or alkoxy group (e.g., C). 1-3 alkoxy), C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 Cycloalkyl, cyano, nitro, amino, NH(C) 1-6 alkyl), N(C) 1-6 Alkyl)2, wherein the alkyl, cycloalkyl, and alkoxy moieties may be further independently converted by one or more halogens, hydroxyl groups, amino groups, cyano groups, C 1-3 Alkoxy, C 1-5 Alkyl substitution;

[0029] p can be 0, 1, 2, 3, or 4.

[0030] In some embodiments, R1 is selected from C. 1-6 Alkyl, (C 0-3 Alkyl)C 3-6 cycloalkyl, (C 0-3 Alkyl)C 3-6 Heterocyclic alkyl; wherein the heteroatom is at least one of nitrogen and oxygen, and the alkyl, cycloalkyl, and heterocyclic alkyl moieties may be further independently atomized by 1, 2, 3, or 4 R atoms. a Substitution; and multiple R groups exist on the same group. a At that time, multiple R a They are independent of each other and can be the same or different; R a Selected from hydrogen, halogen, hydroxyl, C 1-3 Alkyl, wherein C 1-3 Alkyl groups can be further replaced by halogens or hydroxyl groups;

[0031] Preferably, R1 can be selected from More Optimal Selection

[0032] Preferably, in some embodiments, the structure described in general formula (Ⅰa) is provided:

[0033]

[0034] The variables are as defined in this paper.

[0035] R2 is selected from -C 1-6 Alkyl-NH-, -C 0-3 Alkyl-C 3-6 cycloalkyl-NH-,-C 0-3 Alkyl-C 3-6 Heterocyclic alkyl-, -C 0-3 Alkyl-C3-6 Heterocyclic alkyl groups -NH-, -C 7-18 Spirocycloalkyl-NH-, -heterocyclic alkyl-, wherein the heteroatom is at least one of nitrogen and oxygen, and the alkyl, cycloalkyl, heterocyclic alkyl, and spirocycloalkyl moieties may be further independently and independently bound by 1, 2, 3, or 4 R-. b Substitution; and multiple R groups exist on the same group. b At that time, multiple R b They are independent of each other and can be the same or different;

[0036] R b Selected from hydrogen, halogen, hydroxyl, amino, cyano, C 1-3 Alkoxy, C 1-5 Alkyl, wherein C 1-5 Alkyl groups may be further substituted with halogens, hydroxyl groups, or cyano groups;

[0037] The linker is either nonexistent or -(CH2). m - a hydrocarbon chain, wherein any one or more CH2 units in the hydrocarbon chain may further optionally be independently selected from -CH2=CH2-, -C≡C-, -O-, -CO-, -NR L -, -SO-, -SO2- and / or 3-12 membered cyclic groups are substituted, and any CH2 unit or substitution group on the linker may be further replaced by one or more independent groups selected from deuterium, halogen, cyano, hydroxyl, amino, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl groups, 4-8 membered heterocyclic hydrocarbon groups containing oxygen, nitrogen, or sulfur atoms, and C 3-8 Cycloalkyl substituents, wherein R L For hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl; and wherein, when two cyclic groups in the Linker are adjacent, they optionally form together a fused ring, a spiro ring, or a bridged ring structure; the cyclic groups include cycloalkyl and heterocyclic alkyl containing oxygen, nitrogen, or sulfur atoms;

[0038] m can be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

[0039] Degron is the E3 ubiquitin ligase-binding moiety. The Degron moiety is selected from the following E3 ubiquitin ligase moieties:

[0040]

[0041] V is CRc Or N, where R c C is hydrogen or optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups. 1-6 alkyl;

[0042] W represents a single bond, NH, or NR. d CO-NH, O, R d , where R d For unreplaced C 1-6 Alkylene or C-aryl groups optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups 1-6 Alkylene;

[0043] U represents CO and CR. e R f ;R e R f They can be the same or different;

[0044] X1 and X2 are each independently selected from CR e Or N;

[0045] R e R f Each is independently selected from hydrogen, halogen, and C. 1-3 Alkoxy, C 1-6 alkyl;

[0046] Each of the p R4 groups is independently a hydrogen, halogen, hydroxyl, or C group. 1-3 Alkoxy, C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 Cycloalkyl, cyano, nitro, amino, NH(C) 1-6 alkyl), N(C) 1-6 Alkyl)2, wherein the alkyl, cycloalkyl, and alkoxy moieties may be further independently converted by one or more halogens, hydroxyl groups, amino groups, cyano groups, C 1-3 Alkoxy, C 1-5 Alkyl substitution.

[0047] p can be 0, 1, 2, 3, or 4.

[0048] Preferably, R2 is selected from -C 0-3 Alkyl-C 4-6 cycloalkyl-NH-,-C 0-3 Alkyl-C 4-5 Heterocyclic alkyl-, wherein the heteroatom is at least one of nitrogen and oxygen, and the alkyl, cycloalkyl, and heterocyclic alkyl moieties may be further substituted by 1, 2, 3, or 4 hydrogens or halogens, respectively.

[0049] As a further preferred option, R2 is selected from -C 0-3 Alkyl-C 4-6cycloalkyl-NH-,-C 0-3 Alkyl-C 4-5 Heterocyclic alkyl-, wherein the heteroatom is preferably a nitrogen atom, which can be understood as the linker being attached from the nitrogen atom; more preferably, the R2 is selected from any of the following structures: Optimal Selection

[0050] As a specific preferred solution, R1 is R2 is That is, equation (I) has the structure of equation (II):

[0051]

[0052] The variables are as defined in this article.

[0053] In some implementations, the linker is -(CH2). m - hydrocarbon chain, preferably C 1-12 A hydrocarbon chain, wherein any one or more -CH2- units in the hydrocarbon chain are optionally and independently replaced by units of -CH2=CH2-, -C≡C-, -O-, -CO-, -NH- and / or 3- to 10-membered cyclic groups, and wherein any -CH2- unit or replacing group on the linker is optionally replaced by one or more units independently selected from deuterium, halogen, cyano, hydroxyl, amino, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl groups, 4-6 membered heterocyclic hydrocarbon groups containing oxygen or nitrogen atoms, and C 3-6 The cycloalkyl group is substituted; and wherein, when the two cyclic groups in the linker are adjacent, they optionally form a fused ring, spiro ring, or bridged ring structure together; the cyclic groups include cycloalkyl groups and heterocyclic alkyl groups containing oxygen atoms and / or nitrogen atoms.

[0054] m can be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

[0055] Preferably, the cyclic groups that form part of the Linker are each independently derived from divalent groups of the following groups: C 3-8 Cycloalkanes, 3-8 member nitrogen-containing heterocyclic alkanes, C 6-10 Aromatic rings (including benzene rings and fused aromatic rings, such as naphthalene rings), 5-7 membered nitrogen-containing heteroaromatic rings; wherein when two cyclic groups are adjacent, they may optionally form a fused ring, spiro ring or bridged ring structure together; and the cyclic groups may optionally be further substituted by one or more substituents independently selected from deuterium, halogen, hydroxyl, and amino groups.

[0056] More preferably, the cyclic groups that are part of the Linker are each independently selected from divalent groups derived from the following groups: cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclohexane, aziridine, aziridine, pyrrole, dihydropyrrole, pyrrolidine, piperidine, piperazine; wherein when two cyclic groups are adjacent, they may optionally form a fused ring, a spiro ring, or a bridged ring together.

[0057] In some implementations, the linker is a saturated / or partially unsaturated divalent carbon. 1-12 A hydrocarbon chain, wherein one or more hydrocarbon chain units of the hydrocarbon chain are optionally independently replaced by O, CO-, NH and / or 4-6 nitrogen-containing heterocyclic alkanes (e.g., piperidine ring, piperazine ring, tetrahydropyrrole, aziridine butane), and wherein when two cyclic groups in the linker are adjacent, they optionally form together a fused ring, a spiro ring or a bridged ring.

[0058] It is understandable that when there are more than one unit used to replace a hydrocarbon chain unit, they can be adjacent or not adjacent.

[0059] It can also be understood that optional substituents on the linker can be on any chain unit, including on hydrocarbon chain units and / or on substitution units used to replace hydrocarbon chain units.

[0060] The Linker can be -L1-L2-L3-L4-L5-, where L1, L2, L3, L4, and L5 are each independent:

[0061] a) Key;

[0062] b) Randomly selected by 1-3 R g Replacement C 1-12 Alkylene chain;

[0063] c) Randomly selected by 1-3 R g Replacement C 3-12 cycloalkyl;

[0064] d) Randomly selected by 1-3 R g Substituted 3-12 membered heterocyclic groups;

[0065] e) Randomly selected by 1-3 R g Replacement C 2-12 alkenyl chain;

[0066] f) Randomly selected by 1-3 R g Replacement C 2-12 Hypo-alkynyl chain;

[0067] g) 1, 2, 3, 4, 5 or 6 ethylene glycol units

[0068] h) Randomly selected by 1-3 Rg Substituted 7-11 membered spiroheterocyclic alkyl groups; wherein the heteroatom is a nitrogen atom;

[0069] i)-CO-,-C(O)O-,O,-NR L -,S,-C(O)-NR L -;

[0070] Each R g It can be independently a halogen, nitro, cyano, hydroxyl, or C group. 1-6 Alkoxy, C 1-6 Alkyl, amino; R L For hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl.

[0071] In some implementations, the Linker is composed of one or more fragments from (i) to (vii):

[0072] (i)-CO-(CH2) a -NH-, where a is an integer from 0 to 12;

[0073] (ii)CO-[(CH2)2-O-] n -CH2CH2NH, where n is an integer from 0 to 8 (i.e., n is an integer of 0, 1, 2, 3, 4, 5, 6, 7, 8);

[0074] (iii)-CO-(CH2) b -(4-7-membered nitrogen-containing heterocyclic alkanes)-, where b is an integer from 0 to 5 (i.e., b is an integer of 0, 1, 2, 3, 4, or 5);

[0075] (iv)-(CH2) c -(4-7 nitrogen-containing heterocyclic alkanes)-, where c is an integer from 0 to 5 (i.e., c is an integer of 0, 1, 2, 3, 4, or 5);

[0076] (v)-CO-(CH2) d -(4-7 member nitrogen-containing heterocyclic alkanes)-(CH2) e -(4-7 nitrogen-containing heterocyclic alkanes)-, where d and e are integers from 0 to 5 (i.e., d and e are integers of 0, 1, 2, 3, 4, and 5), and d+e≤10;

[0077] (vi)-(CH2) e -7-11 membered spirocycloalkyl containing nitrogen atoms-; where e is an integer of 0, 1, 2, 3, 4, or 5;

[0078] (vii) key;

[0079] The 4-7 nitrogen-containing heterocyclic alkanes can each be independently and optionally substituted with hydroxyl groups or halogens.

