Parp7 inhibitors and uses thereof

Abstract

The present application relates to pyridinone or pyrimidinone derivatives having PARP7 inhibitory activity represented by Formula (I), and isotopic forms, stereoisomers, tautomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs and polymorphs thereof, and describes methods for their preparation and their use in cancer and immune-related diseases.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to a small molecule inhibitor of PARP7, its preparation method, and its application. Background Technology

[0002] PARP7, also known as TIPARP, is a gene located on chromosome 3q25.31 that encodes the protein poly-ADP-ribose polymerase 7, which is located in the cell nucleus. PARP7 belongs to the poly-ADP-ribose polymerase superfamily, which consists of 17 members, all of which contain a common catalytic domain of approximately 230 amino acids. PARP1, 2, 5a, and 5b catalyze the transfer of multiple ADP-ribose units to a substrate; these members are called polyPARPs. PARP3, 4, 6, 7, 8, 9, 10, 11, 12, 14, 15, and 16 catalyze the transfer of a single ADP-ribose unit to a substrate; these members are called monoPARPs. Currently, only the catalytic activity of PARP13 has not been confirmed. PARPs are involved in multiple cellular processes, including DNA damage repair, cell proliferation, apoptosis, DNA methylation, transcriptional regulation, and the WNT signaling pathway. PARP 1 has been developed as a tumor-associated target related to cellular stress caused by DNA damage, gene mutation, or chemotherapy toxicity, and four drugs targeting it have been approved for clinical trials.

[0003] Aromatic hydrocarbon receptor (AHR) ligands upregulate PARP7 expression. Against the backdrop of AHR signaling, PARP7 acts as a negative feedback mechanism to regulate the gene expression of cytochrome P4501A1 and P4501B1. TBK1 is activated at the initiation of pathogen-associated molecular pattern pathways, regulating type I interferon response and antiviral immune activation. PARP7 can bind to TBK1 via the marylate tank, thereby inhibiting TBK1 activation and downregulating downstream type I interferon secretion. Studies have found that PARP7 overactivity in tumors allows cancer cells to escape the immune system, thus promoting tumor growth. Clinical studies indicate that PARP7 is frequently amplified in squamous cell carcinoma, and the chromosome 3q25 where PARP7 is located is a susceptibility site for ovarian cancer, suggesting that PARP7 also plays an important role in ovarian cancer. These clinical findings and preclinical studies provide a strong biomedical basis for the development of PARP7-targeting inhibitors.

[0004] The monoPARP protein family plays a crucial role in multiple stress responses associated with the development of inflammatory diseases, cancer, and neurodegenerative diseases (Nizi et al. J. Med. Chem. 2022, 65, 7532.; Al Ghamdi, Aet. al. Mini. Rev. Med. Chem. 2022, 22, 1597.). PARP7, a member of the monoPARP family, has been shown to be overactive in tumors and plays a key role in cancer cell survival. Inhibition of PARP7 can effectively suppress cancer cell growth, restore interferon signaling, and effectively prevent cancer cells from escaping the immune system (Rodriguez, et al. Elife 2021, 10, No. e60480; Curtin et al. Nat. Rev. Drug Discovery 2020, 19, 711-736; O'Sullivan, Nat. Commun. 2019, 10, 1182.

[0005] Inhibition of PARP7 can activate the innate immune pathway STING, promote the secretion of IFN-β by tumor immune cytokines, restore the interferon signaling pathway, and upregulate phosphorylated STAT1, thereby inhibiting T cell-mediated anti-tumor immunity and inhibiting tumor growth. PARP7 inhibitors can prevent tumor immune escape, induce tumor-specific adaptive immune memory, thereby enhancing immune system signals, relieving innate and adaptive immunosuppression, and ultimately inhibiting tumor growth for a long time. Mechanistically, PARP7 inhibitors can play a synergistic role when used in combination with PD1 monoclonal antibodies, which can more effectively inhibit tumor growth and expand the clinical application of PARP7. In some cancer models, PARP7 inhibitors have shown sustained tumor growth inhibition and effective anti-proliferative effects. And restore interferon signaling (Gozgit et al., Cancer Cell. 2021 Sep13; 39(9):1214-1226). In recent years, some PARP7 inhibitors have been found to have the potential to treat cancer (Gu, H. et. al. J. Med. Chem. 2023, 66, 1, 473-490; Kargbo, R. ACS Med. Chem. Lett. 2022, 13, 1688-1690).

[0006] Current research on PARP7 targeting mainly focuses on activating T cell-mediated anti-tumor immune responses and acquiring adaptive immunity. Studies have revealed that it can be used in combination with anti-tumor drugs of other mechanisms (such as PD1 monoclonal antibodies) to have a synergistic effect, and PARP7 targeting inhibitors have significant clinical value. Summary of the Invention

[0007] The purpose of this invention is to provide a novel small molecule compound with PARP7 inhibitory activity and its applications.

[0008] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0009] The first aspect of this invention provides compounds of formula (I) or their isotopic forms, stereoisomers, tautomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs, and polymorphs thereof.

[0010]

[0011] in,

[0012] X is CH or N;

[0013] R 1 R 2 C104 and C204 are selected independently, either substituted or unsubstituted. 1-10 Alkyl, substituted or unsubstituted C 3-10 cycloalkyl, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted C 2-5 alkenyl, substituted or unsubstituted C 2-3 One of the alkynyl groups;

[0014] A is a chiral or achiral amine unit, and is connected to a pyridinone or pyrimidinone via nitrogen;

[0015] L is selected from one of the following structures:

[0016]

[0017] B is a C that contains at least one N atom. 4-12 Heterocyclic rings, which are single-ring or double-ring, wherein double-ring rings are heterocyclic rings containing bridging, fusion, spiral, or self-connecting bridging units;

[0018] Y is N, C, or CH;

[0019] Z represents N, CH, or CR. 3 , where R 3 For substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 3-10 cycloalkyl, C 1-10 Alkoxy;

[0020] Furthermore, the R 1 R 2 Each of the following groups is independently selected from methyl, ethyl, propyl, fluoromethyl, trifluoromethyl, deuterated methyl, bromine, chlorine, fluorine, cyano, and C. 3-10 One of the cycloalkyl groups; in some preferred embodiments of the invention, R1 R 2 It can be independently selected from -F, -Br, -CN, -CO2Me, -CONH2, or one of the following structural groups:

[0021]

[0022] Furthermore, A is more preferably one of the following structures:

[0023]

[0024] Furthermore, B is more preferably one of the following structures:

[0025]

[0026] Furthermore, the compound represented by formula (I) is a compound represented by the following structural formulas I-1 to I-57:

[0027]

[0028]

[0029]

[0030]

[0031] A second aspect of the present invention provides a pharmaceutical composition comprising one or more of the following: a compound of formula (I) as described in the first aspect, its isotopic form, stereoisomer, tautomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug, and polymorph.

