USP inhibitor, and preparation method therefor and use thereof

Abstract

The present invention relates to the field of medicines, and provides a USP inhibitor, and a preparation method therefor and a use thereof. Specifically provided are a compound having an inhibitory effect on a USP, and a pharmaceutically acceptable salt, a stereoisomer, a solvate or a prodrug thereof, as shown in formula (I). In addition, also provided are a pharmaceutical composition comprising the compound, and a use thereof in the treatment of cancers, immune diseases, cardiovascular diseases, neurological diseases, etc. or in the preparation of a kit for cancers, immune diseases, cardiovascular diseases, neurological diseases or prognosis evaluation of patients.

Description

USP inhibitors, preparation methods and applications thereof

[0001] This application claims priority to Chinese patent application 202311251694.1, filed on September 26, 2023, and Chinese patent application 202410171830.4, filed on February 6, 2024, and cites the full text of the above-mentioned Chinese patent applications. Technical Field

[0002] The present disclosure relates to the field of medicine, and in particular to a compound having USP inhibitory effect, and its use and preparation method. Background Art

[0003] The ubiquitin-proteasome system (USP) is a dynamic, bidirectional protein modification and regulatory system within cells, involved in the degradation and modification of over 80% of proteins within cells. Through polyubiquitination of substrate proteins and subsequent proteasome degradation, USP participates in a variety of life processes, including cell cycle regulation, cell receptor function, gene transcription, immune response, and tumor growth. This pathway is mediated by an enzymatic conjugation cascade, involving the continuous activation of ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), and ubiquitin ligase (E3), completing the ubiquitination modification of substrate proteins (The ubiquitin-proteasome pathway: on protein death and cell life, Ciechanover, EMBO J. 1998 Dec 15; 17(24): 7151-60.), and the degradation or modification of target proteins is completed by the proteasome; on the other hand, the deubiquitinating enzyme family (DUBs) is responsible for hydrolyzing the ester bond, peptide bond or isopeptide bond at the carboxyl terminus of ubiquitin, specifically hydrolyzing ubiquitin molecules from ubiquitin-linked proteins or precursor proteins, playing a role in deubiquitination, reversely regulating protein degradation, and thus affecting protein function.

[0004] Currently, approximately 100 DUBs have been discovered in the human genome. They act as monitors in the regulation of the ubiquitination system and play an important role in regulating various intracellular activities such as cell cycle, DNA repair, immunity and protein homeostasis (Regulation of proteolysis by human deubiquitinating enzymes, Ziad M Eletr, Keith D Wilkinson, Biochim Biophys Acta. 2014 Jan; 1843(1): 114-28.). Dysregulation of DUBs often leads to various human diseases such as cardiovascular disease, neurological disorders and tumors (Development of inhibitors in the ubiquitination cascade, Wei Zhang, Sachdev S Sidhu, FEBS Lett. 2014 Jan 21; 588(2): 356-67.). DUBs are divided into seven major families based on sequence and structural similarities, namely, ubiquitin C-terminal hydrolases (UCHs), ubiquitin-specific proteases (USPs), ovarian tumor domain proteases (OTUs), Machado-Josephin domain proteases (MJDs), Jab1 / MPN domain associated metalloproteases (JAMMs), UFM1-specific zinc finger peptidases (ZUFSPs), and the MIU-containing novel DUB family (MINDY) (Jinhong et al., 2020).

[0005] USPs are the largest and most structurally diverse family of DUBs, comprising approximately 60 members. This family of cysteine ​​proteases contains two short, conserved sequences: an N-terminal Cys-box and a C-terminal His-box, consisting of catalytic cysteine ​​and histidine residues. These sequences, including the catalytic tripartite residues of cysteine, histidine, and aspartic acid / asparagine, enable the removal of ubiquitin molecules from larger proteins. Due to the protease activity of the USP family and its regulatory role in human biological processes, USPs have become potential drug targets. For example, overexpression of USP2 has been associated with prostate cancer (Priolo et al., 2006; Stevenson et al., 2007) and breast cancer (Qu et al., 2015). Aberrant expression of USP7 is found in a variety of cancers, including prostate cancer, lung cancer, brain cancer, colon cancer, breast cancer, epithelial ovarian cancer, liver cancer, and leukemia (Emerging insights into HAUSP (USP7) in physiology cancer and other diseases, Seemana Bhattacharya, Dipankar Chakraborty, Malini Basu, Mrinal K Ghosh, Signal Transduct Target Ther. 2018 Jun 29; 3:17).

[0006] USP21, a member of the USP family, is involved in regulating numerous intracellular activities. USP21 deubiquitinates several key pathway oncogenes and genes with oncogenic potential, including HH / Gli, MEK2, MARK, stem cell renewal / Nanog / GATA3, and WNT. USP21 plays a role in various tumor types, including pancreatic cancer, bile duct cancer, liver cancer, breast cancer (triple-negative breast cancer), renal cancer, bladder cancer, lung cancer, and esophageal squamous cell carcinoma (CN116676391A). USP21 is also associated with clinical prognosis. Furthermore, recent studies have shown that USP21 plays a crucial role in negatively regulating the antiviral innate immune response, primarily through deubiquitination of retinoic acid (RA)-induced gene protein I (RIG-I) and interferon-stimulated gene 15 (ISG15) proteins (TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity, Michaela U Gack 1, Young C Shin, Chul-Hyun Joo, Tomohiko Urano et al. Nature. 2007 Apr 19;446(7138):916-920). Recent studies have found that USP21 gene amplification is common in human pancreatic cancer (22% of PDACs), and high USP21 expression is associated with PDAC progression. In a mouse pancreatic cancer model, exogenous high expression of USP21 accelerated tumor growth, while USP21 knockout inhibited tumor growth. In vitro and in vivo studies have demonstrated that USP21 plays a driving role in cancer. The study further revealed that USP21 plays a role in pancreatic cancer through the WNT pathway.

[0007] Inhibiting USP21 can inhibit cell proliferation and tumor growth. Currently, many research institutions and pharmaceutical companies internationally are researching inhibitors targeting DUBs, but no drugs targeting USP21 have been discovered. There is a significant clinical need for the development of USP21 inhibitors. Therefore, USP21 is a first-in-class cancer target with great potential and market value. The development of specific inhibitors targeting USP21 is of great significance for molecular targeted therapy of related tumors and other immune and other diseases associated with USP21 abnormalities.

[0008] Summary of the Invention

[0009] The present invention provides a novel substituted fused-ring aromatic compound, which has high inhibitory activity as a USP enzyme inhibitor.

[0010] In one aspect of the present invention, there is provided a compound of formula (I), a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof:

[0011] In formula (I), R 1 Selected from -R 4 -C(O)-C 2-6 Alkenyl, -C 2-6 Alkenyl, -N=NR a R b 、-R 4 -C(O)-C 2-6 Alkynyl, -R 4 -SO2-C 2-6 Alkenyl, -R 4 -C(O)-C 1-3 Alkyl, -R 4 -C 2-6 Alkynyl, -R 4 -C(O)-C 3-9 Cycloalkyl, -R 4 -C(O)-3 to 9-membered heterocycloalkyl, -R 4 -C 1-3 Alkyl-C 3-9 Cycloalkyl, -R 4 -C 3-9 Cycloalkyl, -C(O)-NH-C 2-6 Alkynyl, -C(O)-NH-C 2-6 Alkenyl, -C(O)-NH-C 1-3 Alkyl or The C 2-6 Alkenyl, C 2-6 Alkynyl is optionally substituted by 1, 2 or 3 groups independently selected from hydrogen, halogen, cyano, -CF3, C 1-6 alkyl, 3- to 9-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 6-10 Aryl, -C 1-3 Alkyl-NR a R b 、-C 1-3 Alkyl-hydroxy, -C(O)OR a 、-C(O)NR a R b 、-NR a R b Substituents substituted, the C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl, C 1-6 The alkyl group is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b, hydroxyl, carboxyl, -CF3, C 1-6 alkoxy, -O-3 to 9 membered heterocycloalkyl, -O-5 to 10 membered heteroaryl, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2-C 1-3 Alkyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 alkoxy, 5- to 10-membered heteroaryl, 6- to 10-membered aryl, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Substitution of alkynyl groups by substituents;

[0012] In one embodiment, R 4 Selected from NH, NR a or CR a R b ;

[0013] In one embodiment, R a 、R b are each independently selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, the C 1-3 Alkyl, C 3-9 The cycloalkyl group is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1- 3 alkyl, -SO2C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Substitution of alkoxy groups;

[0014] In one embodiment, R 5 、R 6 are independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl, the C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3Alkyl, -SO2-C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1- 6 Alkoxy substituents are substituted;

[0015] In one embodiment, R 2 、R 3 Each independently selected from hydrogen, halogen, cyano, hydroxyl, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, -NR a R b 、-C(O)-C 1-3 Alkyl, -C(O)NR a R b 、-SO2-C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-C 2-6 Alkynyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -C 1-3 Alkyl-NR a R b 、-C 2-6 Alkenyl, -C 2-6 Alkynyl, 5- to 10-membered heteroaryl, C 6-10 Aryl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-3 Alkyl, C 2-6 Alkynyl, C 2-6 alkenyl, 5- to 10-membered heteroaryl, C 6-10 Aryl is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3 substituents;

[0016] In one embodiment, the halogen is selected from fluorine, chlorine or bromine;

[0017] In one embodiment, U 1 、U 2 、U 3 Each independently selected from CR 7 or CH, or adjacent U 2 、U 3 Connected to form a 6- to 10-membered aromatic group;

[0018] In one embodiment, the R7 Selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl, -C 1-3 Alkyl-3 to 9 membered heterocycloalkyl, the C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Substitution of alkynyl groups by substituents;

[0019] In one embodiment, the heterocycloalkyl or heteroaryl radical has at least one heteroatom selected from N, O and S as a ring atom.

[0020] In some embodiments, the R 1 Selected from -R 4 -C(O)-C(R 8 )=CR 9 R 10 、-C(R 8 )=CR 9 R 10 、-C(R 8 )=C(R 9 )-C(O)-NH2、-R 4 -C(O)-C(R 8 )=C(R 9 )-C(O)-NH2、-R 4 -C(O)-C(R 8 )=C(R 9 )-C(O)-OR 10 、-R 4 -C(O)-C(R 8 )=C(R 9 )-CH2-NR a R b 、-N=NR a 、-R 4-C(O)-C≡C-CH3、-R 4 -C(O)-C≡CH, -R 4 -C(O)-CH2-C≡CH、-R 4 -SO2-C(R 8 )=CR 9 R 10 、-R 4 -C(O)-R 5 、-R 4 -CH2-C≡CH, -C(O)-NH-C 1-3 Alkyl, -R 4 -C(O)-C 1-3 Alkyl-R 4 -C(O)-C(R 8 )=CR 9 R 10 、-C(O)-R 4 -C 1-3 Alkyl-R 4 -C(O)-C(R 8 )=CR 9 R 10 、-R 4 -CH2-C 3-9 Cycloalkyl or The C 1-3 Alkyl, C 3-9 Cycloalkyl is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 Substitution of alkoxy groups;

[0021] In one embodiment, R 4 Selected from NH or NR a ;

[0022] In one embodiment, R 5 、R 6 are independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl, the C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2-C 1-3Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1- 6 Alkoxy substituents are substituted;

[0023] In one embodiment, R 8 、R 9 、R 10 are independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, 3- to 9-membered heterocycloalkyl, -C 1-3 Alkyl-C 6-10 Aryl, -C 1-3 Alkyl-5 to 10 membered heteroaryl, -C 1-3 Alkyl-3 to 9 membered heterocycloalkyl, said C 1-3 Alkyl, C 1-6 Alkyl, C 6-10 Aryl, 5 to 10 membered heteroaryl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, -C 1-6 Substitution of alkoxy groups;

[0024] In one embodiment, R a 、R b are each independently selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, the C 1-3 Alkyl, C 3-9 The cycloalkyl group is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1- 3 alkyl, -SO2C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Substitution of alkoxy groups;

[0025] In one embodiment, R 2 、R 3 Each independently selected from hydrogen, halogen, cyano, hydroxyl, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, -NR a Rb , the C 1-6 Alkyl, C 1-6 The alkoxy group is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3 substituents;

[0026] In one embodiment, the halogen is selected from fluorine or chlorine;

[0027] In one embodiment, U 1 、U 2 、U 3 Each independently selected from CR 7 or CH, or adjacent U 2 、U 3 Connected to form a 6- to 10-membered aromatic group;

[0028] In one embodiment, the R 7 Selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl, -C 1-3 Alkyl-3 to 9 membered heterocycloalkyl, the C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Substitution of alkynyl groups by substituents;

[0029] In one embodiment, the heterocycloalkyl or heteroaryl radical has at least one heteroatom selected from N, O and S as a ring atom.

[0030] In some embodiments, R 1 Selected from

[0031] In one embodiment, the R 2 、R 3 Each independently selected from H, CF3;

[0032] In one embodiment, the U 1 、U 2 、U 3 All selected from CH, or adjacent U 2 、U 3 They connect with each other to form a benzene ring;

[0033] In one embodiment, the R 7 Selected from -H, -CH3,

[0034] In some embodiments, Formula (I) is represented by Formula (II-1), Formula (II-2), Formula (II-3), Formula (II-4), Formula (II-5), Formula (II-6), Formula (II-7), Formula (II-8), Formula (II-9), Formula (II-10), Formula (II-11), Formula (II-12), Formula (II-13), Formula (II-14), Formula (II-15), Formula (II-16) or Formula (II-17):

[0035] R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 、R 8 、R 9 、R 10 、R a The definition of is as mentioned above.

[0036] In some embodiments, the compound described by Formula (I), Formula (II-1), Formula (II-2), Formula (II-3), Formula (II-4), Formula (II-5), Formula (II-6), Formula (II-7), Formula (II-8), Formula (II-9), Formula (II-10), Formula (II-11), Formula (II-12), Formula (II-13), Formula (II-14), Formula (II-15), Formula (II-16) or Formula (II-17), its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof is selected from the following group:

[0037] Another aspect of the present invention provides a method for preparing some of the above compounds, their pharmaceutically acceptable salts, stereoisomers, solvates or prodrugs thereof, comprising the following steps:

[0038] Compound (III-1) undergoes an amidation reaction with compound (Y) to generate compound (III-2);

[0039] The compound (Y) includes any one of acyl chloride, carboxylic acid or sulfonyl chloride;

[0040] The R 1 、R 2 、R 3 、R 7 The definition of is as mentioned above.

[0041] Another aspect of the present invention provides a pharmaceutical composition comprising the above-mentioned compound, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the above-mentioned method and at least one pharmaceutically acceptable excipient.

[0042] Another aspect of the present invention provides a use of the above-mentioned compound, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the above-mentioned method, or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating cardiovascular disease, neurological disorder, cancer or immune disease, or in the preparation of a kit for assessing cardiovascular disease, neurological disorder, cancer, immune disease or patient prognosis;

[0043] In a preferred embodiment, the pharmaceutical composition further comprises another drug for treating cardiovascular disease, neurological disorder, cancer or immune disease;

[0044] In a preferred embodiment, use in the preparation of a method for treating a disease associated with USP21 activity;

[0045] In a preferred embodiment, the cancer includes prostate cancer, bile duct cancer, pancreatic cancer, breast cancer, lung cancer, brain cancer, colon cancer, kidney cancer, bladder cancer, epithelial ovarian cancer, esophageal squamous cell carcinoma, liver cancer and leukemia;

[0046] In a more preferred embodiment, the cancer includes pancreatic cancer, bile duct cancer, esophageal squamous cell carcinoma, liver cancer, breast cancer, kidney cancer, bladder cancer and lung cancer.

[0047] Another aspect of the present invention provides a use of the above-mentioned compound, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the above-mentioned method, or the above-mentioned pharmaceutical composition in the preparation of a USP inhibitor; preferably, use in the preparation of a USP21 inhibitor.

[0048] Another aspect of the present invention provides a method for treating a disease associated with a USP mutation or amplification in a patient in need thereof, comprising administering to the patient the above-mentioned compound, a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, or the above-mentioned pharmaceutical composition;

[0049] The USP mutation or amplification includes USP21 mutation or amplification;

[0050] Preferably, the diseases associated with USP mutation or amplification include cardiovascular diseases, neurological disorders, cancer, and immune diseases;

[0051] Preferably, the cancer includes prostate cancer, bile duct cancer, pancreatic cancer, breast cancer, lung cancer, brain cancer, colon cancer, kidney cancer, bladder cancer, epithelial ovarian cancer, liver cancer, esophageal squamous cell carcinoma and leukemia;

[0052] More preferably, the cancer includes pancreatic cancer, bile duct cancer, esophageal squamous cell carcinoma, liver cancer, breast cancer, kidney cancer, bladder cancer and lung cancer.

