WO2023241640A1 - Small molecule inhibitor targeting deubiquitinating enzymes usp25 and usp28 - Google Patents

Abstract

Disclosed in the present application are small molecule inhibitor targeting deubiquitinating enzymes USP25 and USP28. Specifically disclosed are a compound as represented by formula I, or a racemate, a stereoisomer, a tautomer, a solvate, a polymorph, a pharmaceutically acceptable salt or a prodrug thereof. The specific substituents are as defined in the description. The inhibitor can be used for preventing or treating diseases related to ubiquitin-specific proteases USP25 and/or USP28.

Description

A class of small molecule inhibitors targeting deubiquitinating enzymes USP25 and USP28 Technical field

The present invention relates to the field of synthesis of chemical drugs and pharmacological applications, and to inhibitors of ubiquitin-specific protease 25 (USP25) and ubiquitin-specific protease 28 (USP28) and their uses. The inhibitors can be used to prevent or treat diseases or disorders associated with USP25 and/or USP28.

Specifically, the present invention relates to a class of ubiquitin-specific protease USP25/28 inhibitors and their compositions, preparation methods and uses. Such inhibitors can be used to prevent or treat diseases related to USP25 and/or USP28.

Background technique

Ubiquitin is a small molecule regulatory protein (8.5KDa) widely present in biological cells. Ubiquitination refers to the process in which ubiquitin proteins specifically modify target proteins under the action of a series of special enzymes. Typically, polyubiquitinated proteins are degraded by the proteasome, while monoubiquitinated proteins participate in the regulation of cellular pathways. The ubiquitination process can be reversed by a class of proteases called deubiquitinating enzymes (DUBs), which regulate various cellular processes by removing ubiquitin proteins from target proteins.

DUBs are encoded by approximately 100 human genes and are divided into 7 families (USP, UCH, SENP, JAMM, OUT, MJD, MINDY). The largest family is the ubiquitin-specific protease (USP) with more than 50 protein members. . Paper reports have pointed out that some DUBs play a role in the occurrence of tumor cells or inflammation. Andrew Kovalenko et al. pointed out that CYLD in the DUBs family has a negative correlation with tumors through the NF-κB pathway (Kovalenko, A. et al. Nature 2003, 424 (6950), 801–805.), Lim, Seung-Oe et al. published a study on the impact of CSN5 in the DUBs family on PD-L1 (Lim, S.-O. et al. Cancer Cell 2016, 30(6), 925–939.). Junli Liu et al. reported that USP10 and USP13 in the DUBs family affect tumorigenesis by regulating the structure of Beclin1 in the Vps34 complex (Liu, J. et al. Cell 2011, 147(1), 223– 234.). Zhu, Dan et al. reported that OTUB1 in the DUBs family can regulate the degradation of PD-L1 in the endoplasmic reticulum through the ubiquitination process, revealing the key role of the OTUB1-PD-L1 signaling pathway in regulating immune escape of tumor cells ( Zhu, D. et al. Cell Death Differ. 2021, 28(6), 1773–1789.). These studies indicate that specifically inhibiting the activity and function of a certain DUBS may become a potential target for tumor immunotherapy, and therefore the development of appropriate deubiquitinating enzyme inhibitors has become a feasible solution for these symptoms. Related inhibitor research work has also been reported. Yiwei Wang et al. revealed the molecular mechanism of the small molecule compound IU1 that selectively inhibits the deubiquitinating enzyme USP14, and optimized the small molecule IU1-248 with a 10-fold increase in activity based on its structure (Wang, Y.; Jiang et al. Cell Res .2018,28(12),1186–1194.).

The structures of USP28 and USP25 are similar, and their amino acid sequences are highly homologous (Sauer, F. et al. Mol. Cell 2019, 74(3), 421-435.e10.). In cells, USP25 and USP28 can regulate the expression level and half-life of c-Myc oncoprotein, inhibit the viability of a series of cancer cells, and ultimately induce apoptosis (Wrigley, J.D. et al. ACS Chem. Biol. 2017, 12(12),3113–3125).

USP25 is one of the key members of the deubiquitinase system. The protein structure of USP25 contains two peptidase regions, a region that can bind ubiquitin, and two motifs that can interact with ubiquitin. The region associated with ubiquitin hydrolase activity is the peptidase region. USP25 plays a regulatory role in a variety of cellular physiological processes. The Wnt signaling pathway plays an important role in the development of various cancers (Zhan, T. et al. Oncogene 2017, 36(11), 1461–1473.). In the Wnt/β-catenin signaling pathway, USP25 By accelerating the deubiquitination process of Tankyrases to stabilize them, it plays a positive regulatory role in this process (Xu, D. et al. Genes Dev. 2017, 31(10), 1024–1035.). In the process of fighting against viral infection in vivo, USP25 can negatively regulate this process by simultaneously regulating the NF-κB and IRF3 signaling pathways. In the activation process of NF-κB and IRF3, it is necessary to utilize the activated TRAF2, TRAF6, etc. of the polyubiquitin chain at K63 position. The negative regulation of β-interferon expression by USP25 can be achieved by down-regulating the phosphorylation of NF-κB subunit p65 and IRF3; at the same time, USP25 can also promote the deubiquitination process of RIG-I, TRAF2 and TRAF6. , further inhibiting the activation of IRF3 and NF-κB, thereby negatively regulating the antiviral process (Lin, D. et al. Proc. Natl. Acad. Sci. USA 2015, 112 (43), E5901.). In addition, some studies have pointed out the relationship between overexpression of USP25 and Alzheimer's disease (Zheng, Q. et al. Sci. Adv. 2021, 7(1), 1–14.)

Mutations in the epidermal growth factor receptor EGFR can lead to abnormal cell proliferation, which often occurs in the early stages of tumor cells and plays an important role in tumor growth and development. With the assistance of USP25, the E3 ubiquitin ligase Cb1 can bind to EGFR to inhibit the degradation of EGFR. In this process, USP25 plays a negative regulatory role in the down-regulation of EGFR. In drug development, there is hope that USP25 can be used as a target to intervene in the growth and development of EGFR-dependent tumors ( CA et al. Biomolecules 2020,10(11),1–16.).

Research in recent years has found that USP28 in the deubiquitinating hydrolase family can be used as a potential anti-tumor target molecule (Chakravorty, D. et al. Comput. Biol. Chem. 2020, 85 (December 2019), 107208.). In the process of tumor development, the oncoprotein MYC plays an important role in the development of tumors by affecting cell metabolism. The MYC protein family mainly includes four types: C-MYC, N-MYC, L-MYC, and R-MYC. Further research found that USP28 can regulate the stability of MYC and the cancer-promoting protein LSD1 (Lysine-specific demethylase1, LSD1) in cells. (Liu, Z. et al. Acta Pharm. Sin. B 2020, 10(8), 1476–1491.). Therefore, USP28 has the potential to become a drug target for cancer. Squamous cell carcinoma (SCC) promotes DNA repair by expressing ΔNp63, which is a key function in maintaining SCC tumor survival. USP28 acts as a stabilizing effect on ΔNp63 protein. In mouse models, inhibitors of USP28 inhibited tumor growth (Prieto-Garcia, C. et al. EMBO Mol. Med. 2020, 12 (4), 1–25.).

In animal experiments, it has been found that USP28 can resist c-MYC-dependent degradation in the intestine, keeping it stable. It also inhibits the degradation of two other oncogenic protein factors, c-JUN and NOTCH1. USP28, as a target gene of c-MYC, is highly expressed in mouse and human intestinal tumors. Since USP28 and c-MYC form a positive feedback loop, the high expression of USP28 can maintain the high protein level of c-MYC in tumors (Wang, H. et al. FEBS J. 2020, 288, 1325–1342.) .

Animal experiments showed that knocking out the USP28 gene did not cause obvious adverse phenotypes in mice. Further studies found that cell proliferation in the intestinal part was significantly reduced and the differentiation of some cells was impaired. In colorectal cancer-related mouse models, mice knocking out the USP28 gene have fewer intestinal tumors, smaller tumor sizes, and the lifespan of the mice is significantly extended. The deletion animals had fewer intestinal tumors. More importantly, USP28 deletion can reduce tumor volume and significantly extend the life of mice. At the cellular level, loss of USP28 promotes tumor cell differentiation and reduces cell proliferation. These experiments indicate that inhibiting USP28 activity is a potential therapeutic target for colorectal cancer ((Diefenbacher, M.E. et al. J. Clin. Invest. 2014, 124(8), 3407–3418.).

Taken together, small molecule inhibitors targeting USP25 and USP28 and their use in compositions have the potential to become treatments for cancer and other USP25- and USP28-related diseases.

Contents of the invention

One of the purposes of the present application is to provide a class of compounds with USP25 and/or USP28 inhibitory activity.

According to the present invention, there is provided a compound represented by the following formula I or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug:

in,

Ar is selected from a substituted or unsubstituted C6-C14 aryl group, a substituted or unsubstituted 9-10 membered bicyclic fused heterocyclic group containing one or more heteroatoms selected from N, O, and S, where used The substituted substituent is selected from halogen, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl;

Preferably, Ar is selected from substituted or unsubstituted phenyl, substituted or unsubstituted thienopyridine, substituted or unsubstituted furopyridine, substituted or unsubstituted pyrrolopyridine, substituted or unsubstituted pyrazopyridine , substituted or unsubstituted pyrrolopyrimidines, substituted or unsubstituted thienopyrimidines, substituted or unsubstituted benzopyrroles, substituted or unsubstituted benzofurans, substituted or unsubstituted benzothiophenes, substituted or unsubstituted benzoxazole, substituted or unsubstituted benzothiazole, substituted or unsubstituted benzopyrazole, substituted or unsubstituted benzotriazole, substituted or unsubstituted benzopyridine, where substituted The group is selected from halogen, amino, cyano, C1-C6 alkyl, and halogenated C1-C6 alkyl;

Preferably, the above-mentioned substituents for substitution can be 1, 2 or 3 selected from halogen, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl, more preferably, selected from halogen 1, 2 or 3 of , amino, cyano, methyl, ethyl, n-propyl, isopropyl, trifluoromethyl and 2,2,2-trifluoroethyl;

For example, Ar can be selected from the following structures:

m and n are each independently 1 or 2; preferably, m and n are the same;

R ₁ is selected from hydrogen; nitro; halogen; substituted or unsubstituted C1-C6 alkoxy, wherein the substituent used for substitution is selected from C1-C6 alkylamino; substituted or unsubstituted C1-C6 alkyl Carbonylamino, wherein the substituent used for substitution is selected from halogen, amino, C1-C6 alkylamino, 5-7 membered heterocyclyl; substituted or unsubstituted 5-7 membered heterocyclyl, wherein the substituent used for substitution The group is selected from cyano, C1-C6 alkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, halogen; wherein, the 5-7 membered heterocyclic group contains one selected from N, O, S or multiple heteroatoms;

Preferably, R ₁ is selected from hydrogen; nitro; halogen; substituted or unsubstituted C1-C4 alkoxy, wherein the substituent used for substitution is selected from C1-C4 alkylamino; substituted or unsubstituted C1- C4 alkylcarbonylamino, wherein the substituent used for substitution is selected from halogen, amino, C1-C4 alkylamino, piperazinyl, piperidinyl; substituted or unsubstituted piperazinyl; substituted or unsubstituted morpholine base; substituted or unsubstituted furyl; substituted or unsubstituted thienyl; substituted or unsubstituted pyrazolyl; substituted or unsubstituted pyrrolyl; substituted or unsubstituted pyridyl; substituted or unsubstituted oxazole base, wherein the substituents used for substitution are selected from cyano group, C1-C4 alkyl group, C1-C4 alkylcarbonyl group, C1-C4 alkoxycarbonyl group, and halogen;

For example, _R1 is selected from hydrogen, methylaminoethoxy, methoxy, nitro, chloromethylcarbonylamino, methylaminomethylcarbonylamino, piperazinylmethylcarbonylamino, piperidylmethylcarbonylamino, piperazinylmethylcarbonylamino, Azinyl, methylcarbonylpiperazinyl, methylpiperazinyl, morpholinyl, Br, furyl, methylfuryl, methoxycarbonylfuryl, thienyl, chlorothienyl, methylthienyl, cyano Thienyl, pyrazolyl, tert-butoxycarbonylpyrrolyl, oxazolyl, pyridyl;

R ₂ is selected from hydrogen; C1-C6 alkyl; halogen; C1-C6 alkoxy; preferably selected from hydrogen; C1-C4 alkyl; halogen; C1-C4 alkoxy; more preferably hydrogen, methyl , methoxy, ethoxy or propoxy;

Alternatively, R ₁ and R ₂ form a 5-7-membered heterocyclic group, wherein the 5-7-membered heterocyclic group contains one or more heteroatoms selected from N, O, and S. Preferably, the heteroatoms is an O atom.