[0032] Furthermore, in the pharmaceutical composition, any of the compounds represented by formula (I) above, or their isotopic forms, stereoisomers, tautomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs, and polymorphs, are used in combination with other drugs.

[0033] The third aspect of the present invention provides the use of a compound of formula (I) as described in the first aspect, or its isotopic form, stereoisomer, tautomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug, and polymorph, or the pharmaceutical composition described in the second aspect, in the preparation of a PARP7 inhibitor.

[0034] Furthermore, the PARP7 inhibitor is administered orally, parenterally, intravenously, or transdermally.

[0035] The fourth aspect of the present invention provides the use of a compound of formula (I) as described in the first aspect or its isotopic form, stereoisomer, tautomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug, and polymorph, or the pharmaceutical composition described in the second aspect, in the preparation of a medicament for treating a patient’s PARP7-mediated condition.

[0036] Furthermore, the PARP7-mediated diseases include breast cancer, ovarian cancer, and colorectal cancer.

[0037] Furthermore, the drug is administered orally, parenterally, intravenously, or transdermally.

[0038] Furthermore, in the drug, any compound represented by formula (I) above, or its isotopic form, stereoisomer, tautomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug, or polymorph thereof, is used in combination with other drugs.

[0039] As used herein, unless otherwise stated, the following definitions and terms shall apply.

[0040] "R" and "S" are terms used to describe isomers and are descriptors of the stereochemical configuration of asymmetrically substituted carbon atoms. Naming an asymmetrically substituted carbon atom "R" or "S" is accomplished by applying the Cahn-Ingold-Prelog priority rule, which is well known to those skilled in the art and described in Section E, Stereochemistry, of the International Union of Pure and Applied Chemistry (IUPAC) Rules of Nomenclature for Organic Chemistry.

[0041] The term C used here i-j This means that this part has ij carbon atoms. For example, "C 1-10 "Alkyl" refers to an alkyl unit having any number of carbon atoms between 1 and 10.

[0042] As used herein, "alkyl" refers to a fully saturated straight-chain, branched alkane group. In some embodiments, the alkyl group contains 1-6 carbon atoms. In some embodiments, the alkyl group contains 1-4 carbon atoms. In some embodiments, the alkyl group contains 1-3 carbon atoms. In other embodiments, the alkyl group contains 2-3 carbon atoms, and in still other embodiments, the alkyl group contains 1-2 carbon atoms. Non-limiting examples of exemplary alkyl groups include alkyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-heptyl, n-octyl, etc. Additionally, the term "cycloalkyl" refers to a monocyclic or bicyclic saturated carbon ring, each ring having 3 to 10 carbon atoms. A "fused analogue" of cycloalkyl refers to a monocyclic ring fused with an aryl or heteroaryl group, wherein the attachment point is on the non-aryl portion. Examples belonging to cycloalkyl and fused analogues include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydronaphthyl, decahydronaphthyl, indene, etc.

[0043] The term "alkoxy" refers to a straight-chain or branched alkoxy group having the indicated number of carbon atoms. For example, C 1-10 Alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and isopropoxy.

[0044] The term "alkenyl" refers to a carbon chain containing at least one carbon-carbon double bond, which can be straight-chain or branched, or a combination thereof, as described above. 2-5 Alkenyl groups include vinyl, propenyl, 2-methyl-1-propenyl, etc.; the term "alkynyl" refers to a carbon chain containing at least one carbon-carbon triple bond, which can be straight-chain or branched, or a combination thereof. The above C 2-3 Examples of alkenyl groups include ynyl or propynyl.

[0045] "Ar" or "aryl" refers to an aromatic carbon ring moiety having one or more closed rings. Examples include, but are not limited to, phenyl, naphthyl, anthracene, phenanthrene, biphenyl, and pyrene.

[0046] "Heteroaryl" refers to a cyclic moiety having one or more closed rings, having at least one ring with one or more heteroatoms (e.g., oxygen, nitrogen, or sulfur), wherein at least one ring is aromatic, and wherein one or more rings can be independently melted or bridged. Examples include, but are not limited to, pyridyl, pyrrole, pyrazolyl, quinolinyl, isoquinolinyl, indolyl, furanyl, thienyl, quinolinyl, indazolyl, thio[2,3-c]pyrazolyl, benzofuranyl, thienylpyrazolyl, and benzothiazolyl.

[0047] Unless otherwise stated, the term "halogen" or "halogen element" itself, or as part of another substituent, refers to a fluorine, chlorine, bromine, or iodine atom. Furthermore, the term "haloalkyl" means including both monohaloalkyl and polyhaloalkyl. For example, "haloC" 1-4"Alkyl" refers to, but is not limited to, trifluoromethyl, 2,2,2-trifluoromethyl, 4-chlorobutyl, 3-bromopropyl, etc.

[0048] Optical isomers, diastereomers, geometric isomers, and tautomers: Some compounds of formula (I) may contain one or more ring systems, and therefore may have cis and trans isomers. This invention is intended to cover all of these cis and trans isomers. The inclusion of an olefinic double bond, unless otherwise specified, means the inclusion of E and Z geometric isomers.

[0049] Any enantiomer of a compound of general formula (I) can be obtained by stereo-oriented synthesis using optically pure starting materials or reagents with known configurations.

[0050] Furthermore, compounds of formula (I) may also include a series of stable isotope-labeled analogs. For example, one or more protons in a compound of formula (I) may be substituted with deuterium atoms, thereby providing deuterated analogs with improved pharmacological activity.

[0051] "Pharmaceutically acceptable salt" refers to the acid salt or base salt of the compounds of this invention, which has the desired pharmacological activity and is neither biologically desirable nor otherwise desirable. The salt can form with acids, including but not limited to acetic acid, adipic acid, benzoate, citric acid, camphoric acid, camphor sulfonate, dicarboxylate, dodecyl sulfate, ethanesulfonate, fumarate, glucono-heptate, glycerol phosphate, hemisulfate, heptanate, hexanoate, hydrobromide hydrochloride, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, and oxalate.

[0052] The term "PARP-mediated" refers to any disease or other harmful condition in which PARP or its mutants are known to play a role.

[0053] By employing the above technical solution, the present invention has at least the following advantages:

[0054] This invention provides a class of pyridone or pyrimidinone derivatives with PARP7 inhibitory activity. These compounds have high PARP7 inhibitory activity and low cardiotoxicity, and can be used to prepare PARP7 inhibitors and drugs for PARP7-mediated diseases such as breast cancer, ovarian cancer, and colorectal cancer, and have extremely high application value. Detailed Implementation

[0055] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0056] The present invention will be further described below with reference to specific embodiments, so that those skilled in the art can better understand and implement the present invention, but the embodiments are not intended to limit the present invention.