[0053] In another aspect of the present invention, a method for inhibiting USP mutations in a biological sample for non-disease diagnosis or treatment purposes is provided, comprising contacting the biological sample with the above-mentioned compound, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the above-mentioned method, or the above-mentioned pharmaceutical composition. DETAILED DESCRIPTION

[0054] I. Definition

[0055] Based on the above contents of the present disclosure, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present disclosure, other various forms of modifications, replacements or changes can be made.

[0056] definition

[0057] Unless expressly stated otherwise, throughout the specification and claims, the term "comprise" or variations such as "include" or "comprising", etc., will be understood to include the stated elements or components but not to exclude other elements or other components.

[0058] The compounds of the present disclosure may be asymmetric, for example, having one or more stereoisomers. Unless otherwise indicated, all stereoisomers are included, such as enantiomers and diastereomers. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure forms or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents. Racemates, diastereomers, and enantiomers are all included within the scope of the present disclosure.

[0059] The compounds of the present disclosure also include tautomeric forms.Tautomeric forms arise from the exchange of a single bond with an adjacent double bond accompanied by the migration of a proton.

[0060] The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both occurring and not occurring.

[0061] The numerical ranges herein refer to the individual integers within the given range. For example, “C 1-6 " means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms; "C 1-3 ” means that the group can have 1 carbon atom, 2 carbon atoms, or 3 carbon atoms.

[0062] The term "substituted" means that any one or more hydrogen atoms on a particular atom or group are replaced by a substituent, as long as the valence state of the particular atom or group is normal and the substituted compound is stable. When the substituent is a keto group (i.e., =0), it means that two hydrogen atoms are replaced. Unless otherwise specified, the type and number of substituents can be any on the basis of chemical practicability.

[0063] When any variable (such as R n ) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 1-5 R, then the group may be optionally substituted with up to 5 R, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or variants thereof are permissible only if such combinations result in stable compounds.

[0064] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 2,2-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting embodiments of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups, with methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl being preferred in the present disclosure.

[0065] The term "heterocycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen, or S(O)m (wherein m is an integer from 0 to 2), but excluding the ring portion of -OO-, -OS-, or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1-4 are heteroatoms; more preferably, it contains 3 to 8 ring atoms; most preferably, it contains 3 to 8 ring atoms; further preferably, it contains 3-8 membered heterocyclic groups containing 1-3 nitrogen atoms, optionally substituted by 1-2 oxygen atoms, sulfur atoms, or oxo groups, including nitrogen-containing monocyclic heterocyclic groups or nitrogen-containing spiroheterocyclic groups.

[0066] Heterocycloalkyl may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, substituted amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2-C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Alkynyl.

[0067] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. More preferably, phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, including benzo 5- to 10-membered heteroaryl, benzo 3- to 8-membered cycloalkyl, and benzo 3- to 8-membered heteroalkyl, preferably benzo 5- to 6-membered heteroaryl, benzo 3- to 6-membered cycloalkyl, and benzo 3- to 6-membered heteroalkyl, wherein the heterocyclic group is a heterocyclic group containing 1-3 nitrogen atoms, oxygen atoms, or sulfur atoms; or further comprises a three-membered nitrogen-containing fused ring containing a benzene ring.

[0068] Aryl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, substituted amino, hydroxy, carboxyl, -CF3.

[0069] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 12-membered, more preferably 5-membered or 6-membered, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl and the like, preferably triazolyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, pyrimidinyl or thiazolyl; more preferably pyrazolyl, pyrrolyl and oxazolyl.

[0070] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrido, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0071] "Alkenyl" refers to a chain alkenyl group, also known as an alkene group, wherein the alkenyl group can be further substituted with other related groups, for example: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0072] "Alkynyl" refers to (CH≡C-), wherein the alkynyl can be further substituted by other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, cyano, nitro, phenol, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0073] The term "alkoxy" refers to-O-(alkyl) and-O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above. The limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, chloro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0074] The term "cycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon substituent, wherein the ring atoms are all carbon atoms, preferably a 3-6 membered cycloalkyl, and the heteroaryl group may be optionally substituted or unsubstituted.

[0075] The term "adjacent U 2 、U3 are connected to form a 6- to 10-membered aromatic group" 2 、U 3 The carbon atoms of the alkyl group are connected to form a 6- to 10-membered aromatic group, preferably a benzene ring. Non-limiting examples of the compounds formed include wait.

[0076] HATU: O-(7-Azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate

[0077] DIEA or DIPEA: N,N-diisopropylethylamine

[0078] Pd2(dba)3: tris dibenzylideneacetone dipalladium

[0079] xantphos:4,5-bis(diphenylphosphine)-9,9-dimethylxanthene

[0080] Dioxane: 1,4-dioxane

[0081] DMSO: dimethyl sulfoxide

[0082] TCFH:N,N,N',N'-Tetramethylchlorouronium hexafluorophosphate

[0083] NMI:N-Methylimidazole

[0084] DBU:1,8-diazabicycloundec-7-ene

[0085] DCM: dichloromethane

[0086] DMF:N,N-dimethylformamide

[0087] Cinnamic acid:

[0088] TEA: Methacryloyl chloride

[0089] THF: Tetrahydrofuran

[0090] Furan-2,5-dione:

[0091] The hydrogen atoms described in the present disclosure may all be replaced by their isotope deuterium.

[0092] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0093] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1-3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0094] Refers to the chemical bond connection.

[0095] Drug or pharmaceutical composition

[0096] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0097] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acids and bases of the specified compound without adverse biological effects, such as acid addition salts (including organic acids and inorganic acids) or base addition salts (including organic bases and inorganic bases).

[0098] The pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent or a mixture of the two.

[0099] The medicaments or pharmaceutical compositions of the present disclosure can be administered orally, topically, parenterally, or mucosally (e.g., buccally, by inhalation, or rectally) in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers. The oral route is generally desirable. The active agent can be administered orally in the form of capsules, tablets, and the like (see Remington: The Science and Practice of Pharmacy, 20th Edition).

[0100] For oral administration in the form of tablets or capsules, the active drug component can be mixed with non-toxic, pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, sucrose, glucose, mannitol, sorbitol and other reducing and non-reducing sugars, microcrystalline cellulose, calcium sulfate or dibasic calcium phosphate); lubricants (e.g., magnesium stearate, talc or silica, stearic acid, sodium stearyl fumarate, glyceryl behenate, calcium stearate, etc.); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate), coloring and flavoring agents, gelatin, sweeteners, natural and synthetic gums (such as acacia, tragacanth or alginates), buffer salts, carboxymethylcellulose, polyethylene glycol, waxes, etc. For oral administration in liquid form, the drug component can be combined with a non-toxic, pharmaceutically acceptable inert carrier (e.g., ethanol, glycerol, water), an anti-settling agent (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats), an emulsifier (e.g., lecithin or gum arabic), a non-aqueous carrier (e.g., almond oil, oily esters, ethanol or fractionated vegetable oils), a preservative (e.g., methyl or propyl p-hydroxybenzoate or sorbic acid), etc. Stabilizers such as antioxidants (BHA, BHT, propyl citric acid, sodium ascorbate, citric acid) can also be added to stabilize the dosage form.

[0101] Tablets comprising the active compound can be coated by methods well known in the art. The compositions of the present disclosure comprising the compound of formula I as the active compound can also be introduced into beads, microspheres or microcapsules, for example, constructed from polyglycolic acid / lactic acid (PGLA). Liquid preparations for oral administration can take the form of, for example, solutions, syrups, emulsions or suspensions, or they can be presented as dry products reconstituted with water or other suitable excipients before use. Preparations for oral administration can be suitably formulated to release the active compound in a controlled or delayed manner.

[0102] The medicine or pharmaceutical composition of the present disclosure can be delivered parenterally, that is, administered intravenously (iv), intracerebroventricularly (icv), subcutaneously (sc), intraperitoneally (ip), intramuscularly (im), subcutaneously (sd) or intradermally (id), by direct injection, for example, through rapid bolus injection or continuous infusion. The formulation for injection can be presented in unit dosage form, for example, in an ampoule or multi-dose container with an added preservative. The composition can be in the form of an excipient, a suspension, a solution or an emulsion in an oil or aqueous vehicle, and can include formulation agents such as anti-settling agents, stabilizers and / or dispersants. Alternatively, the active ingredient can be reconstituted with a suitable carrier (e.g., sterile pyrogen-free water) before use in powder form.

[0103] The medicaments or pharmaceutical compositions of the present disclosure may also be formulated for rectal administration, for example, as suppositories or retention enemas (eg, containing conventional suppository bases such as cocoa butter or other glycerides).

[0104] The term "treating" includes inhibiting, alleviating, preventing or eliminating one or more symptoms or side effects associated with the disease, condition or disorder being treated.

[0105] The terms "reduce," "inhibit," "alleviate," or "reduce" are used relative to a control. One skilled in the art will readily determine the appropriate control for each experiment. For example, a reduced response in a subject or cell treated with a compound is compared to a response in a subject or cell not treated with the compound.

[0106] The term "excipient" is used herein to include any other compound that may be included in or on the microparticles that is not a therapeutic or biologically active compound. Thus, an excipient should be pharmaceutically or biologically acceptable or relevant, e.g., an excipient is generally non-toxic to a subject. "Excipient" includes a single such compound and is also intended to include a plurality of compounds.

[0107] The term "pharmaceutical composition" means a composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable ingredient selected from the following depending on the mode of administration and the nature of the dosage form, including but not limited to: carriers, diluents, adjuvants, excipients, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, dispersants, temperature-sensitive materials, temperature regulators, adhesives, stabilizers, suspending agents, etc.

[0108] Uses and treatments

[0109] The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal or cell thereof amenable to the methods described herein, whether in vitro or in situ. In some non-limiting embodiments, the patient, subject, or individual is a human.

[0110] The term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof. Inhibition of enzymes in biological samples can be used to achieve a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, bioanalysis, gene expression studies, and identification of biological targets.

[0111] As used herein, the term "USP21-mediated" disorders, diseases and / or conditions as used herein refers to any disease or other deleterious condition in which USP21 or a mutant thereof is known to play a role. Accordingly, another embodiment of the present disclosure relates to treating or lessening the severity of one or more diseases in which USP21 or a mutant thereof is known to play a role.

[0112] The cancer treated by the compounds or compositions of the present disclosure is melanoma, liposarcoma, lung cancer, breast cancer, prostate cancer, leukemia, kidney cancer, esophageal cancer, brain cancer, lymphoma or colon cancer. In certain embodiments, the cancer is primary effusion lymphoma (PEL).

[0113] The compounds of the present disclosure can be used to treat a proliferative disease selected from the group consisting of: a benign or malignant tumor, carcinoma of the brain, kidney, liver, adrenal gland, bladder, breast, stomach, gastric tumor, ovary, colon, rectum, prostate, pancreas, lung, vagina, cervix, testis, genitourinary tract, esophagus, larynx, skin, bone or thyroid; a sarcoma, glioblastoma, neuroblastoma, multiple myeloma or gastrointestinal cancer (particularly colon cancer or colorectal adenoma) or tumors of the neck and head, epidermal hyperplasia, psoriasis, prostatic hyperplasia, neoplasia, neoplasia of epithelial character, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, Hodgkin's and non-Hodgkin's lymphoma, breast cancer, follicular carcinoma, undifferentiated tumor, papillary carcinoma, seminoma, melanoma, MYD88 driven disorders, DLBCL, ABC DLBCL, IL-1 driven disorders, rheumatic or indolent multiple myeloma or leukemia.

[0114] Cancers described herein include, but are not limited to, leukemias (e.g., acute leukemias, acute lymphocytic leukemias, acute myelocytic leukemias, acute myeloblastic leukemias, acute promyelocytic leukemias, acute myelomonocytic leukemias, acute monocytic leukemias, acute erythroleukemias, chronic leukemias, chronic myelocytic leukemias, chronic lymphocytic leukemias), polycythemia vera, lymphomas (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain diseases, and solid tumors, such as sarcomas and carcinomas (e.g., fibrosarcomas, myxosarcomas, liposarcomas, chondrosarcomas, osteogenic sarcomas, chordomas, angiosarcomas, endotheliosarcomas, lymphangiosarcomas, lymphoendotheliosarcomas, synoviomas, mesotheliomas, Ewing's tumors, and sarcomas). (tumor), leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct cancer, choriocarcinoma, seminoma, embryonal tumor, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma, and retinoblastoma).

[0115] Combination therapy

[0116] The present disclosure provides a combination therapy using compounds as described in the present disclosure and other therapeutic drugs. The term "combination therapy" used in the present disclosure includes administering these agents in a sequential manner, i.e., each therapeutic agent is administered at different times, and administering these therapeutic agents, or at least two agents, substantially simultaneously. The order of each agent, or substantially simultaneous administration, can be affected by any appropriate route, including but not limited to, oral route, intravenous route, intramuscular route, subcutaneous route, and direct absorption by mucosal tissue. The agents can be administered by the same route or different routes. For example, the first agent can be administered orally, while the second agent can be administered intravenously. In addition, the combination of agents selected can be administered by intravenous injection, while the other agents of the combination can be administered orally. Alternatively, for example, two or more agents can be administered by intravenous or subcutaneous injection.

[0117] II. Example

[0118] The present disclosure is further explained below with reference to the embodiments. The description of specific exemplary embodiments of the present disclosure is for the purpose of illustration and description. These descriptions are not intended to limit the present disclosure to the precise form disclosed, and it is obvious that many changes and variations are possible based on the teachings of this application specification. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present disclosure and their practical application, so that those skilled in the art can realize and utilize the various exemplary embodiments of the present disclosure and various selections and variations.

[0119] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods.

[0120] Unless otherwise specified, the materials, reagents, etc. used in the following embodiments can be obtained from commercial sources.

[0121] Instruments and Reagents: NMR analysis was performed using an Agilent 400MR DD2 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3), with tetramethylsilane (TMS) as the internal standard. LC-MS analysis was performed using an Agilent 1260 Infinity II–InfinityLab LC / MSD mass spectrometer. HPLC analysis was performed using an Agilent 1260 Infinity II high-pressure liquid chromatograph (Sunfire C18 5 μm, 150 x 4.6 mm column).

[0122] Thin-layer chromatography silica gel plates: HSGF254 silica gel plates (Yantai Jiangyou Silica Gel Development Co., Ltd.), sizes 0.9 mm–1 mm. TLC silica gel plates: GF254 silica gel plates (Yucheng Chemical (Shanghai) Co., Ltd.), sizes 0.2 mm–0.25 mm. Column chromatography: Support: 300–400 mesh silica gel (Qingdao Hailang Silica Gel Desiccant Co., Ltd.), Flash column (Aiger Finomei Claricep Flash amorphous silica gel purification column). Reagents: cis-3-chloroacrylic acid (Biodex Pharmaceuticals), trans-3-chloroacrylic acid (Thermo Fisher Scientific), 2-chloroacrylic acid, vinylsulfonyl chloride (Shanghai Adamas Reagent Co., Ltd.), 5-bromo-2-trifluoromethylpyrimidine, tetrahydrofuran, triisopropyl borate, n-butyllithium, 2-bromo-1,3-dimethyl-5-nitrobenzene, carbon tetrachloride, N-bromosuccinimide, benzoyl peroxide, acetonitrile, 1,2-dichloroethane, N-bromosuccinimide, benzoyl peroxide, 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride, acryloyl chloride, N,N-dimethylformamide, acrylonitrile, 1,4-dioxane, 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, tris(dibenzylideneacetone)dipalladium, dimethyl sulfoxide, sodium sulfite, but-2-ynoic acid, N-methylimidazole, N,N,N',N'-tetramethylchlorouronium hexafluorophosphate, ethylenesulfonyl chloride, dichloromethane, 1,8-diazabicycloundec-7-ene, ethyl acetate, cinnamic acid, N,N-dimethylformamide, (Z)-4-amino-4-oxobut-2-enoic acid, methacryloyl chloride, lithium hydroxide, furan-2,5-dihydro-1,2-dihydro-2,4-dioxobut-2-enoic acid ketone, concentrated sulfuric acid, 2-phenylacrylic acid, 7-amino-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-9-one, NN-diisopropylethylamine, 2-(trifluoromethyl)acrylic acid, 2-chloroethanesulfonyl chloride, (E)-4-(dimethylamino)but-2-enoic acid hydrochloride, trifluoroacetic acid, 2-(hydroxymethyl)acrylic acid, pyridine, phosphorus oxychloride, propiolic acid, dichloromethane, triethylamine, 2-chloroacetyl chloride, 2-benzylacrylic acid, 1-methylimidazole, 3-bromoprop-1-yne, (E)-4-ethoxy-4-oxobut-2-enoic acid, hydroxide Lithium chloride monohydrate, (Z)-3-cyanoacrylic acid, isobutyl chloroformate, 2-fluoropropionic acid, 2-chloro-2,2-difluoroacetic acid, iodomethane, 2-fluoroacrylic acid, 1-fluorocyclopropane-1-carboxylic acid, reduced iron powder, oxalyl chloride, triethylamine, 1-(tert-butoxycarbonyl)-3-fluoroazetidine-3-carboxylic acid, 2-fluoro-2-methylpropionic acid, 1-methylpiperazine, stannous chloride, N-(chloro(dimethylamino)methylene)-N-methylmethylammonium hexafluorophosphate, morpholine, anhydrous magnesium sulfate, tert-butyl piperazine-1-carboxylate, or synthesized using methods known in the art.