For example, in one embodiment,

Ar is substituted or unsubstituted phenyl, thienopyridine, furopyridine, pyrrolopyridine, pyrazopyridine, pyrrolopyrimidine, thienopyrimidine, benzopyrrole, benzofuran, benzothiophene, benzox Azole, benzothiazole, benzopyrazole, benzotriazole, benzopyridine, wherein the substituent used for substitution is selected from halogen, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl; Preferably, the substituents used for substitution may be 1, 2 or 3 selected from halogen, amino, cyano, C1-C6 alkyl, and halogenated C1-C6 alkyl;

m and n are each independently 1 or 2;

R ₁ is selected from hydrogen; nitro; halogen; substituted or unsubstituted C1-C4 alkoxy, wherein the substituent used for substitution is selected from C1-C4 alkylamino; substituted or unsubstituted C1-C4 alkyl Carbonylamino, wherein the substituent for substitution is selected from halogen, amino, C1-C4 alkylamino, piperazinyl, piperidinyl; substituted or unsubstituted piperazinyl; substituted or unsubstituted morpholinyl; substituted or unsubstituted furyl; substituted or unsubstituted thienyl; substituted or unsubstituted pyrazolyl; substituted or unsubstituted pyrrolyl; substituted or unsubstituted pyridyl; substituted or unsubstituted oxazolyl, wherein The substituents used for substitution are selected from cyano group, C1-C4 alkyl group, C1-C4 alkylcarbonyl group, C1-C4 alkoxycarbonyl group, and halogen;

R ₂ is selected from hydrogen; C1-C4 alkyl; halogen; C1-C4 alkoxy.

In another embodiment,

Ar is selected from the following structures:

m and n are each independently 1 or 2;

R ₁ is selected from hydrogen, methylaminoethoxy, methoxy, nitro, chloromethylcarbonylamino, methylaminomethylcarbonyl Amino, piperazinylmethylcarbonylamino, piperidylmethylcarbonylamino, piperazinyl, methylcarbonylpiperazinyl, methylpiperazinyl, morpholinyl, Br, furyl, methylfuryl, methoxy Carbonylfuryl, thienyl, chlorothienyl, methylthienyl, cyanothienyl, pyrazolyl, tert-butoxycarbonylpyrrolyl, oxazolyl, pyridyl;

R ₂ is selected from hydrogen, methyl, methoxy, ethoxy or propoxy.

In another embodiment,

Ar is selected from a substituted or unsubstituted C6-C14 aryl group, a substituted or unsubstituted 9-10 membered bicyclic fused heterocyclic group containing one or more heteroatoms selected from N, O, and S, where used The substituted substituent is selected from halogen, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl;

m and n are each independently 1 or 2;

R ₁ is selected from hydrogen; nitro; halogen; substituted or unsubstituted C1-C6 alkoxy, wherein the substituent used for substitution is selected from C1-C6 alkylamino; substituted or unsubstituted C1-C6 alkyl Carbonylamino, wherein the substituent used for substitution is selected from halogen, amino, C1-C6 alkylamino, 5-7 membered heterocyclyl; substituted or unsubstituted 5-7 membered heterocyclyl, wherein the substituent used for substitution The group is selected from cyano, C1-C6 alkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, halogen; wherein, the 5-7 membered heterocyclic group contains one selected from N, O, S or multiple heteroatoms;

R ₂ is selected from hydrogen, methyl, methoxy, ethoxy or propoxy.

Specifically, the compound represented by formula I according to the present invention may have the structure shown below:

Another object of the present invention is to provide a pharmaceutical composition, which contains the compound represented by Formula I as described above or its racemate, stereoisomer, tautomer, solvate, Polymorphs, pharmaceutically acceptable salts or prodrugs serve as active ingredients, and optionally pharmaceutically acceptable carriers.

Another object of the present invention is to provide the compound represented by Formula I as described above or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or Use of prodrugs in the preparation of USP25 and/or USP28 inhibitors.

According to one aspect of the present invention, there is provided the compound represented by Formula I as described above or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or The use of prodrugs in the preparation of medicaments for preventing or treating diseases related to USP25 and/or USP28.

Another object of the present invention is to provide a method for preventing or treating diseases related to USP25 and/or USP28, which method includes administering to a subject a therapeutically effective amount of a compound represented by Formula I as described above or a compound thereof. Racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts or prodrugs, or said pharmaceutical compositions.

In the present invention, the diseases related to USP25 and/or USP28 include cancer (such as colorectal cancer), inflammation, autoimmune diseases, and neurodegenerative diseases.

In the present invention, a "pharmaceutically acceptable" ingredient is a substance that is suitable for humans and/or animals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), that is, with a reasonable benefit/risk ratio.

In the present invention, "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or excipient used to deliver the active substance of the present invention or its physiologically acceptable salt to animals or humans. The carrier can be liquid or solid.

The pharmaceutical composition of the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols. The composition may contain one or more ingredients selected from the group consisting of sweeteners, coloring agents and preservatives.

The compounds of the present disclosure can be used as monotherapy or combination therapy. In some embodiments, combination therapy includes treating the subject with a chemotherapeutic agent, therapeutic antibody, radiation, cell therapy, or immunotherapy.

definition

As used herein, the term "alkyl", alone or as part of another group, refers to a straight or branched aliphatic saturated hydrocarbon radical. In some embodiments, the alkyl group may contain 1 to 6 carbon atoms, i.e., C1-C6 alkyl, including C1 alkyl (such as methyl), C2 alkyl (such as ethyl), C3 alkyl (such as propyl) or isopropyl), C4 alkyl, C5 alkyl and C6 alkyl. In one embodiment, the alkyl group is a straight chain C1-C4 alkyl group. In another embodiment, the alkyl group is branched C3-6 alkyl. For example, C1-C4 alkyl as used herein refers to a group selected from the group consisting of methyl, ethyl, propyl (n-propyl), isopropyl, butyl (n-butyl), sec-butyl, tert-butyl and isobutyl. base group. Optionally substituted C1-C4 alkyl refers to a C1-C4 alkyl group as defined, which is optionally substituted with one or more permissible substituents as described herein.

As used herein, the term "alkoxy", alone or as part of another group, refers to a group of the formula -ORa1, wherein Ra1 is an alkyl group.

As used herein, the term "alkylamino", alone or as part of another group, refers to a group of the formula -NHRa1, where Ra1 is an alkyl group.

As used herein, the term "haloalkyl", alone or as part of another group, refers to an alkyl group substituted with one or more fluorine, chlorine, bromine and/or iodine atoms.

As used herein, the term "aryl", when used alone or as part of another group, means having Monocyclic or polycyclic (eg, bicyclic) groups of 6 to 14 carbon atoms and 0 heteroatoms provided in an aromatic ring system ("C6-14 aryl"). In some embodiments, an aryl group has 6 ring carbon atoms ("C6 aryl"; eg, phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C10 aryl"; for example, naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("C14 aryl"; for example, anthracenyl).

"Heterocyclyl" or "heterocycle", when used alone or as part of another group, means a ring having carbon atoms and 1 to 4 ring heteroatoms (wherein each heteroatom is independently selected from nitrogen, oxygen , sulfur), an aromatic or non-aromatic ring system group, for example, a 3-10-membered heterocyclyl group refers to an aromatic or non-aromatic ring system group with a total number of ring carbon atoms and ring heteroatoms from 3 to 10. . Heterocyclyl may be a monocyclic ring ("monocyclic heterocyclyl") or a fused ring system, such as a bicyclic ring system ("bicyclic heterocyclyl"), and may be saturated or may be partially unsaturated. For example, a 9-10-membered heterocyclyl group represents an aromatic or non-aromatic ring system group with a total number of ring carbon atoms and ring heteroatoms of 9 to 10, including a 5,6-membered ring fused with a 6-membered ring. Bicyclic heterocyclyl (9-membered bicyclic fused heterocyclyl) and 6,6-bicyclic heterocyclyl (10-membered bicyclic fused heterocyclyl) in which a 6-membered ring is fused with a 6-membered ring. Heterocyclic bicyclic systems may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocycle as defined above is fused to one or more carbocyclyl groups, wherein the point of attachment is on the carbocyclyl or heterocycle.

Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, furyl, thienyl, and pyrrolyl. Exemplary 5-membered heterocyclyl groups containing 2 heteroatoms include, but are not limited to, pyrazolyl, oxazolyl, and thiazolyl. Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 6-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, pyridyl and piperidinyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, but are not limited to, pyrazinyl, piperazinyl, and morpholinyl. Exemplary 5,6-bicyclic heterocyclyl groups (9-membered heterocyclyl) include, but are not limited to, thienopyridine, furopyridine, pyrrolopyridine, pyrazopyridine, pyrrolopyrimidine, thienopyrimidine, Benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzothiazole, benzopyrazole, benzotriazole. Exemplary 6,6-bicyclic heterocyclyl groups (10-membered heterocyclyl) include, but are not limited to, benzopyridine.

In the heterocyclyl group as defined above, the point of attachment is not limited as long as it meets the valence requirements. For example, in heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, where valency permits. For example, piperazinyl (piperazine), the point of attachment can be a carbon atom or a nitrogen atom. For example, furyl, the point of attachment can be at the ortho or meta position, including In the bicyclic heterocyclyl group as defined above, the position where the 5-membered and 6-membered rings or the 6-membered and 6-membered rings are fused is not limited, as long as it meets the valence requirements; in addition, the connection point of the bicyclic heterocyclyl group is also Not limited to, for example, benzofuryl (benzofuran), its connection point can be on the benzene ring or the furan ring; another example is thienopyridyl (thienopyridine), its connection point can be on the benzene ring It can also be on the thiophene ring, as long as it meets the valence requirements.

As used herein, the term "halogen" includes fluorine, chlorine, bromine, and iodine.

In this article, if the substituent is a combination of multiple groups, such as an ABC group, it means an ABC-group, that is, a group connected to the parent core through C, where A and B are connected and C is connected through B. For example, methoxycarbonylfuryl represents a group connected to the parent core through the furan ring, in which the methoxy group is connected to the carbonyl group and connected to the furan ring through the carbonyl group.

Detailed ways

Preparation example: By way of example, the compound of the present invention can be prepared by the following synthetic route

Preparation of heteroatom aromatic groups referred to in the examples:

Dissolve 2-chloro-5-fluoronicotinonitrile (1eq.) and methyl 2-sulfanylacetate (1eq.) in DMF in a round-bottomed flask, add DBU (1.5eq.), and stir the reaction at room temperature. 3 hours, monitored by thin layer chromatography. After the reaction is completed, water is added to slurry, filtered, the filter cake is collected, and dried to obtain A1 (yield 51%).

Dissolve compound A1 (1eq.) in methanol, add 1M NaOH aqueous solution, raise the temperature to 60°C to react for 3 hours, and detect by thin layer chromatography. After the reaction, the methanol was evaporated to dryness, the pH was adjusted to 4-5 with 10% hydrochloric acid aqueous solution, filtered, the filter cake was collected, and dried to obtain F1 (yield 62%).

Compounds F2, F3, F4, and F5 can be synthesized in a manner similar to the synthetic route of F1; the difference is that the raw material A1 is adapted to be changed, and all raw materials can be purchased commercially.

Reaction raw material A2 can be purchased from commercial channels.