[0080] The 4-7 member nitrogen-containing heterocycles mentioned therein are piperazine rings, piperidine rings, tetrahydropyrrole rings, or azacyclic butanes.

[0081] It is understood that, unless otherwise specified, the L portion mentioned above can be connected to the other two portions of the molecule in any direction, such as from left to right or from right to left.

[0082] In some preferred embodiments, the Linker can be selected from any of the following structures:

[0083]

[0084] In some implementations, the Degron portion has the following structure:

[0085]

[0086] V is N or CR c , where R c C is hydrogen or optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups. 1-6 Alkyl; preferably, V is N, CH, C(C) 1-6 Alkyl); more preferably, V is N or CH;

[0087] W represents a single bond, NH, or NR. d -CO-NH-, O, R d , where R d For unreplaced C 1-6 Alkylene or C-aryl groups optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups 1-6 Alkylene; preferably, W is a single bond, CO-NH;

[0088] U represents CO and CR. e R f ;R e R f They can be the same or different; preferably, U is CO or CH2;

[0089] R e R f Selected from hydrogen, halogens, C 1-6 Alkyl, C 1-3 Alkoxy;

[0090] R4 can be independently hydrogen, halogen, hydroxyl, or C. 1-3 Alkoxy, C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10Cycloalkyl, cyano, nitro, amino, NH(C) 1-6 alkyl), N(C) 1-6 Alkyl)2, wherein the alkyl, cycloalkyl, and alkoxy moieties may be further independently converted by one or more halogens, hydroxyl groups, amino groups, cyano groups, C 1-3 Alkoxy, C 1-5 Alkyl substitution; preferably, R4 is selected from hydrogen, halogen, alkoxy, and amino. p is 0, 1, 2, 3, or 4.

[0091] As a preferred embodiment, the compound of formula (I), (II), or (III) wherein the Degron portion is selected from:

[0092]

[0093] The R4, R c R d The definition of p is the same as above.

[0094] As a further preferred option, the Degron portion is selected from:

[0095]

[0096] Selected from hydrogen or halogen; X1 and X2 are each independent CH or N; and p is 0 or 1.

[0097] The special part E is selected from:

[0098]

[0099]

[0100] In this application, unless otherwise stated, the terms used have the meanings defined below. Terms not explicitly defined in this application have the general meanings commonly understood by those skilled in the art.

[0101] The terms “a,” “an,” “the,” and similar terms used in this application shall be understood to include both the singular and the plural, unless the context otherwise specifically indicates or is obviously contradictory.

[0102] In this article, "CO" generally refers to a carbonyl group, i.e., "C=O"; "SO" generally refers to a sulfinyl group, i.e., "S=O"; and "SO2" generally refers to a sulfonyl group, i.e., "S(=O)2". The short hyphen "-" appearing in this invention indicates the linking site of a substituent. For example, -NR a R b This indicates that the group is connected to the rest of the molecule via a nitrogen atom. However, when the connection site of the substituent is obvious to those skilled in the art (e.g., for halogens, hydroxyl groups, NR...), a Rb (etc.), can be omitted.

[0103] When the valence bond of the group has a wavy line When, it indicates that the group is connected to the rest of the molecule through the valence bond.

[0104] In this invention, the "alkyl" and other alkyl portions of the groups (such as alkoxy or haloalkyl) can be branched or branched.

[0105] In this invention, "cycloalkyl" refers to a monocyclic or polycyclic hydrocarbon cyclic group, generally a 3-10 member cycloalkyl group, including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclohexadienyl, cyclohepttrienyl, borneol, norpinyl, norcaryl, adamantyl, benzoyl, decahydronaphthyl, norbornyl, spiro[4.5]decyl, etc. The cycloalkyl group may be unsubstituted or substituted with one or more suitable substituents.

[0106] In this invention, "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). Preferred halogens are fluorine and chlorine.

[0107] In this invention, "heterocyclic alkyl" refers to a 3-10 carbon ring structure, wherein at least one carbon atom in the ring is substituted by at least one heteroatom, including nitrogen, oxygen, or sulfur. Heterocyclic alkyl groups may have one or more carbon-carbon double bonds or carbon-heteroatom double bonds in the cyclic group, as long as the cyclic group does not become aromatic by its presence.

[0108] In this invention, "heterocyclic group" refers to a 3-15 member aliphatic (e.g., fully or partially saturated heterocycle) or aromatic (e.g., heteroaryl) monocyclic or bicyclic ring system. Examples of monocyclic ring systems are any 5- or 6-membered rings containing 1, 2, 3, or 4 heteroatoms independently selected from oxygen, nitrogen, and sulfur. A 5-membered ring has 0-2 double bonds, and a 6-membered ring has 0-3 double bonds. Representative examples of monocyclic systems include, but are not limited to, azacyclobutane, azacyclohexane, aziridine, diazacyclohexane, 1,3-dioxolane, dioxane, dithiazide, furan, imidazole, imidazoline, imidazoline, isothiazole, isothiazolin, isothiazolin, isoxazole, isoxazoline, isoxazoline, morpholine, oxadiazole, oxadiazolin, oxadiazolin, oxazole, oxazoline, oxazolin, piperazine, piperidine, pyran, pyrazine, pyrazole, pyrazoline, pyrazolidine, pyridine, pyrimidine, pyridazine, pyrrole, pyrroleline, pyrrole, tetrahydrofuran, tetrahydrothiophene, tetrazine, tetrazolium, thiadiazole, thiadiazolin, thiadiazolin, thiazolidine, thiazoline, thiazoline, thiophene, thiomorpholine, thiomorpholine sulfone, thiaran, triazine, triazole, trithiazide, etc. Examples of bicyclic systems are any of the above monocyclic systems fused with an aryl group as defined herein, a cycloalkyl group as defined herein, or another monocyclic system as defined herein. Representative examples of bicyclic ring systems include, but are not limited to, benzimidazole, benzothiazole, benzothiadiazole, benzothiophene, benzoxadiazole, 1,4-benzodioxane, benzoxazole, benzofuran, benzopyran, benzothiaran, benzodioxine, 1,3-benzodioxene, zoline, indazole, indole, indolone, dihydroindole, indazine, diazanaphthalene, isobenzofuran, isobenzothiaphene, isoindole, isodihydroindole, isoquinoline, phthalazine, purine, pyranopyridine, pyridopyrimidine, piperidine, quinoline, quinazine, quinoxaline, quinoxaline, tetrahydroisoquinoline, tetrahydroquinoline, thiopyranopyridine, etc.

[0109] In one embodiment of the present invention, the compound of general formula (I) may be selected from the following specific compounds:

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116] The numbers below the above 59 compounds are their corresponding codes. For ease of description and brevity, these codes will be used directly in the following content of this specification.

[0117] Secondly, the present invention provides the above-mentioned 2-aminopyrimidine derivatives and pharmaceutically acceptable salts.

[0118] In one embodiment of the present invention, the pharmaceutically acceptable salt is an organic acid salt or an inorganic acid salt. The organic acid salts include formate, acetate, propionate, pyruvate, glycolate, oxalate, malate, succinate, glutarate, mandelate, citrate, trifluoroacetate, fumarate, oxalate, malate, L-malate, D-malate, lactate, camphor sulfonate, p-toluenesulfonate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, benzoate, tartrate, L-tartrate, D-tartrate, oxalate, succinate, maleate, ascorbate, and amino acid salts. The inorganic acid salts include hydrochloride, hydrobromide, sulfate, phosphate, nitrate, hydroiodide, or perchlorate.

[0119] Thirdly, the present invention provides a composition containing the above-mentioned 2-aminopyrimidine compound or a pharmaceutically acceptable salt thereof.

[0120] In one embodiment of the present invention, the composition comprises at least one 2-aminopyrimidine compound of general formula (I) or a pharmaceutically acceptable salt thereof, another targeting molecule (preferably a conventional cytotoxic drug, a compound used after chemotherapy, a compound used in stem cell induction maintenance therapy, and a compound used in acute myeloid leukemia), and optionally a pharmaceutically acceptable carrier.

[0121] Fourthly, the present invention provides a pharmaceutical preparation for treating or preventing diseases related to FLT3, CHK1, and FLT3 / CHK1, and also contains an adjuvant component.

[0122] In one embodiment of the present invention, the pharmaceutical preparation is a tablet, capsule, powder, granules, ointment, solution, suspension, injection, inhaler, gel, microsphere, or aerosol, etc. The auxiliary ingredient is such as cyclodextrin, arginine, or meglumine. The cyclodextrin is selected from α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, (C... 1-4 alkyl)-α-cyclodextrin, (C 1-4 alkyl)-β-cyclodextrin, (C 1-4 alkyl)-γ-cyclodextrin, (hydroxy-C) 1-4 alkyl)-α-cyclodextrin, (hydroxy-C) 1-4 alkyl)-β-cyclodextrin, (hydroxy-C) 1-4 alkyl)-γ-cyclodextrin, (carboxyl-C)1-4 alkyl)-α-cyclodextrin, (carboxyl-C) 1-4 alkyl)-β-cyclodextrin, (carboxyl-C) 1-4 Alkyl)-γ-cyclodextrin, α-cyclodextrin glycoethers, β-cyclodextrin glycoethers, γ-cyclodextrin glycoethers, α-cyclodextrin sulfonyl ether, β-cyclodextrin sulfonyl ether, and γ-cyclodextrin sulfonyl ether. The auxiliary components also include medically acceptable carriers, adjuvants, or mediators. Other pharmaceutically acceptable pharmaceutical compositions may include ion exchangers, alumina, aluminum stearate, and lecithin; buffering substances include phosphates, glycine, arginine, sorbic acid, etc.

[0123] Fifthly, the present invention provides the use of 2-aminopyrimidine compounds or pharmaceutically acceptable salts in the preparation of medicaments for the prevention or treatment of clinical diseases related to FLT3 and / or CHK1.

[0124] In one embodiment of the present invention, the FLT3-mediated disease is a hematologic disorder or a solid tumor.

[0125] In a further specific embodiment, the hematologic disease is selected from acute myeloid leukemia (AML), acute T-cell leukemia, myelodysplastic syndrome, mixed lineage leukemia (MLL), T-cell acute leukemia (T-ALL), B-cell acute leukemia (B-ALL), chronic myeloid monocytic leukemia (CMML), chronic lymphocytic leukemia, chronic myeloid leukemia, and chronic neutrophilic leukemia. The solid tumor is selected from breast cancer, ovarian cancer, lung cancer, prostate cancer, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, head and neck cancer, thyroid cancer, liver cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, and skin cancer.

[0126] In a sixth aspect, the present invention provides the use of 2-aminopyrimidine compounds or pharmaceutically acceptable salts thereof, alone or in combination with one or more other therapeutic agents, in clinical diseases related to FLT3 and / or CHK1. The other therapeutic agents are selected from IDH1 inhibitors, IDH2 inhibitors, BCL-2 inhibitors, hypomethylating agents, and antimetabolites.