[0057] To facilitate a thorough understanding of the technical solutions in this invention by those skilled in the art, the separation and purification methods, testing methods, English abbreviations, synthesis methods of raw materials or intermediates, and synthesis methods of the target compounds on which the embodiments of this application are based are described below:

[0058] 1. In the following embodiments of this application, the separation and purification methods and testing methods are as follows:

[0059] In the following embodiments of the present invention, unless otherwise specified, column chromatography was performed using silica gel (100-200 mesh) and different eluents. Solvent removal was performed using a Buchii rotary evaporator or a Genevac centrifugal evaporator. LC / MS was performed under acidic mobile phase conditions using a Waters automated purifier and a 19×100mm XTerra 5-micron MSCI 8 column. Nuclear magnetic resonance spectra were recorded using a Varian 400MHz spectrometer. When the term "inert" is used to describe a reactor (e.g., reaction vessel, flask, glass reactor, etc.), it means that the air in the reactor has been replaced by a substantially anhydrous or dry inert gas (e.g., nitrogen, argon, etc.).

[0060] 2. In the following embodiments of this application, the Chinese names corresponding to the English abbreviations are as follows: TEA: triethylamine; EA: ethyl acetate; THF: tetrahydrofuran; DMF: N,N-dimethylformamide; TFA: trifluoroacetic acid; NMP: N-methylpyrrolidone; DCM: dichloromethane; HATU: 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluracil hexafluorophosphate; DIPEA: N,N-diisopropylethylamine.

[0061] 3. The synthesis methods of the reaction raw materials or reaction intermediates used in the following embodiments of this application are as follows:

[0062] The intermediate 2a-2k was synthesized according to the synthetic method shown in reaction route 1 below:

[0063]

[0064] Reaction route 1

[0065] For example, the preparation process of intermediate 2a is as follows:

[0066] S-(+)-2-amino-1-propanol (2.2 g, 29.9 mmol, 1.35 eq) and TEA (15 mL) were added to a CH3CN (50 mL) solution of compound 1a (5.0 g, 22.1 mmol). The reaction mixture was stirred at 85 °C for 6 h until TLC showed complete reaction of compound 1a. The reaction solution was extracted with EA (20 mL × 3), and the organic phases were combined and washed with water and brine. The organic phase was dried over anhydrous Na2SO4 and distilled under reduced pressure to give crude compound, which was purified by silica gel column chromatography to give intermediate 2a (5.6 g, 95% yield).

[0067] By replacing appropriate reactants, intermediate 2b-2k, as shown in Table 1 below, was prepared according to the synthetic method shown in reaction route 1.

[0068] Table 1

[0069]

[0070]

[0071]

[0072] Intermediate 4a-4i was synthesized according to the synthetic method shown in reaction route 1 above:

[0073] For example, the preparation process of intermediate 4a is as follows:

[0074] Methyl acrylate (2.3 g, 26.4 mmol, 3.5 eq) and Cs₂CO₃ (12.6 g, 38.6 mmol, 5.12 eq) were added to a CH₃CN (50 mL) solution of intermediate 2a (2.0 g, 7.5 mmol). The reaction mixture was stirred at room temperature for 12 h until TLC showed complete reaction of intermediate 2a. The reaction mixture was extracted with EA (20 mL × 3), and the organic phases were combined and washed with water and brine. The organic phases were dried over anhydrous Na₂SO₄ and distilled under reduced pressure to give crude compound, which was purified by silica gel column chromatography to give intermediate 3a (1.7 g, yield: 65%).

[0075] To a mixed solution of intermediate 3a (250 mg, 0.71 mmol) in THF (3 mL) and H₂O (3 mL), NaOH (0.6 g, 14.2 mmol, 20 eq) was added. The reaction mixture was stirred overnight at 100 °C until TLC showed complete reaction of intermediate 3a. After cooling to room temperature, the pH was adjusted to 5 with 2N HCl, and the mixture was extracted with EA (20 mL × 3). The organic phases were combined and washed with water and brine. The organic phase was dried over anhydrous Na₂SO₄ and then distilled under reduced pressure to obtain crude intermediate 4a (200 mg, yield: 88%), which was used directly in the next reaction.

[0076] By replacing appropriate reactants, intermediates 4b-4i, as shown in Table 2 below, were prepared according to the synthetic method shown in reaction route 1.

[0077] Table 2

[0078]

[0079]

[0080] The intermediate 8a-8h was synthesized according to the synthetic method shown in reaction route 2 below:

[0081]

[0082] Reaction route 2

[0083] For example, the preparation process of intermediate 8a is as follows:

[0084] Under N2 protection, NaH (0.5 g, 12.3 mmol, 2.0 eq 60% in oil) was added in portions to a DMF (8 mL) solution of compound 5 (1.0 g, 6.2 mmol). After stirring at 0 °C for 30 min, 6a (1.4 g, 7.4 mmol) was added in portions. After addition, the reaction mixture was stirred at room temperature for 1 h until TLC showed complete reaction of intermediate 6a. The reaction mixture was quenched with saturated NH4Cl solution in an ice bath, extracted with EA (10 mL × 3), and the combined organic phases were washed with water and brine. The crude compound was dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain the crude compound, which was purified by silica gel column chromatography to obtain intermediate 7a (1.2 g, 60% yield). 1 H NMR (400MHz, CDCl3) δ8.40(s,1H),7.64(dd,1H),6.63(d,1H),3.65-3.62(m,4H),3.55-3.53(m,4H),1.48(s,9H).

[0085] TFA (0.5 mL) was added to a mixture of intermediate 7a (1.2 g, 3.6 mmol) and DCM (5 mL). The reaction mixture was stirred at room temperature for 1 h until TLC showed that intermediate 7a was completely reacted. Most of the solvent was removed, and the pH was adjusted to 7 with saturated NaHCO3. The reaction mixture was extracted with EA (10 mL × 3). The combined organic phases were washed with water and brine. The organic phases were dried over anhydrous Na2SO4 and then distilled under reduced pressure to give intermediate 8a (0.71 g, 85% yield).

[0086] By replacing appropriate reactants, intermediates 8a-8h, as shown in Table 3 below, were prepared according to the synthetic method shown in reaction route 2.