Compound A2 (1g, 5.68mmol) was dissolved in dry DMF (26mL), cooled to 0°C, NaH (0.68g, 28.33mmol) was slowly added, stirred at 0°C for 1h, and CH ₃ CH ₂ I (11.35mmol, 908 μL), stir the reaction for 10 hours, add ice water dropwise, stop the reaction, adjust the pH of the solution to 7 with 3N hydrochloric acid, extract the resulting mixture with EtOAc, combine the organic layers, wash with brine, dry over anhydrous Na ₂ SO ₄ , filter, concentrate, and pass Silica gel column chromatography separated F6 (yield 42%).

Dissolve compound F6 (512 mg, 2.69 mmol) in DMF (15 mL), add NCS (395.4 mg, 2.96 mmol), and react at 20°C for 18 hours. After the reaction is completed, add water to quench, extract with EtOAc, back-extract with saturated brine, and collect the organic phase , dried over anhydrous sodium sulfate, concentrated, and separated by silica gel column chromatography to obtain F7 (yield 41%)

Dissolve A3 (1.0eq.) and 4,4-dimethoxy-2-butanone (1.2eq.) in anhydrous toluene, and stir at 80°C overnight. Use thin layer chromatography to monitor the reaction. After the reaction is completed, concentrate and perform silica gel column chromatography to separate A4, which can be used directly in the next step.

Dissolve A4 (1.0eq.) and KOH (5eq.) in H ₂ O and ethanol (v:v=1:2), and stir at room temperature for 2h. After stopping the reaction, spin away the organic solvent, slowly add 1M HCl dropwise until no precipitation occurs, filter and drain to obtain F8 (yield 35.3%), ¹ H NMR (500MHz, D ₂ O) δ 8.60 (d, J＝ 6.6Hz, 1H), 8.26 (d, J = 3.4Hz, 1H), 6.91 (d, J = 6.8Hz, 1H), 2.53 (d, J = 3.0Hz, 3H).

The above fragments F9 to F32 are all commercially available.

General synthesis route for target molecules 1:

Compound A5 (1.0eq.) was dissolved in DMF, N-Boc-bromoethylamine (1.4eq.) and cesium carbonate (6eq.) were added, and the reaction was carried out at room temperature under nitrogen protection for 16 hours. The reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate, and washed three times with brine. The organic phase was dried with anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain A6 (yield 81%).

Dissolve compound A6 (1eq.) in anhydrous tetrahydrofuran, cool to 0°C, slowly add sodium hydride (2.5eq.), stir for 60 minutes, add methyl iodide (2eq.) in batches, return to room temperature and react for 2 hours. Add saturated aqueous ammonium chloride solution to quench, and extract with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and separated by silica gel column chromatography to obtain A7 (yield 95%).

Add A7 (1eq.), A8 (1eq.), X-Phos (0.2eq.), palladium acetate (0.1eq.), cesium carbonate (4eq.) and anhydrous toluene into the reaction bottle, and replace with nitrogen for 15 minutes. React at 110°C for 18 hours. The reaction was monitored by thin layer chromatography. After completion, water was added to quench the reaction, and the reaction solution was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and separated by silica gel column chromatography to obtain A9 (yield 24%).

Dissolve compound A9 (1eq.) in methanol and tetrahydrofuran, add 10% palladium on carbon (0.5eq.) and palladium hydroxide (0.5eq.), replace with hydrogen more than 3 times, and raise the temperature to 50°C in a hydrogen atmosphere for reaction 3 hours, detected by thin layer chromatography. After the reaction, filter, collect the filtrate, concentrate and perform silica gel column chromatography to obtain A10 (yield 56%).

Compounds F1 (1.0eq.), A10 (1eq.), EDCl (1.1eq.), HOBt (1.1eq.) and DIPEA (5eq.) were dissolved in anhydrous dichloromethane and reacted at 40°C for 16 hours. The insoluble matter was filtered off, and the filtrate was concentrated for separation and purification to obtain A11 (yield 75%).

Compound A11 (1eq.) was dissolved in DCM, TFA (5eq.) was added, and the reaction was carried out at room temperature for 1 hour. Adjust the pH to 8 with 10% sodium bicarbonate aqueous solution, extract with DCM three times or more, dry with anhydrous sodium sulfate, filter and evaporate the solvent to dryness, and isolate and purify compound I-1 (yield 70%).

Example 1: Obtained according to general synthetic route 1.

1-1 (20 mg, 70% yield). ¹ H NMR (500MHz, MeOD) δ8.52 (dd, J＝2.7, 1.0Hz, 1H), 8.11 (dd, J＝9.1, 2.7Hz, 1H), 6.87–6.76 (m, 2H), 6.76–6.61 (m,2H),4.56(dq,J＝9.3,6.0Hz,1H),4.53–4.44(m,1H),4.23(t,J＝7.6Hz,1H),3.99(t,J＝5.3Hz, 2H), 3.30–3.23 (m, 2H), 2.90 (t, J = 5.2Hz, 2H), 2.45 (s, 3H)ppm; mass spectrum: C20H22FN5O2S[M+H] ⁺ calculated value: 416.2, measured value: 416.2 .

General synthesis route of target molecule 2:

Dissolve A12 (1eq.) and A13 (1.05eq.) in DMF in a round-bottomed flask, add potassium carbonate (3eq.), stir and react at 65°C for 4 hours, and monitor by thin layer chromatography. After the reaction, filter, collect the filtrate, add EA and saturated brine for extraction, dry the organic phase with anhydrous sodium sulfate, and isolate A14 (yield 96%).

Dissolve compound A14 (1eq.) in DCM, add TFA (10eq.), react at room temperature for 1 hour after addition, and detect by thin layer chromatography. After the reaction, adjust the Ph to 8 with 10% sodium bicarbonate aqueous solution, extract with DCM three times, dry the organic phase with anhydrous sodium sulfate, and purify to obtain A15 (yield 67%).

Compounds A15 (1.0eq.), F1 (1eq.), EDCI (1.1eq.), HOBt (1.1eq.) and DIPEA (5eq.) were dissolved in anhydrous dichloromethane and reacted at 40°C for 16 hours. Filter, rinse the filter cake with DCM, collect the filter cake, and dry it to obtain compound I-2 (yield 50%).

Dissolve I-2 (1eq.) in methanol (I-2 has extremely poor solubility), add (Boc) ₂ O (3eq.) to the system, raise the temperature to 40°C and react for 12 hours. After the reaction is completed, purify A16 was obtained (yield 80%).

Dissolve compound A16 (1.0eq.) in chloroform, add zinc powder (4eq.), add glacial acetic acid (10eq.) dropwise at low temperature, and react at room temperature for 1 hour. Filter out the insoluble matter, adjust the pH to 8 with saturated sodium bicarbonate, extract three times with dichloromethane, dry the organic phase with anhydrous sodium sulfate, filter, concentrate and perform silica gel column chromatography to obtain A17 (yield 80%) .

Compound A17 (1eq.) was dissolved in DCM, triethylamine (2eq.) and chloroacetyl chloride (1.2eq.) were added, and the reaction was carried out at room temperature for 2 hours. Add saturated ammonium chloride aqueous solution to quench, and extract with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and separated by silica gel column chromatography to obtain A18 (yield 83%).

Compound A18 (1eq.) was dissolved in DCM, TFA (10eq.) was added, and the reaction was carried out at room temperature for 1 hour. Adjust the pH to 8 with 10% sodium bicarbonate aqueous solution, extract with DCM three times or more, dry with anhydrous sodium sulfate, filter, and evaporate the solvent to dryness. Compound I-3 is purified and isolated (yield 56%).

Add I-3 (1eq.), methylamine aqueous solution (5eq.), potassium carbonate (3eq.) and acetonitrile into the reaction bottle, and react at 50°C for 2 hours. The reaction was monitored by thin layer chromatography. After completion, water was added to quench the reaction, and the reaction solution was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and separated to obtain compound I-4 (yield 63%).

Example 2:

1-2 (150 mg, 50% yield). ¹ H NMR (500MHz, DMSO) δ8.70(d,J＝2.0Hz,1H),8.43(d,J＝7.0Hz,1H),8.41(dd,J＝9.6,2.8Hz,1H),8.06( d,J＝9.2Hz,2H),7.22(s,2H),6.53–6.44(m,2H),4.96–4.76(m,1H),4.36(t,J＝8.5Hz,2H),4.05(dd , J=9.2, 5.6Hz, 2H)ppm; mass spectrum: C17H14FN5O3S[MH] ^- calculated value: 386.2, measured value: 386.2.

Example 3:

1-3 (11 mg, 56% yield). ¹ H NMR (400MHz, DMSO) δ9.97 (s, 1H), 8.71 (d, J = 2.0Hz, 1H), 8.45 (dd, J = 9.5, 2.7Hz, 1H), 7.33 (d, J = 8.9 Hz,2H),6.70(d,J＝8.6Hz,2H),4.54(d,J＝4.6Hz,2H),4.24(s,1H),4.16(s,2H),3.26(d,J＝5.3 Hz, 2H)ppm; high-resolution mass spectrum: C19H17ClFN5O2S[M+H] ⁺ calculated value: 434.08483, measured value: 434.08484.

Example 4:

1-4 (6.5 mg, 63% yield). ¹ H NMR (500MHz, MeOD) δ8.52 (dd, J＝2.6, 1.0Hz, 1H), 8.11 (dd, J＝9.1, 2.7Hz, 1H), 7.30 (d, J＝8.9Hz, 2H), 6.71(d,J＝8.9Hz,2H),4.57(dq,J＝9.4,6.0Hz,1H),4.54–4.46(m,1H),4.30–4.19(m,1H),3.66(s,2H) ,3.40–3.32(m,2H),2.63(s,3H)ppm; Mass spectrum: C20H20FN5O3S[M+H] ⁺ calculated value: 429.2, measured value: 429.2.

General synthesis route 3 for target molecules:

Dissolve compound A14 (1.0eq.) in chloroform, add zinc powder (4eq.), add glacial acetic acid (10eq.) dropwise at low temperature, and react at room temperature for 1 hour. Filter out the insoluble matter, adjust the pH to 8 with saturated sodium bicarbonate, extract three times with dichloromethane, dry the organic phase with anhydrous sodium sulfate, filter, concentrate and perform silica gel column chromatography to obtain A19 (yield 67%) .

Compound A19 (1eq.) was dissolved in DCM, triethylamine (2eq.) and chloroacetyl chloride (1.2eq.) were added, and the reaction was carried out at room temperature for 2 hours. Add saturated ammonium chloride aqueous solution to quench, and extract with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and separated by silica gel column chromatography to obtain A20 (yield 87%).

Add A20 (1eq.), A21 (5eq.), potassium carbonate (3eq.) and acetonitrile into the reaction bottle and react at 50°C for 2 hours. The reaction was monitored by thin layer chromatography. After completion, water was added to quench the reaction, and the reaction solution was extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and separated to obtain A22 (yield 65%).

Compound A22 (1eq.) was dissolved in DCM, TFA (10eq.) was added, and the reaction was carried out at room temperature for 1 hour. Adjust the pH to 8 with 10% sodium bicarbonate aqueous solution, extract with DCM three times or more, dry with anhydrous sodium sulfate, filter and evaporate the solvent to dryness, and purify and isolate A23 (yield 82%).

Compounds A23 (1.0eq.), F1 (1eq.), EDCI (1.1eq.), HOBt (1.1eq.) and DIPEA (5eq.) were dissolved in anhydrous dichloromethane and reacted at 40°C for 16 hours. Add saturated ammonium chloride aqueous solution to quench The mixture was extinguished and extracted three times with DCM. The organic phase was dried over anhydrous sodium sulfate and purified to obtain A24 (yield 40%).

Dissolve A24 (1eq.) in DCM, add boron tribromide (5eq.) at low temperature, and react at room temperature for 1 hour. After the reaction is completed, add sodium bicarbonate aqueous solution to quench, and extract 3 times with DCM. The organic phase is washed with It was dried over sodium sulfate and purified by Pre-HPLC to obtain compound I-5 (yield 58%).