[0127] The compounds of this invention have FLT3 and CHK1 degradation activities, inhibit proliferation of various tumor cell lines, and are effective against various AML mutations, such as internal tandem repeat mutations in the juxtamembrane domain and D835 point mutations in the activation loop of the kinase domain. They can overcome drug resistance caused by point mutations in clinical practice and can be used in the preparation of antitumor drugs. Attached Figure Description

[0128] Figures 1-2The images show the imaging results of the FLT3 / CHK1 protein degradation experiment using the compounds in the examples. Detailed Implementation

[0129] The preparation of compounds will be further illustrated in the following examples. These examples are for illustrative purposes only and do not limit the invention in any way.

[0130] Example 1: 3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)-N-((1s,4s)-4-((2-(4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)propionamide (Compound 1)

[0131]

[0132] Step 1: Synthesis of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (intermediates 1-2)

[0133] 3-Fluorophthalic anhydride (1-1.3 g, 18.06 mmol) and 3-amino-2,6-piperidinedione hydrochloride (3.27 g, 19.87 mmol) were dissolved in acetic acid (30 mL), and then potassium acetate (5.32 g, 54.2 mmol) was added. The mixture was heated to 120 °C and reacted for 4 h. After cooling to room temperature, the reaction solution was poured into 200 mL of ice water. After the solid precipitated, the solvent was removed by filtration. The solid was washed with ice water to obtain a brownish-gray solid 1-2. 1 H NMR (500MHz, DMSO-d6) δ11.15(s,1H),7.95(td,J=8.0,4.5Hz,1H),7.79(d,J=7.3Hz,1H),7.73(t,J=8.9Hz,1H),5.16(dd,J=12.9,5.4Hz,1H),2. 89(ddd,J=17.2,14.0,5.4Hz,1H),2.61(dd,J=14.2,3.3Hz,1H),2.58–2.50(m,1H),2.07(dtd,J=12.9,5.3,2.2Hz,1H); ESI-MS: m / z=277.1[M+H] + .

[0134] Step 2: Synthesis of 4-chloro-5-(trifluoromethyl)pyrimidin-2-amine (intermediates 1-4)

[0135] See patent document CN111646978A.

[0136] Step 3: Synthesis of tert-butyl ((1S,4S)-4-((2-amino-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)carbamate (intermediates 1-5)

[0137] See patent CN 111646978 A.

[0138] Step 4: Synthesis of 1-(4-bromophenoxy)-2-methyl-2-propanol (intermediates 1-7)

[0139] In a sealed container, p-bromophenol (1-6, 500 mg, 2.91 mmol) was dissolved in DMF (10 mL), followed by the sequential addition of cesium carbonate (1896 mg, 5.82 mmol) and methyl propylene oxide (315 mg, 4.37 mmol). The mixture was stirred at 70 °C for 24 h. After the reaction was complete, 30 mL of water was added, and the mixture was extracted with EA (20 mL × 3). The organic layers were combined, washed three times with 30 mL of saturated brine, dried over anhydrous Na₂SO₄, and the solvent was removed by vacuum distillation. The mixture was purified by silica gel column chromatography using PE:EA (5:1) as the eluent to obtain a colorless oily liquid 1-7. 1 H NMR(500MHz, DMSO-d6)δ7.42(d,J=9.0Hz,2H),6.94–6.88(m,2H),4.64(s,1H),3.69(s,2H),1.18(s,6H); ESI-MS:m / z=267.0[M+Na] + .

[0140] Step 5: Synthesis of 1-(4-((4-(((1s,4s)-4-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenoxy)-2-methyl-2-propanol (intermediates 1-8)

[0141] See patent CN 111646978 A.

[0142] Step 6: Synthesis of 3-amino-N-((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)propionamide (intermediates 1-9)

[0143] Compounds 1-8 (100 mg, 0.228 mmol) were weighed and dissolved in DMF (2 mL). Boc-beta-alanine (52 mg, 0.273 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (65 mg, 0.341 mmol), 1-hydroxybenzotriazole (37 mg, 0.273 mmol), and N,N-diisopropylethylamine (45 mg, 0.341 mmol) were added, and the mixture was stirred overnight at room temperature. After the reaction was complete, 3 mL of water was added, and the mixture was extracted with EA (2 mL × 3). The organic layers were combined, washed three times with 3 mL of saturated brine, dried over anhydrous Na₂SO₄, and the solvent was removed by vacuum distillation. The mixture was purified by silica gel column chromatography using DCM:MeOH (40:1) as the eluent to give a white solid 1-9. 1 H NMR(500MHz,DMSO-d6)δ9.50(s,1H),8.18(s,1H),7.63(d,J=7.5Hz,1H),7.59(d,J=9.5Hz, 2H),6.87(d,J=9.0Hz,2H),6.76(t,J=5.5Hz,1H),5.82(s,1H),4.59(s,1H),4.09(s,1H),3. 97–3.79(m,1H),3.67(s,2H),2.89(dd,J=13.0,6.5Hz,2H),2.08(t,J=7.5Hz,2H),1.73(dd ,J=12.5,8.5Hz,4H),1.68–1.54(m,4H),1.36(s,9H),1.19(s,6H); ESI-MS:m / z=611.3[M+H] + .

[0144] Step 7: Synthesis of 3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)-N-((1s,4s)-4-((2-(4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)propionamide (Compound 1)

[0145] Compounds 1-9 (100 mg, 0.160 mmol) were dissolved in 2 mL of DCM. 1 mL of trifluoroacetic acid was slowly added dropwise with stirring in an ice bath. After the addition was complete, the reaction mixture was moved to room temperature and stirred for 3 h. After the reaction was complete, the solvent was evaporated under reduced pressure to obtain an intermediate. This intermediate was dissolved in 2 mL of DMSO along with compound 1-2 (58 mg, 0.208 mmol). Then, N,N-diisopropylethylamine (104 mg, 0.802 mmol) was added, and the mixture was heated to 100 °C overnight. After the reaction was complete, 3 mL of water was added, and the mixture was extracted with EA (2 mL × 3). The organic layers were combined, washed three times with 3 mL of saturated brine, dried over anhydrous Na₂SO₄, and the solvent was removed by reduced pressure distillation. The mixture was purified by silica gel column chromatography using DCM:MeOH (35:1) as the eluent to give compound 1, a yellow-green solid. Yield: 61%; 1 H NMR (500MHz, DMSO-d6) δ11.07(s,1H),9.51(s,1H),8.18(s,1H),7.82(d,J=7.0Hz,1H),7.64–7.59(m,2H),7.58( d,J=4.0Hz,1H),7.16(d,J=8.5Hz,1H),7.03(d,J=7.0Hz,1H),6.89(d,J=9.0Hz,2H),6.71(t,J=6.0Hz,1H),5.77 (s,1H),5.03(dd,J=13.0,5.5Hz,1H),4.59(s,1H),4.07(s,1H),3.84(s,1H),3.67(s,2H),2.93–2.79(m,1H),2. 55(d,J=18.5Hz,1H),2.51(s,1H),2.02–1.91(m,1H),1.70(s,4H),1.60(s,4H),1.55–1.43(m,4H),1.19(s,6H); 13 C NMR (126MHz, DMSO-d6) δ173.36,172.75,170.03,168.81,167.27,160.83,157.40,154.90(q,J CF =4.5Hz),154.30,146.23,136.25,133.03,132.16,125.26(q,J CF=269.7Hz),121.02(2C),117.27,114.35(2C),110.57,109.29,76.40,68.67,51.14,48.51,44 .21,34.90,33.25,30.95,28.37(2C),26.67(2C),24.40(2C),22.13; ESI-MS: m / z=767.3[M+H] + .

[0146] Preparation Example 2: Synthesis of compounds 2-9, 16, 18, 20, 58, 59

[0147] The preparation method is as described in Example 1:

[0148] Using N-Boc-Gamma-aminobutyric acid, Boc-5-aminopentanoic acid, tert-butoxycarbonyl-6-aminohexanoic acid, 7-(N-tert-butoxycarbonylamino)heptanoic acid, BOC-8-aminooctanoic acid, 9-tert-butoxycarbonylamino-nonanoic acid, 10-((tert-butoxycarbonyl)amino)decanoic acid, BOC-11-aminoundecanoic acid, 1-N-BOC-4-piperidinylpropionic acid, 1-N-Boc-3-acetidinecarboxylic acid, 1-(tert-butoxycarbonyl)azacyclobutane-3-ethyl Acid, BOC-amino-monoethylene glycol-carboxylic acid, and 5,8,11,14-tetraoxa-2-azaheptadecanoic acid 1-tert-butyl ester were used to replace Boc-beta-alanine to obtain the corresponding intermediates. These intermediates were then de-Boc-treated with trifluoroacetic acid and reacted with 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione under alkaline conditions with DMSO as solvent to give compounds 2–9, 16, 18, 20, 58, and 59. The target molecules are shown in Table 1 below.

[0149] Table 1

[0150]

[0151]

[0152] Preparation Example 3: Synthesis of compounds 10-14 and 45 Refer to the preparation method in Example 1:

[0153] Replacing cis-(4-aminocyclohexyl)carbamate with 1-tert-butoxycarbonyl-4-aminopiperidine, the resulting intermediate undergoes Buchwald-Hartwig coupling with 1-(4-bromophenoxy)-2-methyl-2-propanol under conditions of 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene), tris(dibenzylideneacetone)dipalladium, and cesium carbonate. The resulting intermediate undergoes amide condensation with Boc-beta-alanine, Boc-5-aminovaleric acid, 7-(N-tert-butoxycarbonylamino)heptanoic acid, 9-tert-butoxycarbonylamino-nonanoic acid, and BOC-11-aminoundecanoic acid, respectively. After de-Bocing with trifluoroacetic acid, the resulting intermediate undergoes a substitution reaction with 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione under alkaline conditions using DMSO as solvent to give compounds 10–14. Compound 45 was obtained by direct substitution reaction of starting material I-8 with 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione under alkaline conditions using DMSO as solvent. The target molecules are shown in Table 2 below.

[0154] Table 2

[0155]

[0156] Preparation Example 4: Synthesis of compounds 15, 17, 19, 21–24, 47, 50

[0157] The preparation method is as described in Example 1:

[0158] The substitution reaction of 4-fluorophthalic anhydride for 3-fluorophthalic anhydride in a glacial acetic acid / potassium acetate buffer system yields 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione.

[0159] Compounds 1-8 were amide-condensed with intermediates 1-8 by substituting Boc-beta-alanine with 1-N-BOC-4-piperidinylpropionic acid, 1-N-Boc-3-acetidinecarboxylic acid, 1-(tert-butoxycarbonyl)azacyclobutane-3-acetic acid, 1-Boc-4-piperidinylcarboxylic acid, 1-Boc-4-piperidinylacetic acid, 4-(N-Boc-4-piperidinyl)butyric acid, and 4-Boc-piperazinylacetic acid. The resulting intermediate was deBoc-treated with trifluoroacetic acid and then substituted with 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione under alkaline conditions to give compounds 15, 17, 19, 21-24.