[0123] Unless otherwise specified, all reactions disclosed herein were carried out under continuous magnetic stirring in dry nitrogen or argon, with dry solvents and reaction temperatures in degrees Celsius.

[0124] Intermediate I-1: Synthesis of 2-(trifluoromethyl)pyrimidin-5-ylboronic acid (I-1)

[0125] 5-Bromo-2-trifluoromethylpyrimidine (5 g, 22.03 mmol) was dissolved in tetrahydrofuran (50 ml), and triisopropyl borate (7.62 ml, 33.04 mmol) was added. The atmosphere was replaced with nitrogen, and the mixture was cooled in cold hydrazine at -85°C. After the temperature dropped to -78°C, n-butyllithium (11.5 ml, 28.64 mmol, 2.5 M) was slowly added dropwise at a temperature below -70°C. After completion of the addition, the mixture was allowed to react for another hour at this temperature. Saturated ammonium chloride was added to quench the mixture below 0°C, and the pH was adjusted to a weak acidic state with 1N hydrochloric acid. Ethyl acetate was added for extraction, and the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target crude product, 2-(trifluoromethyl)pyrimidin-5-ylboronic acid (I-1, 4.92 g), which was used directly in subsequent reactions. ESI[MH] + =191.1

[0126] Intermediate I-2: Synthesis of 5-(bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2)

[0127] Step 1: Synthesis of 2-bromo-1-(bromomethyl)-3-methyl-5-nitrobenzene (I-2-a)

[0128] Dissolve 2-bromo-1,3-dimethyl-5-nitrobenzene (2 g, 8.7 mmol) in carbon tetrachloride (70 mL), add N-bromosuccinimide (1549 mg, 8.7 mmol) and benzoyl peroxide (105 mg, 0.44 mmol) to displace the nitrogen atmosphere, and react at 80°C overnight. After completion of the reaction, cool to room temperature, concentrate the reaction mixture under reduced pressure, and use the crude product directly in the next step to obtain the target product, 2-bromo-1-(bromomethyl)-3-methyl-5-nitrobenzene (I-2-a, 2.7 g, crude product).

[0129] Step 2: Synthesis of 2-bromo-3-methyl-5-nitrobenzaldehyde (I-2-b)

[0130] 2-Bromo-1-(bromomethyl)-3-methyl-5-nitrobenzene (I-2-a, 2.7 g, 8.7 mmol) was dissolved in acetonitrile and stirred at 0°C for 10 minutes. N-methylmorpholine oxide (3 g, 8.7 mmol) was then added portionwise and stirred at 0°C for 10 minutes. The mixture was then reacted at room temperature overnight. After completion of the reaction, the reaction solution was concentrated under reduced pressure and purified via a flash column (petroleum ether:ethyl acetate = 0-15%) to obtain the desired product, 2-bromo-3-methyl-5-nitrobenzaldehyde (I-2-b, 890 mg, 42.3% yield).

[0131] Step 3: Synthesis of 3-methyl-5-nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (I-2-c)

[0132] 2-Bromo-3-methyl-5-nitrobenzaldehyde (I-2-b, 890 mg, 3.66 mmol) was dissolved in acetonitrile (20 mL), and (2-(trifluoromethyl)pyrimidin-5-yl)boric acid (1 g, 5.86 mmol), potassium phosphate (2.3 g, 10.98 mmol), and 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride (134 mg, 0.18 mmol) were added. The mixture was purged with nitrogen for 1 minute and microwave-dried at 120°C for 1 hour. After completion of the reaction, the reaction solution was diluted with ethyl acetate, filtered through celite, concentrated under reduced pressure, and purified on a flash column (petroleum ether:ethyl acetate = 0-20%) to obtain the desired product, 3-methyl-5-nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (I-2-c, 770 mg, yield 67.6%). ESI [M+H] + =312.2

[0133] Step 4: Synthesis of 5-methyl-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2-d)

[0134] 3-Methyl-5-nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (I-2-c, 770 mg, 2.47 mmol) was dissolved in 1,2-dichloroethane (20 mL), and tetrabutylammonium iodide (46 mg, 0.12 mmol) and 2-hydroperoxide-2-methylpropane (aqueous solution) (1426 μL, 14.89 mmol) were added. The atmosphere was purged with nitrogen three times, and the mixture was sealed and reacted at 100°C for 16 hours. The reaction solution was concentrated under reduced pressure and purified by flash column chromatography (petroleum ether:ethyl acetate = 0-35%) to obtain the target product, 5-methyl-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2-d, 250 mg, yield 32.7%). ESI [M+H] + =310.2

[0135] Step 5: Synthesis of 5-(bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2)

[0136] 5-Methyl-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2-d, 250 mg, 0.8 mmol) was dissolved in carbon tetrachloride (10 mL), and N-bromosuccinimide (288 mg, 1.62 mmol) and benzoyl peroxide (195 mg, 0.81 mmol) were added. The atmosphere was replaced with nitrogen and the reaction was carried out at 100°C for 16 hours. After the reaction, the reaction solution was concentrated under reduced pressure and purified by flash column chromatography (petroleum ether:ethyl acetate = 0-35%) to obtain the target product 5-(bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2, 160 mg, yield 51.1%).

[0137] Example 1: Synthesis of N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (1)

[0138] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0754 mmol) was dissolved in acetonitrile and water (1 ml / 0.2 ml). Acryloyl chloride (5 mg, 0.0905 mmol) and potassium carbonate (20 mg, 0.226 mmol) were added at 0°C and the reaction was continued at 0°C for 1 hour. The reaction solution was filtered and concentrated under reduced pressure. The crude product was purified by thin-layer chromatography on silica gel (dichloromethane:methanol = 10:1) to obtain the desired product, N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (1, 9 mg, yield 37.38%). ESI [M+H] + =320.00 1 H NMR (400MHz, DMSO-d6) δ10.61(s,1H),9.46(s,1H),8.20(s,1H),8.05–7.89(m,2H),6.45–6.31(m,2H),5.89–5.82(m,1H).

[0139] Example 2: Synthesis of 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (2)

[0140] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (28 mg, 0.106 mmol) was dissolved in N,N-dimethylformamide (1 mL), and then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (14 mg, 0.106 mmol), N,N-diisopropylethylamine (15 mg, 0.318 mmol), and 2-fluoroacrylic acid (14.5 mg, 0.127 mmol) were added to the reaction solution in sequence and the reaction was allowed to proceed overnight at room temperature. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic phase was collected and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative plate to obtain the target product, 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (2, 0.32 mg, yield 0.9%). ESI [M+H] + =338.10

[0141] Example 3: Synthesis of (E)-3-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylonitrile (3)

[0142] 7-Bromo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (50 mg, 0.15 mmol) and acrylonitrile (16.1 mg, 0.30 mmol) were dissolved in 1,4-dioxane (3 ml), and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (14 mg, 0.03 mmol), trisdibenzylideneacetone dipalladium (13.7 mg, 0.015 mmol), and potassium carbonate (63 mg, 0.46 mmol) were added and reacted in a microwave at 120°C for 0.5 hour. After the reaction was completed, the mixture was diluted with water and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified on a flash column (ethyl acetate:petroleum ether = 0% to 30%) to obtain the target product (E)-3-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylonitrile (3, 18.2 mg, yield 40.4%). ESI [M+H] + =302.2, 1 H NMR (400MHz, DMSO-d6) δ9.61 (s, 1H), 8.23–8.10 (m, 2H), 8.05 (d, J = 7.8Hz, 1H), 7.78 (d, J = 16.7Hz, 1H), 6.74 (d, J = 16.7Hz, 1H).

[0143] Example 4: Synthesis of (E)-3-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (4)

[0144] (E)-3-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylonitrile (3, 17 mg, 0.05 mmol) and potassium carbonate (2 mg, 0.02 mmol) were added to ethanol (3 ml) and dimethyl sulfoxide (0.2 ml), followed by 35% hydrogen peroxide (12 mg, 0.3 mmol). The reaction was allowed to react at room temperature for 2 hours. The mixture was quenched with saturated sodium sulfite solution, monitored with potassium iodide starch paper, and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative liquid chromatography to obtain the target product, (E)-3-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (4, 0.8 mg, 4.2% yield). ESI [M+H] + =320.3, 1 H NMR (400MHz, DMSO-d6) δ9.59(s,1H),8.46(s,1H),8.10(d,J=8.1Hz,1H),7.99(s,1H),7.53(d,J=16.0Hz,1H),7.22(s,1H),6.82(d,J=15.9Hz,1H).

[0145] Example 5: Synthesis of N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-ynamide (5)

[0146] Dissolve 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (10 mg, 0.038 mmol) and but-2-ynoic acid (6.4 mg, 0.075 mmol) in N,N-dimethylformamide (1 ml), then add N-methylimidazole (12 μl, 0.15 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (21 mg, 0.075 mmol). Allow to react at room temperature overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-ynamide (5, 0.53 mg, 4% yield). ESI [M+H] + =332.20 1 H NMR (400MHz, DMSO-d6) δ11.10(s,1H),9.45(d,J=3.1Hz,1H),8.05(s,1H),7.98(d,J=8.1Hz,1H),7.89(d,J=8.4Hz,1H),2.08(s,3H).

[0147] Example 6: Synthesis of N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)ethylenesulfonamide (6)

[0148] Dissolve 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.076 mmol) and ethylenesulfonyl chloride (12.5 mg, 0.098 mmol) in dichloromethane (2 ml), then add 1,8-diazabicycloundec-7-ene (34 mg, 0.22 mmol) and allow to react at room temperature overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)ethylenesulfonamide (6, 1.21 mg, 4.2% yield). ESI [M+H] + =356.20 1H NMR (400MHz, DMSO-d6) δ9.47(s,1H),8.05(d,J=8.1Hz,1H),7.28(d,J=8.0Hz,1H),7.25(s,1H),4.58(t,J=6.8Hz,2H),3.88(t,J=6.7Hz,2H).

[0149] Example 7: Synthesis of (E)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cinnamamide (7)

[0150] Dissolve 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and cinnamic acid (6.4 mg, 0.075 mmol) in N,N-dimethylformamide (2 ml), then add N-methylimidazole (24 μl, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol). Allow to react at room temperature overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification was then performed using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, (E)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cinnamamide (7, 4.8 mg, yield 16.1%). ESI [M+H] + =396.20 1 H NMR (400MHz, DMSO-d6) δ10.69(s,1H),9.45(s,1H),8.24(s,1H),8.05–7.89(m,2H),7.65(d,J=7.9Hz,3H),7.51–7.34(m,3H),6.82(d,J=15.7Hz,1H).

[0151] Example 8: Synthesis of ethyl (Z)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoate (8)

[0152] Dissolve 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (50 mg, 0.19 mmol) and (Z)-4-ethoxy-4-oxobut-2-enoic acid (55 mg, 0.38 mmol) in N,N-dimethylformamide (4 ml), then add N-methylimidazole (62 mg, 0.76 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (107 mg, 0.38 mmol). Allow to react at room temperature overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, ethyl (Z)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoate (8, 42 mg, 57.3% yield). ESI [M+H] + =392.10 1 H NMR (400MHz, DMSO-d6) δ10.99(s,1H),9.48(s,1H),8.18(d,J=2.0Hz,1H),8.02(d,J=8.2Hz,1H),7.93(d,J =2.0Hz,1H),7.20(d,J=15.5Hz,1H),6.76(d,J=15.4Hz,1H),4.23(q,J=7.1Hz,2H),1.27(t,J=7.1Hz,3H).

[0153] Example 9: N 1 Synthesis of -(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)maleamide (9)

[0154] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and (Z)-4-amino-4-oxobut-2-enoic acid (10 mg, 0.09 mmol) were dissolved in N,N-dimethylformamide (2 ml), followed by the addition of N-methylimidazole (25 mg, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol). The reaction was allowed to react at room temperature overnight. After completion, the reaction was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The product was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product N. 1 -(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)maleamide (9, 0.68 mg, yield 2.5%). ESI [M+H] + =363.10 1 H NMR (400MHz, DMSO-d6) δ11.04(s,1H),9.49(s,1H),8.18(d,J=2.0Hz,1H),8.04(d,J=8 .1Hz,1H),7.91(dd,J=8.2,2.1Hz,1H),7.16(d,J=16.1Hz,1H),6.75(d,J=16.1Hz,1H).

[0155] Example 10: Synthesis of N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)methacrylamide (10)

[0156] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) was dissolved in dichloromethane (2 ml) and methacryloyl chloride (10 mg, 0.09 mmol) was added at 0°C and allowed to react at room temperature for 2 hours. After completion, the reaction was diluted with sodium bicarbonate and extracted with dichloromethane. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The product was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)methacrylamide (10, 7.2 mg, 28.8% yield). ESI [M+H] +=334.10 1 H NMR (400MHz, DMSO-d6) δ10.23(s,1H),9.45(s,1H),8.18(s,1H),8.03(d,J=8.2Hz,1H),7.97(d,J=8.2Hz,1H),5.89(s,1H),5.61(s,1H),1.97(s,3H).

[0157] Example 11: Synthesis of 2-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (11)

[0158] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and 2-chloroacrylic acid (16 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N-methylimidazole (25 mg, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol) were added. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, 2-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (11 mg, 7.2 mg, 2.1% yield). ESI [M+H] + =354.00,355.90 1 H NMR (400MHz, DMSO-d6) δ10.66(s,1H),9.48(s,1H),8.46(s,1H),8.15(s,1H),8.03(s,1H),6.51(d,J=2.0Hz,1H),6.18(d,J=2.1Hz,1H).

[0159] Example 12: Synthesis of (Z)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid (12)

[0160] (Z)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid ethyl ester (8, 33 mg, 0.084 mmol) was dissolved in tetrahydrofuran (2 ml) at 0°C, and lithium hydroxide (5.4 mg, 0.127 mmol) was dissolved in water (0.5 ml) and slowly added dropwise into the reaction system for 0.5 hours. After completion of the reaction, the pH was adjusted to 6, and the reaction was extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The product was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to afford the desired product, (Z)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid (12, 1.00 mg, 3.3% yield). ESI [M+H] + =364.00 1 HNMR (400MHz, DMSO-d6) δ10.94(s,1H),9.47(s,1H),8.18(s,1H),8.01(d,J=8. 2Hz, 1H), 7.93 (d, J = 8.1Hz, 1H), 7.11 (d, J = 15.4Hz, 1H), 6.72 (d, J = 15.4Hz, 1H).

[0161] Example 13: Synthesis of 1-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-1H-pyrrole-2,5-dione (13)

[0162] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (32 mg, 0.121 mmol) was dissolved in acetic acid (1 ml), and furan-2,5-dione (13 mg, 0.133 mmol) was added. The mixture was reacted at 60°C for 6 minutes. Two drops of concentrated sulfuric acid were added dropwise, and the mixture was reacted at 60°C for 5 minutes. The mixture was purified by thin-layer chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the target product, 1-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-1H-pyrrole-2,5-dione (13 mg, 1.93 mg, yield 4.63%). ESI [M+H] + =346.2 1H NMR (600MHz, DMSO-d6) δ9.60(s,1H),8.18(d,J=8.0Hz,1H),7.82(s,1H),7.78(d,J=8.0Hz,1H),7.26(s,2H).