Example 5:

1-5 (6 mg, 58% yield). ¹ H NMR (500MHz, CD2Cl2) δ8.94(s,1H),8.53(dd,J＝2.7,0.9Hz,1H),7.72(dd,J＝8.7,2.7Hz,1H),7.38–7.26(m ,2H),6.74–6.63(m,2H),6.00(s,2H),4.66(dtd,J＝9.6,7.0,5.0Hz,1H),4.52(dd,J＝9.5,8.3Hz,1H), 4.20(t,J＝7.9Hz,1H),4.01(t,J＝6.2Hz,1H),3.52–3.45(m,1H),3.30(dt,J＝15.6,6.2Hz,1H),3.06(s ,2H),2.95–2.89(m,4H),2.55(s,4H)ppm; Mass spectrum: C23H26FN7O2S[M+H] ⁺ calculated value: 484.2, measured value: 484.2.

Example 6: Compound I-6 can be synthesized in a manner similar to synthetic route 3 by adapting the change of raw material A21.

1-6 (10 mg, 42% yield). ¹ H NMR (400MHz, MeOD) δ8.57 (dd, J＝2.7, 1.0Hz, 1H), 8.14 (dd, J＝9.1, 2.7Hz, 1H), 7.38 (d, J＝8.8Hz, 2H), 6.52(d,J＝8.8Hz,2H),4.91(d,J＝6.4Hz,1H),4.23(t,J＝7.5Hz,2H),3.95–3.72(m,2H),3.08(s,2H ), 2.55 (s, 4H), 1.72–1.64 (m, 4H), 1.49 (t, J = 12.2Hz, 2H) ppm; mass spectrum: C24H27FN6O2S[M+H] ⁺ calculated value: 483.2, measured value: 483.2.

General synthesis route 4 for target molecules:

Weigh compound A21 (1eq.), 4-bromophenylboronic acid (2eq.), 4AMS (2eq.), Cu(OAc) ₂ (0.4eq.), vacuum, fill with oxygen, and add Et ₃ N ( 3eq.), solvent-dried CH ₂ Cl ₂ , react at 35°C for 12 hours. After the reaction is completed, filter to remove the solid, extract with CH ₂ Cl ₂ and water, collect the organic phase, dry, concentrate, and separate by silica gel column chromatography to obtain A25 ( Yield 62%).

Weigh compounds A25 (1eq.), A13 (2eq.), Pd(OAc) ₂ (0.1eq.), XPHOS (0.2eq.), Cs ₂ CO ₃ (3eq.), vacuum, fill with nitrogen, and create a nitrogen atmosphere Add solvent-dried toluene at 110°C for 12 hours. After the reaction is completed, filter, extract with CH ₂ Cl ₂ and water, collect the organic phase, dry, concentrate, and separate by silica gel column chromatography to obtain A26 (yield 69%).

Weigh compound A26 (1eq.), dissolve it in dry CH ₂ Cl ₂ , add TFA (3 eq.) dropwise, and react at 25°C for 2 hours. After the reaction is completed, extract CH ₂ Cl ₂ and water, and adjust the solution with saturated sodium bicarbonate solution. The pH is 7-8. The organic phase is collected, dried, concentrated, and separated by silica gel column chromatography to obtain A27 (yield 75%).

Weigh compounds A27 (1eq.), F13 (1eq.), EDCl (1.1eq.), HOBt (1.1eq.), DIPEA (3eq.), dissolve them in dry CH ₂ Cl ₂ , and react at 40°C for 12 hours. After completion, CH ₂ Cl ₂ and water were extracted, the organic phase was collected, dried, concentrated, and separated by silica gel thin layer chromatography to obtain A28 (yield 34%).

Weigh compound A28 (1eq.), dissolve it in dry CH ₂ Cl ₂ , add BBr ₃ CH ₂ Cl ₂ solution dropwise, and react at 25°C for 1 hour. After the reaction is completed, add CH ₃ OH dropwise to quench the reaction, and spin the solvent dry. , purified by reverse-phase semi-preparative HPLC to obtain compound I-20 (yield 28%).

Example 7: Adapt the raw material F13 to F6, and synthesize compound I-7 in a manner similar to that in synthetic route 4.

I-7, yield: 41.38%. ¹ H NMR (300MHz, DMSO) δ9.00(d,J＝7.0Hz,1H),8.77(d,J＝1.9Hz,1H),8.48(d,J＝2.0Hz,1H),7.69(d, J＝3.5Hz,1H),6.83(d,J＝8.8Hz,2H),6.59(d,J＝3.5Hz,1H),6.43(d,J＝8.8Hz,2H),4.85(dd,J＝ 13.5,6.7Hz,1H),4.32(q,J＝7.2Hz,2H),4.12(t,J＝7.3Hz,2H),3.69(t,J＝6.6Hz,2H),2.84(d,J＝ 7.9Hz, 8H), 1.39 (t, J = 7.2Hz, 3H). Mass spectrum: C23H29N6O[M+H] ⁺ calculated value: 405.2, measured value: 405.2.

Example 8: Adapting to changing the raw material A13, compound I-8 was synthesized in a manner similar to the synthesis route I-7.

I-8, 39.30% yield. ¹ H NMR (500MHz, CDCl ₃ ) δ8.73(d,J＝2.0Hz,1H),8.33(d,J＝2.1Hz,1H),7.31(d,J＝3.7Hz,1H),6.94(d ,J＝9.1Hz,2H),6.91–6.88(m,2H),6.54(d,J＝3.5Hz,1H),4.38(q,J＝7.3Hz,2H),4.27(dd,J＝14.2, 7.1Hz,1H),3.54(d,J＝12.6Hz,4H),3.06(d,J＝3.4Hz,8H),2.89(t,J＝10.6Hz,4H), 1.49 (t, J=7.3Hz, 3H). Mass spectrum: C25H33N6O[M+H] ⁺ calculated value: 433.5, measured value: 433.2.

Example 9: Adapt the raw material F13 to F2, and synthesize compound I-9 in a manner similar to that in synthetic route 4.

I-9, yield: 35.82%. ¹ H NMR (500MHz, CDCl ₃ ) δ7.81(d,J＝8.3Hz,1H),7.18(d,J＝8.3Hz,1H),6.89–6.83(m,2H),6.70–6.64(m, 2H),4.64(dt,J＝13.5,5.9Hz,1H),4.48(dd,J＝9.4,8.2Hz,1H),4.20(t,J＝7.9Hz,1H),3.38(dd,J＝12.1 ,5.3Hz,1H),3.27(dd,J＝12.1,6.2Hz,1H),3.02(dq,J＝4.4,2.6Hz,8H),2.69(s,3H).

Mass spectrum: C22H27N6OS[M+H] ⁺ calculated value: 423.2, measured value: 423.2.

Example 10: Adapt the raw material F13 to F8, and synthesize compound I-10 in a manner similar to that in synthetic route 4.

I-10, 35.30% yield. ¹ H NMR (300MHz, D ₂ O) δ8.64 (d, J = 7.2 Hz, 1H), 8.20 (s, 1H), 6.98 (d, J = 7.2 Hz, 1H), 6.83 (d, J = 8.8 Hz,2H),6.74(d,J＝8.8Hz,2H),4.73(s,1H),4.44(dd,J＝15.3,8.5Hz,2H),4.28(t,J＝5.6Hz,2H), 3.26–3.17(m,4H), 3.08(d,J=5.0Hz,4H),2.53(s,3H). Mass spectrum: C21H25N7O[M+H] ⁺ calculated value: 392.2, measured value: 392.1.

Example 11: Using F8 and adapting changes to A13, compound I-11 can be synthesized in a manner similar to that in synthetic route 4.

I-11, yield: 42.23%. ¹ H NMR (300MHz, CD ₃ CN) δ8.74(d,J＝7.2Hz,1H),8.44(s,1H),7.00(d,J＝7.2Hz,1H),6.97–6.85(m,4H ),4.17–4.00(m,1H),3.54–3.38(m,2H),2.97(dd,J＝6.5,2.7Hz,4H),2.93–2.85(m,4H),2.67(s,3H), 2.15–2.03(m,4H),1.76(dtd,J=13.4,9.9,3.8Hz,2H). Mass spectrum: C23H30N7O[M+H] ⁺ calculated value: 420.2, measured value: 420.2.

Example 12: Compound I-12 was synthesized in a manner similar to that in synthetic route 4 by adapting the raw materials F13, A13 and A21.

I-12, yield: 38.78%. ¹ H NMR (500MHz, CDCl ₃ ) δ7.96 (s, 1H), 7.84 (d, J = 8.3Hz, 1H), 6.90 (dd, J = 23.1, 9.1Hz, 4H), 4.03–3.90 (m, 1H),3.88–3.81(m,4H),3.52(d,J＝12.9Hz,2H),3.12–3.03(m,4H),2.82(dd,J＝17.2,6.8Hz,2H),2.72(s ,3H),2.10(d,J＝12.0Hz,2H),1.71(dd,J＝16.0,7.3Hz,2H).

Mass spectrum: C25H31N4O2S[M+H] ⁺ calculated value: 452.2, measured value: 452.2.

Example 13: Compound I-13 was synthesized in a manner similar to that in synthetic route 4 by adapting changes to F13, A13 and A21.

I-13, yield 42.10%. ¹ H NMR (400MHz, CDCl ₃ ) δ7.80 (d, J = 8.2Hz, 2H), 7.18 (d, J = 8.3Hz, 1H), 6.97 (d, J = 8.0Hz, 2H), 6.86 (t ,J＝6.9Hz,1H),4.12(d,J＝7.6Hz,1H),3.69(d,J＝12.5Hz,4H),2.93(t,J＝11.6Hz,4H),2.67(s,3H ), 2.13 (d, J = 12.0Hz, 4H), 1.71 (d, J = 10.9Hz, 2H), 1.46 (d, J = 31.7Hz, 2H), 0.87 (d, J = 6.8Hz, 2H).

Mass spectrum: C23H29N6O2[M+H] ⁺ calculated value: 421.2, measured value: 421.2.

Adapting changes to the starting compounds, the following compounds were synthesized in a manner similar to that in Synthetic Scheme 4.

Example 14:

I-14, yield 38.10%. ¹ H NMR (500MHz, CDCl ₃ ) δ9.21(s,1H),9.02(s,1H),7.03–6.83(m,4H),4.24(s,1H),3.92–3.81(m,4H), 3.46(dd,J＝37.3,10.8Hz,2H),3.09(m,4H),2.98(t,J＝10.2Hz,2H),2.25–2.16(m,2H),2.02(d,J＝5.8Hz ,2H),1.89(d,J＝9.6Hz,2H),1.63(dd,J＝15.1,7.3Hz,2H).

Mass spectrum: C22H24N5O2SCl[M+H] ⁺ calculated value: 458.1, measured value: 458.1.

Example 15:

I-15, yield: 43.20%. ¹ H NMR (500MHz, CDCl ₃ ) δ8.33(d,J＝1.9Hz,1H),8.27(s,J＝1.9Hz,1H),6.97–6.82(m,4H),6.23(s,1H) ,3.91(s,1H),3.87–3.81(m,4H),3.48(d,J＝11.9Hz,2H),3.15–3.08(m,2H),3.07(s,4H),2.80(t,J ＝10.9Hz,2H),2.67(dd,J＝11.4,8.9Hz,2H),2.05(d,J＝10.7Hz,2H),1.99(s,3H).

Mass spectrum: C25H32N5O2[M+H] ⁺ calculated value: 434.3, found value: 434.2.

Example 16:

I-16, yield: 40.23%. ¹ H NMR (400MHz, CD ₃ CN) δ9.41 (s, 1H), 8.12–8.07 (m, 1H), 7.65 (d, J = 4.8Hz, 1H), 7.56 (d, J = 2.7Hz, 1H ),7.21(d,J＝2.7Hz,1H),6.82(d,J＝8.8Hz,2H),6.67(d,J＝8.7Hz,2H),4.52–4.46(m,1H),4.19(d ,J＝5.7Hz,2H),3.29(d,J＝2.6Hz,2H),2.26–2.16(m,4H),2.02(d,J＝6.0Hz,4H).