[0160] The substitution reaction of 4,5-difluorophthalic anhydride for 3-fluorophthalic anhydride in a glacial acetic acid / potassium acetate buffer system yields 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione. The substitution of 1-N-BOC-4-piperidinpropionic acid for Boc-beta-alanine followed by amide condensation with intermediate 1-8, followed by de-Boc removal under acidic conditions, and then a substitution reaction with 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione under alkaline conditions yields compound 50.

[0161] Replacing cis-(4-aminocyclohexyl)carbamate with 1-tert-butoxycarbonyl-4-aminopiperidine, the intermediate 2-methyl-1-(4-((4-(piperidin-4-ylamino)-5-(trifluoromethyl)pyrimidin-2-yl)amino)phenoxy)prop-2-ol was obtained according to steps 3 and 5 of Example 1. Then, it underwent an amide condensation reaction with 1-Boc-4-piperidincaric acid. After de-Boc under acidic conditions, it underwent a substitution reaction with 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione under alkaline conditions to obtain compound 47.

[0162] The target molecules mentioned above are shown in Table 3 below.

[0163] Table 3

[0164]

[0165]

[0166] Preparation Example 5: Synthesis of Compound 25

[0167]

[0168] Step 1: Synthesis of 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione (intermediates 1-10)

[0169] The preparation method of intermediates 1-2 in Example 1 is the same, except that the raw material used is 4-fluorophthalic anhydride, which yields a brown solid (85%). 1H NMR (500MHz, DMSO) δ11.13 (s, 1H), 7.99 (dd, J = 8.2, 4.5 Hz, 1H), 7.83 (dd, J = 7. 4,2.2Hz,1H),7.70(td,J=9.4,2.3Hz,1H),5.15(dd,J=12.9,5.4Hz,1H),2.88( ddd,J=17.2,14.0,5.4Hz,1H),2.59(ddd,J=17.1,4.2,2.2Hz,1H),2.51(dd,J =13.0,3.8Hz,1H),2.05(dtd,J=7.6,5.3,2.2Hz,1H); ESI-MS:m / z=277.1[M+H] + .

[0170] Step 2: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(4-hydroxypiperidin-1-yl)isoindoline-1,3-dione (intermediates 1-12)

[0171] Intermediate 1-11 (300 mg, 1.09 mmol) was dissolved in 2 mL of DMSO, and 4-hydroxymethylpiperidine (110 mg, 109 mmol) and DIPEA (561.5 mg, 4.34 mmol) were added sequentially. The mixture was stirred overnight at 95 °C. After the reaction was complete, 30 mL of water was added, and the mixture was extracted with EA (20 mL × 3). The organic layers were combined, washed twice with 20 mL of saturated brine, dried over anhydrous Na₂SO₄, and the solvent was removed by vacuum distillation. The mixture was purified by silica gel column chromatography with DCM:MeOH = 50:1 as the eluent to give a yellow-green solid 1-12. Yield: 84%; 1 HNMR (500MHz, DMSO-d6) δ11.08(s,1H),7.65(d,J=8.5Hz,1H),7.31(d,J=2.0Hz,1H),7.23(dd,J=8.5,2. 5Hz,1H),5.06(dd,J=13.0,5.5Hz,1H),4.75(d,J=4.0Hz,1H),3.82(dt,J=9.0,4.0Hz,2H),3.78–3.71(m, 1H),3.19(ddd,J=13.0,9.5,3.0Hz,2H),2.88(ddd,J=16.5,13.5,5.0Hz,1H),2.62–2.55(m,1H),2.52(d d,J=14.0,3.0Hz,1H),2.05–1.97(m,1H),1.85–1.77(m,2H),1.46–1.37(m,2H); ESI-MS:m / z=358.1[M+H] + .

[0172] Step 3: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(4-oxopiperidin-1-yl)isoindoline-1,3-dione (intermediates 1-13)

[0173] Compound 1-12 (335 mg, 0.937 mmol) was dissolved in 10 mL of DCM. The mixture was stirred in an ice bath, and DMP (795 mg, 1.87 mmol) was slowly added in portions. After the addition was complete, the reaction mixture was brought to room temperature and reacted for 2 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was filtered under reduced pressure, the solvent was evaporated, and the solution was purified by silica gel column chromatography using DCM:MeOH = 50:1 as the eluent to give a yellow-green solid 1-13. Yield: 74%; ESI-MS: m / z = 356.1 [M+H] + .

[0174] Step 4: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(4-((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)piperidin-1-yl)isoindoline-1,3-dione (compound 25)

[0175] Intermediate 1-8 (75 mg, 0.137 mmol) was dissolved in 2 mL of DMF, and compound 1-13 (63 mg, 0.177 mmol) was added. The pH was then adjusted to 5–6 with a catalytic amount of glacial acetic acid. After stirring at room temperature for 1 h, sodium cyanoborohydride (25.7 mg, 0.41 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC. 10 mL of water was added to the reaction mixture, and the mixture was extracted three times with EA (20 mL). The combined organic layers were washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography using DCM:MeOH = 35:1 as the eluent to give compound 25. Yield: 40%. 1H NMR (500MHz, DMSO-d6) δ11.09(s,1H),9.50(s,1H),8.17(s,1H),7.69(d,J=7.0Hz,1H),7.62–7.58(m,2H),7.58–7.54(m,1H),7.12(d,J =8.5Hz,1H),7.02(d,J=7.0Hz,1H),6.87(d,J=9.0Hz,2H),6.63(t,J=6.0Hz,1H),5.82(s,1H),5.05(dd,J=12.5,5.5Hz,1H),4.59(s,1H ),4.09(d,J=5.5Hz,1H),3.84(s,1H),3.67(s,2H),3.30(s,2H),2.88(ddd,J=17.0,14.0,5.5Hz,1H),2.62–2.55(m,1H),2.55–2.51(m, 1H), 2.21 (t, J = 7.0Hz, 2H), 2.02 (dd, J = 9.0, 3.5Hz, 1H), 1.81 (dt, J = 14.5, 7.5Hz, 2H), 1.74 (t, J = 16.5Hz, 4H), 1.60 (s, 4H), 1.19 (s, 6H); 13 C NMR (126MHz, DMSO-d6) δ172.82,171.06,170.11,168.88,167.31,160.84,157.40,154.90(q,J CF =4.9Hz),154.31,146.39,136.26,133.04,132.24,125.27(q,J CF =269.9Hz),121.00(2C),117.14,114.36(2C),110.40,109.09,76.41,68.68,48.54,44.16,41 .57,32.53(2C),30.99,28.30,26.87(2C),26.67(2C),24.84,22.18; ESI-MS: m / z=781.3[M+H] + .

[0176] Preparation Example 6: Synthesis of compounds 26-32 and 56

[0177] The preparation method is as described in Example 5:

[0178] Substitution reactions were carried out with 4-hydroxymethylpiperidine, 4-piperidineethanol, 4-(3-hydroxypropyl)-piperidine, (R)-pyrrolidine-3-methanol, (S)-pyrrolidine-3-methanol, 3-hydroxyazacyclobutane hydrochloride, 3-methylhydroxyazacyclobutane hydrochloride, and (3-fluoroazadin-3-yl)methanol in place of 4-hydroxypiperidine and 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione. The resulting product was then oxidized by Desmartin oxidation, and the product was reductively amination reaction with intermediates 1-8 to give compounds 26-32 and 56.

[0179] The target molecules mentioned above are shown in Table 4 below.

[0180] Table 4

[0181]

[0182]

[0183] Preparation Example 7: Synthesis of Compound 41

[0184] Step 1: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione

[0185] The preparation method of intermediates 1-2 in Example 1 was followed, except that 4,5-difluorophthalic anhydride was used instead of 3-fluorophthalic anhydride, resulting in a brown solid (70%). 1 H NMR (500MHz, DMSO) δ11.15(s,1H),8.16(t,J=7.7Hz,2H),5.17(dd,J=13.0,5.4Hz,1H),2.89(ddd,J=17.2,14.0,5.5Hz, 1H),2.61(ddd,J=17.0,4.1,2.2Hz,1H),2.56–2.50(m,1H),2.07(dtd,J=12.9,5.3,2.3Hz,1H); ESI-MS:m / z=295.1[M+H] + .

[0186] Step 2: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(3-(hydroxymethyl)azacyclobutane-1-yl)isoindoline-1,3-dione

[0187] Referring to step 2 of Example 5, 3-methylhydroxyazacyclobutane hydrochloride was used instead of 4-hydroxypiperidine, and 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione was used instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-isoindoline-1,3-dione, to obtain a yellow-green solid (76%). 1H NMR (500MHz, DMSO-d6) δ11.07(s,1H),7.57(d,J=11.5Hz,1H),6.88(d,J=7.5Hz,1H),5. 05(dd,J=13.0,5.5Hz,1H),4.84(t,J=5.5Hz,1H),4.18(dd,J=8.0,6.5Hz,2H),3.91(t, J=6.0Hz,2H),3.59(t,J=5.5Hz,2H),2.93–2.85(m,1H),2.84–2.77(m,1H),2.61–2.55( m,1H),2.54–2.51(m,1H),2.01(ddd,J=10.5,5.5,3.0Hz,1H); ESI-MS:m / z=362.1[M+H] + .

[0188] Step 3: Synthesis of 1-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde

[0189] The synthesis method in step 3 of Example 5 was followed, except that the starting material used was 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(3-(hydroxymethyl)azacyclobutane-1-yl)isoindoline-1,3-dione, yielding a yellow-green solid; yield: 42%, ESI-MS: m / z = 360.1 [M+H] + .

[0190] Step 4: 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(3-((((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)methyl)azacyclobutane-1-yl)isoindoline-1,3-dione (Compound 41)

[0191] The preparation method follows step 4 of Example 5, replacing 2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde, followed by reductive amination to obtain green solid compound 41. Yield: 62%. 1H NMR (500MHz, DMSO-d6) δ11.08(s,1H),9.47(s,1H),8.15(s,1H),7.61(d,J=9.0Hz,2H),7.57(s,1H),6.88(d, J=7.5Hz,1H),6.86(d,J=9.0Hz,2H),6.09(s,1H),5.06(dd,J=13.0,5.5Hz,1H),4.59(s,1H),4.25(t,J=7.0H z,2H),4.09(s,1H),3.88(s,2H),3.67(s,2H),2.92–2.85(m,1H),2.85–2.75(m,3H),2.71(s,1H),2.62–2.55 (m,1H),2.55–2.51(m,1H),2.06–1.96(m,1H),1.79(dd,J=21.5,10.5Hz,2H),1.72–1.49(m,6H),1.19(s,6H); 13 CNMR(125MHz,DMSO-d6)δ172.76,170.00,166.84,166.43,160.81,157.33,154.79(q,J CF =5.1Hz),154.51,154.21,152.54,144.19(d,J CF =12.0Hz),133.15,129.38,125.19(q,J CF =269.2Hz),120.90(2C),118.07(d,J CF =8.3Hz),114.31(2C),111.20(d,J CF =22.3Hz), 107.71(d,J CF =7.8Hz),76.40,68.66,57.33,51.62,50.40,48.91,30.96,30.49(2C),2 8.75(2C),26.66,26.60(2C),26.26(2C),22.15; ESI-MS: m / z=783.3[M+H] + .