[0163] Example 14: Synthesis of (E)-3-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (14)

[0164] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in N,N-dimethylformamide (1 ml), and (E)-3-chloroacrylic acid (16.1 mg, 0.151 mmol), N-methylimidazole (24.8 mg, 0.302 mmol), and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42.4 mg, 0.151 mmol) were added. The mixture was reacted overnight at room temperature. The filtrate was filtered and purified by thin-layer chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the target product, (E)-3-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (14, 1.47 mg, yield 7.74%). ESI [M+H] + =354.2,356.2 1 H NMR (600MHz, DMSO-d6) δ10.68(s,1H),9.47(s,1H),8.15(s,1H),8.00(d,J=8.2 Hz,1H),7.87(d,J=8.2Hz,1H),7.52(d,J=13.2Hz,1H),6.64(d,J=13.2Hz,1H).

[0165] Example 15: Synthesis of N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-2-phenylacrylamide (15)

[0166] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in N,N-dimethylformamide (1 ml), and 2-phenylacrylic acid (22.4 mg, 0.151 mmol), N-methylimidazole (24.8 mg, 0.302 mmol), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (42.4 mg, 0.151 mmol) were added. The mixture was reacted at room temperature for 4 hours. The filtrate was filtered and purified by thin-layer chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the target product, N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-2-phenylacrylamide (15, 1.54 mg, yield 5.16%). ESI [M+H] + =396.2 1 H NMR (600MHz, DMSO-d6) δ10.72(s,1H),9.47(s,1H),8.22(s,1H),8.02(dd,J=22. 5,8.2Hz,2H),7.52(d,J=8.0Hz,2H),7.39(dt,J=14.0,7.2Hz,3H),6.03(s,1H), 5.84(s,1H).

[0167] Example 16: Synthesis of (Z)-3-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (16)

[0168] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in N,N-dimethylformamide (1 ml), and (Z)-3-chloroacrylic acid (16.1 mg, 0.151 mmol), N-methylimidazole (24.8 mg, 0.302 mmol), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (42.4 mg, 0.151 mmol) were added. The mixture was reacted overnight at room temperature. The filtrate was filtered and purified by thin-layer chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the target product, (Z)-3-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (16, 1.44 mg, yield 5.4%). ESI [M+H] + =354.2,356.2 1H NMR (600MHz, DMSO-d6) δ10.68(s,1H),9.46(s,1H),8.14(s,1H),8.00(d,J=8. 2Hz, 1H), 7.92 (d, J = 1.5Hz, 1H), 7.01 (d, J = 8.0Hz, 1H), 6.58 (d, J = 8.0Hz, 1H).

[0169] Example 17: Synthesis of (E)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-3-(pyridin-4-yl)acrylamide (17)

[0170] (E)-3-(4-Pyridyl)acrylic acid (11.3 mg, 0.076 mmol) was dissolved in N,N-dimethylformamide (2 ml), and then N-methylimidazole (12.4 mg, 0.151 mmol) and 7-amino-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-9-one (10 mg, 0.038 mmol) were added to the reaction solution in sequence. Finally, N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (21 mg, 0.075 mmol) was added and the mixture was reacted at room temperature overnight. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic phase was collected and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative plate (petroleum ether: ethyl acetate = 1:1) to give the target product (E)-N-(9-oxo-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-7-yl)-3-(4-pyridyl)acrylamide (17, 1.09 mg, yield 7.3%). ESI [M+H] + =397.1 1 H NMR (600MHz, DMSO-d6) δ10.87(s,1H),9.47(s,1H),8.66(d,J=4.9Hz,2H),8.25(s,1H), 8.02(d,J=8.2Hz,1H),7.95(d,J=8.2Hz,1H),7.64–7.59(m,2H),7.04(d,J=15.8Hz,2H).

[0171] Example 18: Synthesis of (E)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-3-(3-pyridyl)acrylamide (18)

[0172] (E)-3-(3-Pyridyl)acrylic acid (11.3 mg, 0.076 mmol) was dissolved in N,N-dimethylformamide (2 ml), and then N-methylimidazole (12.4 mg, 0.151 mmol) and 7-amino-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-9-one (10 mg, 0.038 mmol) were added to the reaction solution in sequence. Finally, N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (21 mg, 0.075 mmol) was added and the mixture was reacted at room temperature overnight. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic phase was collected and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative plate (petroleum ether:ethyl acetate = 1:1) to give the target product (E)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-3-(3-pyridyl)acrylamide (18, 2.78 mg, yield 19.3%). ESI [M+H] + =397.1 1 H NMR (600MHz, DMSO-d6) δ10.79(s,1H),9.47(s,1H),8.86(s,1H),8.60(d,J=4.5Hz,1H),8.25(s,1H ),8.13–7.84(m,3H),7.70(d,J=15.8Hz,1H),7.50(dd,J=7.7,4.9Hz,1H),6.93(d,J=15.8Hz,1H).

[0173] Example 19: Synthesis of (E)-N-9-oxo-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-7-yl)-3-(2-pyridyl)acrylamide (19)

[0174] (E)-3-(2-Pyridyl)acrylic acid (11.3 mg, 0.079 mmol) was dissolved in N,N-dimethylformamide (2 ml), and then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (28.7 mg, 0.075 mmol), N-N-diisopropylethylamine (19.6 mg, 0.15 mmol) were added to the reaction solution in sequence, and finally 7-amino-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-9-one (10 mg, 0.038 mmol) was added and the reaction was allowed to react at room temperature overnight. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic phase was collected and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (E)-N-(9-oxo-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-7-yl)-3-(2-pyridyl)acrylamide (19, 1.04 mg, yield 6.98%). ESI [M+H] + =397.1 1 H NMR (600MHz, DMSO-d6) δ10.86(s,1H),9.47(s,1H),8.66(d,J=4.4Hz,1H),8.25(s,1H),8.01(d,J=8.2Hz,1H),7.9 7(d,J=8.2Hz,1H),7.89(t,J=7.6Hz,1H),7.73–7.61(m,2H),7.42(dd,J=7.3,4.8Hz,1H),7.35(d,J=15.3Hz,1H).

[0175] Example 20: Synthesis of N-(9-oxo-2-trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-2-(trifluoromethyl)acrylamide (20)

[0176] 7-Amino-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-9-one (10 mg, 0.038 mmol) and 2-(trifluoromethyl)acrylic acid (7.9 mg, 0.056 mmol) were dissolved in N,N-dimethylformamide (2 ml). Pyridine (8.9 mg, 0.11 mmol) and phosphorus oxychloride (8.7 mg, 0.056 mmol) were then slowly added dropwise to the reaction solution, and the reaction was carried out at room temperature for 5 min. The reaction mixture was quenched with water and extracted with ethyl acetate. The organic phase was collected and washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative plate (petroleum ether: ethyl acetate = 2:1) to give the target product N-(9-oxo-2-trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)-2-(trifluoromethyl)acrylamide (20, 1.49 mg, yield 10.02%). ESI [M+H] + =388.1 1 H NMR (600MHz, DMSO-d6) δ10.86(s,1H),9.49(s,1H),8.11(d,J=1.8Hz,1H),8.03(d,J=8.2Hz,1H),7.99(dd,J=8.2,1.9Hz,1H),6.67(s,1H),6.61(s,1H).

[0177] Example 21: Synthesis of (E)-N,N-dimethyl-N'-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)carboxamide (21)

[0178] 7-Amino-2-trifluoromethyl-9H-indeno[2,1-d]pyrimidin-9-one (10 mg, 0.0377 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-chloroethanesulfonyl chloride (5 μl, 0.0453 mmol) and potassium carbonate (16 mg, 0.116 mmol) were added. The mixture was allowed to react at room temperature for 1 hour. Saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol = 15:1) to afford the desired product (E)-N,N-dimethyl-N'-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)carboxamide (21, 2.32 mg, 19.2% yield). ESI [M+H] + =321.0 1H NMR (600MHz, DMSO-d6) δ9.18(s,1H),7.96(s,1H),7.67(d,J=7.8Hz,1H),7.20(d,J=49.9Hz,2H),2.88(d,J=65.6Hz,3H),2.32(s,3H).

[0179] Example 22: Synthesis of (E)-4-(dimethylamino)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-enamide (22)

[0180] Dissolve 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.75 mmol) and (E)-4-(dimethylamino)but-2-enoic acid hydrochloride (25 mg, 0.15 mmol) in N,N-dimethylformamide (2 ml), then add N-methylimidazole (24 μL, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol). Allow to react at room temperature overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was then purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 22 min) to obtain the desired product, (E)-4-(dimethylamino)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-enamide (22 mg, 2.82 mg, 7.14% yield). ESI [M+H] + =376.90 1 H NMR (400MHz, DMSO-d6) δ10.82(s,1H),9.46(s,1H),8.21(d,J=2.0Hz,1H),8.00(d,J=8.1Hz,1H),7.91(dd,J =8.2, 2.1Hz, 1H), 6.81 (dt, J = 14.8, 7.0Hz, 1H), 6.47 (d, J = 15.3Hz, 1H), 3.92 (d, J = 7.0Hz, 2H), 2.78 (s, 6H).

[0181] Example 23: Synthesis of 2,2,2-trifluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (23)

[0182] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and trifluoroacetic acid (11 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N-methylimidazole (25 mg, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol) were added and the reaction was allowed to proceed overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2,2,2-trifluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (23 mg, 2.54 mg, 9.4% yield). ESI [M+H] + =362.10 1 H NMR (400MHz, DMSO-d6)δ9.52(s,1H),8.12–8.05(m,2H),8.00(dd,J=8.2,2.0Hz,1H).

[0183] Example 24: Synthesis of 2-(hydroxymethyl)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (24)

[0184] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and 2-(hydroxymethyl)acrylic acid (15.3 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and pyridine (19 μl, 0.225 mmol) was added. Phosphorus oxychloride (14 μl, 0.15 mmol) was then added at 0°C. The reaction was allowed to proceed at room temperature. After completion, the reaction solution was added dropwise to a sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification was then performed using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2-(hydroxymethyl)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (24 mg, 0.89 mg, 3.4% yield). ESI [M+H] + =350.30 1H NMR (400MHz, DMSO-d6) δ10.33(s,1H),9.45(s,1H),8.19(s,1H),8.05–7.94(m,1H),5.96(s,1H),5.75(s,1H),5.18(s,1H),4.24(s,2H).

[0185] Example 25: Synthesis of N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propiolamide (25)

[0186] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and propiolic acid (10 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and pyridine (19 μl, 0.225 mmol) was added. Phosphorus oxychloride (14 μl, 0.15 mmol) was then added at 0°C. The reaction was allowed to proceed at room temperature. After completion, the reaction solution was added dropwise to a sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification was then performed using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propynamide (25 mg, 1.78 mg, 7.4% yield). ESI [M+H] + =318.20 1 H NMR (400MHz, DMSO-d6) δ11.28(s,1H),9.47(s,1H),8.05(d,J=1.6Hz,1H),8.00(d,J=8.2Hz,1H),7.90(dd,J=8.2,1.8Hz,1H),4.56(s,1H).

[0187] Example 26: Synthesis of 2-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (26)

[0188] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in dichloromethane (2 ml), and triethylamine (16 μl, 0.113 mmol) was added. 2-chloroacetyl chloride (12 μl, 0.151 mmol) was slowly added under ice-cooling. The mixture was allowed to react at room temperature for one hour. After completion, the reaction was quenched with saturated aqueous ammonium chloride solution and extracted with dichloromethane. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified twice by column chromatography to obtain the desired product, 2-chloro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (26 mg, 5.21 mg, 20.3% yield). ESI [M+H] + =342.3 1 H NMR (400MHz, DMSO-d6) δ10.85(s,1H),9.47(s,1H),8.10(dd,J=5.7,1.7Hz,1H),8.00(dd,J=8.1,2.9Hz,1H),7.87(dd,J=8.2,1.6Hz,1H),4.34(s,2H).

[0189] Example 27: Synthesis of 2-benzyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (27)

[0190] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-benzyl acrylic acid (24.5 mg, 0.151 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.151 mmol), and 1-methylimidazole (25 μl, 0.3 mmol) were added. The mixture was reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified twice using preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-95% over 40 min) to obtain the desired product, 2-benzyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (27 mg, 1.54 mg, 5.0% yield).

[0191] ESI[M+H] + =410.3 1H NMR (400MHz, DMSO-d6) δ10.37(s,1H),9.44(s,1H),8.15(s,1H),7.98(dt,J=19.4,5.0Hz,2H),7.25(qd, J=14.8,7.5Hz,5H),5.95(s,1H),5.58(s,1H),3.70(s,2H).

[0192] Example 28: Synthesis of 7-(prop-2-yn-1-ylamino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (28)

[0193] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 3-bromoprop-1-yne (13.5 μl, 0.151 mmol) and potassium carbonate (16 mg, 0.113 mmol) were added and reacted at room temperature for 3 hours. The temperature was then raised to 80°C and the reaction was allowed to proceed overnight. After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified twice using preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 35 min) to obtain the desired product, 7-(prop-2-yn-1-ylamino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (28 mg, 1.38 mg, 6.04% yield). ESI [M+H] + =304.2 1 H NMR (600MHz, DMSO-d6) δ9.21 (s, 1H), 7.72 (d, J = 8.3Hz, 1H), 7.03 (t, J = 4.5Hz, 2H) ,6.92(dd,J=8.3,2.3Hz,1H), 4.03(dd,J=5.2,1.8Hz,2H), 3.17(t,J=2.3Hz,1H).

[0194] Example 29: Synthesis of (E)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid (29)

[0195] Step 1: Synthesis of ethyl (E)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoate (29-a)

[0196] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.1132 mmol) was dissolved in N,N-dimethylformamide (2 ml), and (E)-4-ethoxy-4-oxobut-2-enoic acid (33 mg, 0.2264 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (64 mg, 0.2264 mmol), and 1-methylimidazole (38 μl, 0.4528 mmol) were added. The mixture was reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate. After concentration under reduced pressure, the mixture was plated (petroleum ether:ethyl acetate = 1:1) to obtain the target product (E)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid ethyl ester (29-a, 20 mg, yield 45.2%). ESI [M+H] + =392.2.

[0197] Step 2: Synthesis of (E)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid (29)

[0198] (E)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid ethyl ester (29-a, 20 mg, 0.052 mmol) was dissolved in a mixed solution (2 ml) of tetrahydrofuran and water (5:1), and lithium hydroxide monohydrate (3.3 mg, 0.078 mmol) was added at low temperature and reacted at room temperature for one hour. After completion of the reaction, the pH was adjusted to 6 with 1 M aqueous hydrogen chloride solution, followed by extraction with ethyl acetate. The organic phase was collected, washed with brine, and dried over anhydrous sodium sulfate. The phase was concentrated under reduced pressure and purified using a preparative plate (petroleum ether:ethyl acetate = 1:1). The product was then purified using a preparative liquid chromatography column (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B:ACN, B%: 10%-70% over 20 min) to afford the desired product, (E)-4-oxo-4-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)but-2-enoic acid (29 mg, 4.78 mg, yield 25.74%). ESI [M+H] + =363.4 1H NMR (600MHz, DMSO-d6) δ10.94(s,1H),9.47(s,1H),8.17(s,1H),8.01(d,J=8.2 Hz,1H),7.92(d,J=8.1Hz,1H),7.11(d,J=15.4Hz,1H),6.71(d,J=15.4Hz,1H).

[0199] Example 30: Synthesis of (Z)-3-cyano-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (30)

[0200] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.0755 mmol) was dissolved in N,N-dimethylformamide (2 ml), and (Z)-3-cyanoacrylic acid (22 mg, 0.226 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (63.5 mg, 0.226 mmol), and 1-methylimidazole (38 μl, 0.453 mmol) were added. The mixture was reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified once using a preparative plate (petroleum ether:ethyl acetate = 1:1) and then twice using a preparative liquid chromatography column (Pack Column 20 ID*250 mm*10 μm; A:H2O (0.1% FA), B:ACN, B%: 10%-95% over 40 min) to obtain the desired product, (Z)-3-cyano-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (30, 2.89 mg, yield 7.42%). ESI [M+H] + =345.2 1 H NMR (400MHz, DMSO-d6) δ11.05(s,1H),9.47(s,1H),8.16(s,1H),8.02(d,J=8.1 Hz,1H),7.89(d,J=7.9Hz,1H),7.16(d,J=16.1Hz,1H),6.73(d,J=16.1Hz,1H).