Mass spectrum: C21H25N6O[M+H] ⁺ calculated value: 377.2, found value: 377.1.

Example 17:

I-17, yield: 17.54%. Mass spectrum: C22H27N6O2[M+H] ⁺ calculated value: 407.48, measured value: 407.4.

Mass spectrum: C21H25N6O[M+H] ⁺ calculated value: 377.2, measured value: 377.1

Example 18:

I-18, yield: 43.20%. ¹ H NMR (300MHz, CDCl ₃ ) δ8.62 (s, 1H), 8.36 (d, J = 7.3Hz, 1H), 6.93 (d, J = 6.3Hz, 2H), 6.86 (d, J = 7.1Hz ,1H),6.56(d,J＝6.3Hz,2H),5.13(d,J＝6.5Hz,1H),4.40–4.30(m,2H),3.88(d,J＝4.1Hz,4H),3.77 (s,2H),3.07(s,4H),2.71(s,3H).

Mass spectrum: C21H25N7O[M+H] ⁺ calculated value: 393.2, measured value: 392.1.

Example 19:

I-19, (4.5mg, yield 35.10%). ¹ H NMR (400MHz, DMSO) δ10.31(s,1H),8.41(d,J＝6.9Hz,1H),8.27(s,1H),8.19(d,J＝7.9Hz,1H),7.23( d,J＝7.9Hz,1H),6.80(d,J＝8.9Hz,1H),6.37(d,J＝8.8Hz,1H),4.51(dd,J＝13.2,6.1Hz,1H),3.98( t,J＝7.4Hz,1H),3.72–3.64(m,2H),3.46–3.39(m,1H),3.26(m,4H),2.94–2.85(m,2H),2.52(m,4H) .

Mass spectrum: C23H27N5O2SF[M+H] ⁺ calculated value: 456.4, found value: 456.0.

Example 20:

I-20 (2.3mg, 28% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.11 (s, 1H), 8.60 (s, 1H), 8.16 (d, J = 7.2Hz, 1H), 7.47(s,1H),6.85(d,J＝8.2Hz,2H),6.66(d,J＝9.3Hz,2H),4.61(d,J＝6.7Hz,1H),4.29(s,1H), 3.65(s,2H),3.40(s,6H),3.34(s,4H)ppm.

Mass spectrum: C21H24N5OS[M+H] ⁺ theoretical value: 394.2, measured value: 394.2.

Example 21:

I-21, yield 36.35%. ¹ H NMR (400MHz, CDCl ₃ ) δ8.81(d,J＝2.1Hz,1H),8.31(d,J＝2.1Hz,1H),7.28(s,1H),7.25(s,1H),6.88 (d,J＝8.8Hz,2H),6.49(d,J＝8.8Hz,2H),5.02(dd,J＝12.0,4.8Hz,1H),4.34(q,J＝7.3Hz,2H),4.28 (t,J＝7.4Hz,2H),3.88–3.80(m,4H),3.74(dd,J＝7.6,4.9Hz,2H),3.07–2.96(m,4H),1.47(t,J＝7.3 Hz,3H).

Mass spectrum: C23H28N5O2[M+H] ⁺ calculated value: 406.2, measured value: 406.2.

Example 22:

I-22, yield: 41.20%. ¹ H NMR (400MHz, CDCl ₃ ) δ8.81(d,J＝2.1Hz,1H),8.31(d,J＝2.1Hz,1H),7.28(s,1H),6.88(d,J＝8.8Hz ,2H),6.49(d,J＝8.8Hz,2H),5.02(dt,J＝12.0,5.9Hz,1H),4.34(q,J＝7.3Hz,2H),4.28(t,J＝7.4Hz ,2H),3.89–3.81(m,4H),3.74(dd,J＝7.6,4.9Hz,2H),3.06–2.98(m,4H),1.47(t,J＝7.3Hz,3H).

Mass spectrum: C23H27N5O2Cl[M+H] ⁺ calculated value: 440.1, measured value: 440.2

Example 23:

I-23 (9.1 mg, 45% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.83 (d, J = 2.0 Hz, 1H), 8.33 (d, J = 2.0 Hz, 1H), 7.39 ( s,1H),7.28(s,1H),6.84(d,J＝8.9Hz,2H),6.48(d,J＝8.9Hz,2H),5.03(dd,J＝12.1,4.9Hz,1H), 4.35(q,J＝7.3Hz,2H),4.27(t,J＝7.4Hz,2H),3.79–3.73(m,5H),1.48(t,J＝7.3Hz,3H)ppm.

Mass spectrum: C20H22ClN4O2[M+H] ⁺ theoretical value: 385.1, measured value: 385.1.

Example 24:

I-24 (4.7 mg, 36% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ 8.84 (d, J = 1.8 Hz, 1H), 8.34 (d, J = 1.9 Hz, 1H), 7.29 ( s,1H),6.87(s,1H),6.81(d,J＝8.5Hz,1H),6.16(s,1H),6.09(d,J＝7.0Hz,1H),5.05(d,J＝7.1 Hz,1H),4.34(dd,J＝14.2,7.4Hz,4H),3.87(s,3H),3.82(s,3H),3.11(s,2H),1.48(dd,J＝7.3,5.0Hz ,3H).ppm.

Mass spectrum: C21H24ClN4O3[M+H] ⁺ theoretical value: 415.1, measured value: 415.1.

Example 25:

I-25 (6.3 mg, 54% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.86 (s, 1H), 8.37 (s, 1H), 7.27 (s, 1H), 6.88 (s, 2H ),6.48(s,2H),5.06(s,1H),4.34(d,J＝7.2Hz,4H),3.70(s,4H),3.58(s,2H),3.18–2.82(m,4H) ,2.11(s,3H),1.47(t,J＝7.2Hz,3H)ppm.

Mass spectrum: C25H30ClN6O2[M+H] ⁺ theoretical value: 481.2, measured value: 481.2.

Example 26:

I-26 (5.2 mg, 51% yield). ¹ H NMR (300MHz, CDCl ₃ ) δ8.94 (s, 1H), 8.47 (s, 1H), 7.93 (d, J = 6.4Hz, 1H), 7.32(s,1H),6.83(d,J＝7.8Hz,2H),6.42(d,J＝8.1Hz,2H),5.00(d,J＝5.6Hz,1H),4.43–4.11(m,4H ),3.88(s,2H),3.34(s,4H),3.20(s,4H),2.77(s,3H),1.53(d,J＝7.4Hz,3H)ppm.

Mass spectrum: C24H30ClN6O[M+H] ⁺ theoretical value: 453.2, measured value: 453.2.

Example 27: Compound 1-27 was synthesized in a manner similar to that in Synthetic Scheme 4

I-27 (6.5 mg, 39% yield). ¹ H NMR (300MHz, CDCl ₃ ) δ 8.82 (d, J = 2.1 Hz, 1H), 8.31 (d, J = 2.1 Hz, 1H), 7.30 ( t,J＝2.3Hz,2H),7.01(d,J＝7.3Hz,1H),6.33(d,J＝8.7Hz,2H),5.04(ddd,J＝12.3,7.2,2.1Hz,1H), 4.31(dt,J＝21.5,7.4Hz,4H), 3.77(dd,J＝7.8,5.1Hz,2H), 1.47(t,J＝7.3Hz,3H)ppm.

Mass spectrum: C19H19BrClN4O[M+H] ⁺ theoretical value: 433.0, measured value: 433.0.

General synthetic route for target molecules 5

Weigh compound I-27, 1eq., 3-boronic acid furan (2eq.), K ₂ CO ₃ (3eq.), Pd(dppf) ₂ Cl ₂ (0.2eq.), vacuum, fill with nitrogen, and under nitrogen atmosphere Add dioxane and water, and react at 90°C for 12 hours. After the reaction is completed, extract with CH ₂ Cl ₂ and water, collect the organic phase, dry, concentrate, and pass through a thin layer of silica gel Compound I-32 was obtained by analytical isolation (yield 42%).

General synthesis route for target molecules 6

Weigh compound A13 (1eq.), 4-bromophenylboronic acid (2eq.), 4AMS (2eq.), Cu(OAc) ₂ (0.4eq.), vacuum, fill with oxygen, and add Et ₃ N ( 3eq.), solvent-dried CH ₂ Cl ₂ , react at 35°C for 12 hours. After the reaction is completed, filter to remove the solid, extract with CH ₂ Cl ₂ and water, collect the organic phase, dry, concentrate, and separate by silica gel column chromatography to obtain A26 ( Yield 34%).

Weigh compound A26 (1eq.), furan-2-boronic acid (2eq.), K ₂ CO ₃ (3eq.), Pd(dppf) ₂ Cl ₂ (0.2eq.), vacuum, fill with nitrogen, and under nitrogen atmosphere Add the solvent 1,4-dioxane and water, and react at 90°C for 12 hours. After the reaction is completed, extract with CH ₂ Cl ₂ and water, collect the organic phase, dry, concentrate, and separate by silica gel column chromatography to obtain A27 (yield 59 %)

Weigh compound A27 (1eq.), dissolve it in dry CH ₂ Cl ₂ , add TFA (3 eq.) dropwise, and react at 25°C for 2 hours. After the reaction is completed, extract CH ₂ Cl ₂ and water, and adjust the solution with saturated sodium bicarbonate solution. The pH is 7-8. The organic phase is collected, dried, concentrated, and separated by silica gel column chromatography to obtain A28 (yield 71%).

Weigh compounds F14 (1eq.), A28 (1.1eq.), EDCI (1.1eq.), HOBt (1.1eq.), DIPEA (3eq.), dissolve them in dry CH ₂ Cl ₂ , and react at 40°C for 12 hours. After the reaction is completed, CH ₂ Cl ₂ and water are extracted, the organic phase is collected, dried, concentrated, and separated by silica gel thin layer chromatography to obtain I-44 (yield 35%).

Example 28:

I-28 (2.1 mg, 23% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ7.86 (d, J = 5.6 Hz, 2H), 7.49–7.45 (m, 3H), 7.39 (d, J ＝6.4Hz,2H),7.07–7.03(m,2H),6.51(d,J＝8.0Hz,1H),6.47(s,1H),6.43(d,J＝3.9Hz,1H),5.30(s ,1H),4.40(dd,J＝1.9,0.7Hz,2H),4.17(d,J＝0.8Hz,2H)ppm.

Mass spectrum: C23H19N4O2[M+H] ⁺ theoretical value: 383.1, measured value: 383.1.

Example 29:

I-29 (3.7mg, 42% yield). ¹ H NMR (300MHz, CDCl ₃ ) δ8.58 (d, J = 2.0Hz, 1H), 7.64 (dd, J = 8.2, 2.7Hz, 2H), 7.46(t,J＝1.6Hz,1H),7.39(d,J＝8.6Hz,2H),6.66(d,J＝0.9Hz,1H),6.55(d,J＝8.4Hz,2H),5.00( dt,J＝12.4,6.2Hz,1H),4.35(t,J＝7.6Hz,2H),3.88–3.77(m,2H)ppm.

Mass spectrum: C21H18FN4O2S[M+H] ⁺ theoretical value: 409.1, measured value: 409.1.

Example 30:

I-30 (7.5 mg, 59% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.83 (d, J = 2.0 Hz, 1H), 8.32 (d, J = 2.1 Hz, 1H), 7.48 ( d,J＝8.6Hz,2H),7.27(s,1H),7.17(t,J＝4.8Hz,2H),7.04(dd,J＝5.0,3.6Hz,1H),6.83(s,1H), 6.49(d,J＝8.4Hz,2H),5.12–5.02(m,1H),4.44–4.29(m,4H),3.83(dd,J＝7.4,5.1Hz,2H),1.47(t,J＝ 7.3Hz,3H)ppm.

Mass spectrum: C23H22ClN4OS[M+H] ⁺ theoretical value: 437.1, measured value: 437.1.