[0192] Preparation Example 8: Synthesis of Compound 51

[0193] The preparation method is similar to that in Example 7, except that 3-hydroxyaziridine hydrochloride is used instead of 3-methylhydroxyaziridine hydrochloride. After the substitution reaction, the ketone is oxidized to a ketone by Desmartin and then subjected to a reductive amination reaction with 1-8 to obtain compound 51. The target molecules are shown in Table 5 below.

[0194] Table 5

[0195]

[0196] Preparation Example 9: Synthesis of compounds 48-49 and 55

[0197] The preparation method of Example 5 is as follows: 3-hydroxyazacyclobutane hydrochloride is used to replace 4-hydroxypiperidine and undergoes a substitution reaction with 2-(2,6-dioxo-piperidin-3-yl)-5-fluoro-isoindoline-1,3-dione, followed by a Desmartin oxidation reaction to obtain the intermediate 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde. Referring to steps 3 and 5 of Example 1: Methylpropane was substituted with (3-methyloxetane-3-yl)methylmethanesulfonyl ester, N-(2-chloroethyl)pyrrolidine hydrochloride, and cyclopropylmethylchloromethyl ester respectively in the reaction with p-bromophenol. The resulting intermediates were then coupled with intermediates 1-5 in a Buchwald-Hartwig coupling reaction under the conditions of 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene), tris(dibenzylacetone)dipalladium, and cesium carbonate. After undergoing a Boc removal reaction, the resulting intermediates were further subjected to reductive amination with 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde to obtain compounds 48-50 and 55. The target molecules are shown in Table 6 below.

[0198] Table 6

[0199]

[0200] Preparation Example 10: Synthesis of Compound 46

[0201] The preparation method is as described in Example 5:

[0202] Replacing cis-(4-aminocyclohexyl)carbamate with 1-tert-butyloxycarbonyl-4-aminopiperidine in a reductive amination reaction with 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidine-4-carboxaldehyde yields compound 46. The target molecules are shown in Table 7 below.

[0203] Table 7

[0204]

[0205] Preparation Example 11: Synthesis of Compound 35

[0206]

[0207] Step 1: Synthesis of tert-butyl 4-(2-(((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)-2-oxoethyl)piperidine-1-carboxylic acid

[0208] Refer to step 6 of Example 1. The amide condensation reaction was carried out using 1-Boc-4-piperidineacetic acid instead of Boc-beta-alanine to give a white solid. Yield: 82%. 1 H NMR(500MHz,DMSO-d6)δ9.51(s,1H),8.18(s,1H),7.65(d,J=7.5Hz,1H),7.60(d,J= 9.0Hz,2H),6.87(d,J=9.0Hz,2H),5.82(s,1H),4.59(s,1H),4.09(s,1H),3.97–3.7 9(m,3H),3.67(s,2H),2.69(s,2H),2.02(t,J=14.5Hz,2H),1.90–1.79(m,1H),1.72 (s,4H),1.58(d,J=12.0Hz,8H),1.38(s,9H),1.19(s,6H); ESI-MS:m / z=665.4[M+H] + .

[0209] Step 2: Synthesis of 2-(1'-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-[1,4'-bipiperidin]-4-yl)-N-((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)acetamide

[0210] Synthesis of compound 25, referring to step 4 of Example 5. Yield: 65%; 1H NMR (500MHz, DMSO-d6) δ11.08(s,1H),9.52(s,1H),8.19(s,1H),7.76–7.69(m,1H),7.69–7.64(m,1H),7.60(d,J=9.0Hz,2H),7.38(s,1H ),7.29(s,1H),6.87(d,J=9.0Hz,2H),5.75(s,1H),5.07(dd,J=13.0,5.5Hz,1H),4.59(s,1H),4.22(t,J=6.5Hz,2H),4.09(dd,J=10.5,5. 0Hz,2H),3.84(s,1H),3.67(s,2H),3.17(d,J=5.0Hz,3H),2.97(t,J=11.5Hz,3H),2.93–2.81(m,2H),2.59(d,J=19.0Hz,1H),2.56–2.51 (m,1H),2.07(s,3H),2.03–1.96(m,2H),1.74(d,J=8.5Hz,5H),1.63(dd,J=19.0,11.0Hz,7H),1.41–1.31(m,2H),1.20(d,J=3.5Hz,6H).; 13 C NMR(125MHz,DMSO-d6)δ173.28,170.97,170.57,168.07,167.42,161.30,157.87,155.36(q,J CF =5.3Hz),155.19,154.77,134.49,133.49,125.74(q,J CF =269.3Hz),125.46,121.50(2C),118.82,118.11,118.06,114.81(2C),108.22,76.86,69.14,61.53,52.22,51.71(2C),49.33,49 .21,47.24(2C),44.41,33.82,32.41(2C),31.45(2C),30.96,28.88(2C),27.34(2C),27.12(2C),22.66; ESI-MS: m / z=904.4[M+H] + .

[0211] Preparation Example 12: Synthesis of Compounds 36-40

[0212] The synthesis method is described in Example 11.

[0213] Compounds 36 and 37 were obtained by reductive amination of 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-carboxaldehyde and 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde, respectively, instead of 2-(2,6-dioxopiperidin-3-yl)-5-(4-oxopiperidin-1-yl)isoindoline-1,3-dione.

[0214] Replacing 1-Boc-4-piperidinic acid with 1-(tert-butoxycarbonyl)azacyclobutane-3-acetic acid, compounds 1-8 undergo amide condensation. After debonding under acidic conditions, the compounds are reductively amination reactions with (2,6-dioxoperidin-3-yl)-5-(4-oxoperidin-1-yl)isoindoline-1,3-dione, 1-(2-(2,6-dioxoperidin-3-yl)-1,3-dioxoperidindoline-5-yl)azacyclobutane-3-carboxaldehyde, and 1-(2-(2,6-dioxoperidin-3-yl)-1,3-dioxoperidindoline-5-yl)piperidin-4-carboxaldehyde to yield compounds 38-40. The target molecules are shown in Table 8 below.

[0215] Table 8

[0216]

[0217] Preparation Example 13: Synthesis of Compound 33

[0218]

[0219] Step 1: Synthesis of tert-butyl 2-(((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)-7-azaspiro[3.5]nonane-7-carboxylate

[0220] Intermediate 1-8 (100 mg, 0.228 mmol) and tert-butyl 2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (81.85 mg, 0.342 mmol) were dissolved in 2 mL of DMF, and the pH was adjusted to 5-6 with catalytic amount of glacial acetic acid. The mixture was stirred at room temperature for 60 min, then sodium cyanoborohydride (44.5 mg, 0.684 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC. 10 mL of water was added to the reaction mixture, and the mixture was extracted three times with EA (20 mL). The organic layers were combined, washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography with DCM:MeOH = 50:1 as the eluent to give a white solid 1-14. Yield: 62%; 1H NMR(500MHz,DMSO-d6)δ9.53(s,1H),8.18(s,1H),7.59(d,J=9.0Hz,2H),6.86(d,J=9.0Hz,2H),5.79(s,1H),4.60(s,1H),4.09(s,1H),3.8 7–3.83(s,4H),3.76(s,2H),3.67(s,2H),2.64(s,2H),2.36(s,2H),1.78–1.56(m,12H),1.36(s,9H),1.19(s,6H); ESI-MS: m / z=663.4[M+H] + .

[0221] Step 2: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(2-((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)-7-azaspiro[3.5]nonane-7-yl)isoindoline-1,3-dione (compound 33)

[0222] 2-(((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (1-14) (100 mg, 0.151 mmol) was dissolved in 2 mL of DCM. 1 mL of trifluoroacetic acid was slowly added dropwise with stirring in an ice bath. After the addition was complete, the reaction mixture was moved to room temperature and stirred for 3 h. After the reaction was complete, the solvent was evaporated under reduced pressure to obtain an intermediate. The obtained intermediate and compound 1-11 (62.5 mg, 0.226 mmol) were dissolved in DMSO (2 mL), and then N,N-diisopropylethylamine (91.3 mg, 0.754 mmol) was added. The mixture was heated to 100 °C and reacted overnight. After the reaction was complete, 3 mL of water was added, and the mixture was extracted with EA (2 mL × 3). The organic layers were combined, washed three times with 3 mL of saturated brine, dried over anhydrous Na₂SO₄, and the solvent was removed by vacuum distillation. The mixture was then purified by silica gel column chromatography with DCM:MeOH (20:1) as the eluent to give a yellow-green solid compound 33. 1H NMR (500MHz, DMSO-d6) δ11.07(s,1H),9.47(s,1H),8.16(s,1H),7.64(d,J=8.5Hz,1H),7.60(d,J=9.0Hz,2H),7.31(s,1H ),7.23(dd,J=8.5,2.0Hz,1H),6.86(d,J=9.0Hz,2H),6.08(s,1H),5.06(dd,J=13.0,5.5Hz,1H),4.59(s,1H),4.10(s,1H) ,3.66(s,2H),3.46(s,2H),3.38(s,2H),2.88(ddd,J=16.5,13.5,5.0Hz,1H),2.78(s,1H),2.58(dd,J=14.0,3.5Hz,1H), 2.55–2.51(m,1H),2.15(s,2H),2.01(dd,J=9.0,4.0Hz,1H),1.82(d,J=8.5Hz,2H),1.60(d,J=18.0Hz,13H),1.19(s,6H); 13 C NMR (125MHz, DMSO-d6) δ173.28,170.57,168.10,167.42,161.28,157.81,155.37,155.26(q,J CF =5.0Hz),154.70,134.50,133.60,130.08,125.64(q,J CF =267.5Hz),125.45,121.40(2C),118.03,117.84,114.78(2C),108.20,76.86,69.14,55.36,50.61(2C),49.20,46. 26,45.20,44.97(2C),38.73,32.23(2C),31.45,29.48(2C),27.13(2C),26.77(2C),22.67; ESI-MS: m / z=819.4[M+H] + .