[0201] Examples 31 and 32: Synthesis of 2-(Morpholinomethyl)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (31), Synthesis of (E)-4-Morpholinomethyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-enamide (32)

[0202] 2-(Morpholinomethyl)acrylic acid (96.9 mg, 0.5657 mmol) was dissolved in tetrahydrofuran (6 ml) and cooled to 0°C under a nitrogen atmosphere. N-methylmorpholine (75 μl, 0.679 mmol) and isobutyl chloroformate (58.5 μl, 0.226 mmol) were added, and the mixture was stirred at 0°C for 1 hour. A solution of 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60 mg, 0.226 mmol) in pyridine (2 ml) was then added. The reaction was allowed to react at room temperature overnight. After the reaction, the reaction mixture was poured into an ice-cold saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and then washed with saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified three times using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: CAN, B%: 10%-95% over 25 min) to obtain the desired product, 2-(morpholinomethyl)-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (31 mg, 1.68 mg, yield 3.55%). ESI [M+H] + =419.20 1 H NMR (400 MHz, DMSO-d6) δ 9.18 (s, 1H), 7.68 (d, J = 8.2 Hz, 1H), 7.12 (t, J = 5.7 Hz, 1H), 6.98 (s, 1H), 6.89 (d, J = 7.9 Hz, 1H), 5.43 (s, 1H), 5.19 (s, 1H), 4.02 (d, J = 5.4 Hz, 2H), 3.44 (d, J = 19.1 Hz, 8H). The target product (E)-4-morpholino-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-enamide (32, 2.14 mg, yield 4.52%). ESI [M+H] + =419.20 1 H NMR (400MHz, DMSO-d6) δ11.23(s,1H),9.47(s,1H),8.19(s,1H),7.99(d,J=8.2Hz ,1H),7.90(d,J=8.2Hz,1H),6.10(s,1H),5.69(s,1H),3.65(s,2H),2.50(s,8H).

[0203] Example 33: Synthesis of 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (33)

[0204] Dissolve 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) in N,N-dimethylformamide (2 ml), add 2-fluoropropionic acid (18 μl, 0.226 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (63.5 mg, 0.2264 mmol), and 1-methylimidazole (38 μl, 0.453 mmol), and allow to react at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product, 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (33, 5.91 mg, yield 15.4%). ESI [M+H] + =340.6 1 H NMR (400MHz, DMSO-d6) δ10.75(s,1H),9.47(s,1H),8.19(s,1H),8.00(q,J=8.5Hz,2H),5.27(dq,J=48.5,6.7Hz,1H),1.55(dd,J=24.5,6.7Hz,3H).

[0205] Example 34: Synthesis of 2-(dimethylamino)methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (34)

[0206] Step 1: Synthesis of 2-(dimethylaminomethyl)acrylic acid (34-a)

[0207] Dissolve 2-(Bromomethyl)acrylic acid (100 mg, 0.61 mmol), dimethylamine hydrochloride (49 mg, 0.6 mmol), and N,N-diisopropylethylamine (211 μl, 1.21 mmol) in dichloromethane (5 ml) and react at room temperature for 1 hour. Filter and concentrate under reduced pressure to obtain the desired product, 2-(dimethylaminomethyl)acrylic acid (34-a, 60 mg, yield 60.1%). ESI [M+H] + =130.7

[0208] Step 2: Synthesis of 2-(dimethylamino)methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (34)

[0209] 2-(Dimethylaminomethyl)acrylic acid (34-a, 49 mg, 0.38 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 7-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (50 mg, 0.19 mmol) and pyridine (76 μl, 0.94 mmol) were added. Phosphorus oxychloride (53 μl, 0.57 mmol) was added at 0°C under nitrogen protection and the reaction was carried out for 1.5 hours. The mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2-(dimethylamino)methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (34 mg, 2.46 mg, 3.5% yield). ESI [M+H] + =377.2 1 H NMR (400MHz, DMSO-d6) δ11.41(s,1H),9.47(s,1H),8.15(s,1H),8.00(d,J=8.2Hz,1H),7.92(d,J=9.8Hz,1H),6.87(s,1H), 6.66(s,1H),6.08(s,1H),5.68(s,1H),2.28(s,6H).

[0210] Example 35: Synthesis of 3-cyano-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (35)

[0211] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.08 mmol), 3-cyanopropionic acid (15 mg, 0.15 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol) and NMI (25 μl, 0.31 mmol) were dissolved in N,N-dimethylformamide (2 ml) and reacted at room temperature overnight. The mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the target product, 3-cyano-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (35 mg, 2.34 mg, 9% yield). ESI [M+H] + =346.9 1 H NMR (400MHz, DMSO-d6) δ10.61(s,1H),9.45(s,1H),8.09(s,1H),7.98(d,J=8.2Hz,1H),7.85(d,J=8.2Hz,1H),2.76(s,4H).

[0212] Example 36: Synthesis of 2,2-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (36)

[0213] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2,2-difluoropropionic acid (25 mg, 0.226 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product, 2,2-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (36, 4.72 mg, yield 11.68%). ESI [M+H] + =358.00 1H NMR (400MHz, DMSO-d6) δ11.02(s,1H),9.50(s,1H),8.17(s,1H),8.10–7.97(m,2H),1.92(dd,J=35.4,15.7Hz,3H).

[0214] Example 37: Synthesis of (E)-4,4,4-trifluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-enamide (37)

[0215] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and (E)-4,4,4-trifluorobut-2-enoic acid (31.7 mg, 0.226 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (E)-4,4,4-trifluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)but-2-enamide (37, 5.06 mg, yield 11.55%). ESI [M+H] + =388.00 1 H NMR (400MHz, DMSO-d6) δ11.04(s,1H),9.47(s,1H),8.16(d,J=1.5Hz,1H),8.01(d,J=8.2Hz,1H),7.87(dd,J=8.2,1.7Hz,1H),7.07–6.91(m,2H).

[0216] Example 38: Synthesis of 2-chloro-2,2-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (38)

[0217] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and 2-chloro-2,2-difluoroacetic acid (20 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N-methylimidazole (25 mg, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol) were added and the reaction was allowed to proceed overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2-chloro-2,2-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (38 mg, 8.24 mg, 29.4% yield). ESI [M+H] + =378.10, 380.10 1 H NMR (400MHz, DMSO-d6) δ11.53(s,1H),9.52(s,1H),8.13–8.05(m,2H),8.01(dd,J=8.2,2.0Hz,1H).

[0218] Example 39: Synthesis of 9-oxo-N-(prop-2-yn-1-yl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (39)

[0219] 9-Oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxylic acid (20 mg, 0.058 mmol), prop-2-yn-1-amine (5 mg, 0.08 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (39 mg, 0.10 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N,N-diisopropylethylamine (38 μl, 0.20 mmol) was added and the reaction was allowed to react overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-80% over 23 min) to obtain the desired product, 9-oxo-N-(prop-2-yn-1-yl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (39 mg, 1.78 mg, 8.09% yield). ESI [M+H] + =332.10 1 H NMR (400MHz, DMSO-d6) δ9.65 (s, 1H), 9.26 (t, J = 5.3Hz, 1H), 8.28 (d, J = 9.8Hz, 2H),8.18(d,J=7.7Hz,1H),4.10(dd,J=5.3,2.3Hz,2H),3.17(t,J=2.4Hz,1H)

[0220] Example 40: Synthesis of 2-cyano-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (40)

[0221] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and 2-cyanopropionic acid (11 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N-methylimidazole (25 mg, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol) were added and the reaction was allowed to proceed overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2-cyano-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (40 mg, 11.23 mg, 43.2% yield). ESI [M+H] + =347.20 1 H NMR (400MHz, DMSO-d6) δ10.84(s,1H),9.47(s,1H),8.05(d,J=1.1Hz,1H),8.01(d,J= 8.2Hz, 1H), 7.85 (dd, J=8.2, 1.5Hz, 1H), 4.00 (q, J=7.2Hz, 1H), 1.54 (d, J=7.2Hz, 3H).

[0222] Example 41: Synthesis of 2,2-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (41)

[0223] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) and 2,2-difluoroacetic acid (11 μl, 0.15 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N-methylimidazole (25 mg, 0.3 mmol) and N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.15 mmol) were added and the reaction was allowed to proceed overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2,2-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acetamide (41 mg, 7.09 mg, 31.6% yield). ESI [M+H] + =344.20 1 H NMR (400MHz, DMSO-d6) δ9.49 (s, 1H), 8.13 (d, J = 1.1Hz, 1H), 8.03 (d, J = 8.2Hz, 1H), 7.97 (dd, J = 8.2, 1.4Hz, 1H), 6.44 (t, J = 53.5Hz, 1H).

[0224] Example 42: Synthesis of N-(cyanomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (42)

[0225] 9-Oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxylic acid (20 mg, 0.058 mmol), 2-aminoacetonitrile (3 mg, 0.08 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (39 mg, 0.10 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N,N-diisopropylethylamine (38 μl, 0.20 mmol) was added and the reaction was allowed to react overnight. After completion of the reaction, the mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-80% over 23 min) to obtain the desired product, N-(cyanomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (42 mg, 2.21 mg, 9.3% yield). ESI [M+H] + =333.10 1 H NMR (400MHz, DMSO-d6) δ9.66 (s, 1H), 9.55 (t, J = 5.2Hz, 1H), 8.32–8.25 (m, 2H), 8.21 (d, J = 7.8Hz, 1H), 4.37 (d, J = 5.3Hz, 2H).

[0226] Example 43: Synthesis of 2-fluoro-N-(3-oxo-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)propyl)acrylamide (43)

[0227] Step 1: Synthesis of tert-butyl (3-oxo-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)propyl)carbamate (43-a)

[0228] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (50 mg, 0.19 mmol) and 3-((tert-butoxycarbonyl)amino)propionic acid (71 mg, 0.377 mmol) were dissolved in N,N-dimethylformamide (3 ml), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (144 mg, 0.377 mmol) and N-methylimidazole (62 μl, 0.756 mmol) and reacted at room temperature overnight. After completion of the reaction, the mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (tert-butyl 3-oxo-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)propyl)carbamate (43-a, 80 mg, yield 97.6%). ESI [M+H] + =437.1

[0229] Step 2: Synthesis of 3-amino-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (43-b)

[0230] Tert-butyl (3-oxo-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)propyl)carbamate (43-a, 30 mg, 0.069 mmol) was dissolved in ethyl acetate (2 ml). 4 M hydrochloric acid in ethyl acetate (2 ml) was added at 0°C and reacted for 1.5 hours. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the target product, 3-amino-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (43-b, 45 mg). ESI [M+H] + =337.2

[0231] Step 3: Synthesis of 2-fluoro-N-(3-oxo-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)propyl)acrylamide (43)

[0232] 3-Amino-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (43-b, 20 mg, 0.059 mmol) and 2-fluoroacrylic acid (8 mg, 0.089 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N,N-diisopropylethylamine (33 μl, 0.18 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (34 mg, 0.089 mmol) were added and reacted at room temperature overnight. The mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the desired product, 2-fluoro-N-(3-oxo-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)amino)propyl)acrylamide (43 mg, 3.56 mg, 14.7% yield). ESI [M+H] + =409.02 1 H NMR (400MHz, DMSO-d6) δ10.46(s,1H),9.44(s,1H),8.65(s,1H),8.11(d,J=1.2Hz,1H),7.96(d,J=8.2Hz,1H),7.84(dd,J=8.2,1. 5Hz, 1H), 5.52 (dd, J = 48.2, 3.3Hz, 1H), 5.31 (ddd, J = 19.0, 10.9, 2.5Hz, 1H), 3.47 (dd, J = 12.6, 6.5Hz, 2H), 2.62 (t, J = 6.8Hz, 2H).

[0233] Example 44: Synthesis of (S)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (44)

[0234] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) was dissolved in N,N-dimethylformamide (2 ml), and (S)-2-fluoropropionic acid (10 mg, 0.11 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.226 mmol) and N-methylimidazole (25 μl, 0.315 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (S)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (44 mg, 7.96 mg, yield 31.1%). ESI [M+H] + =340.10 1 H NMR (400MHz, DMSO-d6) δ10.54(s,1H),9.45(s,1H),8.16(s,1H),8.01–7.96(m,2H),5.24(dq,J=48.4,6.7Hz,1H),1.55(dd,J=24.5,6.7Hz,3H).

[0235] Example 45: Synthesis of N-(2-(2-fluoroacrylamido)ethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (45)

[0236] Step 1: Synthesis of tert-butyl (2-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamido)ethyl)carbamate (45-a)

[0237] 9-Oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxylic acid (30 mg, 0.10 mmol) and tert-butyl (2-aminoethyl)carbamate (24 mg, 0.15 mmol) were dissolved in N,N-dimethylformamide (3 ml), N,N-diisopropylethylamine (55 μl, 0.3 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (57 mg, 0.15 mmol) and reacted at room temperature overnight. After completion of the reaction, the mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (tert-butyl 2-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamido)ethyl)carbamate (45-a, 40 mg, yield 90.9%). ESI [M+H] + =437.1

[0238] Step 2: Synthesis of N-(2-aminoethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (45-b)

[0239] Dissolve tert-butyl (2-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamido)ethyl)carbamate (45-a, 40 mg, 0.092 mmol) in ethyl acetate (2 ml). Add 4M hydrochloric acid in ethyl acetate (2 ml) at 0°C and react for 1.5 hours. After completion of the reaction, concentrate under reduced pressure to obtain the target product, N-(2-aminoethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (45-b, 38 mg). ESI [M+H] + =337.2

[0240] Step 3: Synthesis of N-(2-(2-fluoroacrylamido)ethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (45)

[0241] N-(2-aminoethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (45-b, 20 mg, 0.0059 mmol), 2-fluoroacrylic acid (8 mg, 0.089 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (34 mg, 0.089 mmol) were dissolved in N,N-dimethylformamide (2 ml), and N,N-diisopropylethylamine (33 μl, 0.18 mmol) was added, and the reaction was allowed to react at room temperature overnight. The mixture was diluted with sodium bicarbonate and extracted with ethyl acetate. The organic phase was collected and repeatedly washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to obtain the target product, N-(2-(2-fluoroacrylamido)ethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidine-7-carboxamide (45 mg, 0.83 mg, 3.4% yield). ESI [M+H] + =409.02 1 H NMR (400MHz, DMSO-d6) δ11.69(s,1H),9.64(s,1H),8.89–8.85(m,1H),8.64(s,1H),8.25(dd,J=4.0,2.4Hz,1H),8.17(d,J=8.2Hz,1H),5.39– 5.34(m,1H),5.25(dd,J=15.7,3.3Hz,1H),4.28–4.24(m,1H),4.16(dd,J=14.7,6.8Hz,1H),4.08(dd,J=13.2,6.9Hz,1H),4.05–4.01(m,1H).

[0242] Example 46: Synthesis of 2-fluoro-N-methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (46)

[0243] Step 1: Synthesis of 7-(methylamino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (46-a)

[0244] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60 mg, 0.23 mmol) was dissolved in acetonitrile (3 ml), potassium carbonate (47 mg, 0.34 mmol) and iodomethane (48 mg, 0.34 mmol) were added, and the mixture was sealed and reacted at 90°C overnight. After completion of the reaction, the mixture was cooled to room temperature, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 2:1) to obtain the desired product, 7-(methylamino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (46-a, 14 mg, yield 22.22%). ESI [M+H] + =280.3

[0245] Step 2: Synthesis of 2-fluoro-N-methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (46)

[0246] 7-(Methylamino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (46-a, 14 mg, 0.05 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-fluoroacrylic acid (9 mg, 0.10 mmol), N,N,N',N'-tetramethylchlorouronium hexafluorophosphate (28 mg, 0.10 mmol), and N-methylimidazole (17 μl, 0.20 mmol) were added and reacted at room temperature for 12 h. 2-Fluoroacrylic acid (9 mg, 0.10 mmol) and N-(chloro(dimethylamino)methylene)-N-methylmethylammonium hexafluorophosphate (28 mg, 0.10 mmol) were then added and the temperature was raised to 40°C for 12 h. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified twice using a preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: CAN, B%: 10%-95% over 35 min) to obtain the desired product, 2-fluoro-N-methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (46 mg, 1.18 mg, 6.7% yield). ESI [M+H] + =352.2 1 H NMR (600MHz, DMSO-d6) δ9.58(s,1H),8.11(d,J=7.9Hz,1H),7.84(s,1H),7.76(d,J=7.9Hz,1H),5.35–5.21(m,2H),3.33(s,3H).