Example 31:

I-31 (8.2 mg, 64% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.83 (d, J = 2.0 Hz, 1H), 8.32 (d, J = 2.1 Hz, 1H), 7.54 ( d,J＝8.7Hz,2H),7.40(d,J＝1.1Hz,1H),7.27(s,1H),6.85(d,J＝7.3Hz,1H),6.52–6.42(m,4H), 5.13–4.98(m,1H),4.40–4.30(m,4H),3.83(dd,J＝7.8,5.0Hz,2H),1.47(t,J＝7.3Hz,3H)ppm.

Mass spectrum: C23H22ClN4O2[M+H] ⁺ theoretical value: 421.1, measured value: 421.1.

Example 32:

I-32 (7.3mg, 66% yield). ¹ H NMR (300MHz, CDCl ₃ ) δ 8.83 (d, J = 2.0Hz, 1H), 8.33 (d, J = 2.1Hz, 1H), 7.64 ( s,1H),7.45(t,J＝1.7Hz,1H),7.36(d,J＝8.6Hz,2H),7.28(s,1H),6.85(d,J＝7.4Hz,1H),6.64( dd,J＝1.8,0.8Hz,1H),6.52(d,J＝8.5Hz,2H),5.07(ddd,J＝12.2,7.2,2.3Hz,1H),4.34(q,J＝7.2Hz,4H ), 3.83 (dd, J = 7.8, 5.0 Hz, 2H), 1.48 (t, J = 7.3 Hz, 3H) ppm.

Mass spectrum: C23H22ClN4O2[M+H] ⁺ theoretical value: 421.1, measured value: 421.1.

Example 33:

I-33 (3.9 mg, 39% yield). ¹ H NMR (300MHz, CDCl ₃ ) δ 8.92 (s, 1H), 8.45 (s, 1H), 7.97 (s, 1H), 7.65 (s, 1H) ),7.46(d,J＝6.4Hz,2H),7.24(s,1H),6.41(d,J＝7.0Hz,3H),5.05(s,1H),4.35–4.22(m,4H),3.85 (s,2H),1.45(t,J=7.2Hz,3H)ppm.

Mass spectrum: C22H22ClN6O[M+H] ⁺ theoretical value: 421.1, measured value: 421.1.

Example 34:

I-34 (7.9 mg, 72% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.56 (d, J = 2.2 Hz, 1H), 7.73 (dd, J = 8.4, 2.6 Hz, 1H), 7.49(d,J＝8.5Hz,2H),7.18–7.15(m,2H),7.04(dd,J＝4.9,3.7Hz,1H),6.50(d,J＝8.5Hz,2H),6.16–6.04 (m,3H),5.03–4.95(m,1H),4.34(t,J＝7.5Hz,2H),3.83(dd,J＝7.6,5.2Hz,2H)ppm.

Mass spectrum: C21H18FN4OS2[M+H] ⁺ theoretical value: 425.1, measured value: 425.1.

Example 35:

I-35 (8.1 mg, 70% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.56 (d, J = 2.1 Hz, 1H), 7.69 (dd, J = 8.4, 2.7 Hz, 1H), 7.54(d,J＝8.6Hz,2H),7.40(d,J＝1.1Hz,1H),6.50(d,J＝8.6Hz,2H),6.47–6.41(m,2H),6.10(d,J ＝7.2Hz,3H),5.03–4.95(m,1H),4.33(t,J＝7.5Hz,2H),3.82(dd,J＝7.8,5.2Hz,2H)ppm.

Mass spectrum: C21H18FN4O2S[M+H] ⁺ theoretical value: 409.1, measured value: 409.1.

Example 36:

I-36 (2.3mg, 31% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.56 (s, 1H), 7.80 (d, J = 8.4Hz, 1H), 7.60 (s, 2H), 7.48(s,1H),7.40(s,2H),6.53(d,J＝8.1Hz,2H),6.18(d,J＝18.2Hz,3H),5.00(s,1H),4.35(t,J ＝7.5Hz,2H),3.90–3.81(m,2H)ppm.

Mass spectrum: C20H18FN6OS[M+H] ⁺ theoretical value: 409.1, measured value: 409.1.

Example 37:

I-37 (6.1 mg, 50% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.57 (d, J = 2.1 Hz, 1H), 7.69 (dd, J = 8.4, 2.6 Hz, 1H), 7.53–7.46(m,2H),7.37–7.31(m,3H),6.54(d,J＝8.5Hz,2H),6.19–5.97(m,3H),5.00(d,J＝7.3Hz,1H) ,4.35(t,J＝7.5Hz,2H),3.82(dd,J＝7.8,5.2Hz,2H)ppm.

Mass spectrum: C21H18FN4OS2[M+H] ⁺ theoretical value: 425.1, measured value: 425.1.

Example 38:

I-38 (9.7 mg, 48% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ 8.83 (d, J = 1.9 Hz, 1H), 8.33 (d, J = 2.0 Hz, 1H), 7.48 ( d,J＝8.5Hz,2H),7.34(qd,J＝5.0,2.1Hz,2H),7.31–7.27(m,2H),6.79(d,J＝7.4Hz,1H),6.53(d,J ＝8.5Hz,2H),5.12–5.03(m,1H),4.35(dt,J＝14.5,7.4Hz,4H),3.84(dd,J＝7.8,5.0Hz,2H),1.48(t,J＝ 7.3Hz,3H)ppm.

Mass spectrum: C23H22ClN4OS[M+H] ⁺ theoretical value: 437.1, measured value: 437.1.

Example 39:

I-39 (9.7mg, 48% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.56 (d, J = 2.0Hz, 1H), 7.72 (dd, J = 8.5, 2.7Hz, 1H), 7.30(dd,J＝3.3,1.8Hz,1H),7.22(d,J＝8.5Hz,2H),6.43(d,J＝8.6Hz,2H),6.20(t,J＝3.3Hz,1H), 6.18–6.04(m,4H),4.97(dd,J＝12.4,5.2Hz,1H),4.29(t,J＝7.5Hz,2H),3.72(dd,J＝7.4,4.9Hz,2H),1.42 (s,9H)ppm.

Mass spectrum: C26H27FN5O3S[M+H] ⁺ theoretical value: 508.2, measured value: 508.2.

Example 40:

I-40 (4.2 mg, 38% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.55 (d, J = 2.6 Hz, 1H), 7.84 (dd, J = 8.5, 2.7 Hz, 1H), 7.48(s,1H),7.17(d,J＝8.5Hz,1H),6.48(d,J＝8.5Hz,2H),6.24(s,2H),6.12(d,J＝6.9Hz,1H), 4.99(d,J＝7.0Hz,1H), 4.31(t,J＝7.5Hz,2H), 3.81(dd,J＝7.8,5.2Hz,2H)ppm.

Mass spectrum: C20H17FN5O2S[M+H] ⁺ theoretical value: 410.1, measured value: 410.1.

Example 41:

I-41 (8.1 mg, 52% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.54 (d, J = 2.1 Hz, 1H), 7.91 (dd, J = 8.5, 2.6 Hz, 1H), 7.32(s,2H),6.50–6.26(m,4H),6.22(d,J＝6.8Hz, 1H), 4.97 (dd, J＝12.5, 5.4Hz, 1H), 4.28 (t, J＝7.5Hz, 2H), 3.80 (dd, J＝7.6, 5.4Hz, 2H) ppm.

Mass spectrum: C21H17ClFN4OS2[M+H] ⁺ theoretical value: 459.0, measured value: 459.0.

Example 42:

I-42 (9.2 mg, 81% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.55 (d, J = 1.9 Hz, 1H), 7.83 (dd, J = 8.5, 2.7 Hz, 1H), 7.66(d,J＝8.7Hz,2H),7.23(d,J＝3.6Hz,1H),6.55(d,J＝3.6Hz,1H),6.50(d,J＝8.7Hz,2H),6.33– 6.07(m,3H),5.00(dt,J＝12.6,6.4Hz,1H),4.37(t,J＝7.7Hz,2H),3.97–3.80(m,5H)ppm.

Mass spectrum: C23H20FN4O4S[M+H] ⁺ theoretical value: 467.1, measured value: 467.1.

Example 43:

I-43 (5.1 mg, 61% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.51 (d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.5, 2.5 Hz, 1H), 7.47(d,J＝8.5Hz,2H),6.46(d,J＝8.5Hz,2H),6.39–6.24(m,3H),6.21(d,J＝7.2Hz,1H),5.98(d,J ＝2.1Hz,1H),4.96(dd,J＝12.3,5.3Hz,1H),4.30(t,J＝7.5Hz,2H),3.81(dd,J＝7.5,5.4Hz,2H),2.32(s ,3H)ppm.

Mass spectrum::C22H20FN4O2S[M+H] ⁺ theoretical value: 423.1, measured value: 423.1.

Example 44:

I-44 (4.2 mg, 35% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.45 (d, J = 1.2 Hz, 1H), 8.04 (dd, J = 8.6, 1.6 Hz, 1H), 7.95(d,J＝8.5Hz,1H),7.80(d,J＝7.2Hz,1H),7.52(d,J＝8.6Hz,2H),7.40(s,1H),6.52–6.36(m,4H ),5.13–4.97(m,1H),4.32(t,J＝7.5Hz,2H),3.94(dd,J＝7.5,5.4Hz,2H)ppm.

Mass spectrum: C21H17ClN3O2S[M+H] ⁺ theoretical value: 410.1, measured value: 410.1.

Example 45:

I-45 (7.4 mg, 54% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ8.09 (s, 1H), 7.85 (d, J = 8.2 Hz, 1H), 7.66 (d, J = 8.4 Hz,1H),7.58–7.31(m,4H),6.58–6.31(m,4H),5.11–4.99(m,1H),4.33(t,J＝7.4Hz,2H),3.94–3.84(m, 2H),2.66(s,3H)ppm.

Mass spectrum: C22H20N3O3[M+H] ⁺ theoretical value: 374.1, measured value: 374.1.

Example 46:

I-46 (4.0 mg, 43% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.27 (d, J = 6.0 Hz, 1H), 7.79 (d, J = 2.2 Hz, 1H), 7.56 ( d,J＝8.7Hz,2H),7.41(d,J＝1.2Hz,1H),7.33(d,J＝12.2Hz,2H),6.80(dd,J＝2.1,0.9Hz,1H),6.53( d,J＝8.5Hz,2H),6.48–6.39(m,2H),5.09(td,J＝7.2,5.2Hz,1H),4.38(t,J＝7.6Hz,2H),3.83(dd,J =7.6,5.4Hz,2H)ppm.

Mass spectrum: C22H18FN2O3[M+H] ⁺ theoretical value: 377.1, measured value: 377.1.

Example 47:

I-47 (3.2 mg, 29% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ9.17 (d, J = 1.7 Hz, 1H), 7.91–7.77 (m, 2H), 7.67 (dd, J ＝8.7,1.8Hz,1H),7.56(d,J＝8.7Hz,2H),7.41(d,J＝1.1Hz,1H),6.52(d,J＝8.6Hz,2H),6.48–6.37(m ,2H),5.08(tt,J＝12.6,6.3Hz,1H),4.39(t,J＝7.5Hz,2H),3.87(dd,J＝7.7,5.3Hz,2H)ppm.

Mass spectrum: C21H17BrN3O2S[M+H] ⁺ theoretical value: 454.0, measured value: 454.0.

Example 48:

I-48 (3.7 mg, 33% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.01 (d, J = 2.7 Hz, 1H), 8.35 (s, 1H), 8.27 (d, J = 8.4 Hz,1H),8.20(d,J＝8.8Hz,1H),8.09(dd,J＝8.8,1.9Hz,1H),7.57(d,J＝8.6Hz,2H),7.50(dd,J＝8.3 ,4.2Hz,1H),7.41(s,1H),6.84(d,J＝6.7Hz,1H),6.53(d,J＝8.6Hz,2H),6.47(d,J＝3.3Hz,1H), 6.44(dd,J＝3.3,1.8Hz,1H),5.10(d,J＝7.4Hz,1H),4.40(t,J＝7.5Hz,2H),3.86(dd,J＝7.9,4.8Hz,2H )ppm.

Mass spectrum: C23H20N3O2[M+H] ⁺ theoretical value: 370.1, measured value: 370.2.

