Non-classical ferroptosis inhibitor as well as preparation method and application thereof

Abstract

The invention relates to the field of medicinal chemistry, and particularly discloses indolizine derivatives with a structure as shown in a general formula (I), pharmaceutically acceptable salts and pharmaceutical compositions of the indolizine derivatives. Wherein a part of the compounds have known structures in the literature, but no biological activity is disclosed; the rest compounds are novel structures disclosed for the first time. The compound shown in the general formula (I) has excellent ferroptosis inhibition activity, and the action mechanism of the compound is to up-regulate expression of Nrf2 and / or GPX4 in cells. The compound has a remarkable protection effect in cell models and animal models (such as an MCAO (Macrophage-Amplified Antigen)-induced cerebral apoplexy model and an APAP (Aminogen Protein Protein)-induced liver injury model). Therefore, the compound and the composition provided by the invention can be used for preparing medicines for treating or preventing ferroptosis related diseases (such as cerebral arterial thrombosis and drug-induced liver injury). Formula I

Description

Technical Field

[0001] This invention relates to the fields of medicinal chemistry and pharmaceutical technology, and more specifically, to a class of novel heterocyclic derivatives with ferroptosis-inhibiting activity, their pharmaceutically acceptable salts, their preparation methods, and their applications. Background Technology

[0002] Ferroptosis is a recently discovered iron-dependent, regulatory cell death mechanism caused by the excessive accumulation of lipid peroxides. It differs from apoptosis, necrosis, and autophagy in morphology, biochemistry, and genetics. The core mechanisms of ferroptosis involve the dysfunction of the intracellular antioxidant defense system (especially the glutathione (GSH)-glutathione peroxidase 4 (GPX4) axis), as well as reactive oxygen species (ROS) bursts and lipid peroxidation caused by iron metabolism disorders. A growing body of research indicates that ferroptosis plays a crucial role in the pathological processes of a variety of major diseases, including but not limited to ischemia-reperfusion injury (such as ischemic stroke and myocardial infarction), neurodegenerative diseases (such as Alzheimer's disease and Parkinson's disease), drug-induced liver injury, acute kidney injury, and various cancers (Frontiers in Neuroscience, 2025, 19, 1623485; Molecular medicine reports, 2021, 23(3), 225; Frontiers in Cellular Neuroscience, 2024, 18, 1475934.).

[0003] In the field of ischemic stroke, models such as middle cerebral artery occlusion (MCAO) have confirmed significant ferroptosis characteristics in the ischemic core and penumbra, such as decreased GPX4 activity and accumulation of lipid peroxidation products. Currently, clinical treatments for ischemic stroke (such as thrombolysis and thrombectomy) have a narrow time window and carry risks such as hemorrhagic transformation. There is a lack of effective neuroprotective agents that can specifically protect neurons from ferroptosis damage. Developing drugs that can penetrate the blood-brain barrier and effectively inhibit neuronal ferroptosis is of great significance for prolonging the treatment time window and improving patient prognosis.

[0004] In the field of drug-induced liver injury, acetaminophen (APAP) overdose is one of the main causes of acute liver failure. Recent studies have found that APAP metabolites, after depleting hepatic GSH, directly inhibit GPX4 activity, triggering hepatocyte ferroptosis, which is one of the key mechanisms of its hepatotoxicity. Currently, the first-line antidote, N-acetylcysteine ​​(NAC), mainly works by supplementing GSH precursors, but its therapeutic window is also limited, and it is not effective for existing severe liver injury. Therefore, there is an urgent need to develop novel hepatoprotective drugs that directly target the core pathway of ferroptosis.

[0005] Currently, the reported ferroptosis inhibitors mainly include the following categories: (1) Iron chelators, such as deferoxamine (DFO). They reduce reactive oxygen species produced by iron catalysis at the source by chelating free iron ions (Neural Regen Res 2019, 14(3):532-5). However, these drugs usually have low bioavailability, require continuous administration, and long-term use may lead to side effects such as systemic iron deficiency.