[0223] Preparation Example 14: Synthesis of Compound 34

[0224] Refer to Example 13. 2-oxo-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester was substituted for 2-oxo-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester in a reductive amination reaction. The resulting intermediate underwent a deBoc reaction and then a substitution reaction with compound 1-11 under basic conditions to obtain compound 33. 1H NMR (500MHz, DMSO-d6) δ11.07(s,1H),9.50(s,1H),8.18(s,1H),7.70–7.65(m,1H),7.65–7.54(m,2H),7.31(d,J=2.0 Hz,1H),7.23(dd,J=8.5,2.0Hz,1H),6.87(d,J=9.0Hz,2H),5.82(s,1H),5.06(dd,J=13.0,5.5Hz,1H),4.60(s,1H),4 .18–3.97(m,3H),3.86(s,1H),3.67(s,2H),2.97(t,J=12.0Hz,2H),2.88(ddd,J=16.5,13.5,5.5Hz,1H),2.58(d,J=1 8.0Hz,1H),2.55–2.51(m,1H),2.07(d,J=7.0Hz,2H),2.04–1.94(m,2H),1.86–1.64(m,6H),1.60(s,4H),1.19(s,6H); 13 C NMR (125MHz, DMSO-d6) δ173.16,170.23,168.01,167.38,161.10,157.62,155.26,154.86(q,J CF =5.1Hz),154.61,134.22,133.47,128.74,125.46(q,J CF =269.4Hz),125.27,121.18(2C),116.86,114.28(2C),114.02,104.30,76.48,68.69,59.62(2C),54.96,49.7 1,47.52(2C),46.16,39.81,36.15,31.14,29.17(2C),26.83(2C),26.37(2C),22.36; ESI-MS: m / z=791.3[M+H] + .

[0225] Preparation Example 15: Synthesis of Compound 42

[0226]

[0227] Step 1: Synthesis of methyl 5-(3-(hydroxymethyl)azacyclobutane-1-yl)pyridinecarboxylate

[0228] Methyl 5-fluoropyridine-2-carboxylate (600 mg, 3.87 mmol) was dissolved in 3 mL of DMSO, followed by the addition of 3-methylhydroxyazine hydrochloride (573.6 mg, 4.64 mmol) and K₂CO₃ (641 mg, 4.64 mmol). The reaction mixture was allowed to react overnight at 90 °C. The reaction solution was diluted with water and then extracted with DCM (10 mL × 3). The organic layers were combined and washed three times with saturated brine, then dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to give a white crude product. Yield: 85%; 1 H NMR (500MHz, DMSO-d6) δ8.61(d,J=8.5Hz,1H),7.85(d,J=8.5Hz,1H),7.53(d,J=3.0Hz,1H),4.26(s,1H),3.92(q,J=8. 0Hz, 2H), 3.75 (s, 3H), 3.65 (dd, J=8.0, 5.5Hz, 2H), 3.58 (d, J=6.5Hz, 2H), 2.86–2.76 (m, 1H); ESI-MS: m / z=223.1[M+H] + .

[0229] Step 2: Synthesis of 5-(3-(hydroxymethyl)azacyclobutan-1-yl)pyridinecarboxylic acid

[0230] Methyl 5-(3-(hydroxymethyl)azacyclobutan-1-yl)pyridinecarboxylate (821 mg, 3.69 mmol) was dissolved in 15 mL of methanol-water solution (MeOH:H₂O = 4:1). NaOH (443 mg, 11.08 mmol) was added in portions. After the NaOH was completely dissolved, the reaction solution was heated to 60 °C and reacted for 3 h. TLC monitoring showed that the starting material was completely consumed. The reaction solution was placed in an ice bath, and 1 M dilute HCl solution was slowly added dropwise to adjust the pH to 5, resulting in solid precipitation. The reaction solution was filtered, the filter cake was collected, and dried to obtain a yellowish-brown crude product. This product was used directly in the next reaction without purification. Yield: 62%; ESI-MS: m / z = 231 [M+Na] + .

[0231] Step 3: Synthesis of N-(2,6-dioxopiperidin-3-yl)-5-(3-(hydroxymethyl)azacyclobutane-1-yl)pyridine amide

[0232] 5-(3-(hydroxymethyl)azacyclobutan-1-yl)pyridinecarboxylic acid (500 mg, 2.4 mmol) was dissolved in 5 mL of DMF, and DIPEA (1862 mg, 14.4 mmol) and HATU (1096 mg, 2.88 mmol) were added sequentially. The mixture was stirred at room temperature for 3 min. Then, 3-amino-2,6-piperidinidone hydrochloride (473 mg, 2.88 mmol) was added to the reaction mixture, and the mixture was reacted at room temperature for 2 h. The reaction was monitored by TLC until the starting material was consumed. The reaction mixture was diluted with water and extracted three times (20 mL × 3) with a dichloromethane-methanol (DCM:MeOH = 10:1) mixture. The organic layers were combined, washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The solid was purified by column chromatography to give a white solid. Yield: 46%; 1 H NMR (500MHz, DMSO-d6) δ10.83(s,1H),8.65(d,J=8.5Hz,1H),7.82(d,J=8.5Hz,1H),7.77(d,J=2.5Hz,1H ),6.87(dd,J=8.5,3.0Hz,1H),4.82(t,J=5.0Hz,1H),4.77–4.67(m,1H),4.00(t,J=8.0Hz,2H),3.72(dd ,J=8.0,5.5Hz,2H),3.59(t,J=5.5Hz,2H),2.91–2.82(m,1H),2.78(ddd,J=17.5,13.5,5.5Hz,1H),2.56 –2.51(m,1H),2.16(qd,J=13.0,4.5Hz,1H),2.00(ddd,J=10.5,5.5,3.0Hz,1H); ESI-MS:m / z=319.1[M+H] + .

[0233] Step 4: Synthesis of N-(2,6-dioxopiperidin-3-yl)-5-(3-formylazetane-1-yl)pyridine amide

[0234] The synthesis method in step 3 of Example 5 was followed, except that the starting material used was N-(2,6-dioxopiperidin-3-yl)-5-(3-(hydroxymethyl)azacyclobutane-1-yl)pyridine amide, yielding a white solid; yield: 35%, ESI-MS: m / z = 317.2 [M+H] + .

[0235] Step 5: Synthesis of N-(2,6-dioxopiperidin-3-yl)-5-(3-((((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)methyl)azacyclobutane-1-yl)pyridine amide (compound 42)

[0236] The synthesis method in step 4 of Example 5 is the same as that used in Example 5, except that the starting material is N-(2,6-dioxopiperidin-3-yl)-5-(3-formylazetane-1-yl)pyridine amide, which yields a white solid; yield: 31%; 1 H NMR (500MHz, DMSO-d6) δ10.84(s,1H),9.49(s,1H),8.66(d,J=8.5Hz,1H),8.17(s,1H),7.84(d,J=8.5Hz,1H),7.78(d,J=3.0H z,1H),7.60(d,J=9.0Hz,2H),6.90–6.87(m,1H),6.86(d,J=9.0Hz,2H),5.99(s,1H),4.78–4.67(m,1H),4.59(s,1H),4.13(s, 1H),4.08(t,J=7.5Hz,2H),3.71(s,2H),3.65(d,J=12.0Hz,2H),2.91(s,3H),2.78(ddd,J=17.5,13.5,5.5Hz,2H),2.55–2.51 (m,1H),2.16(qd,J=13.0,4.5Hz,1H),2.01(dd,J=9.0,4.0Hz,1H),1.90(s,1H),1.88–1.77(m,2H),1.65(s,6H),1.19(s,6H); 13 C NMR(125MHz,DMSO-d6)δ173.00,172.49,164.29,160.82,157.36,154.83(q,J CF =5.5Hz),154.24,148.64,137.73,133.12,131.65,126.42,125.22(q,J CF =269.3Hz),122.65,120.93(2C),117.01,114.32(2C),76.40,68.67,55.59,52.05,50.21,49.3 6,31.05,29.03(2C),26.67(2C),26.61(2C),26.30,24.24(2C),22.09; ESI-MS: m / z=740.3[M+H] +.

[0237] Preparation Example 16: Synthesis of Compounds 44 and 52

[0238] The synthesis was performed according to Example 15, replacing methyl 5-fluoropyridine-2-carboxylate with methyl p-fluorobenzoate and methyl 6-fluoronicotinate. The resulting intermediates were then reductively aminationd to give compounds 44 and 52. The target molecules are shown in Table 9 below.

[0239] Table 9

[0240]

[0241] Preparation Example 17: Synthesis of Compound 43

[0242]

[0243] Step 1: Synthesis of (1-(4-bromophenyl)azacyclobutane-3-yl)methanol

[0244] Under nitrogen protection, p-bromoiodobenzene (2000 mg, 7.07 mmol) was dissolved in anhydrous DMSO, and L-proline (326 mg, 2.83 mmol), potassium phosphate (4502 mg, 21.2 mmol), aziridine-3-ylmethanol, and cuprous iodide (471 mg, 2.47 mmol) were added sequentially. The reaction was carried out overnight at 90 °C under nitrogen atmosphere. The reaction solution was filtered, the filtrate was diluted with ethyl acetate and washed three times with saturated brine, the organic layer was dried over anhydrous sodium sulfate and concentrated, and purified by column chromatography using PE:EA = 3:1 as the eluent to give a yellow oily liquid. Yield: 25%; 1 H NMR (500MHz, DMSO-d6) δ7.31–7.22(m,2H),6.37–6.29(m,2H),4.74(t,J=5.5Hz,1H),3.80(t,J=7.5 Hz,2H),3.59–3.54(m,2H),3.51(dd,J=7.0,5.5Hz,2H),2.81–2.71(m,1H); ESI-MS:m / z=242.0[M+H] + .

[0245] Step 2: Synthesis of (1-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)azacyclobutane-3-yl)methanol

[0246] Compounds (1-(4-bromophenyl)azacyclobutane-3-yl)methanol (350 mg, 1.45 mmol), 2,6-bis(benzyloxy)-3-bromopyridine (905 mg, 2.17 mg), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) chloride (106 mg, 0.15 mmol), and cesium carbonate (1413 mg, 4.34 mmol) were dissolved in dioxane aqueous solution (dioxane:water = 4:1). The solution was purged with nitrogen three times and stirred overnight at 100 °C. The reaction mixture was evaporated to dryness and purified by silica gel column chromatography using PE:EA = 3:1 as the eluent to give a white solid. Yield: 56%; 1 H NMR (500MHz, DMSO) δ7.64(d,J=8.0Hz,1H),7.50–7.19(m,12H),6.50(d,J=8.0Hz,1H),6.41(d,J=8.7Hz,2H),5.37(d,J=1 6.0Hz,4H),4.74(t,J=5.5Hz,1H),3.83(t,J=7.5Hz,2H),3.61–3.50(m,4H),2.84–2.72(m,1H); ESI-MS:m / z=453.2[M+H] + .