Example 49:

I-49 (5.5 mg, 34% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.46 (s, 1H), 8.02–7.98 (m, 1H), 7.55 (t, J = 9.1Hz, 3H ),7.40(d,J＝0.9Hz,1H),6.54–6.39(m,4H),5.07(dd,J＝12.4,5.1Hz,1H),4.34(t,J＝7.6Hz,2H),3.92 (dd,J＝7.7,5.2Hz,2H)ppm.

Mass spectrum: C21H17BrN3O2S[M+H] ⁺ theoretical value: 454.0, measured value: 454.0.

Example 50:

I-50 (2.4 mg, 27% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.12 (s, 1H), 8.59 (d, J = 1.3 Hz, 1H), 8.17 (d, J = 8.5 Hz,1H),8.01(dd,J＝8.6,1.6Hz,1H),7.54(d,J＝8.6Hz,2H),7.40(s,2H),6.50(d,J＝8.6Hz,2H), 6.47–6.36(m,3H),5.11–5.04(m,1H),4.35(t,J＝7.5Hz,2H),3.91(dd,J＝7.7,5.2Hz,2H)ppm.

Mass spectrum: C21H19N4O2[M+H] ⁺ theoretical value: 359.1, measured value: 359.2.

Example 51:

I-51 (8.3mg, 49% yield). ¹ H NMR (400MHz, DMSO) δ11.59 (s, 1H), 8.45 (d, J = 7.3Hz, 1H), 8.09 (d, J = 2.9Hz ,1H),7.96(d,J＝7.8Hz,1H),7.60(d,J＝1.2Hz,1H),7.51(d,J＝8.6Hz,2H),7.03–6.88(m,2H),6.64 (d,J＝3.2Hz,1H),6.57–6.41(m,3H),4.95–4.81(m,1H),4.21(t,J＝7.5Hz,2H),3.84–3.70(m,2H), 2.45(s,3H)ppm.

Mass spectrum: C23H22N3O2[M+H] ⁺ theoretical value: 372.2, measured value: 372.2.

Example 52:

I-52 (2.3 mg, 26% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.33 (s, 1H), 7.90 (d, J = 8.5 Hz, 1H), 7.83 (d, J = 8.4 Hz,1H),7.54(d,J＝11.6Hz,1H),7.25–7.22(m,2H),7.00(s,1H),6.87–6.74(m,2H),6.53(d,J＝7.5Hz ,2H),5.06(dd,J＝11.1,5.0Hz,1H),4.35(t,J＝7.6Hz,2H),3.84–3.77(m,2H),2.87(s,3H)ppm.

Mass spectrum: C22H20N3O2S[M+H] ⁺ theoretical value: 390.1, measured value: 390.1.

Example 53:

I-53 (7.2mg, 33% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.70 (s, 1H), 9.62 (d, J = 5.8Hz, 1H), 9.09 (s, 1H), 8.02(d,J＝9.5Hz,1H),7.67(d,J＝9.4Hz,1H),7.45(d,J＝8.6Hz,2H),7.39(s,1H),6.43(d,J＝13.1 Hz, 4H), 5.02 (d, J = 6.2Hz, 1H), 4.28 (t, J = 7.0Hz, 2H), 4.14 (t, J = 6.1Hz, 2H) ppm.

Mass spectrum: C20H18N5O2[M+H] ⁺ theoretical value: 360.1, measured value: 360.1.

Example 54:

I-54 (6.4 mg, 38% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.47 (s, 1H), 9.23 (d, J = 5.8 Hz, 1H), 7.97 (d, J = 9.4 Hz,1H),7.75(s,1H),7.62(d,J＝13.7Hz,2H),7.48(d,J＝8.3Hz,2H),7.39(s,1H),6.46(d,J＝8.3 Hz,2H),6.43(s,2H),5.04(dd,J＝12.8,6.4Hz,1H),4.30(t,J＝7.3Hz,2H),4.14(t,J＝6.5Hz,2H)ppm .

Mass spectrum: C21H19N4O2[M+H] ⁺ theoretical value: 359.1, measured value: 359.1.

Example 55:

I-55 (5.1 mg, 36% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.60 (s, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.91 (d, J = 8.2 Hz,1H),7.82(t,J＝7.1Hz,1H),7.63(t,J＝7.1Hz,1H),7.54(t,J＝6.1Hz,3H),7.40(s,1H),6.51( d,J＝8.7Hz,2H),6.48–6.38(m,2H),5.09(dd,J＝12.4,5.1Hz,1H),4.38(t,J＝7.5Hz,2H),3.91(dd,J =7.8,5.1Hz,2H)ppm.

Mass spectrum: C23H19ClN3O2[M+H] ⁺ theoretical value: 404.1, measured value: 404.1.

Example 56:

I-56 (3.2 mg, 30% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.65–8.58 (m, 1H), 8.56 (s, 1H), 8.29 (d, J = 3.5Hz, 1H ),8.18(d,J＝6.6Hz,1H),7.51(d,J＝8.6Hz,2H),7.39(s,1H),7.17(dd,J＝7.9,4.8Hz,1H),6.49(d ,J＝8.6Hz,2H),6.43(s,2H),5.06(dd,J＝13.0,6.0Hz,1H),4.33(t,J＝7.5Hz,2H),4.10–3.97(m,2H) ppm.

Mass spectrum: C21H19N4O2[M+H] ⁺ theoretical value: 359.1, measured value: 359.1.

Example 57:

I-57 (6.2 mg, 40% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.81 (s, 1H), 8.45 (d, J = 7.4 Hz, 1H), 7.57 (d, J = 1.7 Hz,1H),7.52(d,J＝8.7Hz,2H),7.41(dd,J＝6.6,1.3Hz,2H),7.32(d,J＝1.6Hz,1H),6.49(d,J＝8.7 Hz,2H),6.46–6.40(m,2H),5.12–5.04(m,1H),4.32(t,J＝7.5Hz,2H),4.01(dd,J＝7.6,5.4Hz,2H)ppm.

Mass spectrum: C22H18BrClN3O2[M+H] ⁺ theoretical value: 470.0, measured value: 470.0.

Example 58:

I-58 (9.4 mg, 47% yield). ¹ H NMR (500MHz, CDCl ₃ ) δ9.52 (d, J = 7.6 Hz, 1H), 8.43 (dd, J = 4.7, 1.0 Hz, 1H), 8.19(s,1H),7.97(d,J＝8.0Hz,1H),7.54(d,J＝8.5Hz,2H),7.40(d,J＝1.1Hz,1H),7.14(dd,J＝8.2 ,4.8Hz,1H),6.52(d,J＝8.6Hz,2H),6.47–6.41(m,2H),5.16(dd,J＝13.3,6.1Hz,1H),4.40–4.30(m,2H) ,3.99–3.86(m,2H)ppm.

Mass spectrum: C21H19N4O2[M+H] ⁺ theoretical value: 359.1, measured value: 359.1.

Example 59:

I-59 (8.2 mg, 51% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.33 (d, J = 8.1 Hz, 1H), 7.60 (d, J = 8.4 Hz, 2H), 7.53 ( d,J＝8.7Hz,2H),7.38(t,J＝7.3Hz,2H),7.24(s,1H),6.50(d,J＝8.6Hz,2H),6.43(dd,J＝9.3,2.5 Hz, 2H), 5.10 (d, J = 7.3Hz, 1H), 4.36 (t, J = 7.5Hz, 2H), 3.83 (dd, J = 7.6, 5.3Hz, 2H) ppm.

Mass spectrum: C21H19N4O2[M+H] ⁺ theoretical value: 359.1, measured value: 359.1.

Example 60:

I-60 (7.2 mg, 44% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ9.12 (s, 1H), 8.62 (d, J = 1.4 Hz, 1H), 8.16 (d, J = 8.5 Hz,1H),8.04(dd,J＝8.6,1.7Hz,1H),7.62(d,J＝7.2 Hz,1H),7.53(d,J＝8.7Hz,2H),7.40(d,J＝1.0Hz,1H),6.49(d,J＝8.7Hz,2H),6.45–6.38(m,2H), 5.14–5.01(m,1H),4.35(t,J＝7.5Hz,2H),3.93(dd,J＝7.7,5.3Hz,2H)ppm.

Mass spectrum: C21H18N3O2S[M+H] ⁺ theoretical value: 376.1, measured value: 376.1.

Example 61:

I-61 (5.2 mg, 33% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ8.63 (s, 1H), 8.35 (d, J = 5.5 Hz, 1H), 7.96 (d, J = 8.7 Hz,1H),7.85(d,J＝6.1Hz,1H),7.44(d,J＝8.2Hz,2H),7.33(s,1H),6.42(d,J＝8.3Hz,2H),6.37( s,2H),4.98(d,J＝5.3Hz,1H),4.26(t,J＝7.0Hz,2H),4.01–3.92(m,2H)ppm.

Mass spectrum: C20H18N5O2[M+H] ⁺ theoretical value: 360.1, measured value: 360.1.

Example I-62:

I-62: Yield: 15.00%. ¹ H NMR (500MHz, CDCl ₃ ) δ8.08–7.77(m,1H),7.21–7.10(m,1H),6.96–6.83(m,4H),3.91(s,1H),3.89–3.81(m ,4H),3.48(d,J＝11.9Hz,2H),3.07(s,4H),2.80(t,J＝10.9Hz,2H),2.05(d,J＝10.7Hz,3H),1.99(s , 4H), 1.42 (dd, J = 11.8, 4.4Hz, 2H). Mass spectrum: C24H28N5O3 [M-1] ^- calculated value: 433.2, measured value: 433.1.

Example I-63:

I-63: Yield 42.10%. ¹ H NMR (500MHz, CDCl ₃ ) δ7.86(s,1H),6.94–6.81(m,4H),4.03–3.89(m,1H),3.89–3.81(m,2H),3.55–3.46(m ,2H),3.19(s,1H),3.07(d,J＝4.8Hz,4H),2.87–2.81(m,2H),2.62(s,3H),2.58(s,3H),2.51(d, J=21.5Hz, 2H), 2.08 (d, J=12.8Hz, 4H). Mass spectrum: C25H31N5O2S[M+H] ⁺ calculated value: 466.2, measured value: 466.2.

Example I-64:

I-64: Yield: 14.86%. Mass spectrum: C23H29N6SO[M+H] ⁺ calculated value: 437.28, found value: 437.2. Example I-65:

I-65: Yield: 18.11%. 1H NMR (500MHz, CDCl ₃ ) δ8.57 (s, 1H), 7.64 (d, J = 5.9Hz, 1H), 6.90 (d, J = 8.5Hz, 2H), 6.50 (d, J = 8.5Hz, 2H),6.02(s,3H),5.05-4.90(m,1H),4.28(t,J＝5.0Hz,2H),3.87–3.79(m,4H),3.73(t,J＝5.0Hz,2H ), 3.05 (m, 4H). Mass spectrum: C21H23N5O2SF[M+H] ⁺ calculated value: 428.1, measured value: 428.1.

Example I-66: BR-003006-NX-1

I-66, yield: 33.91%. Mass spectrum: C21H24FN6OS[M+H] ⁺ calculated value: 476.28, measured value: 476.2.

Example I-67: BR-003517-NX-1

I-67, (1.1 mg, 15% yield) mass spectrum: C22H26N5OS[M+H] ⁺ theoretical value: 408.2, measured value: 408.2.

Example I-68: BR-003519-NX-1

I-68: (2.1 mg, 32% yield) Mass spectrum: C22H19FN5OS[M+H] ⁺ theoretical value: 420.1, measured value: 420.1.

Example I-69: BR-003693-NX-1

I-69: (5.0mg, 15.18% yield) ¹ H NMR (500MHz, Chloroform-d) δ7.46 (s, 1H), 7.24 (d, J = 8.6Hz, 2H), 6.84–6.77 (m, 1H),6.60–6.47(m,4H),5.98(d,J＝7.4Hz,1H),4.99(qt,J＝7.3,5.0Hz,1H),4.32(t,J＝7.5Hz,2H), 3.78(dd,J=7.8,5.0Hz,2H),2.74(s,3H).

Mass spectrum: C21H22N4OF3S[M+H] ⁺ theoretical value: 435.1, measured value: 435.2.