[0006] (2) Lipophilic antioxidants / free radical scavengers, such as Ferrostatin-1 (Fer-1) and Liproxstatin-1. They can directly neutralize lipid free radicals and interrupt the chain reaction of lipid peroxidation. However, these compounds often have limitations in terms of metabolic stability, pharmacokinetic properties (such as short half-life) and tissue targeting in vivo, which limits their clinical translation potential (J. Med. Chem. 2018, 61, 22, 10126–10140; Bioorganic & MedicinalChemistry, 2025, 124: 118195.).

[0007] Both of the aforementioned inhibitors belong to the "exogenous intervention" strategy. In contrast, activating the cell's intrinsic antioxidant defense system, particularly the nuclear factor E2-related factor 2 (Nrf2)-GPX4 axis, is considered a more fundamental and broad-spectrum "endogenous enhancement" strategy. Nrf2 is a key transcription factor regulating cellular antioxidant stress responses. Under oxidative stress or activation by certain compounds, Nrf2 enters the nucleus and initiates the expression of a series of cytoprotective genes, including GPX4 and glutamate-cysteine ​​ligase (GCLC), thereby enhancing the cell's ability to resist ferroptosis at its source. Therefore, developing small molecule compounds that can efficiently and specifically activate the Nrf2 / GPX4 pathway is currently a cutting-edge direction in the development of antiferroptosis drugs, but such novel and well-defined lead compounds are still very scarce.

[0008] Chemically, indoleazine derivatives, as an important class of nitrogen heterocyclic compounds, have had their biological activities extensively studied. In the prior art, some literature (e.g., Org. Biomol. Chem., 2015, 13, 10236-10243) discloses the structures of some of the indoleazine derivatives of this invention. However, this literature only discloses the chemical synthesis and structural characterization of such compounds, without any research or testing of their biological activities, nor does it suggest or imply in any way that these compounds have the potential to inhibit ferroptosis, let alone their potential application in the treatment of ischemic stroke, drug-induced liver injury, and other ferroptosis-related diseases. For those skilled in the art, this known compound represents only isolated chemical structural information.

[0009] In summary, there is an urgent need in the art to develop a class of novel ferroptosis inhibitors that have been validated in models of major diseases such as ischemic brain injury and drug-induced liver injury. Although there are known compounds with disclosed structural portions, they do not offer any technical guidance related to the technical problem of this invention (inhibiting ferroptosis to treat related diseases). This invention aims to fill this technical gap by providing a novel class of ferroptosis inhibitors that demonstrate significant efficacy in the aforementioned disease models, possessing important scientific value and application prospects. Summary of the Invention

[0010] The purpose of this invention is to provide a class of novel indoleazine derivatives with excellent ferroptosis inhibitory activity, and their pharmaceutical applications.

[0011] A first aspect of the invention provides the use of compounds of Formula I or pharmaceutically acceptable salts, solvates, stereoisomers, isotope-labeled substances, or prodrugs thereof in the preparation of medicaments for inhibiting ferroptosis: Formula I in: X is -C(O)-; R 1 Selected from: C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, 3, or 4 substituents selected from the group consisting of: halogen, amino, hydroxyl, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy; R 2 Selected from: hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, carboxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; R3 R 4 R 5 R 6 Each is independently selected from: hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, -C(O)OR′, wherein R′ is independently hydrogen or C1-C3 alkyl, and R 3 R 4 R 5 R 6 At least two of them are hydrogen; A is selected from: C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, or 3 substituents selected from Ra; each Ra is independently selected from: hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, -C(O)OR b -O-CH2-phenyl; wherein R b It is a C1-C4 alkyl group.

[0012] In another preferred embodiment, Ra is selected from: hydrogen, deuterium, fluorine, chlorine, bromine, methyl, ethyl, trifluoromethyl, methoxy, nitro, amino, cyano, benzyloxy, -C(O)OCH3 or -C(O)OC2H5.

[0013] In another preferred embodiment, R 1 Selected from: C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-8 membered heterocyclic, phenyl, 5-6 membered heteroaryl.

[0014] In another preferred embodiment, R 1 Selected from: methyl, ethyl, n-propyl, isopropyl, phenyl.