[0247] Example 47: Synthesis of 1-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (47)

[0248] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 1-fluorocyclopropane-1-carboxylic acid (24 mg, 0.226 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product, 1-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (47, 4.45 mg, yield 10.07%). ESI [M+H] + =352.20 1 H NMR (600MHz, DMSO-d6) δ10.69(s,1H),9.47(s,1H),8.21(d,J=1.3Hz,1H),8.08(dd,J=8.2 ,1.6Hz,1H),8.00(d,J=8.2Hz,1H),1.48(ddd,J=18.1,8.5,5.6Hz,2H),1.40–1.31(m,2H).

[0249] Example 48: Synthesis of (R)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (48)

[0250] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and (R)-2-fluoropropionic acid (20.8 mg, 0.226 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (64 mg, 0.226 mmol) and N-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (R)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (48 mg, 4.42 mg, yield 11.50%). ESI [M+H] + =340.3 1 H NMR (600MHz, DMSO-d6) δ10.76(s,1H),9.46(s,1H),8.19(s,1H),8.00(q,J=8.2Hz,2H),5.27(dq,J=48.5,6.7Hz,1H),1.55(dd,J=24.6,6.7Hz,3H).

[0251] Example 49: Synthesis of 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-6-yl)acrylamide (49)

[0252] Step 1: Synthesis of 4-nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (49-a)

[0253] 2-Bromo-4-nitrobenzaldehyde (300 mg, 1.30 mmol) was dissolved in acetonitrile (10 ml), and potassium phosphate (827 mg, 3.9 mmol), (2-(trifluoromethyl)pyrimidin-5-yl)boronic acid (I-1, 345 mg, 1.82 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (47.6 mg, 0.065 mmol) were added. The mixture was allowed to react at 100°C overnight. After completion of the reaction, the mixture was cooled to room temperature, filtered, concentrated under reduced pressure, and purified by normal phase chromatography (petroleum ether:ethyl acetate = 2:1) to obtain the desired product, 4-nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (49-a, 340 mg, 87.63% yield). ESI [M+H] + =298.1.

[0254] Step 2: Synthesis of 4-amino-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (49-b)

[0255] 4-Nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (49-a, 340 mg, 1.14 mmol) was dissolved in a mixture of 1,4-dioxane (8 ml) and water (8 ml) and stirred at 15°C for 10 minutes. Sodium sulfide nonahydrate (907 mg, 3.76 mmol) was then added and stirred at 15°C for 10 minutes. The mixture was then allowed to react at room temperature for 2 hours. After completion of the reaction, the mixture was extracted with ethyl acetate, and the organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure and purified on a preparative plate (petroleum ether:ethyl acetate = 2:1) to obtain the desired product, 4-amino-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (49-b, 180 mg, 59.02% yield). ESI [M+H] + =268.1

[0256] Step 3: Synthesis of tert-butyl (4-formyl-3-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)carbamate (49-c)

[0257] 4-Amino-2-(2-(trifluoromethyl)pyrimidin-5-yl)benzaldehyde (49-b, 180 mg, 0.674 mmol) was dissolved in tetrahydrofuran (6 ml), and 4-dimethylaminopyridine (4.2 mg, 0.0337 mmol) and di-tert-butyl dicarbonate (4.2 mg, 0.169 mmol) were added. The mixture was allowed to react at room temperature for 5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain the target product, tert-butyl (4-formyl-3-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)carbamate (49-c, 320 mg, crude product). ESI [M+H] + =368.2

[0258] Step 4: Synthesis of tert-butyl (9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-6-yl)carbamate (49-d)

[0259] Tert-butyl (4-formyl-3-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)carbamate (49-c, 320 mg, 0.685 mmol) was dissolved in 1,2-dichloroethane (10 ml). Tetrabutylammonium iodide (12.6 mg, 0.034 mmol) and 2-methylpropane hydroperoxide (aqueous solution) (394 μl, 4.11 mmol) were added to the reaction mixture. The atmosphere was purged with nitrogen three times, and the reaction was allowed to proceed overnight at 100°C in a sealed container. The reaction mixture was filtered to remove insoluble matter. The filtrate was concentrated under reduced pressure and purified on a preparative plate (petroleum ether:ethyl acetate = 3:1) to obtain the desired product, tert-butyl (9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-6-yl)carbamate (49-d, 50 mg, yield 15.72%). ESI [M+H]+ =366.2

[0260] Step 5: Synthesis of 6-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (49-e)

[0261] Dissolve tert-butyl (9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-6-yl)carbamate (49-d, 50 mg, 0.674 mmol) in ethyl acetate (1 ml). Add hydrogen chloride in ethyl acetate (3 ml, 4 M) at 0°C and allow to react overnight at room temperature. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain the target product, 6-amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (49-e, 240 mg, crude product). ESI [M+H] + =266.1

[0262] Step 6: Synthesis of 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-6-yl)acrylamide (49)

[0263] 6-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (49-e, 24 mg, 0.091 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-fluoroacrylic acid (18 mg, 0.0109 mmol), pyridine (23 μl, 0.273 mmol) and phosphorus oxychloride (12.5 μl, 0.1365 mmol) were added and reacted at room temperature for 48 h. After completion of the reaction, the reaction was quenched with ice water and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified once using a preparative plate (petroleum ether:ethyl acetate = 1.5:1). The product was then purified a second time using a preparative liquid chromatography column (Pack Column 20 ID*250mm*10um; A:H2O (0.1% FA), B:ACN, B%: 10%-90% over 40 min) to obtain the desired product, 2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-6-yl)acrylamide (49 mg, 0.77 mg, 2.50% yield). ESI [M+H] + =338.1 1 H NMR (600MHz, DMSO-d6) δ9.51(s,1H),8.44(s,1H),8.41(s,1H),7.83(q,J=8.3Hz,2H),5.83(dd,J=47.4,2.8Hz,1H),5.55(dd,J=15.3,2.9Hz,1H).

[0264] Example 50: Synthesis of 1-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclobutane-1-carboxamide (50)

[0265] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 1-fluorocyclobutane-1-carboxylic acid (13 mg, 0.11 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.226 mmol) and 1-methylimidazole (25 μl, 0.315 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product, 1-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclobutane-1-carboxamide (50, 7.71 mg, yield 28.6%). ESI [M+H] + ,=366.10 1 H NMR (400MHz, DMSO-d6) δ10.44(d,J=2.9Hz,1H),9.46(s,1H),8.21(d,J=2.0Hz,1H),8.06(dd,J=8.2,2.0Hz,1H ),7.98(d,J=8.2Hz,1H),2.69–2.58(m,2H),2.49–2.37(m,2H),2.01–1.90(m,1H),1.79(q,J=11.1,9.9Hz,1H).

[0266] Example 51: Synthesis of 2-fluoro-2-methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propionamide (51)

[0267] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-fluoro-2-methylpropionic acid (24 mg, 0.226 mmol), N,N,N',N'-tetramethylchlorouronium hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified once using a preparative plate (petroleum ether:ethyl acetate = 1:1). The product was then purified a second time using a preparative liquid chromatography column (Pack Column 20 ID*250mm*10um; A:H2O (0.1% FA), B:ACN, B%: 10%-90% over 30 min) to obtain the desired product, 2-fluoro-2-methyl-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)propanamide (51, 2.06 mg, 40% yield). ESI [M+H] + =354.30 1 H NMR (400MHz, DMSO-d6) δ10.43(d,J=2.6Hz,1H),9.45(s,1H),8.20(s,1H),8.12–7.89(m,2H),1.60(d,J=21.9Hz,6H).

[0268] Example 52: Synthesis of (1R,2R)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (52)

[0269] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (20 mg, 0.075 mmol) was dissolved in N,N-dimethylformamide (2 ml), and (1R,2R)-2-fluorocyclopropane-1-carboxylic acid (16 mg, 0.11 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (42 mg, 0.226 mmol) and 1-methylimidazole (25 μl, 0.315 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (1R,2R)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (52, 3.60 mg, yield 13.8%). ESI [M+H]+ =352.20 1 H NMR(400MHz,DMSO-d6)δ10.76(s,1H),9.44(s,1H),8.14–8.06(m,1H),7.96(d,J=8.2Hz,1H),7 .91–7.82(m,1H),4.99(dtd,J=66.1,6.2,3.6Hz,1H),2.04(q,J=7.3Hz,1H),1.82–1.61(m,2H).

[0270] Example 53: Synthesis of 2-fluoro-N-(7-oxo-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-5-yl)acrylamide (53)

[0271] Step 1: Synthesis of N-(2-methylnaphthalen-1-yl)acetamide (53-a)

[0272] 2-Methylnaphthyl-1-amine (3 g, 19.1 mmol) was dissolved in dichloromethane (30 ml). Acetyl chloride (2 ml, 28.65 mmol) and triethylamine (5.3 ml, 38.2 mmol) were added at 0°C and allowed to react at room temperature for 3 hours. After completion of the reaction, the mixture was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a flash column (petroleum ether:ethyl acetate = 0-25%) to obtain the desired product, N-(2-methylnaphthyl)acetamide (53-a, 3.7 g, yield 97.4%). ESI [M+H] + =200.20

[0273] Step 2: Synthesis of N-(2-methyl-4-nitronaphthalen-1-yl)acetamide (53-b)

[0274] N-(2-methylnaphthalen-1-yl)acetamide (53-a, 3 g, 15 mmol) was dissolved in acetic anhydride (50 ml). Nitric acid (1.48 ml, 16.5 mmol) was slowly added dropwise at 0°C and allowed to react at room temperature for 3 hours. After completion of the reaction, the pH was adjusted to 8 with sodium hydroxide solution, and the reaction was quenched with sodium sulfite. The mixture was extracted with ethyl acetate, and the organic phase was collected and dried over anhydrous sodium sulfate. After concentration under reduced pressure, the mixture was purified on a flash column (petroleum ether:ethyl acetate = 0-25%) to obtain the desired product, N-(2-methyl-4-nitronaphthalen-1-yl)acetamide (53-b, 1.78 g, yield 48.6%). ESI [M+H] + =245.10

[0275] Step 3: Synthesis of 2-methyl-4-nitronaphthalene-1-amine (53-c)

[0276] N-(2-methyl-4-nitronaphthalen-1-yl)acetamide (53-b, 1.78 g, 7.3 mmol) was dissolved in ethanol (30 ml), and 6 M hydrochloric acid solution (6 ml) was added. The mixture was reacted at 100°C for 16 hours. After completion of the reaction, the pH was adjusted to 8 with sodium hydroxide solution, and the mixture was extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a flash column (petroleum ether:ethyl acetate = 0-35%) to obtain the desired product, 2-methyl-4-nitronaphthalen-1-amine (53-c, 1.3 g, yield 91.5%). ESI [M+H] + =203.10

[0277] Step 4: Synthesis of 1-iodo-2-methyl-4-nitronaphthalene (53-d)

[0278] 2-Methyl-4-nitronaphthalene-1-amine (53-c, 1.3 g, 6.43 mmol) and cuprous iodide (3.66 g, 19.3 mmol) were dissolved in acetonitrile (30 ml). Isoamyl nitrite (2.58 ml, 19.3 mmol) was added dropwise at 0°C. The mixture was allowed to react overnight at room temperature. After completion of the reaction, the reaction solution was diluted with water and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a flash column (petroleum ether:ethyl acetate = 0-5%) to obtain the desired product, 1-iodo-2-methyl-4-nitronaphthalene (53-d, 1 g, 50.1% yield). ESI [M+H] + =314.10,316.10

[0279] Step 5: Synthesis of 5-(2-methyl-4-nitronaphthalen-1-yl)-2-(trifluoromethyl)pyrimidine (53-e)

[0280] 1-Iodo-2-methyl-4-nitronaphthalene (53-d, 1 g, 3.19 mmol), (2-(trifluoromethyl)pyrimidin-5-yl)boric acid (920 mg, 4.79 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (117 mg, 0.16 mmol), and potassium phosphate (2 g, 9.58 mmol) were dissolved in acetonitrile (20 ml). The mixture was microwave-dried at 120°C for 60 minutes under nitrogen protection. The reaction mixture was filtered, concentrated under reduced pressure, and then flash column chromatography (petroleum ether:ethyl acetate = 0-35%) was used to obtain the target product, 5-(2-methyl-4-nitronaphthalene-1-yl)-2-(trifluoromethyl)pyrimidine (53-e, 780 mg, yield 73.4%). ESI [M+H] + =334.20

[0281] Step 6: Synthesis of 5-nitro-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-7-one (53-f)

[0282] 5-(2-Methyl-4-nitronaphthalen-1-yl)-2-(trifluoromethyl)pyrimidine (53-e, 780 mg, 2.34 mmol) was dissolved in dichloroethane (40 ml), and 2-methylpropane-2-hydroperoxide (1.8 ml, 18.7 mmol) and tetrabutylammonium iodide (43 mg, 0.12 mmol) were added. The mixture was reacted at 120°C overnight under nitrogen protection. After concentration under reduced pressure, the product was purified by flash column chromatography (petroleum ether:ethyl acetate = 0-35%) to obtain the target product 5-nitro-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-7-one (53-f, 35 mg, yield 4.3%). ESI [M+H] + =346.20

[0283] Step 7: Synthesis of 5-amino-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-7-one (53-g)

[0284] 5-Nitro-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-7-one (53-f, 35 mg, 0.1 mmol) was dissolved in ethanol (5 ml) and water (6 ml), and reduced iron powder (28 mg, 0.5 mmol) and ammonium chloride (53 mg, 1 mmol) were added. The mixture was reacted at 80°C under nitrogen for 1 hour. The reaction solution was filtered while hot, concentrated under reduced pressure, and then purified by flash column chromatography (petroleum ether:ethyl acetate = 0-35%) to obtain the target product 5-amino-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-7-one (53-g, 20 mg, yield 62.5%). ESI [M+H] + =315.20

[0285] Step 8: Synthesis of 2-fluoro-N-(7-oxo-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-5-yl)acrylamide (53)

[0286] 2-Fluoroacrylic acid (11 mg, 0.12 mmol) was dissolved in dichloromethane (5 ml). Oxalyl chloride (11 μl, 0.12 mmol) and a catalytic amount of NN-dimethylformamide were added dropwise at 0°C under a nitrogen atmosphere. The mixture was allowed to react for 3 hours. Triethylamine (42 μl, 0.17 mmol) and 5-amino-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-7-one (2-h, 20 mg, 0.058 mmol) were then added and allowed to react for one hour. After completion of the reaction, the mixture was diluted with water and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and then purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product, 2-fluoro-N-(7-oxo-9-(trifluoromethyl)-7H-benzo[6,7]indeno[2,1-d]pyrimidin-5-yl)acrylamide (53, 4.53 mg, yield 20.6%). ESI [M+H] + =388.20 1 H NMR(400MHz,DMSO-d6)δ10.84(s,1H),9.97(s,1H),8.77–8.68(m,1H),8.20–8.10(m,1H),7 .92(s,1H),7.90–7.84(m,2H),5.82(dd,J=47.7,3.8Hz,1H),5.55(dd,J=15.6,3.8Hz,1H).

[0287] Example 54: Synthesis of 3-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)azetidine-3-carboxamide (54)

[0288] Step 1: Synthesis of tert-butyl 3-fluoro-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)carbamoyl)azetidine-1-carboxylate (54-a)

[0289] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (80 mg, 0.30 mmol) was dissolved in N,N-dimethylformamide (5 ml), and 1-(tert-butoxycarbonyl)-3-fluoroazetidine-3-carboxylic acid (99 mg, 0.45 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (168 mg, 0.60 mmol) and 1-methylimidazole (99 μl, 1.2 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product, tert-butyl 3-fluoro-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)carbamoyl)azetidine-1-carboxylate (54-a, 140 mg, yield 99%). ESI [M+H] + =467.20

[0290] Step 2: Synthesis of 3-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)azetidine-3-carboxamide (54)

[0291] tert-Butyl 3-fluoro-3-((9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)carbamoyl)azetidine-1-carboxylate (54-a, 30 mg, 0.064 mmol) was dissolved in ethyl acetate (3 ml) and a 4 M hydrochloric acid ethyl acetate solution (3 ml) was added at room temperature. The reaction was allowed to react at room temperature for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and purified by preparative liquid chromatography (Pack Column 20 ID*250 mm*10 μm; A: H2O (0.1% FA), B: ACN, B%: 10%-70% over 22 min) to afford 3-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)azetidine-3-carboxamide (54, 1.89 mg, yield 8.2%). ESI [M+H] + =367.20 1 H NMR (400MHz, DMSO-d6) δ10.73(d,J=2.3Hz,1H),9.49(s,1H),8.24(s,1H),8.04(s,2H),4.54(d,J=18.2Hz,4H).

