Isobenzofuran-1 (3H)-ketone derivative and application thereof

Abstract

The invention discloses an isobenzofuran-1 (3H)-ketone derivative and application thereof, and relates to the isobenzofuran-1 (3H)-ketone derivative as shown in a formula (I), and an isotope form, a stereoisomer, a pharmaceutically acceptable salt, a pharmaceutically acceptable solvate, a hydrate, a prodrug and a polymorphic substance of the isobenzofuran-1 (3H)-ketone derivative, the invention also discloses a preparation method of the compound and application of the compound in cardiovascular ischemic diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and particularly to an isobenzofuran-1(3H)-one derivative and its application. Background Art

[0002] "Stroke" is an acute cerebrovascular disease, mainly divided into hemorrhagic stroke (cerebral hemorrhage or subarachnoid hemorrhage) and ischemic stroke (cerebral infarction, cerebral thrombosis), and is listed as one of the three major diseases threatening human beings together with coronary heart disease and cancer. Among them, the incidence of ischemic stroke (IS) is higher than that of hemorrhagic stroke, accounting for more than half of the total incidence of stroke. The pathogenesis of IS is caused by multiple reasons leading to cerebral ischemia, and then resulting in cerebral functional disorders. The damaged brain tissues are mainly the ischemic penumbra and the ischemic core region (Widimsky et al., 2023. European Heart Journal 44, 1205-1215.). The reason for the damage of the ischemic penumbra is that collateral vessels are difficult to perform normal blood perfusion function, and there are still a large number of surviving neurons in it, which is a reversible injury; the brain cells in the ischemic core region undergo necrosis due to energy depletion and blood supply interruption, usually an irreversible injury (Tun et al., 2017. Diabetes mellitus and stroke: A clinical update. WJD 8, 235.). Multiple factors may induce the occurrence of stroke, such as diseases like atherosclerosis, hypertension, inflammation invading the meninges and cerebral blood vessels, blood diseases, metabolic diseases, brain tumors, etc. Research shows that BMI, history of hypertension, history of hyperlipidemia, history of diabetes, history of heart disease, history of stroke, age, and smoking history are related to the occurrence of IS (Hurford et al., 2020. Pract Neurol 20, 304-316).

[0003] The pathogenesis of ischemic stroke is complex and can be classified into early events and late events according to the onset time (Qin et al., 2022. Sig Transduct Target Ther 7, 215.). The main influencing factors after early (from a few minutes to several hours) ischemic injury include glutamate-induced excitotoxicity, Ca 2+ overload, and ROS-mediated oxidative stress response, etc. The late stage (from several hours to several days) is mainly manifested by inflammatory response and apoptosis. Ischemic brain injury is an enzymatic cascade process of multi-factor, multi-link, and multi-pathway injury. Factors such as cell energy depletion, calcium overload, excitotoxicity, excessive generation of oxygen free radicals, inflammation, and apoptosis are all closely related to neuronal injury after cerebral ischemia.

[0004] In recent years, some synthetic and natural products have been discovered to combat IS. Among them, the racemic or enantiopure forms of 3-n-butylphthalide (NBP) and its derivatives or analogs have shown potential pharmacological properties and good therapeutic effects (Marco-Contelles, Zhang, 2020. J. Med. Chem. 63, 12485-12510.). The disadvantages of this drug are poor PK properties and low bioavailability. Therefore, many researchers are trying to improve PK by derivatization or introduction of hydrophilic functional groups (Edaravone and the ring-opening derivatives of 3-n-butylphthalide can effectively reduce cerebral ischemic injury in rats (Hua et al., 2015. Acta Pharmacol Sin 36, 917-927.)).

[0005] The beneficial effects of NBP on acute or chronic stroke have been well studied and reported in animal models and clinical patients. In 2002, the China Food and Drug Administration (CFDA) approved this drug for clinical phase I-II trials of stroke. NBP shows various protective effects on ischemic stroke, including improving energy metabolism, reducing oxidative damage, improving arteriolar microcirculation, and lowering blood pressure (Liu et al., 2007. Journal of the Neurological Sciences 260, 106-113; Ma et al., 2009. Naunyn-Schmied Arch Pharmacol 379, 565-574.). However, whether NBP has a cardioprotective effect on ischemic injury remains to be studied. Later, (±)-sodium 5-bromo-2-(α-hydroxyamyl) benzoate (BZP) was developed as an effective anti-ischemic stroke drug (Xiao et al., 2019. Journal of Molecular and Cellular Cardiology 135, 52-66). Compared with NBP, although BZP sodium salt shows good water solubility, effective in vitro and in vivo activities, and less hepatocyte toxicity, its drug bioavailability is not very good, and the AUC and hERG of the drug are not good enough. When applied at higher doses, it may also cause toxicity and cardiac arrest. Summary of the Invention

[0006] The object of the present invention is to provide an isobenzofuran-1(3H)-one derivative and its application in the preparation of a drug for preventing and treating cardiovascular ischemic diseases.

[0007] To solve the above technical problems, the present invention provides the following technical solutions:

[0008] The first aspect of the present invention provides an isobenzofuran-1(3H)-one derivative having a structure of Formula I or its isotopic forms, stereoisomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs and polymorphs,

[0009]

[0010] in,

[0011] A is one of H, halogen, and 3-10 membered heterocyclic group;

[0012] E is -CR 1 R 2 -or-O-CR 1 R 2 -;

[0013] Z is one of H, deuterium, halogen, hydroxyl, saturated or unsaturated C1-C4 alkyl, C4-C7 cycloalkyl, and 3-10 membered heterocyclic group;

[0014] The heterocyclic ring in the 3-10 membered heterocyclic group is a heterocarbocyclic ring, heteroaromatic ring, heteroalicyclic ring, heterospirocyclic ring, heterobridged ring or heterocyclic ring having one or more heteroatoms, wherein the heteroatoms are one or more of N, S and O;

[0015] R 1 、R 2 Independently selected from one of H, deuterium, halogen, double bond, triple bond, and C1-C10 alkyl;

[0016] The above heterocyclic groups, alkyl groups, and cycloalkyl groups are unsubstituted or substituted by one or more of the following substituents: halogen, hydroxyl, C1-C10 alkyl, halogen-substituted C1-C10 alkyl, deuterium-substituted C1-C10 alkyl, -CO-R 3 、-NH-CO-R 4 、-COOR 5 ;

[0017] R 3 、R 4 、R 5 Independently selected from unsubstituted or halogen-substituted C1-C10 alkyl groups.

[0018] Furthermore, A is preferably bromine or a 3-10 membered heterocyclic group containing at least one N heteroatom, wherein the heterocyclic group is unsubstituted or substituted by one or more of the following substituents: hydroxyl, chlorine, bromine, methyl, deuterated methyl, -COCH3, -COCF3, -NH-COCH3, -COOC(CH3)3.

[0019] Furthermore, E is preferably -CR 1 R 2 -, R1 、R 2 Independently selected from one of H and C1-C10 alkyl, wherein the C1-C10 alkyl includes but is not limited to methyl, ethyl, propyl, butyl, tert-butyl and the like.

[0020] Furthermore, Z is selected from one of hydrogen, hydroxyl, fluorine, vinyl, C1-C4 alkyl, C3-C6 cycloalkyl, and 3-10 membered heterocyclic group containing N and / or O heteroatoms, wherein the alkyl and heterocyclic groups are unsubstituted or substituted by one or more of the following groups: fluorine, hydroxyl, methyl, deuterated methyl, -COCH3, -COCF3, -NH-COCH3, and -COOC(CH3)3.

[0021] Furthermore, the isobenzofuran-1(3H)-one derivative preferably has the general structural formula shown in the following formulas Ia to Ic:

[0022]

[0023] Wherein, n is 1, 2 or 3;

[0024] A is bromine or a 3-10 membered heterocyclic group containing at least one nitrogen heteroatom, wherein the heterocyclic group is unsubstituted or substituted with one or more of the following substituents: hydroxyl, chlorine, bromine, methyl, deuterated methyl, -COCH3, -COCF3, -NH-COCH3, -COOC(CH3)3;

[0025] Z is selected from one of hydrogen, hydroxyl, fluorine, vinyl, C1-C4 alkyl, C3-C6 cycloalkyl, and 3-10 membered heterocyclic group containing N and / or O heteroatoms, wherein the alkyl and heterocyclic groups are unsubstituted or substituted by one or more of the following groups: fluorine, hydroxyl, methyl, deuterated methyl, -COCH3, -COCF3, -NH-COCH3, -COOC(CH3)3.

[0026] Furthermore, the A is selected from one of the following structures:

[0027]

[0028] Furthermore, the E is selected from one of the following structures:

[0029]

[0030] Furthermore, Z is selected from one of the following structures:

[0031]

[0032] Furthermore, the isobenzofuran-1(3H)-one derivative is a compound represented by the following structural formulas I-01 to I-121:

[0033]

[0034]

[0035]

[0036]

[0037]

[0038]

[0039] The second aspect of the present invention provides a pharmaceutical composition comprising one or more of the isobenzofuran-1(3H)-one derivatives described in the first aspect, their isotopic forms, stereoisomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs and polymorphs.

[0040] Furthermore, in the pharmaceutical composition, the above-mentioned isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph is used in combination with other drugs.

[0041] Furthermore, in the pharmaceutical composition, the isobenzofuran-1(3H)-one derivative includes but is not limited to one or more of the compounds represented by the above formulas I-19, I-36, I-40, I-50, I-61, I-80, I-81, I-99, I-101, I-102, I-104, I-113, I-114, I-118, I-119, I-120, and I-121, and more preferably the compounds represented by formulas I-40, I-61, I-80, I-104, I-118, and I-121.

[0042] The third aspect of the present invention provides the use of the isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph described in the first aspect or the pharmaceutical composition described in the second aspect in the preparation of a drug for preventing and treating cardiovascular ischemic diseases.

[0043] Furthermore, the cardiovascular ischemic disease includes but is not limited to ischemic stroke.

[0044] Furthermore, the drug is administered orally, parenterally, intravenously or transdermally.

[0045] As used herein, the following definitions and terms shall apply unless otherwise indicated.

[0046] The terms "R" and "S", as used to describe enantiomers, are descriptors of the stereochemical configuration of an asymmetrically substituted carbon atom. Naming an asymmetrically substituted carbon atom as "R" or "S" is done by applying the Cahn-Ingold-Prelog priority rules, which are well known to those skilled in the art and are described in Section E, Stereochemistry, of the IUPAC (International Union of Pure and Applied Chemistry) rules for the nomenclature of organic chemistry.

[0047] As used herein, the term Ci-Cj means that the moiety has i - j carbon atoms. For example, "C1-C10 alkyl" means that the alkyl moiety has any number of carbon atoms between 1 and 10.

[0048] As used herein, "saturated alkyl" means a straight-chain or branched-chain alkane group that is fully saturated, and "unsaturated alkyl" means a straight-chain or branched-chain alkane group that contains a carbon-carbon double bond and / or a carbon-carbon triple bond. Non-limiting examples of exemplary alkyl groups include methyl, 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 carbocyclic ring, each ring having 3 to 10 carbon atoms.

[0049] The term "3- to 10-membered heterocyclic group" refers to one or more closed rings having one or more heteroatoms (e.g., oxygen, nitrogen, or sulfur), including heterocyclic carbocycles, heteroaromatic rings, heterocyclic alicyclics, heterospirocycles, heterobridged rings, hetero-fused rings, etc. Exemplarily, and without limitation, it includes oxiranyl, pyridyl, pyrrolyl, pyrazolyl, quinolinyl, isoquinolinyl, indolyl, furyl, thienyl, thiazolyl, benzofuranyl, benzothiazolyl, etc.

[0050] Unless otherwise specified, the term "halogen" or "halo element" by itself or as part of another substituent refers to a fluorine, chlorine, bromine, or iodine atom. Additionally, the term "halogen-substituted alkyl" means including monohalogenated alkyls and polyhalogenated alkyls. For example, "halogen-substituted C1-C4 alkyl" refers to including, but not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, etc.

[0051] Optical isomers, diastereoisomers, geometric isomers, and tautomers: The above isobenzofuran-1(3H)-one derivatives contain one or more ring systems and thus may exist as cis and trans isomers. The present invention is intended to cover all such cis and trans isomers. The presence of an olefinic double bond, unless otherwise specified, means including E and Z geometric isomers.

[0052] Any enantiomer of the isobenzofuran-1(3H)-one derivative can be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration.

[0053] In addition, isobenzofuran-1(3H)-one derivatives may also include a series of stable isotope-labeled analogs. For example, one or more protons in the isobenzofuran-1(3H)-one derivatives can be replaced by deuterium atoms, thereby providing deuterated analogs with improved pharmacological activities.

[0054] "Pharmaceutically acceptable salts" refer to acid salts or base salts of the compounds of the present invention, which salts have the desired pharmacological activities and are neither biologically desirable nor otherwise desirable. The salts can be formed with acids including but not limited to acetic acid, adipic acid, benzoate, citric acid, camphoric acid, camphorsulfonate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrobromide hydrochloride, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, oxalate.

[0055] By the above technical solutions, the present invention has at least the following advantages:

[0056] The present invention provides a series of isobenzofuran-1(3H)-one derivatives. The in vitro cell experiment results show that such compounds have an inhibitory effect on H2O2-induced PC12 cell damage, and compared with edaravone, butylphthalide, and BZP, some of the isobenzofuran-1(3H)-one derivatives provided by the present invention have better inhibitory effects; in addition, the hERG safety evaluation and animal experiment results show that such isobenzofuran-1(3H)-one derivatives have low cardiotoxicity and better drug PK properties, and can be used in the preparation of drugs for preventing cardiovascular ischemic diseases, having extremely high potential application values. Detailed implementation manners

[0057] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the description of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0058] The present invention will be further described below in conjunction with specific embodiments, so that those skilled in the art can better understand the present invention and be able to implement it, but the exemplified embodiments are not used as limitations to the present invention.

[0059] For the convenience of those skilled in the art to fully understand the technical solutions in the present invention, the separation and purification methods and testing methods, English abbreviations, synthesis methods of raw materials or intermediates, and synthesis methods of target compounds based on the embodiments of this application are described as follows:

[0060] 1. In the following embodiments of the present application, the separation and purification methods and testing methods are as follows:

[0061] In the following embodiments of the present invention, unless otherwise specified, for column chromatography, silica gel (60 - 120, 100 - 200 mesh) and eluents with different solvent gradients (ethyl acetate / petroleum ether) are used. Solvent removal is carried out using a Buchii rotary evaporator or a Genevac centrifugal evaporator. Under acidic mobile phase conditions, LC / MS is performed using a Waters automatic purifier and a 19×100mm XTerra 5 micron MS C18 column. Nuclear magnetic resonance spectra are 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 an inert gas (e.g., nitrogen, argon, etc.) that is substantially free of water or dry.