Example I-70: BR-003694-NX-1

I-70: (11.0mg, 2.72% yield) ¹ H NMR (500MHz, Chloroform-d) δ7.46 (s, 1H), 7.24 (d, J = 8.6Hz, 2H), 6.84–6.77 (m, 1H),6.60–6.47(m,4H),5.98(d,J＝7.4Hz,1H),4.99(qt,J＝7.3,5.0Hz,1H),4.32(t,J＝7.5Hz,2H), 3.78(dd,J=7.8,5.0Hz,2H),2.74(s,3H).

Mass spectrum: C21H23N4OF3S[M+H] ⁺ theoretical value: 407.1, measured value: 407.0.

Example I-71: BR-003891-NX-1

I-71: (11.0 mg, 22.4% yield) ¹ H NMR (500 MHz, Chloroform-d) δ 8.82 (d, J = 2.1 Hz, 1H), 8.33 (d, J = 2.2 Hz, 1H), 7.29 (s,1H),6.70(d,J＝8.4Hz,1H),6.47(d,J＝4.2Hz,1H),6.32(s,1H),5.04(q,J＝5.6,5.0Hz,1H) ,4.52(t,J＝8.6Hz,2H),4.36 (q,J＝7.3Hz,2H),4.27(t,J＝7.4Hz,2H),3.75(s,2H),3.18(t,J＝8.6Hz,2H),1.48(d,J＝7.3Hz ,3H).

Mass spectrum: C21H22N4O2Cl[M+H] ⁺ theoretical value: 397.1, measured value: 397.0.

Example I-72: BR-003892-NX-1

I-72: Mass spectrum: C23H20N4O2F3S[M+H] ⁺ theoretical value: 472.2, measured value: 473.0.

Example I-73: BR-003893-NX-1

I-73: Mass spectrum: C21H17N2O2F4[M+H] ⁺ theoretical value: 405.11, measured value: 405.0.

Example I-74: BR-003894-NX-1

I-74: (43.0mg, 25.03% yield) ¹ H NMR (500MHz, Chloroform-d) δ8.07 (dd, J=6.7, 2.3Hz, 1H), 8.02 (ddd, J=8.6, 4.6, 2.4 Hz,1H),7.30(t,J＝9.2Hz,1H),7.25–7.22(m,2H),6.82(tt,J＝7.4,1.1Hz,1H),6.63(d,J＝5.5Hz,1H ),6.54–6.49(m,2H),4.99(tdd,J＝7.3,4.6,2.6Hz,1H),4.32(ddd,J＝7.9,7.2,0.7Hz,2H),3.78(ddd,J＝7.9 ,4.7,0.8Hz,2H).

Mass spectrum: C17H15N2OF4[M+H] ⁺ theoretical value: 339.10, measured value: 339.0.

Example I-75: BR-008207-NX-1

I-75: (3.9 mg, 35% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.83 (d, J = 2.0 Hz, 1H), 8.33 (d, J = 2.1 Hz, 1H), 7.43 (d,J＝8.6Hz,2H),7.29(s,1H),6.95(d,J＝3.5Hz,1H),6.74(d,J＝7.6Hz,1H),6.68(dd,J＝3.5, 1.1Hz,1H),6.49(d,J＝8.6Hz,2H),5.12–5.02(m,1H),4.35(q,J＝7.3Hz,4H),3.83(dd,J＝7.8,5.0Hz, 2H), 2.48 (d, J = 0.7Hz, 3H), 1.48 (t, J = 7.3Hz, 3H) ppm.

Mass spectrum: C24H24ClN4OS[M+H] ⁺ theoretical value: 451.1, measured value: 451.1.

Example I-76: BR-008208-NX-1

I-76: (2.4 mg, 25% yield). ¹ H NMR (400MHz, CDCl ₃ ) δ 8.85 (d, J = 2.1 Hz, 1H), 8.35 (d, J = 2.1 Hz, 1H), 7.59 –7.53(m,1H),7.50(d,J＝8.7Hz,2H),7.32(s,1H),7.14(d,J＝3.9Hz,1H),6.70(s,1H),6.58–6.46( m,2H),5.15–5.10(m,1H),4.46–4.33(m,4H),3.96–3.88(m,2H),1.50(d,J=7.3Hz,3H)ppm.

Mass spectrum: C24H21ClN5OS[M+H] ⁺ theoretical value: 462.1, measured value: 462.1.

Test example

Enzyme activity inhibition test method:

USP25 enzyme activity inhibition test method: The reaction system solution is 50mM Tris (pH 7.5), 150mM NaCl, 1mM DTT, and 0.05% Tween-20. Pipette 10ul of 20nM USP25 (50mM Tris pH 7.5, 150mM NaCl, 1mM DTT, 20% glycerol) into a 96-well plate at room temperature; add 10ul of compounds of different concentrations and incubate for 20 minutes; finally add 10ul of 20uM substrate Ub- AMC reacted for 30 minutes. The final concentration of USP25 protein in the reaction system is 4nM, the final concentration of substrate Ub-AMC is 4uM, and the final concentration of DMSO is 1%. The fluorescence signal was detected in kinetic mode on a multifunctional microplate reader Synergy Neo2 (BioTek) (excitation wavelength 360 nm and emission wavelength 460 nm); the reaction rate was calculated using the change in fluorescence signal within the first 30 minutes, which was within the linear interval of the assay Inside. In order to verify that the compound does not interfere with the detection system, no protein was added to the above reaction as a control to test the impact of the compound on the reaction.

K _i , the change value of the fluorescence signal of USP25 treated with the compound for 30 minutes; K ₀ , the change value of the fluorescence signal of USP25 treated with DMSO for 30 minutes.

USP28 enzyme activity inhibition test method:

The reaction system solution is 50mM Tris (pH 7.5), 150mM NaCl, 1mM DTT, and 0.05% Tween-20. Pipette 10ul of 10nM USP28 (50mM Tris pH 7.5, 150mM NaCl, 1mM DTT, 20% glycerol) into a 96-well plate at room temperature; add 10ul of compounds of different concentrations and incubate for 20 minutes; finally add 10ul of 5uM substrate Ub- AMC reacts for 15 minutes. The final concentration of USP28 protein in the reaction system was 2nM, the final concentration of substrate Ub-AMC was 1uM, and the final concentration of DMSO was 1%. The fluorescence signal was detected in kinetic mode on a multifunctional microplate reader Synergy Neo2 (BioTek) (excitation wavelength 360 nm and emission wavelength 460 nm); the reaction rate was calculated using the change in fluorescence signal within the first 15 minutes, which was within the linear interval of the assay Inside. In order to verify that the compound does not interfere with the detection system, no protein was added to the above reaction as a control to test the impact of the compound on the reaction.

K _i , the change value of the fluorescence signal of USP28 treated with the compound for 30 minutes; K ₀ , the change value of the fluorescence signal of USP28 treated with DMSO for 30 minutes.

Compound activity inhibition test results:

Activity inhibition test results of compound 100uM single point concentration

Gradient activity inhibition test results of some compounds:

The symbols in the table correspond to the IC50 results as follows:

“++++”: 1uM–10uM; “+++”: 10uM–30uM; “++”: 30uM–50uM; “+”: 50uM–100uM; “-”: >100uM

Claims (10)

The compound represented by formula I, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug:
in, Ar is selected from a substituted or unsubstituted C6-C14 aryl group, a substituted or unsubstituted 9-10 membered bicyclic fused heterocyclic group containing one or more heteroatoms selected from N, O, and S, where used The substituted substituent is selected from halogen, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl; m and n are each independently 1 or 2; R ₁ is selected from hydrogen; nitro; halogen; substituted or unsubstituted C1-C6 alkoxy, wherein the substituent used for substitution is selected from C1-C6 alkylamino; substituted or unsubstituted C1-C6 alkyl Carbonylamino, wherein the substituent used for substitution is selected from halogen, amino, C1-C6 alkylamino, 5-7 membered heterocyclyl; substituted or unsubstituted 5-7 membered heterocyclyl, wherein the substituent used for substitution The group is selected from cyano, C1-C6 alkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, halogen; wherein, the 5-7 membered heterocyclic group contains one selected from N, O, S or multiple heteroatoms; R ₂ is selected from hydrogen; C1-C6 alkyl; halogen; C1-C6 alkoxy; Alternatively, R ₁ and R ₂ form a 5-7-membered heterocyclic group, wherein the 5-7-membered heterocyclic group contains one or more heteroatoms selected from N, O, and S. Preferably, the heteroatoms is an O atom. The compound of formula I according to claim 1, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug, wherein , Ar is selected from substituted or unsubstituted phenyl, substituted or unsubstituted thienopyridine, substituted or unsubstituted furopyridine, substituted or unsubstituted pyrrolopyridine, substituted or unsubstituted pyrazopyridine, substituted or unsubstituted pyrrolopyrimidine, substituted or unsubstituted thienopyrimidine, substituted or unsubstituted benzopyrrole, substituted or unsubstituted benzofuran, substituted or unsubstituted benzothiophene, substituted or unsubstituted benzene Oxazoles, substituted or unsubstituted benzothiazoles, substituted or unsubstituted benzopyrazoles, substituted or unsubstituted benzotriazoles, substituted or unsubstituted benzopyridines, wherein the substituents used for substitution are each Independently selected from halogen, amino, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl; preferably, the substituent used for substitution is selected from halogen, amino, cyano, C1-C6 alkyl, 1, 2 or 3 of the halogenated C1-C6 alkyl groups. The compound of formula I according to claim 1, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug, wherein , Ar is selected from the following structures:
The compound represented by formula I according to any one of claims 1 to 3, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug, which, R ₁ is selected from hydrogen; nitro; halogen; substituted or unsubstituted C1-C4 alkoxy, wherein the substituent used for substitution is selected from C1-C4 alkylamino; substituted or unsubstituted C1-C4 alkyl Carbonylamino, wherein the substituent for substitution is selected from halogen, amino, C1-C4 alkylamino, piperazinyl, piperidinyl; substituted or unsubstituted piperazinyl; substituted or unsubstituted morpholinyl; substituted or unsubstituted furyl; substituted or unsubstituted thienyl; substituted or unsubstituted pyrazolyl; substituted or unsubstituted pyrrolyl; substituted or unsubstituted pyridyl; substituted or unsubstituted oxazolyl, wherein The substituents used for substitution are respectively selected from cyano group, C1-C4 alkyl group, C1-C4 alkylcarbonyl group, C1-C4 alkoxycarbonyl group, and halogen. The compound represented by formula I according to any one of claims 1 to 3, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable Salt or prodrug, wherein R ₁ is selected from hydrogen, methylaminoethoxy, methoxy, nitro, chloromethylcarbonylamino, methylaminomethylcarbonylamino, piperazinylmethylcarbonylamino, piperidylmethyl Carbonylamino, piperazinyl, methylcarbonylpiperazinyl, methylpiperazinyl, morpholinyl, Br, furyl, methylfuryl, methoxycarbonylfuranyl, thienyl, chlorothienyl, methyl Thienyl, cyanothienyl, pyrazolyl, tert-butoxycarbonylpyrrolyl, oxazolyl, pyridyl. The compound represented by formula I according to any one of claims 1 to 5, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable Salt or prodrug, wherein R ₂ is selected from hydrogen; C1-C4 alkyl; halogen; C1-C4 alkoxy. The compound represented by formula I according to any one of claims 1 to 5, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable Salt or prodrug, wherein R ₂ is hydrogen, methyl, methoxy, ethoxy or propoxy. The compound of formula I according to claim 1, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt or prodrug, wherein , the compound Has the structure shown below:






The compound represented by formula I according to any one of claims 1 to 8, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable Use of salts or prodrugs in the preparation of USP25 and/or USP28 inhibitors. The compound represented by formula I according to any one of claims 1 to 8, or its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable Salts or prodrugs for use in the preparation of medicaments for the prevention or treatment of diseases associated with USP25 and/or USP28, Preferably, the diseases associated with USP25 and/or USP28 include cancer, inflammation, autoimmune diseases, and neurodegenerative diseases.