[0015] In another preferred embodiment, R 2 It is hydrogen.

[0016] In another preferred embodiment, A is selected from: phenyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0017] In another preferred embodiment, the compound has the structure shown in Formula II: II n is 0, 1, 2, or 3; R 1 R 2 R 3 R 4 R 5 R 6 X and Ra are defined as above.

[0018] In another preferred embodiment, A is selected from: , , The definition of Ra is as described above.

[0019] In another preferred embodiment, R 3 R 4 R 5 R 6 It is hydrogen.

[0020] In another preferred embodiment, R 3 R 4 R 5 It is hydrogen; R 6 It is a halogen or a C1-C6 alkyl group.

[0021] In another preferred embodiment, R 3 R 4 R 6 It is hydrogen; R 5 It is a halogen or a C1-C6 alkyl group.

[0022] In another preferred embodiment, R 3 R 4 R 6 It is hydrogen; R 4 It is a halogen or a C1-C6 alkyl group.

[0023] In another preferred embodiment, R 3 R 4 R 5 R 6 In the middle, R 5 Or R 6 One of them is methyl or bromine, and the rest are hydrogen.

[0024] In another preferred embodiment, the compound is selected from compounds with the following structures: .

[0025] In another preferred embodiment, the ferroptosis is ferroptosis associated with ischemic brain injury or drug-induced liver injury.

[0026] In another preferred embodiment, the drug is used to treat or prevent acetaminophen (APAP)-induced liver injury.

[0027] In another preferred embodiment, the drug is used to reduce serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and malondialdehyde (MDA), and / or increase glutathione (GSH) levels.

[0028] A second aspect of the invention provides a compound of Formula I or a pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug thereof: Formula I in: X is -C(O)-; R 1 Selected from: C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, 3, or 4 substituents selected from the group consisting of: halogen, amino, hydroxyl, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy; R 2 It is hydrogen; R 3 R 4 R 5 R 6 Each is independently selected from: hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, -C(O)OR′, wherein R′ is independently hydrogen or C1-C3 alkyl, and R 3 R 4 R 5 R 6 At least two of them are hydrogen; A is selected from: C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, or 3 substituents selected from Ra; each Ra is independently selected from: hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, -C(O)OR b -O-CH2-phenyl; wherein R b It is a C1-C4 alkyl group; An additional condition is that the compound represented by Formula I is not selected from the following compounds: .

[0029] In another preferred embodiment, Ra is selected from: hydrogen, deuterium, fluorine, chlorine, bromine, methyl, ethyl, trifluoromethyl, methoxy, nitro, amino, cyano, benzyloxy, -C(O)OCH3 or -C(O)OC2H5.

[0030] In another preferred embodiment, R 1Selected from: C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-8 membered heterocyclic, phenyl, 5-6 membered heteroaryl.

[0031] In another preferred embodiment, R 1 Selected from: methyl, ethyl, n-propyl, isopropyl, phenyl.

[0032] In another preferred embodiment, A is selected from: phenyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0033] In another preferred embodiment, the compound has the structure shown in Formula II: II n is 0, 1, 2, or 3; R 1 R 2 R 3 R 4 R 5 R 6 X and Ra are as defined above.

[0034] In another preferred embodiment, A is selected from: , , The definition of Ra is as described above.

[0035] In another preferred embodiment, R 3 R 4 R 5 R 6 It is hydrogen.

[0036] In another preferred embodiment, R 3 R 4 R 5 It is hydrogen; R 6 It is a halogen or a C1-C6 alkyl group.

[0037] In another preferred embodiment, R 3 R 4 R 6 It is hydrogen; R 5 It is a halogen or a C1-C6 alkyl group.

[0038] In another preferred embodiment, R 3 R 4 R 6 It is hydrogen; R 4 It is a halogen or a C1-C6 alkyl group.

[0039] In another preferred embodiment, R 3 R 4 R 5 R 6 In the middle, R5 Or R 6 One of them is methyl or bromine, and the rest are hydrogen.

[0040] In another preferred embodiment, the compound is selected from compounds with the following structures: .