[0062] 2. In the following embodiments of the present application, the Chinese names corresponding to the English abbreviations are as follows: HATU: 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DPCI: N,N'-diisopropylcarbodiimide; DIEA: N,N-diisopropylethylamine; TEA: triethylamine; DMAP: 4-dimethylaminopyridine; DMF: N,N-dimethylformamide; NMP: N-methylpyrrolidone; THF: tetrahydrofuran; DMA: N,N-dimethylacetamide; PTSA: p-toluenesulfonic acid; NIS: N-iodosuccinimide; DMSO-d6: deuterated dimethyl sulfoxide; TLC: thin layer chromatography; eq: equivalent; mol: mole; mmol: millimole; L: liter; mL: milliliter; MHz: megahertz; δ: chemical shift; Hrs, hr, h: hour; Ms: mass spectrometry; m / z: mass-to-charge ratio.

[0063] 3. In the following embodiments of the present application, the starting materials or intermediates used are commercially available or prepared according to known literature methods, as follows:

[0064] The structure of intermediate 2-4 is shown in the following reaction route 1:

[0065]

[0066] Reaction Route 1

[0067] The preparation process of intermediate 2 is as follows:

[0068] Compound 1 (4.9 g, 25.76 mmol) was added to H2SO4 (50 mL) at 0 °C, and then KNO3 (2.6 g, 25.76 mmol) was added in small portions. The mixture was stirred for 0.5 h, and then the reaction mixture was poured into H2O and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give a crude mixture. The residue was purified by silica gel column chromatography to give 3-butyl-6-nitroisobenzofuran-1(3H)-one (Intermediate 2, 5.0 g, yield: 95%).

[0069] The preparation process of Intermediate 3 is as follows:

[0070] Air was removed from the mixture of Compound 2 (5.0 g, 21.28 mmol) and 10% Pd / C (0.5 g) in EtOH (50 mL) under vacuum, and then a hydrogen balloon was placed. After 2 h, the mixture was filtered through diatomaceous earth and evaporated. The crude mixture was purified by silica gel column chromatography to give 6-amino-3-butylisobenzofuran-1(3H)-one (Intermediate 3, 4.5 g, yield: 92%).

[0071] The preparation process of Intermediate 4 is as follows:

[0072] A solution of sodium nitrite (1.6 g, 24.15 mmol) in H2O (10 mL) was added to a mixture of Compound 3 (4.5 g, 21.95 mmol) in 48% hydrobromic acid (45 mL) and H2O (15 mL), and the mixture was stirred for 1 h. Then CuBr (1.9 g, 13.39 mmol) was added. The reaction mixture was stirred at 75 °C for 3 - 8 h until TLC showed complete consumption of the starting material and cooled to room temperature. The reaction mixture was extracted with ethyl acetate (50 mL × 3), and the combined organic layers were washed with an aqueous NaCl solution, dried over anhydrous Na2SO4 and evaporated to give a crude mixture. The residue was purified by silica gel column chromatography to give 6-bromo-3-butylisobenzofuran-1(3H)-one (Intermediate 4, 3.8 g, yield: 85%). The product was characterized by NMR and mass spectrometry, and the results are as follows:

[0073] 1 H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 1.8 Hz, 1H), 7.78 (dd, J = 8.1, 1.8 Hz, 1H), 7.32 (d, J = 8.1 Hz, 1H), 5.44 (dd, J = 7.9, 4.1 Hz, 1H), 2.09–1.96 (m, 1H), 1.82–1.68 (m, 1H), 1.54 - 1.29 (m, 4H), 0.90 (t, J = 7.0 Hz, 3H).

[0074] LCMS(ESI): 270.1 [M+H] + 。

[0075] The structure of Intermediate 5-11 is shown in the following Reaction Route 2:

[0076]

[0077] Reaction Route 2

[0078] The preparation process of Intermediate 5 (2-(1-hydroxypentyl)benzoic acid) is as follows:

[0079] Dissolve Intermediate 4 (10.0 g, 52.56 mmol) in MeOH / H2O solution (100 mL, 1:1 v / v). Then heat the solution at 70 °C for 3 hours. Remove the solvent under reduced pressure from the mixture, and dissolve it in water with 2M HCl at 0 °C to pH 4-5. Extract the mixture with cold Et2O (15 mL × 3) and use it for the next step without any purification.

[0080] The preparation process of Intermediate 6 ((S)-1-phenylethane-1-ammonium (S)-2-(1-hydroxypentyl)benzoate) is as follows:

[0081] Dropwise add (S)-α-methylbenzylamine (6.37 g, 52.56 mmol) to a stirred solution of Compound 5 (10.0 g, 52.56 mmol) in cold Et2O below -20 °C. Stir the reaction mixture at -10 °C for 30 minutes, then raise the temperature to room temperature for 10 hours and filter to obtain a crude product in the form of a white solid (4.3 g, two-step yield 25%).

[0082] The preparation processes of Intermediate 8 ((S)-3-butylisobenzofuran-1(3H)-one) and Intermediate 9 ((R)-3-butylisobenzofuran-1(3H)-one) are as follows:

[0083] Add 2M NaOH to Compound 6 (1.10 g, 3.6 mmol) in H2O (20 mL) and stir at RT for 1 h. Extract the reaction solution with Et2O (10 mL × 3). Acidify the aqueous phase to pH 2 with 3M HCl and stir at room temperature for 0.5 hour. Then extract this solution with Et2O (10 mL × 3). Dry the combined organic layers, filter and concentrate to obtain the target compound as a pale yellow oil (0.70 g, 72% yield).

[0084] HPLC conditions: Chiral analytical column: CHIRALPAK IC-3 (150×4.6 mm 3um), mobile phase: n-hexane:IPA = 85:15; temperature: 35 °C; flow rate: 1.0 mL / min; wavelength: 224 nm; retention time: 8.60 min (main) and 9.03 min (minor), to obtain Intermediate 8.

[0085] HPLC conditions: Chiral analytical column: CHIRALPAK IC-3 (150×4.6 mm 3um), mobile phase: n-hexane:IPA = 90:10; temperature: 35 °C; mobile phase: 1.0 mL / min; wavelength: 224 nm; retention time: 10.758 min (minor) and 11.248 min (main), to obtain Intermediate 9.

[0086] The preparation processes of Intermediate 10 ((S)-6-bromo-3-butylisobenzofuran-1(3H)-one) and Intermediate 11 ((R)-6-bromo-3-butylisobenzofuran-1(3H)-one) are as follows:

[0087] Using the method for preparing Intermediate 3 from Intermediate 1 in the reaction route, Intermediate 10 was synthesized from (S)-3-butylisobenzofuran-1(3H)-one (Intermediate 8).

[0088] Chiral analytical column: CHIRALPAK IC-3 (150×4.6 mm 3um); mobile phase: n-hexane:IPA = 90:10; temperature: 35 °C; mobile phase: 1.0 mL / min; wavelength: 224 nm; retention time: 10.119 min (minor) and 11.082 min (main), to obtain Intermediate 10.

[0089] Using the method for preparing Intermediate 3 from Intermediate 1 in the reaction route, Intermediate 10 was synthesized from (R)-3-butylisobenzofuran-1(3H)-one (Intermediate 9).

[0090] Chiral analytical column: CHIRALPAK IC-3 (150×4.6 mm 3um); mobile phase: n-hexane:IPA = 90:10; temperature: 35 °C; flow rate: 1.0 mL / min; wavelength: 224 nm; retention time: 9.891 min (main) and 10.874 min (minor), to obtain Intermediate 11.

[0091] Compounds I-01 to I-12 were prepared according to the method shown in Reaction Route 3, as follows:

[0092]

[0093] Reaction Route 3

[0094] The preparation process is as follows:

[0095] Pd(dppf)Cl2 (32.00 mg, 0.04 mmol) and Na2CO3 (189.09 mg, 1.78 mmol) were added to a degassed mixture of 6-bromo-3-butylisobenzofuran-1(3H)-one (Intermediate 4) (120 mg, 0.37 mmol), a boronic acid derivative (0.67 mmol) in 1,4-dioxane (1.5 mL) and H2O (1.5 mL), and the mixture was stirred at 80°C under a N2 atmosphere for 2 hours. The mixture was then cooled to room temperature, diluted with water, and extracted with ethyl acetate (20 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and filtered. The filtrate was purified by chromatography to obtain the target compound.

[0096] Example 1

[0097] 3-Butyl-6-(pyridin-3-yl)isobenzofuran-1(3H)-one (I-01), the structure is shown below:

[0098]

[0099] The compound was prepared from intermediate 4 and pyridin-3-ylboronic acid (A-01) according to reaction route 3. The product was characterized, and the characterization results are as follows:

[0100] 1 H NMR (400MHz, CDCl3) δ8.89(s,1H),8.67(s,1H),8.09(s,1H),7.93–7.87(m,2H),7.56(d,J=7.9Hz,1H),7.44–7.41( m,1H),5.54(dd,J=7.8,4.2Hz,1H),2.13–2.04(m,1H),1.86–1.77(m,1H),1.54–1.38(m,4H),0.93(t,J=7.0Hz,3H).

[0101] LCMS (ESI): 268.4 [M+H] + .

[0102] Example 2

[0103] 3-Butyl-6-(pyridin-2-yl)isobenzofuran-1(3H)-one (I-02), the structure is shown below:

[0104]

[0105] The compound was prepared from intermediate 4 and pyridin-2-ylboronic acid (A-02) according to reaction scheme 3. The product was characterized. The characterization results are as follows: LCMS (ESI): 268.1 [M+H] + .

[0106] Example 3

[0107] 3-Butyl-6-(pyridin-4-yl)isobenzofuran-1(3H)-one (I-03), the structure is shown below:

[0108]

[0109] The compound was prepared from intermediate 4 and pyridin-4-ylboronic acid (A-03) according to reaction route 3. The product was characterized, and the characterization results are as follows:

[0110] 1 H NMR (400MHz, CDCl3) δ8.72(d,J=5.3Hz,2H),8.15(d,J=1.6Hz,1H),7.93(dd,J=8.0,1.7Hz,1H),7.60– 7.51(m,3H),5.55(dd,J=7.8,4.2Hz,1H),2.13–2.04(m,1H),1.87–1.75(m,1H),0.93(t,J=7.1Hz,3H).

[0111] LCMS (ESI): 268.5 [M+H] + .

[0112] Example 4

[0113] 3-Butyl-6-(3-methylpyridin-4-yl)isobenzofuran-1(3H)-one (I-04), the structure is shown below:

[0114]

[0115] The compound was prepared from intermediate 4 and (3-methylpyridin-4-yl)boronic acid (A-04) according to reaction scheme 3. The product was characterized. The characterization results are as follows: LCMS (ESI): 282.1 [M+H] + .

[0116] Example 5

[0117] 3-Butyl-6-(5-chloropyridin-3-yl)isobenzofuran-1(3H)-one (I-05), the structure is shown below:

[0118]

[0119] This compound was prepared from Intermediate 4 and (5-chloropyridin-3-yl)boronic acid (A-05) according to Reaction Route 3. The product was characterized, and the characterization results are as follows: LCMS(ESI): 303.7 [M+H] + 。

[0120] Example 6

[0121] 3-Butyl-6-(5-fluoropyridin-3-yl)isoindoline-1(3H)-one (I-06), the structure is shown as follows:

[0122]

[0123] This compound was prepared from Intermediate 4 and (5-fluoropyridin-3-yl)boronic acid (A-06) according to Reaction Route 3. The product was characterized, and the characterization results are as follows: LCMS(ESI): 268.1 [M+H] + 。

[0124] Example 7

[0125] 3-Butyl-6-(pyrimidin-5-yl)isoindoline-1(3H)-one (I-07), the structure is shown as follows:

[0126]

[0127] This compound was prepared from Intermediate 4 and pyrimidin-5-ylboronic acid (A-07) according to Reaction Route 3. The product was characterized, and the characterization results are as follows: LCMS(ESI): 269.1 [M+H] + 。

[0128] Example 8

[0129] 3-Butyl-6-(2-methylpyrimidin-5-yl)isoindoline-1(3H)-one (I-08), the structure is shown as follows:

[0130]

[0131] This compound was prepared from Intermediate 4 and (2-methylpyrimidin-5-yl)boronic acid (A-08) according to Reaction Route 3. The product was characterized, and the characterization results are as follows: LCMS(ESI): 283.1 [M+H] + 。

[0132] Example 9

[0133] 3-Butyl-6-(1-methyl-1H-pyrazol-4-yl)isoindoline-1(3H)-one (I-09), the structure is shown as follows:

[0134]

[0135] This compound is prepared from intermediate 4 and (1-methyl-1H-pyrazol-4-yl)boronic acid (A-09) according to Reaction Route 3, and the product is characterized with the following results:

[0136] 1 H NMR(400MHz,CDCl3)δ7.94(s,1H),7.80(s,1H),7.76(dd,J=8.0,1.6Hz,1H),7.68(s,1H),7.41(d,J=7.9Hz,1H),5.47(dd,J=7.8,4.2Hz,1H),3.97(s,3H),2.11–1.98(m,1H),1.84–1.71(m,1H),1.54–1.34(m,4H),0.91(t,J=7.1Hz,3H).

[0137] LCMS(ESI):271.2[M+H] + 。

[0138] Example 10

[0139] 3-Butyl-6-(1H-pyrazol-4-yl)isobenzofuran-1(3H)-one (I-10), the structure is shown as follows:

[0140]

[0141] This compound is prepared from intermediate 4 and (1H-pyrazol-4-yl)boronic acid (A-10) according to Reaction Route 3, and the product is characterized with the following result: 257.1[M+H] + 。

[0142] Example 11

[0143] 3-Butyl-6-(1-(methyl-d3)-1H-pyrazol-4-yl)isobenzofuran-1(3H)-one (I-11), the structure is shown as follows:

[0144]

[0145] This compound is prepared from intermediate 4 and (1-(methyl-d3)-1H-pyrazol-4-yl)boronic acid (A-11) according to Reaction Route 3, and the product is characterized with the following result: 274.1[M+H] + 。

[0146] Example 12

[0147] 3-Butyl-6-(1H-imidazol-4-yl)isobenzofuran-1(3H)-one (I-12), the structure is shown as follows:

[0148]

[0149] The compound was prepared from intermediate 4 and (1H-imidazol-4-yl)boronic acid (A-12) according to reaction scheme 3. The product was characterized and the results were as follows: 257.1 [M+H] + .

[0150] Compounds I-13 to I-16 were prepared according to the method shown in Reaction Scheme 4, as shown below:

[0151]

[0152] Reaction Scheme 4

[0153] Example 13

[0154] 3-Butyl-6-(1,2,3,6-tetrahydropyridin-4-yl)iso-benzofuran-1(3H)-one (I-13), the structure of which is shown below:

[0155]

[0156] The preparation process is as follows:

[0157] (1) Intermediate 4 (120 mg, 0.37 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (A-13) were reacted by Suzuki reaction to synthesize intermediate 13;

[0158] (2) TFA (5 eq) was added to a solution of Boc-protected compound 13 in anhydrous dichloromethane, and the reaction mixture was stirred at room temperature for 25 minutes (monitored by TLC). The reaction was then quenched by adding a saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and the solvent was removed by distillation under reduced pressure to obtain the deprotected product. The product was characterized, and the characterization results are as follows:

[0159] 1H NMR (400MHz, CDCl3) δ7.87(s,1H),7.69(dd,J=8.0,1.7Hz,1H),7.43(d,J=8.0Hz,1H),6.14(d,J=1.8Hz,1H),5.48(dd,J=7.8,4.2Hz,1H),3.91 (d,J=3.0Hz,2H),3.48(t,J=6.0Hz,2H),2.91(d,J=6.9Hz,2H),2.11–1.98(m,1H),1.83–1.70(m,1H),1.52–1.34(m,4H),0.91(t,J=7.0Hz,3H).