[0292] Examples 55 and 56: Synthesis of 3-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclobutane-1-carboxamide (55) and 7-((1-(3-fluorocyclobutyl)ethyl)amino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (56)

[0293] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-fluoro-2-methylpropionic acid (24 mg, 0.226 mmol), N,N,N',N'-tetramethylchlorouronium hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and the mixture was stirred at room temperature for 4 hours. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain 7-((1-(3-fluorocyclobutyl)ethyl)amino)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (55, 2.60 mg, yield 5.8%). ESI [M+H] + =366.30 1 H NMR (400 MHz, DMSO-d6) δ 10.46 (s, 1H), 9.44 (s, 1H), 8.11 (d, J = 1.4 Hz, 1H), 7.95 (d, J = 8.2 Hz, 1H), 7.86 (dd, J = 8.2, 1.7 Hz, 1H), 5.11 (dt, J = 14.1, 7.1 Hz, 1H), 4.97 (dt, J = 14.2, 7.1 Hz, 1H), 4.13 (q, J = 5.2 Hz, 1H), 3.16 (d, J = 5.2 Hz, 3H), 2.78–2.66 (m, 1H), 2.64–2.53 (m, 2H), 2.42–2.26 (m, 2H). Column 20 ID*250mm*10um; A: H2O (0.1% FA), B: ACN, B%: 10%-90% over 25 min] The target product, 3-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclobutane-1-carboxamide (56 mg, 0.77 mg, 1.7% yield), was purified for the second time. ESI [M+H] + =366.20 1H NMR (600MHz, DMSO-d6) δ10.44(s,1H),9.42(s,1H),8.10(s,1H),7.94(d,J=8.2Hz,1H),7.85(d,J=7.6Hz,1H),5.31– 5.25(m,1H),5.21–5.16(m,1H),3.25(dd,J=12.9,9.7Hz,1H),2.57(ddd,J=13.0,9.4,3.0Hz,2H),2.49–2.40(m,2H).

[0294] Example 57: Synthesis of 3,3-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclobutane-1-carboxamide (57)

[0295] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-fluoro-2-methylpropionic acid (24 mg, 0.226 mmol), N,N,N',N'-tetramethylchlorouronium hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified once using a preparative plate (petroleum ether:ethyl acetate = 1:1). The product was then purified a second time using a preparative liquid chromatography column (Pack Column 20 ID*250mm*10um; A:H2O (0.1% FA), B:ACN, B%: 10%-90% over 30 min) to obtain the desired product, 3,3-difluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclobutane-1-carboxamide (57 mg, 1.13 mg, 2.5% yield). ESI [M+H] + =384.30 1 H NMR (400MHz, DMSO-d6) δ10.56(s,1H),9.42(s,1H),8.07(d,J=1.2Hz,1H),7.93(d,J=8. 2Hz,1H),7.82(dd,J=8.2,1.5Hz,1H),3.18–3.06(m,1H),2.83(dt,J=17.0,8.5Hz,4H).

[0296] Example 58: Synthesis of (1R,2S)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (58)

[0297] 7-Amino-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (30 mg, 0.113 mmol) was dissolved in N,N-dimethylformamide (2 ml), and 2-fluoro-2-methylpropionic acid (24 mg, 0.226 mmol), N,N,N',N'-tetramethylchlorouronium hexafluorophosphate (64 mg, 0.226 mmol) and 1-methylimidazole (38 μl, 0.452 mmol) were added and reacted at room temperature for 2 hours. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified on a preparative plate (petroleum ether:ethyl acetate = 1:1) to obtain the target product (1R,2S)-2-fluoro-N-(9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (58, 4.69 mg, yield 11.8%). ESI [M+H] + =352.30 1 H NMR (400MHz, DMSO-d6) δ10.92(s,1H),9.41(s,1H),8.03(s,1H),7.92(d,J=8.2Hz,1H),7.83–7.73(m,1H),4.91 (d,J=65.1Hz,1H),2.34(ddd,J=17.4,10.0,7.1Hz,1H),1.63–1.49(m,1H),1.26(ddd,J=21.3,13.8,7.3Hz,1H).

[0298] Example 59: Synthesis of 2-fluoro-N-(5-((4-methylpiperazin-1-yl)methyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (59)

[0299] Step 1: Synthesis of 5-((4-methylpiperazin-1-yl)methyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-a)

[0300] 5-(Bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2, 140 mg, 0.36 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 1-methylpiperazine (48 μL, 0.43 mmol) and diisopropylethylamine (94 μL, 0.54 mmol) were added. The reaction was allowed to react at room temperature for 1 hour. After completion, the reaction solution was diluted with water and extracted three times with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (dichloromethane:methanol = 0-15%) to obtain the target product, 5-((4-methylpiperazin-1-yl)methyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-a, 74 mg, 50.5% yield). ESI[M+H] + =408.3

[0301] Step 2: Synthesis of 7-amino-5-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-b)

[0302] 5-((4-methylpiperazin-1-yl)methyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-a, 74 mg, 0.201 mmol) was dissolved in ethanol (5 mL). 1M aqueous hydrochloric acid (5 mL) and stannous chloride (85 mg, 0.602 mmol) were added. The nitrogen atmosphere was replaced three times and the reaction was allowed to proceed at 50°C for 2 hours. After completion, the pH was adjusted to approximately 8 with saturated aqueous sodium bicarbonate solution. The reaction was extracted three times with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified on a preparative plate to obtain the target product, 7-amino-5-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-b, 58 mg, 76.5% yield). ESI [M+H] + =378.2

[0303] Step 3: Synthesis of 2-fluoro-N-(5-((4-methylpiperazin-1-yl)methyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (59)

[0304] 7-Amino-5-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-b, 35 mg, 0.093 mmol) was dissolved in acetonitrile (2 mL), and 2-fluoroacrylic acid (17 mg, 0.186 mmol), N-(chloro(dimethylamino)methylene)-N-methylmethylammonium hexafluorophosphate (52 mg, 0.186 mmol) and 1-methylimidazole (46 μL, 0.37 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification was then performed using a preparative liquid chromatography (HPLC) column (Pack Column 30 ID*250 mm*10 μm; A: H2O (0.01% TFA), B: ACN, B%: 10%-80% over 33 min) to afford the desired product, 2-fluoro-N-(5-((4-methylpiperazin-1-yl)methyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (59 mg, 2.69 mg, 6.4% yield). ESI [M+H] + =450.2 1 H NMR (400MHz, DMSO-d6) δ9.26(s,1H),8.61(s,1H),8.17(d,J=1.9Hz,1H),8.05(d,J=1.7Hz,1H),5.79(dd,J=47.5,3. 9Hz,1H),5.53(dd,J=15.5,3.9Hz,1H),3.87(s,2H),3.07(s,3H),2.69(s,3H),2.53–2.51(m,2H),2.50–2.48(m,3H).

[0305] Example 60: Synthesis of 2-fluoro-N-(5-(morpholinomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (60)

[0306] Step 1: Synthesis of 5-(morpholinomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-a)

[0307] 5-(Bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2, 240 mg, 0.62 mmol) was dissolved in N,N-dimethylformamide (5 mL), and morpholine (59 μL, 0.68 mmol) and diisopropylethylamine (108 μL, 0.93 mmol) were added. The reaction was allowed to react at room temperature for 1 hour. After completion, the reaction solution was diluted with water and extracted three times with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 0-40%) to obtain the target product, 5-(morpholinomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-a, 110 mg, 45% yield). ESI [M+H] + =394.3

[0308] Step 2: Synthesis of 7-amino-5-(morpholinomethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-b)

[0309] 5-(Morpholinomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-a, 110 mg, 0.28 mmol) was dissolved in ethanol (5 mL). 1M aqueous hydrochloric acid (5 mL) and stannous chloride (117 mg, 0.84 mmol) were added. The nitrogen atmosphere was replaced three times and the reaction was allowed to proceed at 50°C for 2 hours. After completion, the pH was adjusted to 8 with saturated sodium bicarbonate solution. The reaction was extracted three times with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by preparative thin-layer chromatography to obtain the target product, 7-amino-5-(morpholinomethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-b, 60 mg, 58.8% yield). ESI [M+H] + =365.2

[0310] Step 3: Synthesis of 2-fluoro-N-(5-(morpholinomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (60)

[0311] 7-Amino-5-(morpholinomethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-b, 20 mg, 0.066 mmol) was dissolved in acetonitrile (2 mL), and 2-fluoroacrylic acid (12 mg, 0.132 mmol), N,N,N',N'-tetramethyluronium chloride hexafluorophosphate (37 mg, 0.132 mmol) and 1-methylimidazole (22 μL, 0.264 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification was then performed using a preparative liquid chromatography (HPLC) column (Pack Column 30 ID*250 mm*10 μm; A: H2O (0.01% TFA), B: ACN, B%: 10%-80% over 33 min) to afford the desired product, 2-fluoro-N-(5-(morpholinomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (60 mg, 2.46 mg, 8.5% yield). ESI [M+H] + =437.2 1 H NMR(400MHz,DMSO-d6)δ10.78(s,1H),9.38(s,1H),8.18(s,1H),8.09(s,1H),5.80(dd,J =47.5,3.9Hz,1H),5.53(dd,J=15.5,3.9Hz,1H),3.61(s,4H),2.51(s,2H),2.50(s,4H).

[0312] Example 61: Synthesis of 2-fluoro-N-(5-methyl-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (61)

[0313] Step 1: Synthesis of 7-amino-5-methyl-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (61-a)

[0314] 5-Methyl-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2-d, 30 mg, 0.097 mmol) was dissolved in dioxane (2 mL) and water (2 mL). Sodium sulfide nonahydrate (77 mg, 0.32 mmol) was added at 15°C, and the nitrogen atmosphere was replaced three times. The reaction was allowed to react at room temperature for 3 hours. After completion, the reaction was diluted with water and extracted three times with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by thin-layer chromatography on silica gel (ethyl acetate:petroleum ether = 1:1) to obtain the desired product, 7-amino-5-methyl-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (61-a, 17 mg, 62.5% yield). ESI [M+H] + =280.2

[0315] Step 2: Synthesis of 2-fluoro-N-(5-methyl-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (61)

[0316] 7-Amino-5-methyl-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (61-a, 17 mg, 0.061 mmol) was dissolved in acetonitrile (2 mL), and 2-fluoroacrylic acid (10 mg, 0.12 mmol), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (34 mg, 0.12 mmol) and 1-methylimidazole (20 μL, 0.24 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified via preparative liquid chromatography (Pack Column 30 ID*250 mm*10 μm; A: H2O (0.01% TFA), B: ACN, B%: 10%-80% over 27 min) to afford the desired product, 2-fluoro-N-(5-methyl-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (61 mg, 1.7 mg, yield 7.9%). ESI [M+H] + =352.2 1 H NMR (400MHz, DMSO-d6) δ10.66(s,1H),9.39(s,1H),8.06(s,1H),7.90(s,1H),5.78(dd,J=47.4,3.5Hz,1H),5.51(dd,J=15.5,3.7Hz,1H),2.62(s,3H).

[0317] Example 62: Synthesis of 1-fluoro-N-(5-(morpholinomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (62)

[0318] 7-Amino-5-(morpholinomethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (60-b, 40 mg, 0.11 mmol) was dissolved in acetonitrile (3 mL), and 1-fluorocyclopropane-1-carboxylic acid (23 mg, 0.22 mmol), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (62 mg, 0.22 mmol) and 1-methylimidazole (36 μL, 0.44 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified via preparative liquid chromatography (Pack Column 30 ID*250 mm*10 μm; A: H2O (0.01% TFA), B: ACN, B%: 10%-80% over 33 min) to afford the desired product, 1-fluoro-N-(5-(morpholinomethyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (62 mg, 8.11 mg, 16.5% yield). ESI [M+H] + =451.2 1 H NMR (400MHz, DMSO-d6) δ10.71(s,1H),9.26(s,1H),8.16(d,J=2.0Hz,1H),8.02(d,J=2.2Hz,1H),3.75(s,2 H),3.55(t,J=4.6Hz,4H),2.54–2.48(m,2H),2.48–2.46(m,2H),1.50–1.44(m,2H),1.36(tt,J=9.2,5.0Hz, 2H).

[0319] Example 63: Synthesis of 1-fluoro-N-(5-((4-methylpiperazin-1-yl)methyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (63)

[0320] 7-Amino-5-((4-methylpiperazin-1-yl)methyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (59-b, 20 mg, 0.053 mmol) was dissolved in acetonitrile (2 mL), and 2-fluoroacrylic acid (11 mg, 0.11 mmol), N-(chloro(dimethylamino)methylene)-N-methylmethylammonium hexafluorophosphate (31 mg, 0.11 mmol) and 1-methylimidazole (26 μL, 0.22 mmol) were added and reacted at room temperature overnight. After completion of the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified via preparative liquid chromatography (Pack Column 30 ID*250 mm*10 μm; A: H2O (0.01% TFA), B: ACN, B%: 10%-80% over 33 min) to afford the desired product, 1-fluoro-N-(5-((4-methylpiperazin-1-yl)methyl)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)cyclopropane-1-carboxamide (63 mg, 2.69 mg, 6.4% yield). ESI [M+H] + =464.2 1 H NMR (400MHz, DMSO-d6) δ9.46(s,1H),9.25(s,1H),8.15(d,J=1.9Hz,1H),8.05(d,J=1.9Hz,1H),3.85(s,2H),3.39(d,J=11.9Hz,2H),3.06(d, J=12.8Hz,2H),2.97(d,J=10.9Hz,2H),2.77(s,3H),2.40(t,J=11.7Hz,2H),1.49(ddd,J=18.1,8.6,5.5Hz,2H),1.36(dt,J=14.7,7.3Hz,2H).

[0321] Example 64: Synthesis of 2-fluoro-N-(9-oxo-5-(piperazin-1-ylmethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (64)

[0322] Step 1: Synthesis of 5-(bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (64-a)

[0323] 5-(Bromomethyl)-7-nitro-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (I-2, 150 mg, 0.39 mmol) was dissolved in ethanol (5 mL), and 1M aqueous hydrochloric acid (5 mL) and stannous chloride (163 mg, 1.16 mmol) were added. The nitrogen atmosphere was replaced three times, and the reaction was carried out at 50°C for 2 hours. After completion, the pH was adjusted to 8 with saturated sodium bicarbonate solution, and the mixture was extracted three times with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified on a preparative plate to obtain the target product, 7-amino-5-(morpholinomethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (64-a, 80 mg, 57.6% yield).