[0041] A third aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compound described in the second aspect of the present invention, or a pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0042] A fourth aspect of the present invention provides a method for inhibiting ferroptosis in vitro, comprising the steps of: Inhibiting ferroptosis in cells by contacting them with the compound of Formula I or its pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug: Formula I in, R 1 R 2 R 3 R 4 R 5 R 6 X and A are as defined above.

[0043] A fifth aspect of the present invention provides a method for treating ferroptosis-related diseases, comprising: Administer the compound of Formula I or a pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug thereof to a subject in need: Formula I in, R 1 R 2 R 3 R 4 R 5 R 6 X and A are as defined above.

[0044] In another preferred embodiment, the iron death-related diseases include ischemic brain injury or drug-induced liver injury.

[0045] The compounds of general formula I of this invention can be prepared by organic synthesis methods known in the art. Those skilled in the art can make reasonable designs based on known organic synthesis methods.

[0046] An exemplary method includes the following steps: compound a reacts with substituted or unsubstituted methylpyridine b to give intermediate c; intermediate c undergoes intramolecular cyclization to give intermediate d, which is then subjected to an acylation reaction to obtain the D series compounds. Compounds D15 and D18 are reduced to give compounds D19–D20. Among them, R 1 R 2 R 3 R 4 R 5 R 6 As stated above, A.

[0047] Further, the method includes the following steps: the reaction of compound a with substituted or unsubstituted methylpyridine b is carried out in an organic solvent; the molar ratio of compound a to methylpyridine is 1:1 to 2; the reaction temperature is 10 to 40 °C, and the time is 6 to 18 hours; The intramolecular cyclization reaction of intermediate c includes the following steps: the reaction is carried out under the action of DMF-Me2SO4 and a base; the volume ratio of DMF-Me2SO4 to base is 1:0.8-1.5; the solvent for the reaction is an organic solvent; the reaction temperature is 0-30 °C and the time is 1-6 hours. The acylation reaction of intermediate d includes the following steps: carried out under the action of an oxidant and a catalyst; the molar ratio of intermediate i, alkyl acid, oxidant and catalyst is 1:1 to 6:1 to 2:0.001 to 0.01; the reaction temperature is 50 to 100 °C and the time is 4 to 10 hours.

[0048] The reduction reaction of compound D includes the following steps: the compound is reacted under the action of a reducing agent; the volume ratio of the compound to the reducing agent is 1:0.8 to 1.5; the reaction temperature is 30 to 90°C, and the reaction time is 5 to 12 hours.

[0049] Furthermore, the organic solvent for the reaction of compound a with methylpyridine is acetonitrile; the molar ratio of compound a to methylpyridine b is 1:1.2; the reaction temperature is room temperature and the reaction time is 12 hours. The intramolecular cyclization reaction of intermediate c includes the following steps: the DMF-Me2SO4 is obtained by reacting DMF and Me2SO4 in equimolar amounts at 60 °C; the base is triethylamine; the volume ratio of DMF-Me2SO4 to the base is 1:0.8; the solvent for the reaction is DMF; the reaction temperature is 0~20 °C, and the time is 2 hours; The acylation reaction of the intermediate includes the following steps: the oxidant is iodobenzene acetate; the catalyst is palladium acetate; the intermediates i and R... 1 The molar ratio of OH, oxidant, and catalyst is 1:2:1.5:0.005; the reaction temperature is 90 °C and the reaction time is 6 hours.

[0050] The reduction reaction of the compound D15 / D18 includes the following steps: the reducing agent is SnCl2·2H2O; the volume ratio of the compound to the reducing agent is 1:1.1; the reaction temperature is 70℃ and the time is 7 hours.

[0051] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0052] Figure 1 The antiferroptosis effect of the target compounds is shown in (A). PC12 cell viability was assessed (n=3) after treatment with 1 μM RSL3 and various concentrations of different indoleazine compounds; (B, C) Quantitative analysis of EC50 values ​​for compounds D1 (B) and F2 (C). Data are expressed as mean ± standard deviation (n=3).