[0247] Step 3: Synthesis of 3-(4-(3-(hydroxymethyl)azacyclobutane-1-yl)phenyl)piperidine-2,6-dione

[0248] (1-(4-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)azacyclobutane-3-yl)methanol (470 mg, 1.72 mmol) was dissolved in 8 mL of methanol, and Pd / C (50 mg) was added. After evacuation, hydrogen gas was introduced, and the reaction was stirred at room temperature for 3 h under a hydrogen atmosphere. The reaction was monitored by TLC until the reactants had completely reacted. The reaction solution was filtered, and the filtrate was evaporated to dryness and purified by column chromatography using DCM:MeOH = 70:1 as the eluent to give a white solid. Yield: 86%; 1H NMR (500MHz, DMSO-d6) δ10.74(s,1H),6.99(d,J=8.5Hz,2H),6.36(d,J=8.5Hz,2H),4.73(t,J=5. 5Hz,1H),3.85–3.74(m,2H),3.68(dd,J=11.0,5.0Hz,1H),3.60–3.54(m,2H),3.50(dd,J=7.0,5. 5Hz,2H),2.81–2.70(m,1H),2.62(ddd,J=17.0,11.0,5.5Hz,1H),2.45(dt,J=17.0,4.5Hz,1H),2 .10(dtd,J=15.5,11.0,4.5Hz,1H),1.98(ddt,J=15.5,10.5,5.0Hz,1H); ESI-MS:m / z=275.1[M+H] + .

[0249] Step 4: Synthesis of 1-(4-(2,6-dioxopiperidin-3-yl)phenyl)azacyclobutane-3-carboxaldehyde

[0250] The synthesis method in step 3 of Example 5 was followed, except that the starting material used was 3-(4-(3-(hydroxymethyl)azacyclobutane-1-yl)phenyl)piperidine-2,6-dione, yielding a white solid; yield: 86%, ESI-MS: m / z = 273.1 [M+H] + .

[0251] Step 5: Synthesis of 3-(4-(3-((((1s,4s)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)methyl)azacyclobutane-1-yl)phenyl)piperidine-2,6-dione (compound 43)

[0252] The synthesis method in step 4 of Example 5 is the same as that in which the raw material used is 1-(4-(2,6-dioxopiperidin-3-yl)phenyl)azacyclobutane-3-carboxaldehyde, yield: 22%; 1H NMR (500MHz, DMSO-d6) δ10.76(s,1H),9.55(s,1H),8.21(s,1H),7.59(d,J=9.0Hz,2H),7.02(d,J=8.5Hz,2H),6.8 7(d,J=9.0Hz,2H),6.41(d,J=8.5Hz,2H),5.68(s,1H),4.59(s,1H),4.19(s,1H),3.94(t,J=7.5Hz,2H),3.70(dd,J =10.5,4.0Hz,1H),3.66(s,2H),3.61–3.53(m,2H),3.17(d,J=5.0Hz,1H),3.12(s,1H),2.98(s,1H),2.68–2.62(m ,1H),2.60(d,J=5.5Hz,1H),2.11(d,J=9.0Hz,1H),2.03–1.79(m,6H),1.68(dd,J=34.5,11.5Hz,4H),1.19(s,6H); 13 CNMR(125MHz,DMSO-d6)δ174.73,173.53,160.86,157.53,155.00(q,J CF =5.2Hz),154.36,150.77,139.48,133.00,128.85(2C),127.67,125.27(q,J CF =269.3Hz),121.16(2C),114.38(2C),111.30(2C),76.42,68.70,55.78,53.84,48.37,46.58, 31.26(2C),27.73,26.69(2C),26.43(2C),26.15(2C),25.14,22.12; ESI-MS: m / z=696.3[M+H] + .

[0253] Preparation Example 18: Synthesis of Compound 53

[0254] The synthesis was performed according to Example 17. The intermediate obtained by replacing p-bromoiodobenzene with 1-bromo-3-fluoro-4-iodobenzene was reductively amination to give compound 53. The target molecules are shown in Table 10 below.

[0255] Table 10

[0256]

[0257] Preparation Example 18: Synthesis of Compound 54

[0258]

[0259] Step 1: Synthesis of (1-(4-nitrophenyl)piperidin-4-yl)methanol

[0260] K₂CO₃ (1.96 g) was added to a solution of 1-fluoro-4-nitrobenzene (1.0 g, 7.1 mmol) and 4-piperidinemethanol (980 mg, 8.5 mmol) in DMF (20 mL). The mixture was stirred at 80 °C for 15 hours. The reaction mixture was cooled to room temperature, poured into ice water (40 mL), and stirred for 20 minutes. The solid was filtered off, washed with water (20 mL × 2), and dried to give the product (1.4 g, 83.8%). 1 H NMR(400MHz,DMSO)δ8.03(d,J=9.5Hz,2H),7.01–6.98(m,2H),4.54(t,J=5.5Hz,1H),4.07–4.04(m,2H) ,3.29–3.26(m,2H),3.00–2.93(m,2H),1.76–1.67(m,3H),1.21–1.11(m,2H); ESI-MS: m / z=237.2[M+H] + .

[0261] Step 2: Synthesis of (1-(4-aminophenyl)piperidin-4-yl)methanol

[0262] At room temperature, 1.4 g (0.593 mmol) of (1-(4-nitrophenyl)piperidin-4-yl)methanol was dissolved in 20 mL of MeOH, and 2.8 g of 10% Pd / C was added to the solution. After evacuation, H2 was introduced, and this process was repeated three times. The mixture was stirred at room temperature for 5 h. The mixture was filtered through a diatomaceous earth mat and washed with MeOH. The filtrate was concentrated to give the product (83%). 1 H NMR (400MHz, DMSO) δ6.77–6.61(m,2H),6.54–6.38(m,2H),4.53(brs,2H),4.45(t,J=5.5Hz,1H),3.32–3.27( m,2H),2.46–2.41(m,2H),1.76–1.62(m,2H),1.50–1.31(m,3H),1.27–1.08(m,2H); ESI-MS: m / z=207.2[M+H] + .

[0263] Step 3: Synthesis of (1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)methylacetate

[0264] Acrylic acid (130 mg, 0.182 mmol) was added to a solution of (250 g, 0.121 mmol) in PhMe (4 mL). The mixture was stirred at 90 °C for 15 h. The reaction was cooled to 25 °C, and then HOAc (4.0 mL) and urea (364 g, 0.606 mmol) were added. The mixture was stirred at 110 °C for 24 h. The reaction was monitored by HPLC. The reaction was cooled to 25 °C and concentrated under vacuum. The residue was dissolved in EtOAc (15 mL) and then adjusted to pH 7 with saturated NaHCO3. The resulting solution was extracted with 2 × 200.0 mL of EtOAc, and the organic layers were combined. The organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified on silica gel (PE:EtOAc = 1:1) to give the product (175 g, 74%). 1 H NMR (400MHz, DMSO) δ10.32(s,1H),7.20(d,J=9.0Hz,2H),6.99(d,J=9.0Hz,2H),3.98(d,J=6.0Hz,2H),3.80–3.66(m ,2H),2.74–2.72(m,4H),2.09(s,3H),1.80(d,J=13.5Hz,4H),1.37(dd,J=12.0,3.0Hz,3H); ESI-MS:m / z=346.2[M+H] + .

[0265] Step 4: Synthesis of 1-(4-(4-(hydroxymethyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0266] (350 mg, 0.121 mmol) of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)methyl acetate was added to 2N HCl (2.6 mL) at 25 °C. The mixture was stirred at 100 °C for 15 h. The reaction was monitored by HPLC. The reaction was cooled to 0 °C and then adjusted to pH 7 with saturated NaHCO3. The solid was collected by filtration, washed with water (5 mL), and dried to obtain the product (169 mg, 55%). 1H NMR (400MHz, DMSO) δ10.26(s,1H),7.13(d,J=8.9Hz,2H),6.92(d,J=9.0Hz,2H),4.49(s,1H),3.78–3.61(m,4H),3.3 0–3.28(m,2H),2.70–2.66(m,4H),1.75–1.72(m,2H),1.52–1.49(m,1H),1.28–1.18(m,2H); ESI-MS:m / z=304.2[M+H] + .

[0267] Step 5: Synthesis of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidine-4-carboxaldehyde

[0268] The synthesis method in step 3 of Example 5 was followed, except that the starting material used was 1-(4-(4-(hydroxymethyl)piperidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione, yielding a white solid; yield: 21%, ESI-MS: m / z = 302.1 [M+H] + .

[0269] Step 6: Synthesis of 1-(4-(4-((((1S,4S)-4-((2-((4-(2-hydroxy-2-methylpropoxy)phenyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)cyclohexyl)amino)methyl)piperidin-1-yl)phenyl)dihydropyrimidin-2,4(1H,3H)-dione

[0270] The synthesis method in step 4 of Example 5 is the same as that used in Example 5, except that the raw material used is 1-(4-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)phenyl)piperidine-4-carboxaldehyde, with a yield of 23%. 1 H NMR (500MHz, DMSO-d6) δ10.58(s,1H),9.54(s,1H),8.21(s,1H),7.59(d,J=9.0Hz,2H),7. 12(d,J=8.5Hz,2H),6.94(d,J=9.0Hz,2H),6.87(d,J=9.0Hz,2H),5.71(s,1H),4.59(s,1H ),4.20(s,1H),4.10(d,J=12.5Hz,2H),3.80–3.62(m,4H),3.67(s,2H),3.15(s,1H),2.98 (t,J=12.0Hz,2H),2.82(s,2H),1.96–1.53(m,13H),1.19(s,6H); ESI-MS:m / z=725.4[M+H] + .

[0271] Preparation Example 19: Synthesis of Compound 57

[0272]

[0273] The method for synthesizing 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde is referenced in patent CN 113412259 A.

[0274] The synthesis of compound 57 is based on the method described in step 4 of Example 5, except that the starting material used is 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)azacyclobutane-3-carboxaldehyde, with a yield of 36%. 1 HNMR (500MHz, DMSO-d6) δ11.08(s,1H),9.45(s,1H),8.16(s,1H),7.61(d,J=9.0Hz,2H),7.21(s,1H),6.86(d,J=9.0Hz,2H),6. 76(d,J=2.0Hz,1H),6.63(dd,J=8.5,2.0Hz,1H),6.07(s,1H),4.78–4.67(m,1H),4.59(s,1H),4.22(s,2H),4.13(t,J=8.0Hz,3H ),3.72(dd,J=8.5,5.0Hz,2H),3.67(s,2H),2.92–2.86(m,1H),2.86(d,J=23.0Hz,2H),2.75(s,2H),2.63–2.57(m,1H),2.53(dd ,J=5.5,4.0Hz,1H),2.04–1.98(m,1H),1.83(dd,J=20.5,10.0Hz,2H),1.73–1.54(m,6H),1.20(s,6H); ESI-MS: m / z=751.4[M+H] + .

[0275] Examples of in vitro biological experiments

[0276] Unless otherwise specified, the experimental materials, reagents, operations and methods used in the following in vitro biological experimental examples are all available from commercial sources or can be easily obtained or prepared based on existing technology.