[0087] Table 3

[0088]

[0089]

[0090] Intermediate 11a-11i was synthesized according to the synthetic method shown in reaction route 3 below:

[0091]

[0092] Reaction route 3

[0093] For example, the preparation process of intermediate 11a is as follows:

[0094] A mixture of compound 9a (1.5 g, 8.2 mmol), compound 6a (1.5 g, 8.2 mmol, 1.0 eq), and K₂CO₃ (2.3 g, 16.5 mmol, 2.0 eq) in NMP (20 mL) was stirred at 80 °C for 10 h until TLC showed complete reaction of compound 9a. The reaction mixture was extracted with EA (20 mL × 3), and the combined organic phases were washed with water and brine. The organic phase was dried over anhydrous Na₂SO₄ and distilled under reduced pressure to give crude compound, which was purified by silica gel column chromatography to give intermediate 10a (2.1 g, 77% yield).

[0095] TFA (1.2 mL) was added to a DCM (10 mL) solution of intermediate 10a (2.1 g, 6.3 mmol). The reaction mixture was stirred at room temperature for 1 h until TLC showed that intermediate 10a was completely reacted. Most of the solvent was removed, and the pH was adjusted to 7 with saturated NaHCO3. The reaction mixture was extracted with EA (15 mL × 3). The organic phases were combined and washed with water and brine. The organic phase was dried over anhydrous Na2SO4 and then distilled under reduced pressure to give intermediate 11a (1.4 g, 95% yield).

[0096] By replacing appropriate reactants, intermediates 11a-11i, as shown in Table 4 below, were prepared according to the synthetic method shown in reaction route 3.

[0097] Table 4

[0098]

[0099]

[0100] Reaction route 4:

[0101]

[0102] The specific preparation process for reaction route 4 is as follows:

[0103] A solution of intermediate 4a (0.31 mmol, 1 eq), HATU (131 mg, 0.34 mmol, 1.1 eq), DIPEA (81 mg, 0.63 mmol, 2.0 eq), and intermediate 8a (0.34 mmol, 1.1 eq) in DCM (5 mL) was stirred at room temperature for 2 h until TLC showed complete reaction of intermediate 4a. The reaction mixture was extracted with DCM (8 mL × 3), the organic phases were combined and washed with water and brine, dried over anhydrous Na₂SO₄, and distilled under reduced pressure to give the crude compound, which was purified by TLC to give compound 12.

[0104] Example 1

[0105] 5-Bromo-6-{[(2S)-1-[(3-oxoylide-3-{4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-1)

[0106] The compound was prepared from intermediates 4a (Table 2) and 8a (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0107] 1 H NMR (400MHz, CDCl3) δ13.45(s,1H),8.39(s,1H),7.89(s,1H),7.65(dd,1H),6.62(d,1H),5.58(d,1H),4.40-4.34(m,1H),3. 85-3.82(t,2H),3.86-3.82(m,2H),3.76-3.71(m,4H),3.69-3.59(m,4H),3.54-3.52(m,2H),2.68-2.65(t,2H),1.25(d,3H).

[0108] LCMS: m / z = 534.45[M+l]+.

[0109] Example 2

[0110] 6-[4-(3-{[(2S)-2-[(5-bromo-6-oxoylide-1H-pyrimidin-4-yl)amino]propyl]oxy}propionyl)piperazin-1-yl]pyridine-3-carboxylonitrile(I-2)

[0111] The compound was prepared from intermediates 4a (Table 2) and 8e (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0112] LCMS: m / z = 490.11[M+l]+.

[0113] Example 3

[0114] 5-Bromo-6-{[(2S)-1-({3-[4-(5-fluoropyridin-2-yl)piperazin-1-yl]-3-oxopropyl}oxy)propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-3)

[0115] The compound was prepared from intermediates 4a (Table 2) and 8f (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0116] LCMS: m / z = 483.11[M+l]+.

[0117] Reaction route 5:

[0118]

[0119] Reaction route 5

[0120] Compound 16 was prepared according to reaction route 5. First, compound 13 was reduced with sodium borohydride to obtain compound 14. Compound 14 could be prepared by CBr4, PPh3 or Tosyl to obtain compound 15. Then, it was reacted with intermediate 8 or intermediate 11 to obtain compound 16.

[0121] Example 4

[0122] 5-Bromo-6-{[(2S)-1-[(3-{4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-4)

[0123] This compound was prepared from intermediates 13a and 8a (Table 3) according to reaction route 5. The product was characterized, and the results are as follows:

[0124] LCMS: m / z = 519.13[M+l]+.

[0125] Example 5

[0126] 5-Bromo-6-{[(2S)-1-[(3-{5-[5-(trifluoromethyl)pyridin-2-yl]-2,5-diazabicyclo[2.2.1]hept-2-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-5)

[0127] The compound was prepared from intermediates 13a and 8b (Table 3) according to reaction route 5. The product was characterized, and the results are as follows:

[0128] LCMS: m / z = 531.13[M+l]+.

[0129] Example 6

[0130] 5-Bromo-6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,5-diazabicyclo[2.2.1]hept-5-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-6)

[0131] The compound was prepared from intermediates 4a (Table 2) and 8b (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0132] 1 H NMR (400MHz, CDCl3) δ13.01(s,1H),8.34(s,1H),7.90(d,1H),7.58(dd,1H),6.32(d,1H),5.53(d,1H),4.38-4.24(m,1H),3.83-3.75(m,2H),3. 72-3.68(m,2H),3.65-3.60(m,1H),3.56-3.54(m,2H),3.52-3.48(m,1H ),3.46-3.37(m,2H),3.18-3.12(m,2H),2.04-1.90(m,2H),1.14(d,3H).

[0133] LCMS: m / z = 546.45[M+l]+.

[0134] Reaction route 6:

[0135]

[0136] Reaction route 6

[0137] Example 7

[0138] 5-Bromo-6-{[(2S)-1-[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-7)

[0139] The compound was prepared from intermediate 2a (Table 1) via reaction route 6 to obtain intermediate 20, which was then reacted with 8a (Table 3) via reaction route 4. The products were characterized, and the results are as follows:

[0140] LCMS: m / z = 544.12[M+l]+.

[0141] Example 8

[0142] 5-Bromo-6-{[(2S)-1-[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}methyl)azacyclobut-1-yl]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-8)

[0143] The compound I-7, prepared in Example 7, was obtained by reducing the product under standard amide functional group conditions. The product was characterized, and the results are as follows:

[0144] LCMS: m / z = 530.14[M+l]+.

[0145] Example 9

[0146] 5-Bromo-6-{[(2S)-1-[3-({4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-9)

[0147] This compound was prepared from intermediates 20 and 11a (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0148] LCMS: m / z = 545.12[M+l]+.

[0149] Example 10

[0150] 5-Bromo-6-{[(2S)-1-[(3-{4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-10)

[0151] This compound was prepared from intermediates 13a and 11a (Table 4) according to reaction route 5. The product was characterized, and the results are as follows:

[0152] LCMS: m / z = 520.12[M+l]+.