[0160] LCMS (ESI): 272.4 [M+H] + .

[0161] Example 14

[0162] 3-Butyl-6-(piperidin-4-yl)isobenzofuran-1(3H)-one (I-14), the structure is shown below:

[0163]

[0164] Compound 13 is dissolved in ethanol and a catalytic amount of palladium on carbon (10% w / w) is added. The flask is purged with hydrogen for a few minutes to expel air and is connected to a balloon filled with hydrogen. The mixture is stirred at room temperature for 4 hours (measured by TLC) under a hydrogen atmosphere. The reaction mixture is filtered through diatomaceous earth and the catalyst is washed with ethanol. The organic solvent is evaporated under reduced pressure to obtain compound 14.

[0165] Compound 14 was subjected to Boc-deprotection using TFA according to a standard Boc-deprotection method (refer to step (2) in the preparation of compound I-13). The crude product was purified by column chromatography to obtain compound I-14. The product was characterized as follows: LCMS (ESI): 275.1 [M+H] + .

[0166] Example 15

[0167] 3-Butyl-6-(2,5-dihydro-1H-pyrrol-3-yl)isobenzofuran-1(3H)-one (I-14), the structure of which is shown below:

[0168]

[0169] Compound 4 (120 mg, 0.37 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (A-15) (0.67 mmol) were used to synthesize the target product 15 through the Suzuki reaction, and then Boc-deprotection was carried out to obtain the target product I-15 (refer to step (2) in the preparation of reference compound I-13). The product was characterized, and the characterization results were as follows: LCMS (ESI): 258.1 [M+H] + 。

[0170] Example 16

[0171] 3-Butyl-6-(pyrrolidin-3-yl)isoindoline-1(3H)-one (I-16), the structure is shown as follows:

[0172]

[0173] Using the method described in Example 14, compound I-16 was prepared from compound 15.

[0174] LCMS (ESI): 260.2 [M+H] + 。

[0175] The intermediate amine ((R 1 NHR 2 ) used in the following examples was commercially available or prepared in the laboratory:

[0176]

[0177] Compounds I-17 to I-44 and intermediate amine A40-A44 were prepared according to one of the methods shown in Reaction Routes 5 to 7, as specifically shown below:

[0178]

[0179] Reaction Route 5

[0180] The specific operation is as follows:

[0181] In a dry round-bottom flask equipped with a condenser, a solution of intermediate 4 (100 mg, 0.37 mmol) and amine (0.41 mmol) in dioxane (5 mL) was taken. The flask was evacuated and backfilled with nitrogen three times. Then, Pd(dba)2 (36.63 mg, 0.04 mmol), XantPhos (23.14 mg, 0.04 mmol), and Cs2CO3 (358.60 mg, 1.11 mmol) were added, and the flask was heated to 85 - 90 °C for 4 h. According to TLC analysis, the reaction was completely consumed. The mixture was cooled to room temperature, diluted with ethyl acetate, washed with water, concentrated in vacuo, and purified by flash chromatography on silica gel to obtain the product.

[0182]

[0183] Reaction Route 6

[0184] The specific operation is as follows:

[0185] (1) The Boc-protected intermediate was prepared by the Buchwald reaction of intermediate 4 (120 mg, 0.37 mmol) and Boc-protected amine;

[0186] (2) TFA (5 eq) was added via syringe to an anhydrous dichloromethane solution of the Boc-protected intermediate, and the reaction mixture was stirred at room temperature for 25 minutes (monitored by TLC). Then, the reaction was quenched by adding saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain the deprotected desired product.

[0187]

[0188] Reaction Route 7

[0189] For the specific operation, see patent documents US20130189620 and CN108069892A.

[0190] Example 17

[0191] 3-Butyl-6-(2-azaspiro[3.3]hept-2-yl)isoindoline-1(3H)-one (I-17), the structure is shown as follows:

[0192]

[0193] Using intermediate 4 (100 mg, 0.37 mmol) and 2-azaspiro[3.3]heptane (A-17) (0.41 mmol), the target compound was prepared according to Reaction Route 5 to obtain the product (I-17). LCMS (ESI): 286.2 [M+H] + 。

[0194] Example 18

[0195] 3-Butyl-6-(2-oxa-6-azaspiro[3.3]hept-6-yl)isobenzofuran-1(3H)-one (I-18), the structure of which is shown below:

[0196]

[0197] The target compound (I-18) was prepared using intermediate 4 and 2-oxa-6-azaspiro[3.3]heptane (A-18) according to Reaction Scheme 5. The product was characterized, and the characterization results are as follows:

[0198] 1 H NMR (400MHz, CDCl3) δ7.16(d,J=8.2Hz,1H),6.77(d,J=2.2Hz,1H),6.64(dd,J=8.2,2.3Hz,1H),5.30(dd,J=7.7,4. 2Hz,1H),4.77(s,4H),4.00(s,4H),1.97–1.79(m,1H),1.72–1.55(m,1H),1.40–1.24(m,4H),0.82(t,J=7.0Hz,3H).

[0199] LCMS (ESI): 288.0 [M+H] + .

[0200] Example 19

[0201] 3-Butyl-6-(2,6-diazaspiro[3.3]hept-2-yl)isobenzofuran-1(3H)-one (I-19), the structure of which is shown below:

[0202]

[0203] According to the method shown in Reaction Scheme 6, Intermediate 4 (120 mg, 0.37 mmol) was reacted with tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (A-19) (0.67 mmol) via Buchwald reaction to synthesize Intermediate 19, which was then subjected to Boc-deprotection reaction to obtain the target product. The product was characterized, and the characterization results are as follows:

[0204] 1H NMR (400MHz, CDCl3) δ7.24(s,1H),6.85(d,J=2.1Hz,1H),6.72(d,J=5.9Hz,1H),5.38(dd,J=7.8,4.4Hz,1H), 4.31(s,4H),4.12(s,4H),2.03–1.91(m,1H),1.70(d,J=9.8Hz,1H),1.48–1.30(m,4H),0.89(t,J=6.6Hz,3H).

[0205] LCMS (ESI): 287.2 [M+H] + .

[0206] Example 20

[0207] 3-Butyl-6-(2,7-diazaspiro[3.5]nonan-7-yl)isobenzofuran-1(3H)-one (I-20), the structure of which is shown below:

[0208]

[0209] According to the method shown in Reaction Scheme 6, Intermediate 4 was first reacted with tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (A-20) to obtain a Boc-protected product. The Boc-protection was then removed using trifluoroacetic acid in DCM to obtain the desired product (I-20). LCMS (ESI): 315.2 [M+H] + .

[0210] Example 21

[0211] 3-Butyl-6-(2,7-diazaspiro[3.5]nonan-2-yl)isobenzofuran-1(3H)-one (I-21), the structure of which is shown below:

[0212]

[0213] According to the method shown in Reaction Scheme 6, Intermediate 4 was first reacted with tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate (A-21) to obtain a Boc-protected product. The Boc-protection was then removed using trifluoroacetic acid in DCM to obtain the desired product (I-21). LCMS (ESI): 315.2 [M+H] + .

[0214] Example 22

[0215] 3-Butyl-6-(7-oxa-2-azaspiro[3.5]nonan-2-yl)isobenzofuran-1(3H)-one (I-22), the structure of which is shown below:

[0216]

[0217] The target compound was prepared using intermediate 4 and 7-oxa-2-azaspiro[3.5]nonane (A-22) according to Reaction Route 5 to obtain the product (I-22). LCMS (ESI): 316.1 [M+H] + .

[0218] Example 23

[0219] 3-Butyl-6-(2,7-diazaspiro[3.5]nonan-2-yl)isoindoline-1(3H)-one (I-23), the structure is shown as follows:

[0220]

[0221] According to the method shown in Reaction Route 6, intermediate 4 was first reacted with tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (A-23) to obtain the Boc-protected product. Then the Boc-protection was removed using a DCM solution of trifluoroacetic acid to obtain the desired product (I-23). LCMS (ESI): 315.3 [M+H] + .

[0222] Example 24

[0223] 3-Butyl-6-(2-oxa-7-azaspiro[3.5]nonan-7-yl)isoindoline-1(3H)-one (I-24), the structure is shown as follows:

[0224]

[0225] The target compound was prepared using intermediate 4 and 2-oxa-7-azaspiro[3.5]nonane (A-24) according to Reaction Route 5 to obtain the product (I-24). LCMS (ESI): 316.2 [M+H] + .

[0226] Example 25

[0227] 3-Butyl-6-(octahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)isoindoline-1(3H)-one (I-25), the structure is shown as follows:

[0228]

[0229] According to the method shown in Reaction Scheme 6, Intermediate 4 was first reacted with tert-butyl octahydro-1H-pyrrolo[3,2-c]pyridine-1-carboxylate (A-25) to obtain a Boc-protected product. The Boc-protection was then removed using trifluoroacetic acid in DCM to obtain the desired product (I-25). LCMS (ESI): 315.2 [M+H] + .

[0230] Example 26

[0231] 3-Butyl-6-morpholinoisobenzofuran-1(3H)-one (I-26), the structure is shown below:

[0232]

[0233] The target compound (I-26) was prepared using intermediate 4 and morpholine (A-26) according to Reaction Scheme 5. The product was characterized, and the characterization results are as follows:

[0234] 1 H NMR (400MHz, DMSO-d6) δ7.50(d,J=8.4Hz,1H),7.41(dd,J=8.5,2.4Hz,1H),7.19(d,J=2.3Hz,1H),5.51(dd,J=7.5,4.0Hz,1H), 3.74(dd,J=5.8,3.9Hz,4H),3.22–3.14(m,4H),2.05–1.93(m,1H),1.70–1.58(m,1H),1.39–1.20(m,4H),0.85(t,J=7.0Hz,3H).

[0235] LCMS (ESI): 276.3 [M+H] + .

[0236] Example 27

[0237] 3-Butyl-6-thiomorpholinoisobenzofuran-1(3H)-one (I-27), the structure is shown below:

[0238]

[0239] The target compound was prepared using intermediate 4 and thiomorpholine (A-27) according to reaction scheme 5 to obtain product (I-27). LCMS (ESI): 292.1 [M+H] + .

[0240] Example 28

[0241] 3-Butyl-6-(1,1-sulfonyldi-morpholino)iso-benzofuran-1(3H)-one (I-28), the structure is shown below:

[0242]

[0243] The target compound was prepared using Intermediate 4 and thiomorpholine 1,1-dioxide (A-28) according to Reaction Route 5 to obtain the product (I-28). LCMS (ESI): 324.2 [M+H] + 。

[0244] Example 29

[0245] 3-Butyl-6-(6-methyl-2,6-diazaspiro[3.3]hept-2-yl)iso-benzofuran-1(3H)-one (I-29), the structure is shown below:

[0246]

[0247] The target compound was prepared using Intermediate 4 and 2-methyl-2,6-diazaspiro[3.3]heptane (A-29) according to Reaction Route 5 to obtain the product (I-29). LCMS (ESI): 301.2 [M+H] + 。

[0248] Example 30

[0249] 3-Butyl-6-(6-(methyl-d3)-2,6-diazaspiro[3.3]hept-2-yl)iso-benzofuran-1(3H)-one (I-30), the structure is shown below:

[0250]

[0251] The target compound was prepared using Intermediate 4 and 2-(methyl-d3)-2,6-diazaspiro[3.3]heptane (A-30) according to Reaction Route 5 to obtain the product (I-30). LCMS (ESI): 304.2 [M+H] + 。

[0252] Example 31

[0253] 6-(6-Acetyl-2,6-diazaspiro[3.3]hept-2-yl)-3-butyliso-benzofuran-1(3H)-one (I-31), the structure is shown below:

[0254]

[0255] The target compound was prepared using intermediate 4 and 1-(2,6-diazaspiro[3.3]heptan-2-yl)ethan-1-one (A-31) according to Reaction Route 5 to obtain product (I-31). LCMS (ESI): 329.2 [M+H] + 。

[0256] Example 32

[0257] 3-Butyl-6-(6-(2,2,2-trifluoroacetyl)-2,6-diazaspiro[3.3]heptan-2-yl)isoindoline-1(3H)-one (I-32), the structure is shown below:

[0258]

[0259] The target compound was prepared using intermediate 4 and 2,2,2-trifluoro-1-(2,6-diazaspiro[3.3]heptan-2-yl)ethan-1-one (A-32) according to Reaction Route 5 to obtain product (I-32). LCMS (ESI): 383.2 [M+H] + 。

[0260] Example 33

[0261] 3-Butyl-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isoindoline-1(3H)-one (I-33), the structure is shown below:

[0262]

[0263] The target compound was prepared using intermediate 4 and (3as,6as)-octahydropyrrolo[3,4-c]pyrrole (A-33) according to Reaction Route 5 to obtain product (I-33). The product was characterized, and the characterization results are as follows:

[0264] 11H NMR (400 MHz, CDCl3) δ 7.29 (d, J = 4.6 Hz, 1H), 7.04 (d, J = 2.3 Hz, 1H), 6.94 (dd, J = 8.4, 2.4 Hz, 1H), 5.42 (dd, J = 7.6, 4.2 Hz, 1H), 4.03 (dd, J = 8.9, 6.6 Hz, 2H), 3.72 (dd, J = 9.0, 3.4 Hz, 2H), 3.54 (dd, J = 9.4, 7.3 Hz, 2H), 3.31 (dd, J = 9.7, 2.9 Hz, 2H), 3.17–3.07 (m, 2H), 2.07–1.93 (m, 1H), 1.75 (dd, J = 28.0, 8.6 Hz, 1H), 1.53–1.32 (m, 4H), 0.93 (t, J = 7.1 Hz, 3H).

[0265] LCMS (ESI): 302.2 [M+H] + 。

[0266] Example 34

[0267] N-(1-(1-Butyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)azetidin-3-yl)acetamide (I-34), the structure is shown below:

[0268]

[0269] The target compound was prepared using Intermediate 4 and N-(azetidin-3-yl)acetamide (A-34) according to Reaction Route 5 to obtain the product (I-34). LCMS (ESI): 303.1 [M+H] + 。

[0270] Example 35

[0271] 6-(2-Oxa-5-azabicyclo[2.2.1]hept-5-yl)-3-butylisobenzofuran-1(3H)-one (I-35), the structure is shown below:

[0272]

[0273] The target compound was prepared using Intermediate 4 and 2-oxa-5-azabicyclo[2.2.1]heptane (A-35) according to Reaction Route 5 to obtain the product (I-35). LCMS (ESI): 288.2 [M+H] + 。

[0274] Example 36

[0275] 3-Butyl-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isoindoline-1(3H)-one (I-36), the structure is shown below:

[0276]

[0277] According to the method shown in Reaction Route 6, first react intermediate 4 with tert-butyl (3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (A-36) to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the desired product (I-36). The product was characterized, and the characterization results are as follows:

[0278] 1 H NMR (400 MHz, CDCl3) δ 7.23 (d, J = 8.3 Hz, 1H), 6.99 (d, J = 2.3 Hz, 1H), 6.91 (dd, J = 8.4, 2.3 Hz, 1H), 5.37 (dd, J = 7.6, 4.2 Hz, 1H), 3.44 (dd, J = 9.9, 7.1 Hz, 2H), 3.33 (dd, J = 22.5, 11.8 Hz, 4H), 3.11–2.93 (m, 4H), 2.02–1.88 (m, 1H), 1.77–1.64 (m, 1H), 1.48–1.30 (m, 4H), 0.88 (t, J = 7.0 Hz, 3H).