[0277] Example A: Test of the inhibitory activity of the compound on the proliferation of MV4-11 cells

[0278] (1) Cell line: Human acute myeloid leukemia cell line MV4-11

[0279] (2) Experimental steps and processing methods: Cells in the growth phase were digested with trypsin and then counted, with a result of 1×10⁻⁶. 4 Cell / well seeding density: Cells were seeded in 96-well plates at 80 μL per well and incubated overnight at 37°C with 5% CO2. 20 μL of a pre-set concentration of the compound (six concentration gradients per compound, three replicates per concentration) was added to each well, and the cells were incubated for 72 h (blank wells were in medium containing only 10% FBS, and control wells were in complete medium containing 0.2% DMSO). 20 μL of LMT solution was added to each well and incubated at 37°C for 2 h. The absorbance of the solution at 490 nm and 690 nm was measured using a SpectraMAX 340 microplate reader. Cell viability was calculated using the formula: Cell viability % = (OD of drug-treated cells – ID of blank cells) / (OD of control cells – OD of blank cells) × 100%. IC50 was calculated using GraphpadPrism 8. 50 .

[0280] The inhibitory activity of the compounds of the present invention against MV4-11 cell proliferation is shown in Table 11. Table 11 shows the IC50 values ​​of the compounds of the examples against MV4-11 cell proliferation. 50 The concentrations are in the range of 0.01–2000 nM, preferably 0.01–1000 nM, more preferably 0.01–100 nM, even more preferably 0.01–50 nM, and most preferably 0.01–10 nM. Most of the compounds in the examples exhibited strong activity against MV4-11 cell proliferation inhibition. “++++” in Table 11 indicates the IC50 value of the tested compounds against MV4-11 cell proliferation inhibition activity. 50 Less than 50 nM, "++" indicates the IC50 of the compound tested against the proliferation inhibitory activity of MV4-11 cells. 50 Values ​​range from 50 to 200 nM, with "++" indicating the IC50 value of the tested compound against the proliferation inhibitory activity of MV4-11 cells. 50 A value between 200 and 1000 indicates that the IC50 value of the test substance inhibits the proliferation of MV4-11 cells. "+" indicates that the IC50 value is between 200 and 1000. 50 The value is between 1000 and 2000 nM.

[0281] Table 11: Inhibitory activity of the compounds of the present invention against the proliferation of MV4-11 cells

[0282]

[0283] Example B: FLT3 / CHK1 protein degradation assay (Western Blot)

[0284] The degradation activity of the compounds of this invention on FLT3 / CHK1 protein was detected in MV4-11 cells using Western blotting.

[0285] Experimental Procedure and Processing Methods: Cells were lysed using RIPA cell lysis buffer, and protein samples were collected. The protein concentration of each sample was determined using the BCA method. Samples were transferred to EP tubes, SDS buffer was added, and the tubes were heated in a boiling water bath to fully denature the proteins. After preparing the SDS-PAGE gel, it was loaded with electrolyte and electrophoresis solution. The processed protein samples were then loaded into the wells of the SDS-PAGE gel. SDS-PAGE electrophoresis was performed, and the proteins were then transferred from the gel membrane to a nitrocellulose membrane. After transfer, the protein membrane was washed with TBST washing buffer for 1–2 min, and then blocked in 5% (TBST dissolved) skim milk solution at room temperature for 60 min. After blocking, the membrane was incubated overnight at 4°C with diluted primary antibody and gently shaken. The primary antibody was recovered, and the membrane was washed three times with TBST on a shaker for 5–10 min each time. Then, the membrane was incubated with secondary antibody diluted to an appropriate concentration at room temperature on a shaker for one hour. After incubation, the TBST-washed membrane was subjected to imaging and development analysis three times, and the results are as follows: Figure 1 and Figure 2 The compound shown can degrade FLT3 and CHK1 proteins in MV4-11 cells in a concentration-dependent manner.

[0286] The foregoing examples and descriptions of certain embodiments should be considered illustrative and not limiting of the invention as defined by the claims. It will be readily understood that many variations and combinations of the above features may be used without departing from the invention as set forth in the claims. All such variations are intended to fall within the scope of the invention. All cited references are incorporated herein by reference in their entirety.

Claims

1. A compound of formula (Ⅰ) or a pharmaceutically acceptable salt, stereoisomer, or solvate thereof, in: R1 is selected from C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 cycloalkyl, (C 0-3 Alkyl)C 3-10 Heterocyclic alkyl groups, wherein the heteroatom in the heterocyclic alkyl group is at least one of nitrogen, oxygen, and sulfur, and the alkyl, cycloalkyl, and heterocyclic alkyl moieties may be further independently and independently bound by 1, 2, 3, or 4 R atoms. a Substitution; and multiple R groups exist on the same group. a At that time, multiple R a They are independent of each other and can be the same or different; R2 is selected from -C 1-6 Alkyl-NH-, -C 0-3 Alkyl-C 3-6 cycloalkyl-NH-,-C 0-3 Alkyl-C 3-8 Heterocyclic alkyl-, -C 0-3 Alkyl-C 3-8 Heterocyclic alkyl groups -NH-, -C 7-18 Spirocycloalkyl-NH-, - heteroatom-containing C 6-18 Spirocycloalkyl-, wherein the heteroatom is at least one of nitrogen, oxygen, and sulfur, and the alkyl, cycloalkyl, heterocycloalkyl, and spirocycloalkyl moieties may each be independently further bound by 1, 2, 3, or 4 R- atoms. b Substitution; and multiple R groups exist on the same group. b At that time, multiple R b They are independent of each other and can be the same or different; R a R b Each group is independently selected from hydrogen, halogen, hydroxyl, amino, cyano, and C. 1-3 Alkoxy, C 1-5 Alkyl, wherein C 1-5 Alkyl groups may be further substituted with halogens, hydroxyl groups, or cyano groups; Each of the q R3 groups is independently hydrogen, halogen, hydroxyl, or C. 1-3 Alkoxy, C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 Cycloalkyl, cyano, nitro, amino, wherein the alkyl, cycloalkyl, and alkoxy moieties may be further independently converted by one or more halogens, hydroxyl groups, amino groups, cyano groups, or C. 1-3 Alkoxy, C 1-5 Alkyl substitution; q can be 0, 1, 2, 3, or 4; The linker is either nonexistent or -(CH2). m - a hydrocarbon chain, wherein any one or more -CH2- units in the hydrocarbon chain can be independently converted by -CH2=CH2-, -O-, -CO-, -NR L -, -SO-, -SO2- and / or 3-12 membered cyclic groups are substituted, and any of the above-mentioned -CH2- units or substituted groups on the linker may be further replaced by one or more independent groups selected from deuterium, halogen, cyano, hydroxyl, amino, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl groups, 4-8 membered heterocyclic hydrocarbon groups containing oxygen, nitrogen, or sulfur atoms, and C 3-8 Cycloalkyl substituents, wherein R L For H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl; and wherein, when two cyclic groups in the Linker are adjacent, they optionally form together a fused ring, a spiro ring, or a bridged ring structure; the cyclic groups include cycloalkyl and heterocyclic alkyl containing oxygen, nitrogen, or sulfur atoms; m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20; Degron is the E3 ubiquitin ligase-binding moiety; the Degron moiety is selected from the following structures: V is CR c Or N, where R c C is hydrogen or optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups. 1-6 alkyl; W represents bonds, NH, NR d -CO-NH-, O, R d , where R d For unreplaced C 1-6 Alkylene or C-aryl groups optionally substituted with one or more halogens, hydroxyl groups, and / or cyano groups 1-6 Alkylene; U is -CO-, -CR e R f ;R e、 R f They can be the same or different; X1 and X2 are each independently selected from CR e Or N; R e R f Each is independently selected from hydrogen, halogen, C 1-6 Alkyl, C 1-3 Alkoxy; Each of the p R4 groups is independently a hydrogen, halogen, hydroxyl, or C group. 1-3 Alkoxy, C 1-6 Alkyl, (C 0-3 Alkyl)C 3-10 Cycloalkyl, cyano, nitro, amino, NH(C) 1-6 Alkyl), N(C) 1-6 Alkyl)2, wherein the alkyl, cycloalkyl, and alkoxy moieties may be further independently converted by one or more halogens, hydroxyl groups, amino groups, cyano groups, C 1-3 Alkoxy, C 1-5 Alkyl substitution; p can be 0, 1, 2, 3, or 4.

2. The compound according to claim 1, characterized in that: R1 can be selected from 3. The compound according to claim 1, characterized in that, R2 is selected from: -C 0-3 Alkyl-C 4-6 cycloalkyl NH-, -C 0-3 Alkyl-C 4-5 Heterocyclic alkyl-, wherein the heteroatom in the heterocyclic alkyl group is at least one of nitrogen and oxygen, and the alkyl, cycloalkyl, and heterocyclic alkyl moieties may be further substituted independently by 1, 2, 3, or 4 hydrogens, deuteriums, or halogens.

4. The compound according to claim 3, wherein the R2 portion is selected from any of the following structures:

5. The compound according to claim 4, characterized in that: It has the structure shown in general formula (II): Linker and Degron are defined as in claim 1.

6. The compound according to any one of claims 1 to 5, wherein the Linker is composed of one or more fragments selected from (i) to (vii): (i)-CO-(CH2) a -NH-, where a is an integer from 0 to 12; (ii)-CO-[(CH2)2-O-] n -CH2CH2NH-, where n is an integer of 0, 1, 2, 3, 4, or 5; (iii)-CO-(CH2) b -(4-7-membered nitrogen-containing heterocyclic alkanes)-, where b is an integer of 0, 1, 2, 3, 4, or 5; (iv)-(CH2) c -(4-7-membered nitrogen-containing heterocyclic alkanes)-, where c is an integer of 0, 1, 2, 3, 4, or 5; (v)-CO-(CH2) d -(4-7 member nitrogen-containing heterocyclic alkanes)-(CH2) e -(4-7 nitrogen-containing heterocyclic alkanes)-, where d and e are integers of 0, 1, 2, 3, 4, and 5, and d+e≤10; (vi)-(CH2) e -7-11 membered spirocycloalkyl containing nitrogen atoms-; where e is an integer of 0, 1, 2, 3, 4, or 5; (vii) key; in, Each of the 4-7 nitrogen-containing heterocyclic alkanes may be independently and optionally substituted with a hydroxyl group or a halogen.

7. The compound according to claim 6, characterized in that, The linker is selected from any of the following structures:

8. The compound according to any one of claims 1 to 5, characterized in that, The Degron part is selected from X1 and X2 are independent CH or N; R4 is F, Cl or Br, and p is 0 or 1.

9. The compound according to claim 1, characterized in that, Selected from the following compounds: And optical isomers or pharmaceutically acceptable salts of any of the above compounds.

10. The use of a compound of any one of claims 1-9 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for use alone or in combination for treating tumors, skin diseases, inflammation, autoimmune diseases or neurodegenerative diseases.

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