[0153] Example 11

[0154] 5-Bromo-6-{[(2S)-1-[(3-oxoylide-3-{4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-11)

[0155] This compound was prepared from intermediates 4a (Table 2) and 11a (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0156] 1 H NMR(400MHz, CDCl3)δ13.40(s,1H),8.49(s,2H),7.89(s,1H),5.57(d,1H),4.40-4.34(m,1H),3.92-3.87(m,4H ),3.85-3.82(t,2H),3.72-3.69(m,2H),3.59-3.56(m,6H),3.54-3.52(m,2H),2.69-2.65(t,2H),1.25(d,3H).

[0157] LCMS: m / z = 535.40[M+l]+.

[0158] Example 12

[0159] 5-Bromo-6-{[(2S)-1-[(3-{5-[5-(trifluoromethyl)pyrimidin-2-yl]-2,5-diazabicyclo[2.2.1]hept-2-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-12)

[0160] This compound was prepared from intermediates 4a (Table 2) and 11b (Table 4) according to reaction route 5. The product was characterized, and the results are as follows:

[0161] LCMS: m / z = 532.12[M+l]+.

[0162] Example 13

[0163] 5-Bromo-6-{[(2S)-1-({3-[5-(5-fluoropyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]hept-2-yl]propyl}oxy)propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-13)

[0164] This compound was prepared from intermediates 13a and 11c (Table 4) according to reaction route 5. The product was characterized, and the results are as follows:

[0165] LCMS: m / z = 482.12[M+l]+.

[0166] Example 14

[0167] 2-[2-(3-{[(2S)-2-[(5-bromo-6-oxoylide-1H-pyrimidin-4-yl)amino]propyl]oxy}propyl)-2,5-diazabicyclo[2.2.1]hept-5-yl]pyrimidin-5-carboxynitrile (I-14)

[0168] This compound was prepared from intermediates 13a and 11d (Table 4) according to reaction route 5. The product was characterized, and the results are as follows:

[0169] LCMS: m / z = 489.13[M+l]+.

[0170] Example 15

[0171] 5-Bromo-6-{[(2S)-1-({3-[2-(5-fluoropyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]hept-5-yl]-3-oxopropyl}oxy)propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-15)

[0172] This compound was prepared from intermediates 4a (Table 2) and 11c (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0173] LCMS: m / z = 496.10[M+l]+.

[0174] Example 16

[0175] 5-Bromo-6-{[(2S)-1-({3-[(3aR,6aS)-5-[5-(trifluoromethyl)pyridin-2-yl]octahydropyrrolo[4,3-c]pyrrolo-2-yl]propyl}oxy)propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-16)

[0176] This compound was prepared from intermediates 13a and 8c (Table 3) according to reaction route 5. The product was characterized, and the results are as follows:

[0177] LCMS: m / z = 545.14[M+l]+.

[0178] Example 17

[0179] 5-Bromo-6-{[(2S)-1-({3-oxoylide-3-[(3aS,6aR)-5-[5-(trifluoromethyl)pyridin-2-yl]octahydropyrrolo[4,3-c]pyrrolo-2-yl]propyl}oxy)propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-17)

[0180] The compound was prepared from intermediates 4a (Table 2) and 8c (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0181] 1 H NMR (400MHz, CDCl3) δ13.11(s,1H),8.36(s,1H),7.90(d,1H),7.60(dd,1H),6.34(d,1H),5.60(d,1H),4.36-4.32( m,1H),3.86-3.73(m,6H),3.54-3.48(m,3H),3.45-3.40(m,3H),3.18-3.05(m,2H),2.57-2.54(t,2H),1.22(d,3H).

[0182] LCMS: m / z = 560.50[M+l]+.

[0183] Example 18

[0184] 5-Bromo-6-{[(2S)-1-({3-oxoylide-3-[(3aS,6aR)-5-[5-(trifluoromethyl)pyrimidin-2-yl]octahydropyrrolo[4,3-c]pyrrolo-2-yl]propyl}oxy)propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-18)

[0185] This compound was prepared from intermediates 4a (Table 2) and 11e (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0186] LCMS: m / z = 560.12[M+l]+.

[0187] Example 19

[0188] 5-Bromo-6-{[(2S)-1-[(3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-19)

[0189] This compound was prepared from intermediates 13a and 8d (Table 3) according to reaction route 5. The product was characterized, and the results are as follows:

[0190] LCMS: m / z = 531.13[M+l]+.

[0191] Example 20

[0192] 5-Bromo-6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-20)

[0193] This compound was prepared from intermediates 4a (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0194] 1 H NMR (400MHz, CDCl3) δ13.21(s,1H),8.35(s,1H),7.91(s,1H),7.60(dd,1H),6.27(d,1H),5.59(d,1H),4.41-4. 37(m,1H),4.35(s,2H),4.22-15(m,6H),3.80-3.77(m,2H),3.55-3.48(m,2H),2.38-2.65(t,2H),1.25(d,3H).

[0195] LCMS: m / z = 546.50[M+l]+.

[0196] Example 21

[0197] 6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-5-methyl-3,4-dihydropyrimidin-4-one (I-21)

[0198] This compound was prepared from intermediates 4d (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0199] LCMS: m / z = 481.21[M+l]+.

[0200] Example 22

[0201] 5-Cyclopropyl-6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-22)

[0202] This compound was obtained by reacting compound I-20 prepared in Example 20 with cyclopropanoboric acid under Suzuki reaction conditions. The product was characterized, and the results are as follows:

[0203] LCMS: m / z = 507.23[M+l]+.

[0204] Example 23

[0205] 5-Fluoro-6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-23)

[0206] The compound was prepared from intermediates 4c (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0207] LCMS: m / z = 485.18[M+l]+.

[0208] Example 24

[0209] 5-Chloro-6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-24)

[0210] The compound was prepared from intermediates 4b (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0211] 1 H NMR (400MHz, DMSO) δ12.15(s,1H),8.38(s,1H),7.95(s,1H),7.78(dd,1H),6.48(d,1H),6.39(d,1H),4.37-4.31(m,1H),4.30(s ,2H),4.18-4.13(m,4H),4.02(s,2H),3.61-3.58(t,2H),3.51-3.44(m,1H),3.33-3.18(m,1H),2.25-2.24(t,2H),1.13(d,3H).

[0212] LCMS: m / z = 501.90[M+l]+.

[0213] Reaction route 7:

[0214]

[0215] Reaction route 7

[0216] Example 25

[0217] 6-{[(2S)-1-[(3-oxoylide-3-{6-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-2-yl}propyl)oxy]propyl-2-yl]amino}-5-(trifluoromethyl)-3,4-dihydropyrimidin-4-one (I-25)

[0218] This compound was prepared from compound I-20 prepared in Example 20 according to reaction route 7. The product was characterized, and the characterization results are as follows:

[0219] LCMS: m / z = 535.18[M+l]+.