[0279] 13 C NMR (101 MHz, DMSO) δ 170.90, 148.97, 138.73, 126.70, 123.34, 120.52, 107.23, 81.27, 52.56, 49.70, 41.54, 34.36, 26.74, 22.36, 14.28.

[0280] LCMS (ESI): 301.3 [M+H] + 。

[0281] Example 37

[0282] 3-Butyl-6-(piperazin-1-yl)isoindoline-1(3H)-one (I-37), the structure is shown below:

[0283]

[0284] According to the method shown in Reaction Scheme 6, Intermediate 4 was first reacted with tert-butyl piperazine-1-carboxylate (A-37) to obtain a Boc-protected product. The Boc-protection was then removed using trifluoroacetic acid in DCM to obtain the desired product (I-37). The product was characterized, and the characterization results are as follows:

[0285] 1 H NMR (400MHz, CDCl3) δ7.31–7.23(m,2H),7.19–7.15(m,1H),5.35(dd,J=7.6,4.2Hz,1H),3.37(dd,J=6.7,3.6H z,4H),3.21(t,J=5.0Hz,4H),1.99–1.87(m,1H),1.74–1.60(m,1H),1.42–1.28(m,4H),0.84(t,J=7.0Hz,3H).

[0286] LCMS (ESI): 275.5 [M+H] + .

[0287] Example 38

[0288] 6-(3,6-diazabicyclo[3.1.1]hept-3-yl)-3-butylisobenzofuran-1(3H)-one (I-38), the structure of which is shown below:

[0289]

[0290] According to the method shown in Reaction Scheme 6, Intermediate 4 was first reacted with tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (A-38) to obtain a Boc-protected product. The Boc-protection was then removed using trifluoroacetic acid in DCM to obtain the desired product (I-38). The product was characterized, and the results are as follows:

[0291] 1 H NMR (400MHz, CDCl3) δ7.25(d,J=8.4Hz,1H),7.05(d,J=2.4Hz,1H),6.96(dd,J=8.5,2.5Hz,1H),5.35(dd,J=7.6,4.2Hz,1H),4.13(d,J=6.1Hz,2H), 3.71(d,J=11.1Hz,2H),3.61(d,J=11.1Hz,2H),2.94–2.84(m,1H),1.98– 1.85(m,1H),1.72–1.60(m,2H),1.43–1.27(m,4H),0.84(t,J=7.1Hz,4H).

[0292] LCMS (ESI): 287.5 [M+H] + .

[0293] Example 39

[0294] 6-((1R,4R)-2,5-diazabicyclo[2.2.1]hept-2-yl)-3-butylisobenzofuran-1(3H)-one (I-39), the structure of which is shown below:

[0295]

[0296] According to the method shown in Reaction Scheme 6, Intermediate 4 was first reacted with tert-butyl (1R,4R)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (A-39) to obtain a Boc-protected product. The Boc-protection was then removed using trifluoroacetic acid in DCM to obtain the desired product (I-39). The product was characterized, and the results are as follows:

[0297] 1 H NMR (400MHz, CDCl3) δ7.24(d,J=3.3Hz,1H),6.96(s,1H),6.84(dd,J=8.4,2.4Hz,1H),5.39(dd,J=7.6,4.2Hz,1H),4.43(s,1H),4.13(s,1H),3. 72–3.66(m,1H),3.30(d,J=9.6Hz,1H),3.23(s,2H),2.13–1.92(m,3H), 1.72(dd,J=22.3,9.6Hz,1H),1.51–1.32(m,4H),0.91(t,J=6.7Hz,3H).

[0298] LCMS (ESI): 287.4 [M+H] + .

[0299] Example 40

[0300] 3-Butyl-6-(3-hydroxyazetidin-1-yl)isobenzofuran-1(3H)-one (I-40), the structure of which is shown below:

[0301]

[0302] The target compound (I-40) was prepared using intermediate 4 and azetidine-3-ol (A-40) according to Reaction Scheme 5. The product was characterized, and the characterization results are as follows:

[0303] 1H NMR (400MHz, CDCl3) δ7.22(d,J=8.2Hz,1H),6.84(d,J=2.2Hz,1H),6.72(dd,J=8.3,2.2Hz,1H),5.37(dd,J=7.7,4.2Hz,1H),4.85–4.76 (m,1H),4.25–4.17(m,2H),3.74(dd,J=8.4,4.5Hz,2H),2.02–1.91(m,1H),1.78–1.66(m,1H),1.50–1.28(m,4H),0.90(t,J=7.1Hz,3H).

[0304] 13 C NMR (101MHz, DMSO) δ170.76,152.99,139.00,126.65,123.20,118.83,105.64,81.35,62.26,61.34,34.41,26.84,22.37,14.27.

[0305] LCMS (ESI): 262.1 [M+H] + .

[0306] Example 41

[0307] 3-Butyl-6-(4-hydroxypiperidin-1-yl)isobenzofuran-1(3H)-one (I-41), the structure is shown below:

[0308]

[0309] The target compound was prepared using intermediate 4 and piperidin-4-ol (A-41) according to reaction scheme 5 to obtain product (I-41). LCMS (ESI): 276.1 [M+H] + .

[0310] Example 42

[0311] 3-Butyl-6-(6-hydroxy-2-azaspiro[3.3]hept-2-yl)isobenzofuran-1(3H)-one (I-42), the structure of which is shown below:

[0312]

[0313] The target compound was prepared using intermediate 4 and 2-azaspiro[3.3]heptan-6-ol (A-42) according to reaction scheme 5 to obtain the product (I-42). LCMS (ESI): 302.2 [M+H] + .

[0314] Example 43

[0315] 3-Butyl-6-(3-hydroxy-3-methylazetidin-1-yl)isoindoline-1(3H)-one (I-43), the structure is shown as follows:

[0316]

[0317] The target compound was prepared using Intermediate 4 and 3-methylazetidin-3-ol (A-43) according to Reaction Route 5 to obtain the product (I-43). The product was characterized, and the characterization results are as follows:

[0318] 1 H NMR (400 MHz, CDCl3) δ 7.14 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 2.2 Hz, 1H), 6.65 (dd, J = 8.3, 2.2 Hz, 1H), 5.30 (dd, J = 7.7, 4.2 Hz, 1H), 3.83 (d, J = 7.6 Hz, 2H), 3.73 (d, J = 7.6 Hz, 2H), 1.94–1.84 (m, 1H), 1.69–1.60 (m, 1H), 1.56 (s, 3H), 1.41–1.24 (m, 4H), 0.83 (t, J = 7.0 Hz, 3H).

[0319] LCMS (ESI): 276.2 [M+H] + .

[0320] Example 44

[0321] 3-Butyl-6-(6-hydroxy-6-methyl-2-azaspiro[3.3]hept-2-yl)isoindoline-1(3H)-one (I-44), the structure is shown as follows:

[0322]

[0323] The target compound was prepared using Intermediate 4 and 6-methyl-2-azaspiro[3.3]hept-6-ol (A-44) according to Reaction Route 5 to obtain the product (I-44). The product was characterized, and the characterization results are as follows:

[0324] 11H NMR (400 MHz, CDCl3) δ 7.21 (d, J = 8.2 Hz, 1H), 6.83 (s, 1H), 6.73–6.66 (m, 1H), 5.38 (dd, J = 7.7, 4.2 Hz, 1H), 3.92 (d, J = 7.6 Hz, 4H), 2.36 (s, 4H), 2.04–1.91 (m, 1H), 1.70 (dd, J = 23.1, 9.0 Hz, 1H), 1.52–1.30 (m, 7H), 0.90 (t, J = 6.9 Hz, 3H).

[0325] LCMS (ESI): 316.1 [M+H] + 。

[0326] The intermediates 26a - 26f used in the following examples were prepared according to Reaction Route 8, which is specifically shown as follows:

[0327]

[0328] Reaction Route 8

[0329] Preparation of Intermediate 26a (with the structure shown in Reaction Scheme 26A)

[0330] (1) A mixture of compound 23 (3.0 g, 14.1 mmol), 1,2 - dichloroethane (25 mL), N - bromosuccinimide (3.0 g, 16.9 mmol) and azobisisobutyronitrile (0.09 g, 0.6 mmol) was refluxed for 6 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain compound 24 (3.5 g, yield: 85%). 1 1H NMR (400 MHz, CDCl3) δ 8.05 (d, 1H), 7.89 (dd, 1H), 7.51 (d, 1H), 7.35 (s, 1H); LCMS (ESI): [M+H] + : 292.9。

[0331] Compound 24 (3.5 g, 12.0 mmol) was suspended in 20 mL of H2O and refluxed for 2 h. The reaction mixture was cooled to room temperature and stirred overnight. The two - phase mixture was extracted with ethyl acetate (20 mL × 3), and the combined organic layers were dried over anhydrous Na2SO4 and evaporated to obtain the crude product 25 (2.7 g, yield: 100%).

[0332] 1 1H NMR (400 MHz, CDCl3) δ 8.01 (d, 1H), 7.97 (dd, 1H), 7.65 (d, 1H); LCMS (ESI): [M+H] + : 230.0。

[0333] (2) Add magnesium, 4-bromobut-1-ene (AI-1), dry THF, and iodine. Heat the mixture under reflux for 2 h to generate a Grignard reagent. Then add the Grignard reagent to a THF solution of compound 25 at 0 °C. Stir the mixture at 0 °C for 2 h, quench with HCl, and evaporate THF. Dilute the resulting solution with DCM and acidify to pH 2.0. Continue the reaction overnight, dilute the solution with EtOAc, and dry the organic phase over Na2SO4. Evaporate the solvent, and purify the residue by silica gel column chromatography to obtain compound 26a as a white solid in 28% yield. [M+H] + : 268.0.

[0334] Intermediates 26a - 26f were prepared according to the method shown in Reaction Route 8 as specifically shown in the following table:

[0335]

[0336]

[0337] Intermediates 28a and 28b were prepared according to the literature method (CN 1091,114,15 A, J. Med. Chem. 2020, 63, 21, 12485–12510):

[0338]

[0339] Compounds I-45 to I-60 and intermediates 29a - 33a, 29b - 33b were prepared according to one of the methods shown in Reaction Routes 9 - 11 as specifically shown below:

[0340]

[0341] Reaction Route 9

[0342]

[0343] Reaction Route 10

[0344]

[0345] Reaction Route 11

[0346] Intermediates 33a and 33b were prepared according to the method shown in Reaction Route 11 as specifically shown in the following table:

[0347]

[0348]

[0349] Example 45

[0350] 3-(But-3-en-1-yl)-6-(2,6-diazaspiro[3.3]heptan-2-yl)isoindoline-1(3H)-one (I-45), the structure is shown as follows:

[0351]

[0352] According to the method shown in Reaction Route 9, first react intermediate 26a with intermediate A-19 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc-protection to obtain the desired product (I-45). LCMS(ESI): 285.2[M+H] + 。

[0353] Example 46

[0354] 3-(2-Cyclopropylethyl)-6-(2,6-diazaspiro[3.3]heptan-2-yl)isoindoline-1(3H)-one (I-46), the structure is shown as follows:

[0355]

[0356] According to the method shown in Reaction Route 9, first react intermediate 26b with intermediate A-19 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc-protection to obtain the desired product (I-46). LCMS(ESI): 299.2[M+H] + 。

[0357] Example 47

[0358] 6-((3aR,6aS)-Hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-propylisoindoline-1(3H)-one (I-47), the structure is shown as follows:

[0359]

[0360] According to the method shown in Reaction Route 9, first react intermediate 26c with intermediate A-36 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc-protection to obtain the desired product (I-47). LCMS(ESI): 287.2[M+H] + 。

[0361] Example 48

[0362] 6-((3aR,6aS)-Hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-pentylisoindoline-1(3H)-one (I-48), the structure is shown as follows:

[0363]

[0364] According to the method shown in Reaction Route 9, first react Intermediate 26d with Intermediate A-36 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the desired product (I-48). LCMS (ESI): 315.2 [M+H] + 。

[0365] Example 49

[0366] 6-((3aR,6aS)-Hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-(4-hydroxybutyl)isobenzofuran-1(3H)-one (I-49), the structure is shown as follows:

[0367]

[0368] According to the method shown in Reaction Route 10, first react Intermediate 28b with Intermediate A-36 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the OAc-protected product. Use K2CO3 in MeOH / H2O to remove the OAc protection to obtain the desired product (I-49). LCMS (ESI): 317.1 [M+H] + 。

[0369] Example 50

[0370] 3-(4-Fluorobutyl)-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isobenzofuran-1(3H)-one (I-50), the structure is shown as follows:

[0371]

[0372] According to the method shown in Reaction Route 11, deacetylate Intermediate 28b with K2CO3, then add DAST to the DCM solution to obtain Intermediate 33b. Through the Buchwald coupling reaction between Intermediate 33b and Intermediate A-36, and then perform Boc-deprotection with TFA in DCM to obtain the target product I-50. LCMS (ESI): 317.1 [M+H] + 。

[0373] Example 51

[0374] 6-((3aR,6aS)-Hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-(3-hydroxypropyl)isobenzofuran-1(3H)-one (I-50), the structure is shown as follows:

[0375]

[0376] According to the method shown in reaction route 10, first react intermediate 28a with intermediate A-36 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc-protection to obtain the OAc-protected product. Use K2CO3 in MeOH / H2O to remove the OAc-protection to obtain the desired product (I-51). LCMS (ESI): 303.2 [M+H] + 。

[0377] Example 52

[0378] 3-(3-Hydroxypropyl)-6-(2,6-diazaspiro[3.3]hept-2-yl)isobenzofuran-1(3H)-one (I-50), the structure is shown as follows:

[0379]

[0380] According to the method shown in reaction route 10, first react intermediate 28a with intermediate A-19 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc-protection to obtain the OAc-protected product. Use K2CO3 in MeOH / H2O to remove the OAc-protection to obtain the desired product (I-52). LCMS (ESI): 289.1 [M+H] + 。

[0381] Example 53

[0382] 3-(3-Fluoropropyl)-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isobenzofuran-1(3H)-one (I-53), the structure is shown as follows:

[0383]

[0384] According to the method shown in reaction route 11, deacetylate intermediate 28a with K2CO3, then add DAST to the DCM solution to obtain intermediate 33a. Through the Buchwald coupling reaction between intermediate 33a and intermediate A-36, and then perform Boc-deprotection with TFA in DCM to obtain the target product I-53. LCMS (ESI): 305.2 [M+H] + 。

[0385] Example 54

[0386] 3-(3-Fluoropropyl)-6-(2,6-diazaspiro[3.3]hept-2-yl)isobenzofuran-1(3H)-one (I-54), the structure is shown as follows:

[0387]

[0388] Following the method shown in Reaction Scheme 11, intermediate 16a was deacetylated with KCO, and then DAST was added to a DCM solution to afford intermediate 33a. A Buchwald coupling reaction between intermediate 33a and intermediate A-19, followed by Boc-deprotection with TFA in DCM, afforded the target product I-54. LCMS (ESI): 291.1 [M+H] + .