[0324] Step 2: Synthesis of tert-butyl 4-((7-amino-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-5-yl)methyl)piperazine-1-carboxylate (64-b)

[0325] 7-Amino-5-(morpholinomethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-9-one (64-a, 80 mg, 0.224 mmol) was dissolved in N,N-dimethylformamide (4 mL), and tert-butyl piperazine-1-carboxylate (49 mg, 0.27 mmol) and diisopropylethylamine (57 μL, 0.33 mmol) were added. The mixture was reacted at room temperature for 1 hour. After completion of the reaction, the reaction solution was diluted with water and extracted three times with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 0-40%) to obtain the target product, tert-butyl 4-((7-amino-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-5-yl)methyl)piperazine-1-carboxylate (64-b, 50 mg, yield 49.1%). ESI [M+H] + =464.2

[0326] Step 3: Synthesis of tert-butyl 4-((7-(2-fluoroacrylamide)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-5-yl)methyl)piperazine-1-carboxylate (64-c)

[0327] tert-Butyl 4-((7-amino-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-5-yl)methyl)piperazine-1-carboxylate (64-b, 50 mg, 0.11 mmol) was dissolved in acetonitrile (3 mL), and 1-fluorocyclopropane-1-carboxylic acid (23 mg, 0.22 mmol), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (62 mg, 0.22 mmol) and 1-methylimidazole (36 μL, 0.44 mmol) were added and reacted at room temperature overnight. After the reaction, the reaction was quenched with saturated aqueous sodium bicarbonate solution, and then extracted with ethyl acetate. The organic phase was collected and dried over anhydrous sodium sulfate to obtain a crude product. The crude product was purified by flash column chromatography (petroleum ether:ethyl acetate = 0-40%) to obtain the target product, tert-butyl 4-((7-(2-fluoroacrylamide)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-5-yl)methyl)piperazine-1-carboxylate (64-c, 25 mg, yield 42.1%). ESI [M+H] + =536.2

[0328] Step 4: Synthesis of 2-fluoro-N-(9-oxo-5-(piperazin-1-ylmethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (64)

[0329] Tert-butyl 4-((7-(2-fluoroacrylamide)-9-oxo-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-5-yl)methyl)piperazine-1-carboxylate (64-c, 25 mg, 0.047 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (0.6 mL) was added, and the reaction was carried out at room temperature for 2 hours. After the reaction, the mixture was concentrated under reduced pressure and then purified using a preparative liquid chromatography column (Pack Column 30 ID*250 mm*10 μm; A: H2O (0.01% TFA), B: ACN, B%: 10%-80% over 33 min) to obtain the desired product, 2-fluoro-N-(9-oxo-5-(piperazin-1-ylmethyl)-2-(trifluoromethyl)-9H-indeno[2,1-d]pyrimidin-7-yl)acrylamide (64 mg, 0.98 mg, 4.78% yield). ESI [M+H] + =436.2 1HNMR (400MHz, DMSO-d6) δ10.76(s,1H),9.26(s,1H),8.61(s,1H),8.17(d,J=1.9Hz,1H),8.05(d,J=1.7Hz,1H),5.7 9(dd,J=47.5,3.9Hz,1H),5.53(dd,J=15.5,3.9Hz,1H),3.87(s,2H),3.07(s,3H),2.69(s,3H),2.53–2.51(m,2H).

[0330] Test Example 1: Bioluminescence test of compounds inhibiting USP21 enzyme activity

[0331] Experimental reagents and instruments:

[0332] GST-USP21 protein (Boston Biochem), Ub-AML (aminofluorescein-labeled ubiquitin, Boston Biochem), DUB buffer (50 mM HEPES, pH 7.8, 100 mM NaCl, 0.5 mM EDTA, 0.01% (v / v) Tween-20, 1 mM DTT), luciferin detection reagent (Promega), DMSO. Microplate reader (TECAN), white 384-well microplate, and microplate thermostat shaker.

[0333] Bioluminescence assay to test the inhibitory activity of compounds on USP21 catalytic activity:

[0334] Ub-AML was diluted to 3 μM using 50 mM HEPES, pH 7.5. The diluted Ub-AML solution and luciferin detection reagent were mixed in a ratio of 1:9 to prepare the Ub-AML-LDR reagent. GST-USP21 was diluted to 8 nM using DUB buffer and transferred to a 384-well plate. The diluted compound was incubated with the diluted compound in a microplate shaker for 30 minutes (30°C, 500 rpm). After the Ub-AML-LDR reagent was added to the test wells (final concentration: 2 nM GST-USP21, 150 nM Ub-AML), the bioluminescence signal was immediately measured using a TECAN Spark multi-mode microplate reader. The detection parameters were: kinetic mode detection of bioluminescence, detection time of 40 minutes, detection every two minutes, and integration time of 500 ms. The initial reaction rate (slope of the first-order reaction) of each well was calculated, and the data were analyzed using a 4-parameter logistic model to calculate the IC 50 The results of USP21 enzyme inhibition activity test are shown in Table 1:

[0335] For IC 50 Value, where "+++" means IC50 Between 0nM and 100nM; “++” indicates IC 50 Between 100nM and 1μM (inclusive); "+" indicates IC 50 Between 1 μM and 10 μM (inclusive).

[0336] Table 1

[0337] Test Example 2 Cell Proliferation Inhibitory Activity Test

[0338] PANC-1 human pancreatic cancer cells and JIMT-1 human breast cancer cells were cultured in complete medium at 37°C in a 5% CO2 incubator. Cells in the logarithmic growth phase were digested and counted, and the density was adjusted to 1.11 x 10 cells using complete medium. 4 cells / mL, inoculated into 384-well cell culture plates (Greiner, 781098) at a volume of 45 μL per well, and cultured overnight in an incubator at 37°C and 5% CO2. The next day, the test compound was prepared into a 10mM stock solution with DMSO, and diluted 3-fold with DMSO starting from the highest dose of 10 μM, with a total of 8 concentration points. Then, it was diluted with complete culture medium to form working solutions of different concentrations and added to the corresponding wells. Two parallel wells were set for each well, and a DMSO well was set as a negative control. Finally, the DMSO content of all test wells was consistent at 0.1%. After continuing to culture in an incubator at 37°C and 5% CO2 for 72 hours, the cells were cultured using The luminescence signal was detected by the Luminescence module of TECAN microplate reader, and the curve was fitted by the 4-parameter method of GraphPad Prism software to calculate the IC 50 value.

[0339] The results of the cell proliferation inhibition activity test are shown in Table 2 below:

[0340] For IC 50 Value, where "+++" means IC 50 Between 0nM and 100nM; “++” indicates IC 50 Between 100nM and 1μM (inclusive); "+" indicates IC 50 Between 1 μM and 10 μM (inclusive).

[0341] The results showed that the compound of the present invention exhibited strong cell proliferation inhibition activity on both PANC-1 pancreatic cancer cells and JIMT-1 human breast cancer cells.

[0342] Table 2

[0343] The foregoing descriptions of specific exemplary embodiments of the present invention are for purposes of illustration and description. These descriptions are not intended to limit the invention to the precise forms disclosed, and it is apparent that many modifications and variations are possible in light of the foregoing teachings. The exemplary embodiments have been selected and described for the purpose of explaining the specific principles of the invention and their practical application, thereby enabling those skilled in the art to make and utilize a variety of exemplary embodiments of the invention and various options and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A compound of formula (I), a pharmaceutically acceptable salt, stereoisomer, solvate or a prodrug thereof: In formula (I), R 1 Selected from -R 4 -C(O)-C 2-6 Alkenyl, -C 2-6 Alkenyl, -N=NR a R b , -R 4 -C(O)-C 2-6 Alkynyl, -R 4 -SO2-C 2-6 Alkenyl, -R 4 -C(O)-C 1-3 Alkyl, -R 4 -C 2-6 Alkynyl, -R 4 -C(O)-C 3-9 Cycloalkyl, -R 4 -C(O)-3 to 9-membered heterocycloalkyl, -R 4 -C 1-3 Alkyl-C 3-9 Cycloalkyl, -R 4 -C 3-9 Cycloalkyl, -C(O)-NH-C 2-6 Alkynyl, -C(O)-NH-C 2-6 Alkenyl, -C(O)-NH-C 1-3 Alkyl or The C 2-6 Alkenyl, C 2-6 Alkynyl is optionally substituted by 1, 2 or 3 groups independently selected from hydrogen, halogen, cyano, -CF3, C 1-6 alkyl, 3- to 9-membered heterocycloalkyl, 5- to 10-membered heteroaryl, C 6-10 Aryl, -C 1-3 Alkyl-NR a R b , -C 1-3 Alkyl-hydroxy, -C(O)OR a 、-C(O)NR a R b 、-NR a R b Substituents are substituted, the C 1-3 Alkyl, C 3-9 Cycloalkyl, 3- to 9-membered heterocycloalkyl, C 1-6 The alkyl group is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 alkoxy, -O-3 to 9 membered heterocycloalkyl, -O-5 to 10 membered heteroaryl, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2-C 1-3 Alkyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 alkoxy, 5- to 10-membered heteroaryl, 6- to 10-membered aryl, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Substitution of alkynyl groups by substituents; R 4 Selected from NH, NR a or CR a R b ; R a , R b are each independently selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, the C 1-3 Alkyl, C 3-9 The cycloalkyl radical is optionally substituted by 1, 2 or 3 radicals independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2C 1- 3 alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Substitution of alkoxy groups; R 5 , R 6 are each independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl, the C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxyl, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2-C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Substitution of alkoxy groups; R 2 , R 3 are independently selected from hydrogen, halogen, cyano, hydroxyl, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, -NR a R b 、-C(O)-C 1-3 Alkyl, -C(O)NR a R b 、-SO2-C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-C 2-6 Alkynyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -C 1-3 Alkyl-NR a R b , -C 2-6 Alkenyl, -C 2-6 Alkynyl, 5- to 10-membered heteroaryl, C 6-10 Aryl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-3 Alkyl, C 2-6 Alkynyl, C 2-6 alkenyl, 5- to 10-membered heteroaryl, C 6- 10 Aryl is optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3 substituents; The halogen is selected from fluorine, chlorine or bromine; U 1 , U 2 , U 3 Each independently selected from CR 7 or CH, or adjacent U 2 , U 3 Connected to each other to form a 6- to 10-membered aromatic group; The R 7 Selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, 3- to 9-membered heterocycloalkyl, -C 1-3 Alkyl-3 to 9 membered heterocycloalkyl, the C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Substitution of alkynyl groups by substituents; The heterocycloalkyl or heteroaryl group has at least one heteroatom selected from N, O and S as a ring atom.

2. The compound according to claim 1, its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, wherein: R 1 Selected from -R 4 -C(O)-C(R 8 )=CR 9 R 10 ,-C(R 8 )=CR 9 R 10 ,-C(R 8 )=C(R 9 )-C(O)-NH2、-R 4 -C(O)-C(R 8 )=C(R 9 )-C(O)-NH2、-R 4 -C(O)-C(R 8 )=C(R 9 )-C(O)-OR 10 , -R 4 -C(O)-C(R 8 )=C(R 9 )-CH2-NR a R b 、-N=NR a , -R 4 -C(O)-C≡C-CH3, -R 4 -C(O)-C≡CH, -R 4 -C(O)-CH2-C≡CH, -R 4 -SO2-C(R 8 )=CR 9 R 10 , -R 4 -C(O)-R 5 , -R 4 -CH2-C≡CH, -C(O)-NH-C 1-3 Alkyl, -R 4 -C(O)-C 1-3 Alkyl-R 4 -C(O)-C(R 8 )=CR 9 R 10 、-C(O)-R 4 -C 1-3 Alkyl-R 4 -C(O)-C(R 8 )=CR 9 R 10 , -R 4 -CH2-C 3-9 Cycloalkyl or The C 1-3 Alkyl, C 3-9 The cycloalkyl radical is optionally substituted by 1, 2 or 3 radicals independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 Substitution of alkoxy groups; R 4 Select from NH or NR a ; R 5 , R 6 are each independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl, the C 1-6 Alkyl, C 3-6 Cycloalkyl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxyl, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2-C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Substitution of alkoxy groups; R 8 , R 9 , R 10 are each independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 6-10 aryl, 5- to 10-membered heteroaryl, 3- to 9-membered heterocycloalkyl, -C 1-3 Alkyl-C 6- 10 Aryl, -C 1-3 Alkyl-5 to 10 membered heteroaryl, -C 1-3 Alkyl-3 to 9 membered heterocycloalkyl, the C 1-3 Alkyl, C 1-6 Alkyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, 3- to 9-membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, -C 1-6 Substitution of alkoxy groups; R a , R b are each independently selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, the C 1-3 Alkyl, C 3-9 The cycloalkyl radical is optionally substituted by 1, 2 or 3 radicals independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, hydroxy, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2C 1- 3 alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Substitution of alkoxy groups; R 2 , R 3 are each independently selected from hydrogen, halogen, cyano, hydroxyl, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, -NR a R b , the C 1-6 Alkyl, C 1-6 The alkoxy group is optionally substituted by 1, 2 or 3 groups independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3 substituents; The halogen is selected from fluorine or chlorine; U 1 , U 2 , U 3 Each independently selected from CR 7 or CH, or adjacent U 2 , U 3 Connected to each other to form a 6- to 10-membered aromatic group; The R 7 Selected from hydrogen, C 1-3 Alkyl, C 3-9 Cycloalkyl, 3- to 9-membered heterocycloalkyl, -C 1-3 Alkyl-3 to 9 membered heterocycloalkyl, the C 1-3 Alkyl, C 3-9 Cycloalkyl, 3 to 9 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 independently selected from -H, halogen, methyl, ethyl, propyl, isopropyl, cyano, amino, -NR a R b , hydroxyl, carboxyl, -CF3, C 1-6 Alkoxy, -C(O)OC 1-3 Alkyl, -C(O)-C 1-3 Alkyl, -SO2C 1-3 Alkyl, -C 1-3 Alkyl-hydroxyl, -C 1-3 Alkyl-cyano, -C 1-3 Alkyl-C 1-6 Alkoxy, -NHC(O)-C 2-6 Alkenyl, -NHC(O)-C 2-6 Substitution of alkynyl groups by substituents; The heterocycloalkyl or heteroaryl group has at least one heteroatom selected from N, O and S as a ring atom.

3. The compound according to claim 1-2, its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, wherein: The R 1 Selected from The R 2 , R 3 Each independently selected from H, CF3; The U 1 , U 2 , U 3 All selected from CH, or adjacent U 2 , U 3 They are connected to each other to form a benzene ring; The R 7 Selected from -H, -CH3, 4. The compound according to claim 1-3, its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, wherein formula (I) is represented by formula (II-1), formula (II-2), formula (II-3), formula (II-4), formula (II-5), formula (II-6), formula (II-7), formula (II-8), formula (II-9), formula (II-10), formula (II-11), formula (II-12), formula (II-13), formula (II-14), formula (II-15), formula (II-16) or formula (II-17): R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R a The definition as described in any one of claims 1 to 3.

5. The compound according to any one of claims 1 to 4, its pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, wherein: The compound is selected from the group consisting of:

6. A method for preparing a partial compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt, a stereoisomer, a solvate or a prodrug thereof, comprising the following steps: Compound (III-1) undergoes an amidation reaction with compound (Y) to generate compound (III-2); The compound (Y) includes any one of acyl chloride, carboxylic acid or sulfonyl chloride; The R 1 , R 2 , R 3 , R 7 The definition as described in any one of claims 1-5.

7. A pharmaceutical composition, characterized in that The composition comprises the compound according to any one of claims 1 to 5, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the method according to claim 6 and at least one pharmaceutically acceptable excipient.

8. Use of the compound according to any one of claims 1 to 5, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the method according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a drug for treating cardiovascular disease, neurological disorder, cancer or immune disease, or in the preparation of a kit for evaluating cardiovascular disease, neurological disorder, cancer, immune disease or patient prognosis; Preferably, the pharmaceutical composition further comprises another drug for treating cardiovascular disease, neurological disorder, cancer or immune disease; Preferably, use in the preparation of a method for treating a disease associated with USP21 activity; Preferably, the cancer includes prostate cancer, bile duct cancer, pancreatic cancer, breast cancer, lung cancer, brain cancer, colon cancer, kidney cancer, bladder cancer, epithelial ovarian cancer, liver cancer, esophageal squamous cell carcinoma and leukemia; More preferably, the cancer includes pancreatic cancer, bile duct cancer, esophageal squamous cell carcinoma, liver cancer, breast cancer, kidney cancer, bladder cancer and lung cancer.

9. Use of the compound according to any one of claims 1 to 5, its pharmaceutically acceptable salt, stereoisomer, solvate, prodrug thereof, or the compound prepared by the method according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a USP inhibitor; Preferably, the invention relates to a method for preparing a USP21 inhibitor.

10. A method for treating a USP mutation or amplification-related disease in a patient in need thereof, comprising administering to the patient a compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt, stereoisomer, solvate or prodrug thereof, or a pharmaceutical composition according to claim 7; The USP mutation or amplification includes USP21 mutation or amplification; Preferably, the USP mutation or amplification-related diseases include cardiovascular diseases, neurological disorders, cancer, and immune diseases; Preferably, the cancer includes prostate cancer, bile duct cancer, pancreatic cancer, breast cancer, lung cancer, brain cancer, colon cancer, kidney cancer, bladder cancer, epithelial ovarian cancer, liver cancer, esophageal squamous cell carcinoma and leukemia; More preferably, the cancer includes pancreatic cancer, bile duct cancer, esophageal squamous cell carcinoma, liver cancer, breast cancer, kidney cancer, bladder cancer and lung cancer.

11. A method for inhibiting USP mutations in a biological sample for non-disease diagnosis or treatment purposes, comprising contacting the biological sample with a compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt, stereoisomer, solvate, or prodrug thereof, or a compound prepared by the method according to claim 6, or a pharmaceutical composition according to claim 7.

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