[0220] Example 26

[0221] 6-{[(2S)-1-[(3-{6-[3-methyl-5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-2-yl}-3-oxoylidenepropyl)oxy]propyl-2-yl]amino}-5-methyl-3,4-dihydropyrimidin-4-one (I-26)

[0222] The compound was prepared from intermediate 4d (Table 2) and 8g (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0223] LCMS: m / z = 495.23[M+l]+.

[0224] Example 27

[0225] 6-{[(2S)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyrimidin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-5-methyl-3,4-dihydropyrimidin-4-one (I-27)

[0226] This compound was prepared from intermediates 4d (Table 2) and 11f (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0227] LCMS: m / z = 482.20[M+l]+.

[0228] Example 28

[0229] 5-Bromo-6-{[(2S)-1-[(3-{2-[3-methoxy-5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}-3-oxoylidenepropyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-28)

[0230] The compound was prepared from intermediate 4a (Table 2) and 8h (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0231] LCMS: m / z = 575.12[M+l]+.

[0232] Example 29

[0233] 5-Bromo-6-{[(2R)-1,1,1-trifluoro-3-[(3-oxoylide-3-{6-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-2-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-29)

[0234] This compound was prepared from intermediates 4e (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0235] LCMS: m / z = 599.08[M+l]+.

[0236] Example 30

[0237] 5-Bromo-6-{[(2R)-1,1,1-trifluoro-3-[(3-oxoylide-3-{6-[5-(trifluoromethyl)pyrimidin-2-yl]-2,6-diazaspiro[3.3]hept-2-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-30)

[0238] This compound was prepared from intermediates 4e (Table 2) and 11f (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0239] LCMS: m / z = 600.07[M+l]+.

[0240] Example 31

[0241] 5-Bromo-6-{[(2R)-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl]amino}-3,4-dihydropyrimidin-4-one (I-31)

[0242] This compound was prepared from intermediates 4f (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0243] LCMS: m / z = 545.10[M+l]+.

[0244] Example 32

[0245] 5-Bromo-6-({2-methyl-1-[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]propyl-2-yl}amino)-3,4-dihydropyrimidin-4-one (I-32)

[0246] The compound was prepared from intermediate 4g (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0247] LCMS: m / z = 559.12[M+l]+.

[0248] Example 33

[0249] 5-Bromo-6-[({[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]methyl}cyclopropyl)amino]-3,4-dihydropyrimidin-4-one (I-33)

[0250] The compound was prepared from intermediates 4h (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0251] LCMS: m / z = 557.10[M+l]+.

[0252] Example 34

[0253] 5-Bromo-6-[(2S)-2-{[(3-oxoylide-3-{4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}propyl)oxy]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-34)

[0254] This compound was prepared from intermediate 4i (Table 2) and 11a (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0255] 1 H NMR (400MHz, CDCl3) δ13.25(s,1H),8.50(s,2H),7.84(s,1H),4.73-4.71(m,1H),4.03-3.97(m,1H),3.94-3.88(m,4H),3.82-3 .76(m,2H),3.74-3.65(m,4H),3.59-3.56(m,2H),3.41-3.36(m,1H),2.65-2.61(t,2H),2.00-1.95(m,2H),1.49-1.42(m,2H).

[0256] LCMS: m / z = 561.30[M+l]+.

[0257] Example 35

[0258] 5-Bromo-6-[(2S)-2-{[(3-oxoylide-3-{4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}propyl)oxy]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-35)

[0259] This compound was prepared from intermediate 4i (Table 2) and 8a (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0260] 1 H NMR(400MHz, CDCl3)δ8.39(s,1H),7.83(s,1H),7.66(dd,1H),6.65(d,1H),4.71-4.69(m,1H),4.00-3.95(m,1H),3.83-3.75(m,4H), 3.74-3.72(m,2H),3.70-3.66(m,2H),3.64-3.61(m,4H),3.40-3.36(m,1H),2.64-2.61(t,2H),1.99-1.94(m,2H),1.46-1.43(m,2H).

[0261] LCMS: m / z = 560.30[M+l]+.

[0262] Example 36

[0263] 5-Bromo-6-[(2S)-2-{[(3-{4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}propyl)oxy]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-36)

[0264] This compound was prepared by reducing the amide group of compound I-35 prepared in Example 35 with Ca(BH4)2. The product was characterized, and the characterization results are as follows:

[0265] LCMS: m / z = 545.14[M+l]+.

[0266] Example 37

[0267] 6-[(2S)-2-[({3-[(3aR,6aS)-5-[5-(trifluoromethyl)pyridin-2-yl]octahydropyrrolo[4,3-c]pyrrolo-2-yl]propyl}oxy)methyl]tetrahydro-1H-pyrrolo-1-yl]-5-bromo-3,4-dihydropyrimidin-4-one (I-37)

[0268] This compound was prepared from intermediates 4i (Table 2) and 8c (Table 3) according to reaction route 5. The product was characterized, and the results are as follows:

[0269] LCMS: m / z = 571.16[M+l]+.

[0270] Example 38

[0271] 5-Bromo-6-[(2S)-2-[({3-oxoylide-3-[(3aS,6aR)-5-[5-(trifluoromethyl)pyridin-2-yl]octahydropyrrolo[4,3-c]pyrrolo-2-yl]propyl}oxy)methyl]tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-38)

[0272] This compound was prepared from intermediates 4i (Table 2) and 8c (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0273] LCMS: m / z = 585.14[M+l]+.

[0274] Example 39

[0275] 5-Bromo-6-[(2S)-2-[({3-oxoylide-3-[(3aS,6aR)-5-[5-(trifluoromethyl)pyrimidin-2-yl]octahydropyrrolo[4,3-c]pyrrolo-2-yl]propyl}oxy)methyl]tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-39)

[0276] This compound was prepared from intermediates 4i (Table 2) and 11e (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0277] LCMS: m / z = 586.13[M+l]+.

[0278] Example 40

[0279] 5-Bromo-6-[(2S)-2-{[(3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-40)

[0280] This compound was prepared from intermediate 13i and 8d (Table 3) according to reaction route 5. The product was characterized, and the results are as follows:

[0281] LCMS: m / z = 557.14[M+l]+.

[0282] Example 41

[0283] 5-Bromo-6-[(2S)-2-{[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyridin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-41)

[0284] This compound was prepared from intermediates 4i (Table 2) and 8d (Table 3) according to reaction route 4. The product was characterized, and the results are as follows:

[0285] LCMS: m / z = 571.12[M+l]+.