[0389] Example 55

[0390] 3-(4,4-difluorobutyl)-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isobenzofuran-1(3H)-one (I-55), the structure of which is shown below:

[0391]

[0392] According to the method shown in Reaction Scheme 9, Intermediate 26e is first reacted with Intermediate A-36 to obtain a Boc-protected product. The Boc-protection is then removed using trifluoroacetic acid in DCM to obtain the desired product (I-55). LCMS (ESI): 337.2 [M+H] + .

[0393] Example 56

[0394] 6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-(4,4,4-trifluorobutyl)isobenzofuran-1(3H)-one (I-56), the structure of which is shown below:

[0395]

[0396] According to the method shown in Reaction Scheme 9, Intermediate 26f is first reacted with Intermediate A-36 to obtain a Boc-protected product. The Boc-protection is then removed using trifluoroacetic acid in DCM to obtain the desired product (I-56). LCMS (ESI): 355.2 [M+H] + .

[0397] Example 57

[0398] 6-(3,6-diazabicyclo[3.1.1]hept-3-yl)-3-(but-3-en-1-yl)isobenzofuran-1(3H)-one (I-57), the structure of which is shown below:

[0399]

[0400] According to the method shown in Reaction Route 9, first react Intermediate 26a with Intermediate A-38 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the desired product (I-57). LCMS (ESI): 285.1 [M+H] + .

[0401] Example 58

[0402] 6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)-3-(4-hydroxybutyl)isobenzofuran-1(3H)-one (I-58), the structure is shown as follows:

[0403]

[0404] According to the method shown in Reaction Route 10, first react Intermediate 28b with Intermediate A-38 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the OAc-protected product. Use K2CO3 in MeOH / H2O to remove the OAc protection to obtain the desired product (I-58). LCMS (ESI): 303.3 [M+H] + .

[0405] Example 59

[0406] 6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)-3-(4-fluorobutyl)isobenzofuran-1(3H)-one (I-58), the structure is shown as follows:

[0407]

[0408] According to the method shown in Reaction Route 11, deacetylate Intermediate 28b with K2CO3, then add DAST to the DCM solution to obtain Intermediate 33b. Through the Buchwald coupling reaction between Intermediate 33b and Intermediate A-38, and then perform Boc-deprotection with TFA in DCM to obtain the target product I-59. LCMS (ESI): 305.1 [M+H] + .

[0409] Example 60

[0410] 6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)-3-(3-hydroxypropyl)isobenzofuran-1(3H)-one (I-60), the structure is shown as follows:

[0411]

[0412] According to the method shown in Reaction Route 10, first react intermediate 28a with intermediate A-38 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the OAc-protected product. Use K2CO3 in MeOH / H2O to remove the OAc protection to obtain the desired product (I-60). LCMS (ESI): 289.2 [M+H] + 。

[0413] Compounds I-61 to I-76 were prepared according to the method shown in Reaction Route 12, as follows:

[0414]

[0415] Reaction Route 12

[0416] Example 61

[0417] (S)-3-Butyl-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isobenzofuran-1(3H)-one (I-61), the structure is as follows:

[0418]

[0419] According to the method shown in Reaction Route 12, react intermediate 10 with intermediate A-36 to obtain the Boc-protected product. Then use a DCM solution of trifluoroacetic acid to remove the Boc protection to obtain the desired product (I-61). Characterize the product, and the characterization results are as follows:

[0420] 1 H NMR (400 MHz, CDCl3) δ 7.23 (d, J = 8.4 Hz, 1H), 6.99 (d, J = 2.3 Hz, 1H), 6.91 (dd, J = 8.4, 2.3 Hz, 1H), 5.37 (dd, J = 7.6, 4.2 Hz, 1H), 3.48–3.40 (m, 2H), 3.39–3.27 (m, 4H), 3.10–2.93 (m, 4H), 2.01–1.89 (m, 1H), 1.76–1.64 (m, 1H), 1.46–1.29 (m, 4H), 0.88 (t, J = 7.1 Hz, 3H).

[0421] LCMS (ESI): 301.3 [M+H] + 。

[0422] Example 62

[0423] (R)-3-Butyl-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)isobenzofuran-1(3H)-one (I-62), the structure is shown as follows:

[0424]

[0425] According to the method shown in Reaction Route 12, intermediate 11 was reacted with intermediate A-36 to obtain the Boc-protected product. Then, the Boc-protection was removed using a DCM solution of trifluoroacetic acid to obtain the desired product (I-62). LCMS(ESI): 301.2 [M+H] + 。

[0426] Example 63

[0427] (R)-3-Butyl-6-(2,6-diazaspiro[3.3]hept-2-yl)isobenzofuran-1(3H)-one (I-63), the structure is shown as follows:

[0428]

[0429] According to the method shown in Reaction Route 12, intermediate 11 was reacted with intermediate A-19 to obtain the Boc-protected product. Then, the Boc-protection was removed using a DCM solution of trifluoroacetic acid to obtain the desired product (I-63). LCMS(ESI): 287.2 [M+H] + 。

[0430] Example 64

[0431] (S)-3-Butyl-6-(2,6-diazaspiro[3.3]hept-2-yl)isobenzofuran-1(3H)-one (I-64), the structure is shown as follows:

[0432]

[0433] According to the method shown in Reaction Route 12, intermediate 10 was reacted with intermediate A-19 to obtain the Boc-protected product. Then, the Boc-protection was removed using a DCM solution of trifluoroacetic acid to obtain the desired product (I-64). LCMS(ESI): 287.1 [M+H] + 。

[0434] Example 65

[0435] (S)-3-Butyl-6-morpholinylisobenzofuran-1(3H)-one (I-65), the structure is shown as follows:

[0436]

[0437] According to the method shown in Reaction Scheme 12, Intermediate 10 is reacted with Intermediate A-26 to obtain the desired product (I-65). LCMS (ESI): 276.2 [M+H] + .

[0438] Example 66

[0439] (R)-3-Butyl-6-morpholinoisobenzofuran-1(3H)-one (I-66), the structure of which is shown below:

[0440]

[0441] According to the method shown in Reaction Scheme 12, intermediate 11 is reacted with intermediate A-26 to obtain the desired product (I-66). LCMS (ESI): 276.3 [M+H] + .

[0442] Example 67

[0443] (S)-3-Butyl-6-(4-hydroxy-4-methylpiperidin-1-yl)isobenzofuran-1(3H)-one (I-67), the structure is shown below:

[0444]

[0445] According to the method shown in Reaction Scheme 12, Intermediate 10 is reacted with Intermediate A-45 to obtain the desired product (I-67). LCMS (ESI): 304.2 [M+H] + .

[0446] Example 68

[0447] (R)-3-Butyl-6-(4-hydroxy-4-methylpiperidin-1-yl)isobenzofuran-1(3H)-one (I-68), the structure of which is shown below:

[0448]

[0449] According to the method shown in Reaction Scheme 12, Intermediate 11 is reacted with Intermediate A-45 to obtain the desired product (I-68). LCMS (ESI): 304.1 [M+H] + .

[0450] Example 69

[0451] (S)-3-Butyl-6-(3-hydroxy-3-methylazetidin-1-yl)isobenzofuran-1(3H)-one (I-69), the structure of which is shown below:

[0452]

[0453] According to the method shown in Reaction Route 12, react Intermediate 10 with Intermediate A-43 to obtain the desired product (I-69). LCMS (ESI): 276.2 [M+H] + 。

[0454] Example 70

[0455] (R)-3-Butyl-6-(3-hydroxy-3-methylazetidin-1-yl)isoindoline-1(3H)-one (I-70), the structure is shown as follows:

[0456]

[0457] According to the method shown in Reaction Route 12, react Intermediate 11 with Intermediate A-43 to obtain the desired product (I-70). LCMS (ESI): 276.2 [M+H] + 。

[0458] Example 71

[0459] (S)-3-Butyl-6-(3-hydroxyazetidin-1-yl)isoindoline-1(3H)-one (I-71), the structure is shown as follows:

[0460]

[0461] According to the method shown in Reaction Route 12, react Intermediate 10 with Intermediate A-40 to obtain the desired product (I-71). Characterize the product, and the characterization results are as follows:

[0462] 1 H NMR (400 MHz, CDCl3) δ 7.22 (d, J = 8.2 Hz, 1H), 6.84 (d, J = 2.2 Hz, 1H), 6.72 (dd, J = 8.3, 2.2 Hz, 1H), 5.37 (dd, J = 7.7, 4.2 Hz, 1H), 4.85–4.76 (m, 1H), 4.25–4.18 (m, 2H), 3.74 (dd, J = 8.4, 4.5 Hz, 2H), 2.02–1.91 (m, 1H), 1.76–1.67 (m, 1H), 1.49–1.31 (m, 4H), 0.90 (t, J = 7.1 Hz, 3H).

[0463] LCMS (ESI): 262.1 [M+H] + 。

[0464] Example 72

[0465] (R)-3-Butyl-6-(3-hydroxyazetidin-1-yl)isoindoline-1(3H)-one (I-72), the structure is shown as follows:

[0466]

[0467] According to the method shown in Reaction Route 12, intermediate 11 was reacted with intermediate A-40 to obtain the desired product (I-72). LCMS(ESI): 262.2 [M+H] + .

[0468] Example 73

[0469] (R)-3-Butyl-6-((R)-3-hydroxypyrrolidin-1-yl)isoindoline-1(3H)-one (I-73), the structure is shown as follows:

[0470]

[0471] According to the method shown in Reaction Route 12, intermediate 11 was reacted with (R)-pyrrolidin-3-ol to obtain the desired product (I-73). LCMS(ESI): 276.2 [M+H] + .

[0472] Example 74

[0473] (S)-3-Butyl-6-((R)-3-hydroxypyrrolidin-1-yl)isoindoline-1(3H)-one (I-74), the structure is shown as follows:

[0474]

[0475] According to the method shown in Reaction Route 12, intermediate 10 was reacted with (R)-pyrrolidin-3-ol to obtain the desired product (I-74). LCMS(ESI): 276.2 [M+H] + .

[0476] Example 75

[0477] (R)-3-Butyl-6-((S)-3-hydroxypyrrolidin-1-yl)isoindoline-1(3H)-one (I-75), the structure is shown as follows:

[0478]

[0479] According to the method shown in Reaction Route 12, intermediate 11 was reacted with (S)-pyrrolidin-3-ol to obtain the desired product (I-75). LCMS(ESI): 276.1 [M+H] + .

[0480] Example 76

[0481] (S)-3-Butyl-6-((S)-3-hydroxypyrrolidin-1-yl)isobenzofuran-1(3H)-one (I-76), the structure is shown as follows:

[0482]

[0483] According to the method shown in Reaction Route 12, intermediate 10 was reacted with (S)-pyrrolidin-3-ol to obtain the desired product (I-76). LCMS (ESI): 276.2 [M+H] + 。

[0484] Example 77

[0485] 6-Bromo-3-(3-hydroxypropyl)isobenzofuran-1(3H)-one (I-77) was prepared according to the method shown in Reaction Route 13, specifically as follows:

[0486]

[0487] (1) A mixture of compound 34 (3.0 g, 14.1 mmol), 1,2-dichloroethane (25 mL), N-bromosuccinimide (3.0 g, 16.9 mmol) and azobisisobutyronitrile (0.09 g, 0.6 mmol) was refluxed for 6 hours. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain compound 35 (3.5 g, yield: 85%). 1 1H NMR (400 MHz, CDCl3) δ 8.05 (d, 1H), 7.89 (dd, 1H), 7.51 (d, 1H), 7.35 (s, 1H); LCMS (ESI): 292.9 [M+H] + 。

[0488] (2) Compound 35 (3.5 g, 12.0 mmol) was suspended in 20 mL of water and refluxed for 2 h. The reaction mixture was cooled to room temperature and stirred overnight. The two-phase mixture was extracted with ethyl acetate (20 mL × 3), and the combined organic layers were dried over anhydrous Na2SO4 and evaporated to obtain the crude product 36 (2.7 g, yield: 100%). 1 1H NMR (400 MHz, CDCl3) δ 8.01 (d, 1H), 7.97 (dd, 1H), 7.65 (d, 1H); LCMS (ESI): 230.0 [M+H]+.

[0489] (3) A mixed solution of compound 36 (2.7 g, 11.8 mmol), compound 37 (2.4 g, 14.1 mmol), Bi(OAc)3 (91 mg, 0.24 mmol), H2SO4 (23 mg, 0.24 mmol) and CH3CN (5.0 mL) was stirred at room temperature under argon for 6 - 10 h until TLC showed that compound 36 was completely consumed. Then the reaction mixture was extracted with ethyl acetate (15 mL × 3), and the combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to obtain a crude mixture. Purification by column chromatography gave compound 38 (2.1 g, yield: 70%). 1 H NMR (400 MHz, CDCl3) δ 7.96 (d, 1H), 7.75 (dd, 1H), 7.35 (d, 1H), 5.75 - 5.64 (m, 1H), 5.46 (t, 1H), 5.17 - 5.12 (m, 2H), 2.74 - 2.59 (m, 2H); LCMS (ESI): 254.1 [M + H]+.

[0490] (4) To a stirred solution of compound 38 (0.6 g, 2.4 mmol) in anhydrous THF (5 mL), borane - dimethyl sulfide complex (10.0 M, DMS solution, 0.28 mL, 2.8 mmol) was added while maintaining 0 °C and an argon atmosphere. The reaction mixture was warmed to room temperature and stirred overnight. At room temperature, H2O2 (30%, 2.7 g, 23.7 mmol) and NaOH (0.5 g, 11.9 mmol) were added, and the resulting mixture was stirred for 3 h. The mixture was extracted with ethyl acetate (5 mL × 3), and the combined organic layers were dried over anhydrous Na2SO4. After filtration and evaporation, the crude mixture was purified by silica gel column chromatography to give compound I - 77 (0.21 g, yield: 33%); 1 H NMR (400 MHz, CDCl3) δ 8.00 (d, 1H), 7.97 (dd, 1H), 7.66 (d, 1H), 5.67 - 5.65 (m, 1H), 4.51 (br, 1H), 3.42 (t, 1H), 2.13 - 2.06 (m, 1H), 1.78 - 1.70 (m, 1H), 1.53 - 1.37 (m, 2H); LCMS (ESI): 272.1 [M + H] + 。

[0491] Intermediate 39: Preparation of 3 - (5 - bromo - 3 - oxo - 1,3 - dihydroisobenzofuran - 1 - yl)propyl 4 - methylbenzenesulfonate is as follows:

[0492] Compound I-77 (0.21 g, 0.77 mmol) was dissolved in DCM (7 mL) under N₂, cooled to 0°C, and TsCl (0.22 g, 1.16 mmol), DIPEA (0.2 g, 1.55 mmol), and DMAP (9.5 mg, 0.077 mmol) were added. The reaction mixture was stirred from 0°C to room temperature for 6-10 hours. Upon completion, the mixture was extracted with ethyl acetate (15 mL x 3). The combined organic layers were washed with water and brine, dried over anhydrous Na₂SO₄, and evaporated to obtain a crude mixture. Purification by column chromatography afforded compound 39 (70 mg, yield: 21%).