[0286] Example 42

[0287] 5-Bromo-6-[(2S)-2-{[(3-oxoylide-3-{2-[5-(trifluoromethyl)pyrimidin-2-yl]-2,6-diazaspiro[3.3]hept-6-yl}propyl)oxy]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-42)

[0288] This compound was prepared from intermediate 4i (Table 2) and 11f (Table 4) according to reaction route 4. The product was characterized, and the results are as follows:

[0289] LCMS: m / z = 572.12[M+l]+.

[0290] Example 43

[0291] 5-Bromo-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-43)

[0292] The compound was prepared from intermediates 2i (Table 1) and 8a (Table 3) via reaction routes 6 and 4. The product was characterized, and the results are as follows:

[0293] LCMS: m / z = 570.14[M+l]+.

[0294] Example 44

[0295] 5-Bromo-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}methyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-44)

[0296] This compound was prepared from compound I-43 prepared in Example 43 according to reaction route 7. The product was characterized, and the characterization results are as follows:

[0297] LCMS: m / z = 556.16[M+l]+.

[0298] Example 45

[0299] 5-Bromo-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-45)

[0300] This compound was prepared from intermediates 2i (Table 1) and 11a (Table 4) according to reaction routes 6 and 4. The product was characterized, and the results are as follows:

[0301] LCMS: m / z = 571.13[M+l]+.

[0302] Example 46

[0303] 5-Methyl-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-46)

[0304] The compound was prepared from intermediates 2j (Table 1) and 11a (Table 4) according to reaction routes 6 and 4. The product was characterized, and the results are as follows:

[0305] LCMS: m / z = 507.24[M+l]+.

[0306] Example 47

[0307] 5-(trifluoromethyl)-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-47)

[0308] This compound was prepared from compound I-45 prepared in Example 45 according to reaction route 7. The product was characterized, and the characterization results are as follows:

[0309] LCMS: m / z = 561.21[M+l]+.

[0310] Example 48

[0311] 5-Methyl-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-48)

[0312] The compound was prepared from intermediate 2j (Table 1) and 8a (Table 3) via reaction routes 6 and 4. The product was characterized, and the results are as follows:

[0313] LCMS: m / z = 506.24[M+l]+.

[0314] Example 49

[0315] 5-Cyclopropyl-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-49)

[0316] This compound was obtained by reacting compound I-43 prepared in Example 43 with cyclopropanoboric acid under Suzuki reaction conditions. The product was characterized, and the results are as follows:

[0317] LCMS: m / z = 532.26[M+l]+.

[0318] Example 50

[0319] 5-(trifluoromethyl)-6-[(2S)-2-{[3-({4-[5-(trifluoromethyl)pyridin-2-yl]piperazin-1-yl}carbonyl)azacyclobut-1-yl]methyl}tetrahydro-1H-pyrrolo-1-yl]-3,4-dihydropyrimidin-4-one (I-50)

[0320] This compound was prepared from compound I-43 prepared in Example 43 according to reaction route 7. The product was characterized, and the characterization results are as follows:

[0321] LCMS: m / z = 560.21[M+l]+.

[0322] Example 51

[0323] 6-{4-[(1-{[(2S)-1-(5-methyl-6-oxoylide-1H-pyrimidin-4-yl)tetrahydro-1H-pyrrolo-2-yl]methyl}azacyclobut-3-yl)carbonyl]piperazin-1-yl}pyridin-3-carboxynitrile (I-51)

[0324] The compound was prepared from intermediates 2j (Table 1) and 8e (Table 3) via reaction routes 6 and 4. The product was characterized, and the results are as follows:

[0325] LCMS: m / z = 463.25[M+l]+.

[0326] Reaction route 8:

[0327]

[0328] Reaction route 8

[0329] Example 52

[0330] 6-{[(2S)-1-[3-({4-[5-(trifluoromethyl)pyridin-2-yl]-1,2,3,6-tetrahydropyridin-1-yl}carbonyl)azacyclobut-1-yl]propyl-2-yl]amino}-5-(trifluoromethyl)-3,4-dihydropyrimidin-4-one (I-52)

[0331] The compound was prepared by condensing intermediate 26 obtained from reaction route 8 with intermediate 20 to obtain the product, which was then prepared according to reaction route 7. The product was characterized, and the characterization results are as follows:

[0332] LCMS: m / z = 531.19[M+l]+.

[0333] Example 53

[0334] 6-{[(2S)-1-[3-({4-[5-(trifluoromethyl)pyridin-2-yl]hexahydropyridin-1-yl}carbonyl)azacyclobut-1-yl]propyl-2-yl]amino}-5-(trifluoromethyl)-3,4-dihydropyrimidin-4-one (I-53)

[0335] The compound was prepared by condensing intermediate 27 obtained from reaction route 8 with intermediate 20 to obtain the product, which was then prepared according to reaction route 7. The product was characterized, and the characterization results are as follows:

[0336] LCMS: m / z = 533.20[M+l]+.

[0337] Example 54

[0338] 6-{[(2S)-1-({3-oxoylidene-3-[4-(5-vinylpyrimidin-2-yl)piperazin-1-yl]propyl}oxy)propyl-2-yl]amino}-5-(trifluoromethyl)-3,4-dihydropyrimidin-4-one (I-54)

[0339] The compound was prepared from intermediates 4a (Table 1) and 11j (Table 4) according to reaction routes 4 and 7. The product was characterized, and the results are as follows:

[0340] LCMS: m / z = 482.20[M+l]+.

[0341] Example 55

[0342] 6-{[(2S)-1-({3-[4-(5-ethynylpyrimidin-2-yl)piperazin-1-yl]-3-oxopropyl}oxy)propyl-2-yl]amino}-5-(trifluoromethyl)-3,4-dihydropyrimidin-4-one (I-55)

[0343] This compound was prepared from intermediate 4a (Table 1) and 11h (Table 4) according to reaction routes 4 and 7. The product was characterized, and the results are as follows:

[0344] LCMS: m / z = 480.19[M+l]+.

[0345] Example 56

[0346] 6-{[(2S)-1-({3-[4-(5-cyclopropylpyrimidin-2-yl)piperazin-1-yl]-3-oxopropyl}oxy)propyl-2-yl]amino}-5-(trifluoromethyl)-3,4-dihydropyrimidin-4-one (I-56)

[0347] This compound was prepared from intermediates 4a (Table 1) and 11i (Table 4) according to reaction routes 4 and 7. The product was characterized, and the results are as follows:

[0348] LCMS: m / z = 496.22[M+l]+.

[0349] Reaction route 9:

[0350]

[0351] Reaction route 9

[0352] Example 57

[0353] 4-{[(2S)-1-[(3-oxoylidene-3-{4-[5-(trifluoromethyl)pyrimidin-2-yl]piperazin-1-yl}propyl)oxy]propyl-2-yl]amino}-3-methyl-1,2-dihydropyridin-2-one (I-57)

[0354] The compound was prepared from intermediate 31 obtained by reaction route 9, and then prepared with intermediate 11a (Table 4) according to reaction route 4. The product was characterized, and the characterization results are as follows:

[0355] LCMS: m / z = 469.48[M+l]+.