[0493] 1 H NMR(400MHz, CDCl3)δ8.00(d,1H),7.80-7.75(m,3H),7.35-7.30(m,3H),5.42-5.39(m,1H),4.15-4.11 (m,1H),4.05-4.00(m,1H),2.44(s,3H),2.21-2.16(m,1H),1.87-1.68(m,3H); LCMS(ESI): 426.3[M+H] + .

[0494] Compounds I-78 to I-92 were prepared according to the method shown in reaction scheme 14 or 15, as shown below:

[0495]

[0496] Reaction Scheme 14

[0497] The specific operations are as follows:

[0498] A mixture of compound 39 (0.16 mmol) and amine 40 (0.25 mmol), K2CO3 (57 mg, 0.41 mmol), KI (8.2 mg, 0.049 mmol) in CH3CN (5 mL) was stirred under N2 at 50 to 80°C for 4-8 h until TLC showed complete consumption of compound 39. The reaction mixture was then cooled to room temperature and extracted with DCM (15 mL x 3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give a crude product. The crude product was purified by TLC to give a moderate to excellent yield of the product (60-80%).

[0499]

[0500] Reaction Scheme 15

[0501] The specific operations are as follows:

[0502] (1) Similar to Reaction Route 14, replace amine 40 with amine 41 therein;

[0503] (2) Dissolve compound 42 (100 mg) in DCM (5 ml), and add TFA (0.5 ml). Stir the reaction mixture at room temperature for 0.5 - 1 h until TLC shows that compound 42 is completely consumed. Remove most of the solvent, adjust the pH to 7 with saturated NaHCO3, extract with ethyl acetate (15 mL × 3), and combine the organic layers. Wash each layer with water and brine, dry over anhydrous Na2SO4 and evaporate to obtain a crude mixture. Purify by preparative silica gel plate to obtain the product in moderate to good yield.

[0504] Example 78

[0505] 6 - Bromo - 3-(3-(piperidin - 1 - yl)propyl)iso - benzofuran - 1(3H)-one (I - 78), the structure is shown as follows:

[0506]

[0507] According to the method shown in Reaction Route 14, react intermediate 39 with piperidine (40a) to obtain the desired product (I - 78). LCMS(ESI): 339.2[M + H] + 。

[0508] Example 79

[0509] 6 - Bromo - 3-(3 - morpholinopropyl)iso - benzofuran - 1(3H)-one (I - 79), the structure is shown as follows:

[0510]

[0511] According to the method shown in Reaction Route 14, react intermediate 39 with morpholine (40b) to obtain the desired product (I - 79). LCMS(ESI): 341.2[M + H] + 。

[0512] Example 80

[0513] 3-(3-(2 - Oxo - 6 - azaspiro[3.3]hept - 6 - yl)propyl)-6 - bromoiso - benzofuran - 1(3H)-one (I - 80), the structure is shown as follows:

[0514]

[0515] According to the method shown in Reaction Route 14, react intermediate 39 with 2 - oxo - 6 - azaspiro[3.3]heptane (40d) to obtain the desired product (I - 80).

[0516] 11H NMR (400 MHz, CDCl3) δ 8.01 (s, 1H), 7.78 (dd, 1H), 7.35 (d, 1H), 5.46 - 5.43 (m, 1H), 4.74 (s, 4H), 3.46 (s, 4H), 2.56 - 2.49 (m, 2H), 2.17 - 2.09 (m, 2H), 1.75 - 1.70 (m, 1H), 1.57 - 1.50 (m, 1H).

[0517] LCMS (ESI): 353.3 [M + H] + 。

[0518] Example 81

[0519] 6 - Bromo - 3-(3 - ((3aR,6aS)-tetrahydro - 1H - furo[3,4 - c]pyrrol - 5(3H)-yl)propyl)iso - benzofuran - 1(3H)-one (I - 81), the structure is shown as follows:

[0520]

[0521] According to the method shown in Reaction Route 14, intermediate 39 was reacted with hexahydro - 1H - furo[3,4 - c]pyrrole (40e) to obtain the desired product (I - 81).

[0522] 1 1H NMR (400 MHz, CDCl3) δ 8.01 (s, 1H), 7.79 (d, 1H), 7.40 (d, 1H), 5.50 - 5.47 (m, 1H), 3.69 - 3.64 (m, 4H), 3.28 - 3.21 (m, 2H), 3.04 - 3.01 (m, 2H), 2.79 - 2.73 (m, 2H), 2.44 - 2.40 (m, 2H), 2.27 - 2.19 (m, 1H), 1.90 - 1.83 (m, 1H), 1.81 - 1.73 (m, 2H).

[0523] LCMS (ESI): 367.2 [M + H] + 。

[0524] Example 82

[0525] 3-(3-(2 - Oxabicyclo[2.2.1]hept - 5 - yl)propyl)-6 - bromo - iso - benzofuran - 1(3H)-one (I - 82), the structure is shown as follows:

[0526]

[0527] According to the method shown in Reaction Scheme 14, intermediate 39 is reacted with 2-oxa-5-azabicyclo[2.2.1]heptane (40 g) to obtain the desired product (I-82).

[0528] 1 H NMR(400MHz, CDCl3)δ8.02(d,1H),7.78(dd,1H),7.34(dd,1H),5.51-5.47(m,1H) ,4.40(s,1H),4.02-4.00(m,1H),3.63-3.61(m,1H),3.52-3.51(m,1H),2.95-2.9 2(m,1H),2.78-2.71(m,1H),2.69-2.61(m,1H),2.55-2.52(m,1H),2.18-2.12(m, 1H),1.89-1.87(m,1H),1.84-1.80(m,1H),1.77-1.74(m,1H),1.71-1.52(m,2H).

[0529] LCMS (ESI): 353.0 [M+H] + .

[0530] Example 83

[0531] 6-Bromo-3-(3-(3-hydroxyazetidin-1-yl)propyl)isobenzofuran-1(3H)-one (I-83), the structure of which is shown below:

[0532]

[0533] According to the method shown in reaction scheme 14, intermediate 39 is reacted with azetidin-3-ol (40h) to obtain the desired product (I-83). LCMS (ESI): 327.2 [M+H] + .

[0534] Example 84

[0535] 6-Bromo-3-(3-(3-hydroxy-3-methylazetidin-1-yl)propyl)isobenzofuran-1(3H)-one (I-84), the structure of which is shown below:

[0536]

[0537] According to the method shown in reaction scheme 14, intermediate 39 is reacted with 3-methylazetidin-3-ol (40i) to obtain the desired product (I-84). LCMS (ESI): 341.2 [M+H] + .

[0538] Example 85

[0539] 3-(3-(2-oxa-5-azabicyclo[2.2.1]hept-5-yl)propyl)-6-bromo-isobenzofuran-1(3H)-one (I-85), the structure of which is shown below:

[0540]

[0541] According to the method shown in reaction scheme 14, intermediate 39 is reacted with 2-oxa-5-azabicyclo[2.2.1]heptane (40j) to obtain the desired product (I-85). LCMS (ESI): 355.2 [M+H] + .

[0542] Example 86

[0543] 6-Bromo-3-(3-(6-fluoro-2-azaspiro[3.3]hept-2-yl)propyl)isobenzofuran-1(3H)-one (I-86), the structure of which is shown below:

[0544]

[0545] According to the method shown in reaction scheme 14, intermediate 39 is reacted with 6-fluoro-2-azaspiro[3.3]heptane (40k) to obtain the desired product (I-86). LCMS (ESI): 369.2 [M+H] + .

[0546] Example 87

[0547] 3-(3-(6-azaspiro[2.5]octan-6-yl)propyl)-6-bromoisobenzofuran-1(3H)-one (I-87), the structure of which is shown below:

[0548]

[0549] According to the method shown in reaction scheme 14, intermediate 39 was reacted with 6-azaspiro[2.5]octane (40n) to obtain the desired product (I-87). LCMS (ESI): 365.2 [M+H] + .

[0550] Example 88

[0551] 3-(3-((1R,5S)-3-azabicyclo[3.1.0]hex-3-yl)propyl)-6-bromoisobenzofuran-1(3H)-one (I-88), the structure is shown below:

[0552]

[0553] According to the method shown in reaction scheme 14, intermediate 39 is reacted with 3-azabicyclo[3.1.0]hexane (40o) to obtain the desired product (I-88). LCMS (ESI): 365.2 [M+H] + .

[0554] Example 89

[0555] 6-Bromo-3-(3-(piperazin-1-yl)propyl)isobenzofuran-1(3H)-one (I-89), the structure of which is shown below:

[0556]

[0557] According to the method shown in Reaction Scheme 15, intermediate 39 is reacted with tert-butyl piperazine-1-carboxylate (41a) to obtain the desired product (I-89). LCMS (ESI): 340.2 [M+H] + .

[0558] Example 90

[0559] 6-Bromo-3-(3-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propyl)isobenzofuran-1(3H)-one (I-90), the structure of which is shown below:

[0560]

[0561] According to the method shown in Reaction Scheme 15, intermediate 39 is reacted with hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (41b) to obtain the desired product (I-90).

[0562] 1 H NMR(400MHz, CDCl3)δ8.00(s,1H),7.78(d,1H),7.37(d,1H),5.50-5.47(m,1H),3.41-3.39(m,2H),3.08-3.02(m,2H),2.95-2 .92(m,2H),2.80-2.71(m,2H),2.48-2.46(m,2H),2.30-2.28(m,2H),2.19-2.11(m,1H),1.77-1.71(m,1H),1.64-1.57(m,2H).

[0563] LCMS (ESI): 366.3 [M+H] + .

[0564] Example 91

[0565] 3-(3-(2,5-diazabicyclo[2.2.1]hept-2-yl)propyl)-6-bromoisobenzofuran-1(3H)-one (I-91), the structure is shown below:

[0566]

[0567] According to the method shown in Reaction Scheme 15, intermediate 39 is reacted with tert-butyl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (41c) to obtain the desired product (I-91). LCMS (ESI): 352.2 [M+H] + .

[0568] Example 92

[0569] 3-(3-(3,6-diazabicyclo[3.1.1]hept-3-yl)propyl)-6-bromoisobenzofuran-1(3H)-one (I-92), the structure of which is shown below:

[0570]

[0571] According to the method shown in reaction scheme 15, intermediate 39 is reacted with tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (41d) to obtain the desired product (I-92). LCMS (ESI): 352.2 [M+H] + .

[0572] Example 93

[0573] 6-Bromo-3-(4-hydroxybutyl)isobenzofuran-1(3H)-one (I-92) was prepared according to the method shown in Reaction Scheme 16 as follows:

[0574]

[0575] (1) To a mixture of compound 44 (23.7 g, 0.23 mol) in DCM (150 mL) was added TEA (64.8 g, 0.64 mol) at 0°C under N2, stirred for 30 min, and then compound 43 (30 g, 0.21 mol) was added. After addition, the reaction mixture was stirred at room temperature for 5-8 h until TLC showed that compound 43 was completely consumed. The mixture was quenched with saturated NH4Cl and extracted with DCM (100 mL×3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give a crude product. The residue was purified by silica gel column chromatography to give compound 45 (42.6 g, yield: 97%). 11H NMR (400 MHz, CDCl3) δ 7.30 - 7.21 (m, 5H), 3.73 - 3.46 (m, 2H), 1.2 (d, 12H); LCMS (ESI): 206.3 [M+H] + 。

[0576] (2) At -78 °C under N2, tBuLi (82 mL, 0.11 mol) and N,N,N',N'-tetramethylethylenediamine (12.5 g, 0.11 mol) were added dropwise to a mixture of compound 45 (20 g, 0.097 mol) in anhydrous THF (150 mL), stirred for 1 h, then compound 46 (11.7 g, 0.12 mol) was added dropwise. The reaction mixture was stirred at -78 °C for 5 - 8 h until TLC showed complete consumption of compound 3. It was quenched with saturated NH4Cl, extracted with DCM (100 mL × 3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to obtain the crude product, and the residue was purified by silica gel column chromatography to give compound 47 (19.9 g, yield 67%). 1 1H NMR (400 MHz, CDCl3) δ 7.73 (d, 1H), 7.49 - 7.45 (m, 1H), 7.41 - 7.39 (m, 1H), 7.18 (d, 1H), 4.09 - 4.06 (m, 1H), 3.57 - 3.48 (m, 4H), 2.96 - 2.92 (m, 2H), 1.81 - 1.76 (m, 2H), 1.57 - 1.51 (m, 8H), 1.12 (d, 6H); LCMS (ESI): 306.4 [M+H] + 。

[0577] (3) Compound 47 (19.9 g, 0.065 mol) was dissolved in anhydrous MeOH (80 mL). At 0 °C, NaBH4 (3.7 g, 0.098 mol) was added portionwise to the mixture. After addition, the reaction mixture was stirred at room temperature for 5 - 8 h until TLC showed complete consumption of compound 47. It was cooled to 0 °C, quenched with 2N HCl, extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to obtain the crude product, and the residue was filtered. Purification by silica gel column chromatography gave compound 48 (14.0 g, yield: 70%). 11H NMR (400 MHz, CDCl3) δ 7.48 (d, 1H), 7.40 - 7.35 (m, 1H), 7.33 - 7.31 (m, 1H), 7.29 - 7.27 (d, 1H), 4.61 - 4.57 (m, 1H), 4.53 (t, 1H), 3.74 - 3.70 (m, 2H), 3.50 - 3.44 (m, 4H), 1.90 - 1.71 (m, 1H), 1.74 - 1.69 (m, 1H), 1.52 (d, 6H), 1.46 - 1.42 (m, 2H), 1.11 (d, 6H); LCMS (ESI): 308.2 [M + H] + 。

[0578] (4) To a mixture of compound 48 (14.0 g, 0.046 mol) and toluene (100 mL), TsOH (78 mg, 0.46 mmol) was added. The reaction mixture was stirred at 130 °C for 12 h until TLC showed complete consumption of compound 48, cooled to room temperature, extracted with DCM (100 mL × 3), and the combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give a crude product. The residue was purified by silica gel column chromatography to give compound 49 (7.5 g, yield: 80%). 1 1H NMR (400 MHz, CDCl3) δ 7.81 (d, 1H), 7.62 (t, 1H), 7.48 - 7.44 (t, 1H), 7.41 (d, 1H), 5.45 - 5.42 (m, 1H), 3.60 - 3.56 (m, 2H), 2.47 (s, 1H), 2.07 - 2.01 (m, 1H), 1.75 - 1.70 (m, 1H), 1.57 - 1.51 (m, 4H); LCMS (ESI): 207.2 [M + H] + 。

[0579] (5) At 0 °C under N2, to a mixture of compound 49 (7.5 g, 0.036 mol) in anhydrous THF (80 mL), Et3N (14.7 g, 0.15 mol) was added, and then acetyl chloride (10.3 mL, 0.15 mol) was added dropwise. The reaction mixture was stirred at room temperature for 5 - 8 h until TLC showed complete consumption of compound 7, extracted with EA (80 mL × 3), and the combined organic layers were washed with water and brine, dried over NaOH and anhydrous Na2SO4 and evaporated to give a crude mixture. The residue was purified by silica gel column chromatography to give compound 50 (7.2 g, yield: 80%).

[0580] (6) At 0 °C, a mixture of compound 50 (7.2 g, 0.029 mol) dissolved in H2SO4 (30 mL) was added portionwise with KNO3 (4.4 g, 0.043 mol). The mixture was stirred for 30 min, then the reaction mixture was diluted with water and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give a crude mixture, and the residue was purified by silica gel column chromatography to give compound 51 (6.9 g, yield: 81%). 1 1H NMR (400 MHz, CDCl3) δ 8.73 (d, 1H), 8.56 (dd, 1H), 7.64 (d, 1H), 5.61 - 5.58 (m, 1H), 4.09 - 4.06 (t, 2H), 2.21 - 2.12 (m, 1H), 2.04 (s, 3H), 1.88 - 1.81 (m, 1H), 1.74 - 1.70 (m, 2H), 1.63 - 1.56 (m, 2H); LCMS (ESI): 294.3 [M+H] + 。

[0581] (7) A mixture of compound 51 (6.9 g, 0.024 mol) and 10% Pd / C (0.7 g) in EtOH (50 mL) was purged with hydrogen, and then a hydrogen balloon was placed. After 2 h, the mixture was filtered through diatomaceous earth and evaporated, and the crude product was purified by silica gel column chromatography to give compound 52 (5.6 g, yield: 90%). 1 1H NMR (400 MHz, CDCl3) δ 7.17 (d, 1H), 7.09 (d, 1H), 6.96 (dd, 1H), 5.39 - 5.36 (m, 1H), 4.07 - 4.04 (t, 2H), 3.94 (br, 2H), 2.04 (s, 3H), 2.01 - 1.98 (m, 1H), 1.75 - 1.68 (m, 3H), 1.58 - 1.52 (m, 2H); LCMS (ESI): 264.3. [M+H] + 。

[0582] (8) An aqueous solution (10 mL) of sodium nitrite (1.6 g, 0.023 mol) was added to a mixture of compound 52 (5.6 g, 0.021 mol) in hydrobromic acid (45 mL) and H2O (9 mL). The mixture was stirred for 1 h, and then CuBr (1.9 g, 0.013 mol) was added. After the addition, the reaction mixture was stirred at 75 °C for 3 - 8 h until TLC showed complete consumption of compound 52. It was cooled to room temperature, extracted with ethyl acetate (50 mL × 3), and the combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give a crude mixture. The residue was purified by silica gel column chromatography to give compound 53 (4.9 g, yield: 71%). 1 1H NMR (400 MHz, CDCl3) δ 8.02 (d, 1H), 7.79 (dd, 1H), 7.32 (d, 1H), 5.46 - 5.42 (m, 1H), 4.08 - 4.04 (t, 2H), 2.12 - 2.06 (m, 1H), 2.04 (s, 3H), 1.81 - 1.76 (m, 1H), 1.74 - 1.68 (m, 2H), 1.57 - 1.51 (m, 2H); LCMS (ESI): 328.2 [M + H] + .

[0583] (9) K2CO3 (6.2 g, 0.045 mol) was added to a mixture of compound 53 (4.9 g, 0.015 mol) in MeOH (45 mL) and H2O (9 mL). The reaction mixture was stirred at room temperature. The temperature was maintained for 2 h until TLC showed complete consumption of compound 53. It was extracted with ethyl acetate (50 mL × 3), and the combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give compound I - 93 (3.6 g, yield: 85%). 1 1H NMR (400 MHz, CDCl3) δ 8.02 (d, 1H), 7.78 (dd, 1H), 7.33 (d, 1H), 5.47 - 5.44 (m, 1H), 3.66 (t, 2H), 2.13 - 2.04 (m, 1H), 1.82 - 1.78 (m, 1H), 1.66 - 1.58 (m, 2H), 1.56 - 1.51 (m, 2H); LCMS (ESI): 286.2 [M + H] + .

[0584] Compounds I - 94 to I - 117 were prepared according to the method shown in Reaction Route 17, as follows:

[0585]

[0586] The specific operation is as follows:

[0587] (1) A mixture of compound I-93 (3.6 g, 0.013 mol) was cooled to 0°C in DCM (30 mL) under N2, and TsCl (3.6 g, 0.019 mol), DIPEA (3.3 g, 0.025 mol), and DMAP (0.15 g, 0.0013 mol) were added. The reaction mixture was stirred at 0°C to room temperature for 6-10 hours. The reaction mixture was extracted with ethyl acetate (50 mL×3), and the combined organic layers were washed with water and brine, dried over anhydrous Na2SO4, and evaporated to give the crude product. Purification by flash chromatography gave compound 54 (3.1 g, yield: 56%). 1 H NMR(400MHz, CDCl3)δ8.01(s,1H),7.80-7.75(m,3H),7.35-7.29(m,3H),5.40-5.37(m,1H),4.02(t ,2H),2.45(s,3H),2.07-2.00(m,1H),1.74-1.69(m,3H),1.57-1.47(m,2H); LCMS(ESI): 440.3[M+H] + .

[0588] (2) Compound 54 was prepared according to the method shown in Reaction Scheme 14 to obtain Compound 55;

[0589] (3) Compound 56 was prepared according to the method shown in Reaction Scheme 15 to obtain Compound 57;

[0590] Example 94

[0591] 6-Bromo-3-(4-(pyrrolidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-94), the structure of which is shown below:

[0592]

[0593] According to the method shown in Reaction Scheme 17, intermediate 54 is reacted with pyrrolidine (40p) to obtain the desired product (I-94). LCMS (ESI): 339.2 [M+H] + .

[0594] Example 95

[0595] 6-Bromo-3-(4-(3-hydroxypyrrolidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-95), the structure of which is shown below:

[0596]

[0597] According to the method shown in Reaction Route 17, react intermediate 54 with pyrrolidin-3-ol (40q) to obtain the desired product (I-95). LCMS (ESI): 355.2 [M+H] + 。

[0598] Example 96

[0599] 6-Bromo-3-(4-(3-fluoropyrrolidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-96), the structure is shown as follows:

[0600]

[0601] According to the method shown in Reaction Route 17, react intermediate 54 with 3-fluoropyrrolidine (40r) to obtain the desired product (I-96). LCMS (ESI): 357.2 [M+H] + 。

[0602] Example 97

[0603] 6-Bromo-3-(4-(4-hydroxy-4-methylpiperidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-97), the structure is shown as follows:

[0604]

[0605] According to the method shown in Reaction Route 17, react intermediate 54 with 4-methylpiperidin-4-ol (40s) to obtain the desired product (I-97). LCMS (ESI): 383.3 [M+H] + 。

[0606] Example 98

[0607] 6-Bromo-3-(4-(4,4-difluoropyrrolidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-98), the structure is shown as follows:

[0608]

[0609] According to the method shown in Reaction Route 17, react intermediate 54 with 4,4-difluoropyrrolidine (40t) to obtain the desired product (I-98). LCMS (ESI): 389.3 [M+H] + 。

[0610] Example 99

[0611] 6-Bromo-3-(4-(3-hydroxy-3-methylazetidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-99), the structure is shown as follows:

[0612]

[0613] According to the method shown in Reaction Scheme 17, intermediate 54 is reacted with 3-methylazetidin-3-ol (40i) to obtain the desired product (I-99).

[0614] 1 H NMR(400MHz, CDCl3)δ8.02(d,1H),7.97(dd,1H),7.67(d,1H),5.97(br,1H),5.65-5.63(m,1H),3.95-3.94(m,2H),3.81-3.79(m,2H) ,3.11-3.07(m,2H),2.11-2.08(m,1H),1.76-1.68(m,1H),1.52-1.48(m,2H),1.41(s,3H),1.38-1.36(m,2H); LCMS(ESI): 355.2[M+H] + .

[0615] Example 100

[0616] 6-Bromo-3-(4-(3-hydroxyazetidin-1-yl)butyl)isobenzofuran-1(3H)-one (I-100), the structure of which is shown below:

[0617]

[0618] According to the method shown in reaction scheme 17, intermediate 54 is reacted with azetidin-3-ol (40h) to obtain the desired product (I-100). LCMS (ESI): 341.2 [M+H] + .

[0619] Example 101

[0620] 3-(4-((1R,5S)-3-azabicyclo[3.1.0]hex-3-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-101), the structure is shown below:

[0621]

[0622] According to the method shown in Reaction Scheme 17, intermediate 54 is reacted with 3-azabicyclo[3.1.0]hexane (40o) to obtain the desired product (I-101).

[0623] 11H NMR (400 MHz, CDCl3) δ 7.98 (d, 1H), 7.79 (dd, 1H), 7.37 (d, 1H), 5.47 - 5.45 (m, 1H), 3.74 - 3.72 (m, 2H), 3.12 - 3.10 (m, 2H), 3.03 - 2.96 (m, 2H), 2.18 - 2.15 (m, 1H), 2.03 - 1.97 (m, 2H), 1.75 - 1.73 (m, 3H), 1.55 - 1.46 (m, 2H), 0.86 - 0.81 (m, 2H); LCMS (ESI): 351.1. [M + H] + 。

[0624] Example 102

[0625] 6 - Bromo - 3-(4 - ((1R,5S)-6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexan - 3 - yl)butyl)isobenzofuran - 1(3H) - one (I - 102), the structure is shown as follows:

[0626]

[0627] According to the method shown in Reaction Route 17, intermediate 54 was reacted with 6,6 - dimethyl - 3 - azabicyclo[3.1.0]hexane (40 l) to obtain the desired product (I - 102).

[0628] 1 1H NMR (400 MHz, CDCl3) δ 7.98 (s, 1H), 7.80 (d, 1H), 7.38 (d, 1H), 5.47 - 5.44 (m, 1H), 3.78 - 3.75 (m, 2H), 2.94 - 2.90 (m, 2H), 2.75 - 2.71 (m, 2H), 2.17 - 2.12 (m, 1H), 1.93 - 1.89 (m, 2H), 1.80 - 1.76 (m, 2H), 1.75 - 1.66 (m, 1H), 1.58 - 1.48 (m, 2H), 1.13 (s, 3H), 1.08 (s, 3H); LCMS (ESI): 379.1 [M + H]+.

[0629] Example 103

[0630] 6 - Bromo - 3-(4 - ((3aR,6aS)-hexahydrocyclopenta[c]pyrrol - 2(1H) - yl)butyl)isobenzofuran - 1(3H) - one (I - 103), the structure is shown as follows:

[0631]

[0632] According to the method shown in Reaction Route 17, intermediate 54 was reacted with octahydrocyclopenta[c]pyrrole (40f) to obtain the desired product (I-103).

[0633] Example 104

[0634] 6-Bromo-3-(4-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)butyl)isoindolin-1(3H)-one (I-104), the structure is shown as follows:

[0635]

[0636] According to the method shown in Reaction Route 17, intermediate 54 was reacted with hexahydro-1H-furo[3,4-c]pyrrole (40e) to obtain the desired product (I-104).

[0637] 13 C NMR (101 MHz, DMSO) δ 168.84, 149.58, 137.56, 128.15, 127.93, 125.38, 122.62, 81.52, 72.55, 58.41, 53.86, 42.83, 33.51, 26.22, 22.32; LCMS (ESI): 379.1 [M+H] + 。

[0638] Example 105

[0639] 6-Bromo-3-(4-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)butyl)isoindolin-1(3H)-one (I-105), the structure is shown as follows:

[0640]

[0641] According to the method shown in Reaction Route 17, intermediate 54 was reacted with tert-butyl (3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate to obtain the desired product (I-105). LCMS (ESI): 380.3 [M+H] + 。

[0642] Example 106

[0643] 6-Bromo-3-(4-((3aR,6aR)-hexahydropyrrolo[3,2-b]pyrrol-1(2H)-yl)butyl)isoindolin-1(3H)-one (I-106), the structure is shown as follows:

[0644]

[0645] According to the method shown in Reaction Route 17, react intermediate 54 with tert-butyl hexahydropyrrolo[3,2-b]pyrrole-1(2H)-carboxylate to obtain the desired product (I-106). LCMS(ESI): 380.3 [M+H] + 。

[0646] Example 107

[0647] 6-Bromo-3-(4-(octahydro-1,5-naphthyridin-1(2H)-yl)butyl)isobenzofuran-1(3H)-one (I-107), the structure is shown as follows:

[0648]

[0649] According to the method shown in Reaction Route 17, react intermediate 54 with tert-butyl octahydro-1,5-naphthyridine-1(2H)-carboxylate to obtain the desired product (I-107). LCMS(ESI): 408.3 [M+H] + 。

[0650] Example 108

[0651] 6-Bromo-3-(4-((3aR,7aS)-octahydro-2H-isoindol-2-yl)butyl)isobenzofuran-1(3H)-one (I-108), the structure is shown as follows:

[0652]

[0653] According to the method shown in Reaction Route 17, react intermediate 54 with octahydro-1H-isoindole (40m) to obtain the desired product (I-108). LCMS(ESI): 393.3 [M+H] + 。

[0654] Example 109

[0655] 3-(4-(6-Azaspiro[2.5]octan-6-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-109), the structure is shown as follows:

[0656]

[0657] According to the method shown in Reaction Route 17, react intermediate 54 with 6-azaspiro[2.5]octane (40n) to obtain the desired product (I-109). LCMS(ESI): 379.3 [M+H] + 。

[0658] Example 110

[0659] 3-(4-(2,5-Diazabicyclo[2.2.1]hept-2-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-110), the structure is shown as follows:

[0660]

[0661] According to the method shown in Reaction Route 17, intermediate 54 was reacted with tert-butyl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (41c) to obtain the desired product (I-110). LCMS(ESI): 366.3 [M+H] + .

[0662] Example 111

[0663] 3-(4-(3,6-Diazabicyclo[3.1.1]hept-3-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-111), the structure is shown as follows:

[0664]

[0665] According to the method shown in Reaction Route 17, intermediate 54 was reacted with tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (41d) to obtain the desired product (I-111). LCMS(ESI): 366.3 [M+H] + .

[0666] Example 112

[0667] 3-(4-(2-Oxa-5-azabicyclo[2.2.1]hept-5-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-112), the structure is shown as follows:

[0668]

[0669] According to the method shown in Reaction Route 17, intermediate 54 was reacted with 2-oxa-5-azabicyclo[2.2.1]heptane (40g) to obtain the desired product (I-112).

[0670] 11H NMR (400 MHz, CDCl3) δ 7.94 (s, 1H), 7.74 (d, 1H), 7.32 (dd, 1H), 5.44 - 5.40 (m, 1H), 4.54 (s, 1H), 4.22 - 4.20 (m, 1H), 4.14 - 4.12 (m, 1H), 3.75 - 3.71 (m, 1H), 3.44 - 3.38 (m, 1H), 3.06 - 3.03 (m, 1H), 2.96 - 2.92 (m, 2H), 2.38 - 2.35 (m, 1H), 2.15 - 2.11 (m, 1H), 2.05 - 2.02 (m, 1H), 1.91 - 1.84 (m, 2H), 1.71 - 1.68 (m, 1H), 1.52 - 1.50 (m, 2H); LCMS (ESI): 367.4 [M + H] + 。

[0671] Example 113

[0672] 3-(4-(2-Azaspiro[3.3]heptan-2-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-113), the structure is shown as follows:

[0673]

[0674] According to the method shown in Reaction Route 17, intermediate 54 was reacted with 2-azaspiro[3.3]heptane (40c) to obtain the desired product (I-113).