[0356] Bioactivity Research

[0357] 1. In vitro inhibitory activity study

[0358] The bioactivity of some of the target compounds synthesized in Examples 1-57 above was tested, using commercially available drugs RBN-2397 and Olaparib as control samples. Their in vitro inhibitory activity against PARP7 was tested, and the specific procedures are as follows:

[0359] (1) Add 25 μL of histone (active motif) solution to each well of a 384 microplate and incubate overnight at 4°C.

[0360] (2) Prepare PBST buffer, blocking buffer and analysis buffer.

[0361] (3) Wash the histone-coated 384-well plate three times with PBST buffer, then block with 50 μL of blocking buffer at room temperature for 1 h, and finally wash with PBST buffer three times.

[0362] (4) Prepare 2000×(20mM) compound. Transfer 50nL of the compound from the source plate to a 96-well plate using 19.95uL buffer. Shake well and centrifuge at 1000rpm for 1min. Transfer 5μL of DMSO / compound to each well of a 384-well microplate.

[0363] (5) Except for the minimum control well, add 10 μL of enzyme mixture to each well and incubate with the compound at room temperature for 10 min. Add 10 μL of detection buffer to the minimum control well. Then add 10 μL of 2.5×Biotin-NAD+(R&D) to each well and incubate at 25 °C for 60 min. Finally, wash three times with PBST buffer.

[0364] (6) Add 25 μL of Strep-HRP (Thermo Pierce), incubate at room temperature for 1 h, and wash 3 times with PBS buffer. Then add 25 μL of QuantaRed enhancer mixture. Incubate for 10 min. Finally, add 2.5 μL of QuantaRed (Thermo Pierce) stop solution, shake for 10-30 seconds to stop peroxidase activity.

[0365] (7) Take the reading immediately on Paradigm (Ex550 / Em620).

[0366] (8) First, perform data fitting: Equation (1): inh%=(maximum signal - composite signal) / (maximum signal - minimum signal)×100. Then, calculate according to Equation (2): Y=Bottom+(Top-Bottom) / (1+(IC 50 Calculate IC using ( / X)×HillSlope) 50 In the formula, Y is the inhibition rate, X is the compound concentration, Top and Bottom are the minimum and maximum values ​​observed on the curve, respectively, and HillSlope refers to the absolute value of the maximum slope of the curve, i.e., the midpoint of the curve.

[0367] Table 5. In vitro inhibition test IC50 of different compounds on PARP7 50 Test Results

[0368] compound IC50(nM) RBN-2397 4.80±1.87 Olaparib 134.35 I-1 34.45±0.37 I-6 28.93±4.86 I-20 10.88±0.23 I-34 4.05±1.20 I-35 4.23±2.06 I-57 8.35±0.24

[0369] According to the IC50 test results of the above-mentioned in vitro inhibition assay for PARP7, the new compound provided in this embodiment of the invention has high inhibitory activity for PARP7, which is superior to that of Olaparib. Moreover, the in vitro inhibitory activity of compounds I-34 and I-35 for PARP7 is higher than that of RBN-2397.

[0370] 2. In vitro hERG study

[0371] Taking compound I-35 as an example, this invention tests the hERG IC of this compound. 50 The test results showed that compound I-35 had hERG IC values. 50 The value was 7.14 ± 1.63 μm. This indicates that the compound exhibits low inhibitory activity against hERG potassium channels, but at low doses, it shows minimal cardiovascular toxicity, meeting pharmaceutical requirements.

[0372] 3. Pharmacokinetic studies

[0373] Taking compound I-35 as an example, male SD rats were used as test animals. After a single dose, the blood concentration of the compound was measured and its pharmacokinetic behavior was evaluated. The specific procedures are as follows:

[0374] Three healthy adult male SD rats were selected for the intravenous injection group. The solvent for the intravenous injection group was 10% DMSO + 90% (30% HP-β-CD). An appropriate amount of the compound was weighed, dissolved in the corresponding volume of 10% DMSO, and then the corresponding volume of 90% (30% HP-β-CD) was added sequentially. The mixture was thoroughly mixed and vortexed to dissolve, yielding a clear solution of 0.1 mg / mL. After intravenous injection of 1 mg / kg into the rats, whole blood was collected at a certain time to prepare plasma. The concentration of the parent drug in the plasma was detected by LC-MS / MS, and relevant pharmacokinetic parameters were calculated using WinNonlin software. The experimental results are shown in Table 6 below.

[0375] Table 6. Pharmacokinetic parameters of rats administered 1 mg / kg via intravenous injection.

[0376]

[0377] Three healthy adult male SD rats were selected as the oral administration group. The solvent for the oral administration group was 10% DMSO + 90% (30% HP-β-CD). An appropriate amount of the compound was weighed, dissolved in the corresponding volume of 10% DMSO, and then the corresponding volume of 90% (30% HP-β-CD) was added sequentially. The mixture was thoroughly mixed and vortexed to dissolve, yielding a clear solution of 1 mg / ml. After oral administration of 10 mg / kg to the rats, whole blood was collected at a certain time to prepare plasma. The concentration of the parent drug in the plasma was detected by LC-MS / MS, and relevant pharmacokinetic parameters were calculated using WinNonlin software. The experimental results are shown in Table 7 below.

[0378] Table 7 Pharmacokinetic parameters of rats administered by gavage (n=3, males)

[0379]

[0380] In summary, the novel compounds provided by this invention have high inhibitory activity against PARP7, low inhibitory activity against hERG potassium channels, and good drug-like properties, and have extremely high application value in the preparation of drugs for the prevention and / or treatment of diseases related to elevated PARP7 levels in the body.

[0381] The embodiments described above are merely preferred embodiments for fully illustrating the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. A compound of formula I-34 or formula I-35, or a pharmaceutically acceptable salt thereof. 。 2. A pharmaceutical composition, characterized in that, It comprises one or more of the compounds of formula I-34 or I-35 as claimed in claim 1, or their pharmaceutically acceptable salts.

3. The use of a compound of formula I-34 or I-35 as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a PARP7 inhibitor.

4. The application according to claim 3, characterized in that, The PARP7 inhibitors are administered orally, parenterally, intravenously, or transdermally.

5. The use of a compound of formula I-34 or I-35 as claimed in claim 1, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a medicament for treating a patient’s PARP7-mediated condition.

6. The application according to claim 5, characterized in that, The PARP7-mediated diseases include breast cancer, ovarian cancer, and colorectal cancer.

7. The application according to claim 5, characterized in that, The drug is administered orally, parenterally, intravenously, or transdermally.

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