[0675] 1 1H NMR (400 MHz, CDCl3) δ 7.98 (d, 1H), 7.79 (dd, 1H), 7.39 (d, 1H), 5.47 - 5.44 (m, 1H), 4.01 - 3.95 (m, 4H), 3.03 - 2.99 (m, 2H), 2.32 - 2.28 (m, 4H), 2.18 - 2.11 (m, 1H), 1.90 - 1.84 (m, 2H), 1.82 - 1.76 (m, 2H), 1.72 - 1.67 (m, 1H), 1.59 - 1.52 (m, 2H); LCMS (ESI): 365.4 [M + H] + 。

[0676] Example 114

[0677] 3-(4-(2-Oxa-6-azaspiro[3.3]heptan-6-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-114), the structure is shown as follows:

[0678]

[0679] According to the method shown in Reaction Scheme 17, intermediate 54 is reacted with 2-oxa-6-azaspiro[3.3]heptane (40d) to obtain the desired product (I-114).

[0680] Example 115

[0681] 3-(4-(2,6-diazaspiro[3.3]hept-2-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-115), the structure of which is shown below:

[0682]

[0683] According to the method shown in reaction scheme 17, intermediate 54 is reacted with tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (41h) to obtain the desired product (I-115). LCMS (ESI): 366.3 [M+H] + .

[0684] Example 116

[0685] 6-Bromo-3-(4-(6-hydroxy-2-azaspiro[3.3]hept-2-yl)butyl)isobenzofuran-1(3H)-one (I-116), the structure of which is shown below:

[0686]

[0687] According to the method shown in Reaction Scheme 17, intermediate 54 was reacted with 2-azaspiro[3.3]heptan-6-ol (40u) to obtain the desired product (I-116). LCMS (ESI): 381.3 [M+H] + .

[0688] Example 117

[0689] 6-Bromo-3-(4-(6-fluoro-2-azaspiro[3.3]hept-2-yl)butyl)isobenzofuran-1(3H)-one (I-117), the structure of which is shown below:

[0690]

[0691] According to the method shown in reaction scheme 17, intermediate 54 is reacted with 6-fluoro-2-azaspiro[3.3]heptane (40k) to obtain the desired product (I-117). LCMS (ESI): 383.3 [M+H] + .

[0692] The chirally pure compound was prepared according to one of the methods shown in reaction schemes 18-19, as shown below:

[0693]

[0694] Reaction Scheme 18

[0695] Specific operation: The racemic mixture was separated by SFC to split the mixture of 55 or 57 and isolate the optically pure compound.

[0696]

[0697] Reaction Scheme 19

[0698] Specific operations:

[0699] (2R)-3,3,3-trifluoro-2-methoxy-2-phenylpropionyl chloride was added to a solution of compound I-93 and DMAP in anhydrous pyridine, and the mixture was stirred at 20°C for 6 hours and then concentrated under a stream of N2. The residue was chromatographed on silica gel to give the desired compound. Mosher ester was separated into pure diastereomers by preparative chiral HPLC (using a CHIRALCEL OD column and a 40% iPrOH, hexane solvent system, flow rate 10 mL / min) to give compound 58. Compound 58 was treated with a MeOH solution of K2CO3 and water at 20°C for 3.5 hours. The mixture was cooled and neutralized with a MeOH solution of 0.1M HCl and 7MNH3. The resulting mixture was evaporated to dryness under reduced pressure, and the residue was then chromatographed on silica gel to give compound 60. LCMS (ESI): 286.1 [M+H] + .

[0700] Compound 61 was prepared according to the method shown in Reaction Scheme 19 using Intermediate I-93 and (2S)-3,3,3-trifluoro-2-methoxy-2-phenylpropionyl chloride. LCMS (ESI): 286.3 [M+H] + .

[0701]

[0702] Reaction Scheme 20

[0703] Specific operation: The chiral compounds 60 and 61 were separated from the racemic structure by SFC separation.

[0704] Example 118

[0705] (S)-3-(4-(2-oxa-6-azaspiro[3.3]hept-6-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-118), the structure of which is shown below:

[0706]

[0707] According to the method shown in Reaction Scheme 20, intermediate 61 is reacted with 2-oxa-6-azaspiro[3.3]heptane (40d) to give the product.

[0708] HPLC conditions: Chiral analytical column: ChiralPak AD (50×4.6 mm 3 μm), mobile phase: CO2:Ethanol (0.05% DEA) = 60:40; temperature: 35°C; flow rate: 3.0 mL / min; wavelength: 220 nm; retention time: 2.139 minutes (minor) 2.388 minutes (major), to obtain the target compound I-118, Ee: 98.2%.

[0709] LCMS (ESI): 353.4 [M+H] + .

[0710] Example 119

[0711] (R)-3-(4-(2-oxa-6-azaspiro[3.3]hept-6-yl)butyl)-6-bromoisobenzofuran-1(3H)-one (I-119), the structure of which is shown below:

[0712]

[0713] According to the method shown in Reaction Scheme 20, intermediate 60 is reacted with 2-oxa-6-azaspiro[3.3]heptane (40d) to give the product.

[0714] HPLC conditions: Chiral analytical column: ChiralPakAD (50×4.6mm 3um), mobile phase: CO2:Ethanol (0.05% DEA) = 60:40; temperature: 35°C; flow rate: 3.0 ml / min; wavelength: 220 nm; retention time: 2.116 minutes (main), to obtain the target compound I-119, Ee:>99.5%.

[0715] LCMS (ESI): 353.1 [M+H] + .

[0716] Example 120

[0717] (S)-6-Bromo-3-(4-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)butyl)isobenzofuran-1(3H)-one (I-120), the structure is shown below:

[0718]

[0719] According to the method shown in reaction scheme 20, intermediate 61 is reacted with hexahydro-1H-furo[3,4-c]pyrrole (40e) to obtain the product. LCMS (ESI): 381.2 [M+H] + .

[0720] Example 121

[0721] (R)-6-Bromo-3-(4-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)butyl)isobenzofuran-1(3H)-one (I-121), the structure is shown below:

[0722]

[0723] According to the method shown in reaction scheme 20, intermediate 60 is reacted with hexahydro-1H-furo[3,4-c]pyrrole (40e) to obtain the product. LCMS (ESI): 381.3 [M+H] + .

[0724] Biological activity studies

[0725] 1. Inhibitory effect on H2O2-induced PC12 cell damage

[0726] The biological activity of some of the compounds synthesized in the above examples was tested, and the inhibitory effects of commercially available edaravone, butylphthalide and BZP sodium salt on H2O2-induced PC12 cell damage were tested. The specific procedures are as follows:

[0727] PC12 cells (CellBio, Cat# iCell-r026) were cultured in RPMI-1640 medium containing 10% fetal bovine serum in an incubator at 37°C and 5% CO2. The cells were seeded in 96-well plates at a density of 5 × 10 cells per well. 3 After the cells were plated and stabilized, PC12 cells were treated with the corresponding concentrations of the test sample for 4 hours. H₂O₂ solution (final concentration 400 μM) was then added for 24 hours to establish the injury model. At the indicated time points, CCK-8 assays were performed, and the OD values at 450 nm were measured on a microplate reader (800TS, BIOTEK) for data analysis.

[0728] Injury inhibition rate = (injury model group cell inhibition rate - compound group cell inhibition rate) / injury model group cell inhibition rate × 100%, wherein, cell inhibition rate = (1 - test group OD value / blank group OD value) × 100%.

[0729] Table 1 Inhibitory effects of different compounds on H2O2-induced PC12 cell damage

[0730]

[0731]

[0732] *P < 0.05, **P < 0.01 vs PC12 + H2O2

[0733] According to the test results of the inhibitory effect of the above compounds on H2O2-induced PC12 cell damage, the above isobenzofuran-1(3H)-one derivatives all showed inhibitory effects on H2O2-induced PC12 cell damage, and compared with edaravone, butylphthalide, and BZP, some isobenzofuran-1(3H)-one derivatives had better inhibitory effects, such as compounds I-40, I-61, I-80, I-104, I-118, I-121, etc.

[0734] 2. hERG safety evaluation study

[0735] Test the hERG IC 50 value of some compounds synthesized in the above examples, and the specific operation is as follows:

[0736] Locate HEK293 cells (Invitrogen, Cat#K1236) to the ideal position and use the fine control of the robotic arm to approach the cell surface. Apply gentle suction through the side port of the electrode to form a gigahertz seal. Obtain the morphology of the whole cell by repeated, brief, and strong suction until the patch ruptures. First, depolarize to stimulate the hERG current, then inactivate it, and observe the inactivated tail current. Determine the hERG current amplitude using the maximum tail current magnitude. Perform carrier control on the cells to establish a baseline. Once the hERG current is found to be stable for 5 minutes, apply the working solution. In the presence of the test compound, record the hERG current for about 5 minutes to reach a steady state, and then capture 5 scans. In the dose-response experiment, apply 5 doses of the test compound to the cells cumulatively from low concentration to high concentration. Measure the hERG after-current with the highest concentration of the test compound as the internal low control to normalize the percentage inhibition. Use PatchMaster or Clampfit software to extract the peak current from the raw data. Use GraphpadPrism 8.0 to plot the dose-response curve of the percentage inhibition of the test compound concentration, and fit it to a sigmoidal dose-response curve with a variable slope.

[0737] The test results are shown in Table 2 below:

[0738] Table 2 hERG IC 50 values

[0739] Compound <![CDATA[IC 50 (μM)]]> I-36 1.094 I-40 25.837 I-61 1.712 I-80 5.445 I-104 2.386

[0740] As shown in Table 2, the series of isobenzofuran-1(3H)-one derivatives provided by the present invention have relatively high hERG IC 50 Value, among which the IC of compound I-40 50 The value is as high as 25.837 μM. Therefore, it can be seen that this type of isobenzofuran-1(3H)-one derivatives have low inhibitory activity on hERG potassium channels, and when used in a small amount, they have less cardiovascular toxicity and meet the needs of pharmaceutical use.

[0741] 3. Pharmacokinetic study in rats

[0742] Experimental animals: SD male rats (purchased from SPF Laboratory Animal Technology Co., Ltd.). Age: 6-8 weeks, weight: 180-300 g. Dosage: intravenous (1 mg / kg); PO (10 mg / kg).

[0743] Blood collection time points: IV (0.083, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 10, 24h); PO (0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 10, 24h).

[0744] All blood samples were transferred to plastic microcentrifuge tubes containing anticoagulant and centrifuged at 4000 g at 4°C for 5 min. The supernatant was transferred to a microcentrifuge tube without anticoagulant, and plasma was stored at -75 ± 15°C. After pretreatment, LC-MS / MS analysis was performed. Pharmacokinetic parameters were calculated using WinNonlin 8.3.1 software.

[0745] The test results are shown in Table 3 below:

[0746] Table 3 Pharmacokinetic parameters of different compounds in rats

[0747]

[0748]

[0749] As shown in Table 3, the present invention provides a series of isobenzofuran-1(3H)-one derivatives, which show good safety and tolerability in animal models.

[0750] In summary, the series of isobenzofuran-1(3H)-one derivatives provided by the present invention have a high inhibitory effect on H2O2-induced PC12 cell damage, low inhibitory activity on hERG potassium channels and good drugability, and have extremely high potential application value in the preparation of drugs for the prevention and / or treatment of cardiovascular ischemic diseases.

[0751] The above-described embodiments are merely preferred embodiments cited to fully illustrate the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or transformations made by those skilled in the art on the basis of the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. An isobenzofuran-1(3H)-one derivative having the structure of Formula I, or an isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug, and polymorph thereof, wherein, A is one of H, halogen, and a 3- to 10-membered heterocyclic group; E is -CR 1 R 2 - or -O-CR 1 R 2 -; Z is one of H, deuterium, halogen, hydroxyl, saturated or unsaturated C1-C4 alkyl, C4-C7 cycloalkyl, and a 3- to 10-membered heterocyclic group; wherein, the heterocycle in the 3- to 10-membered heterocyclic group is a heterocarbon ring, heteroaromatic ring, heterocycloaliphatic ring, heterospiro ring, heterobridged ring, or hetero-fused ring having one or more heteroatoms, and the heteroatoms are one or more of N, S, and O; R 1 、R 2 independently selected from one of H, deuterium, halogen, double bond, triple bond, C1-C10 alkyl; The above-mentioned heterocyclic group, alkyl group, cycloalkyl group are unsubstituted or substituted by one or more of the following substituents: halogen, hydroxyl group, C1-C10 alkyl group, halogen-substituted C1-C10 alkyl group, deuterium-substituted C1-C10 alkyl group, -CO-R 3 , -NH-CO-R 4 , -COOR 5 ; R 3 、R 4 、R 5 are independently selected from unsubstituted or halogen-substituted C1-C10 alkyl groups.

2. The isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph according to claim 1, characterized in that, A is bromine or a 3- to 10-membered heterocyclic group containing at least one N heteroatom, and the heterocyclic group is unsubstituted or substituted by one or more of the following substituents: hydroxyl, chlorine, bromine, methyl, deuterated methyl, -COCH3, -COCF3, -NH-COCH3, -COOC(CH3)3.

3. The isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph according to claim 1, characterized in that, E is -CR 1 R 2 -, R 1 、R 2 independently selected from one of H and C1-C10 alkyl groups.

4. The isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph according to claim 1, characterized in that, The compound has a general structural formula shown in Formulas Ia to Ic: wherein, n is 1, 2, or 3.

5. The isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph according to claim 1, characterized in that, Z is selected from one of hydrogen, hydroxyl, fluorine, vinyl, C1-C4 alkyl, C3-C6 cycloalkyl, and a 3- to 10-membered heterocyclic group containing N and / or O heteroatoms, and the alkyl and heterocyclic group are unsubstituted or substituted by one or more of the following groups: fluorine, hydroxyl, methyl, deuterated methyl, -COCH3, -COCF3, -NH-COCH3, -COOC(CH3)3.

6. The isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph according to claim 1, characterized in that, A is selected from one of the following structures: E is selected from one of the following structures: Z is selected from one of the following structures:

7. The isobenzofuran-1(3H)-one derivative or its isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug and polymorph according to claim 1, characterized in that, The isobenzofuran-1(3H)-one derivative is a compound shown in the following structural formulas I-01 to I-121:

8. A pharmaceutical composition, characterized in that, Comprising one or more of the isobenzofuran-1(3H)-one derivatives, isotopic forms, stereoisomers, pharmaceutically acceptable salts, pharmaceutically acceptable solvates, hydrates, prodrugs, and polymorphs described in any one of claims 1-6.

9. Use of an isobenzofuran-1(3H)-one derivative described in any one of claims 1-7, or an isotopic form, stereoisomer, pharmaceutically acceptable salt, pharmaceutically acceptable solvate, hydrate, prodrug, polymorph thereof, and the pharmaceutical composition described in claim 8 in the preparation of a drug for preventing and treating cardiovascular ischemic diseases, wherein the cardiovascular ischemic diseases include ischemic stroke.

10. The application according to claim 9, wherein The drug is administered by oral, parenteral, intravenous injection, or transdermal administration.

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