[0053] Figure 2 The cytotoxicity of compounds D1 and F2 in PC12 cells is shown. Data are presented as mean ± standard deviation (n=3).

[0054] Figure 3 The inhibitory activities of compounds and the classic ferroptosis inhibitors Fer1 and DFP in PC12 cells in a ferroptosis model induced by RSL3 (1 μM) are shown. (A) Activity results of compounds D1-D22; (B) Activity results of compounds D23-D29.

[0055] Figure 4 The safety profile of the preferred compounds and the classic ferroptosis inhibitor Fer-1 was demonstrated in PC12 and 293T cells.

[0056] Figure 5 Transmission electron microscopy morphology of PC12 cells restored by compound D1 (12.5 μM) induced by RSL3 (1 μM).

[0057] Figure 6 This shows the experimental results of iron chelation studies on compounds D1, D12, D14, and D15.

[0058] Figure 7This study presents the experimental results of the free radical scavenging effect of compounds D1, D12, D14, and D15.

[0059] Figure 8 Fluorescence imaging of lipid peroxidation in PC12 cells showing compounds D12, D14, D15 and Fer-1 (1 μM) in an RSL3 (1 μM)-induced ferroptosis model.

[0060] Figure 9 Fluorescence imaging and quantification of lipid peroxidation in PC12 cells with compounds D12, D14, D15 and Fer-1 (1 μM) in an RSL3 (1 μM)-induced ferroptosis model, ***P < 0.001 compared with the control group.

[0061] Figure 10 This demonstrates the effect of Western blot analysis on the levels of key proteins caused by D12 treatment.

[0062] Figure 11 The effects of compound D12 on acetaminophen-induced acute liver injury are shown in the following images: (A) Visual image of mouse liver tissue; (B) Eosin (H&E) staining results of mouse liver tissue; (C) Serum aspartate aminotransferase (ALT) and (D) alanine aminotransferase (AST) analysis in mice; (E) Malondialdehyde (MDA) and (F) glutathione (GSH) level analysis in mouse liver.

[0063] Figure 12 Compound D12 significantly reduced cerebral ischemia-reperfusion injury in MCAO rats. (A) Schematic diagram of the experimental procedure. (B) Representative images of TTC-stained brain sections from mice in the sham-operated group, model group, and D12-treated group. Magenta: normal tissue; white: infarcted tissue. (C) Quantitative analysis of cerebral infarct volume in each group; (D) Longa neurological deficit score. Data are expressed as mean ± standard deviation (n=3). Compared with the model group, p < 0.05; ** p < 0.001. Detailed Implementation

[0064] Through extensive and in-depth research, including numerous screenings and tests, the inventors have for the first time discovered that compounds of Formula I exhibit excellent inhibitory effects on ferroptosis. Furthermore, current experiments demonstrate that this compound inhibits ferroptosis independently of free radical capture and iron chelation pathways, making it suitable for preparing non-chelating, non-free radical capture ferroptosis inhibitors. Based on this, the present invention further confirms the potential application value of this type of compound in the prevention and treatment of cerebral ischemia-reperfusion injury and in the relief of acetaminophen-induced acute liver injury. The following detailed description, in conjunction with examples, illustrates the present invention, but the scope of the invention is not limited thereto.

[0065] the term In this invention, the term "halogen" refers to F, Cl, Br, or I.

[0066] In this invention, "C1-C6 alkyl" refers to a straight-chain or branched alkyl group comprising 1 to 6 (1, 2, 3, 4, 5 or 6) carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl or similar groups.

[0067] In this invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3-8 (1, 2, 3, 4, 5, 6, 7, or 8) carbon atoms on a ring, and non-limitingly includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. The term "C..." 3-6 "Cycloalkyl" has a similar meaning.

[0068] In this invention, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1-6 carbon atoms, and includes, without limitation, methoxy, ethoxy, propoxy, isopropoxy, and butoxy. Preferably, it is a C1-C4 alkoxy group.

[0069] In this invention, the term "haloalkyl" refers to a branched or straight-chain alkyl group substituted with one or more halogen atoms, up to the maximum permissible number of halogen atoms. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, monofluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0070] In this invention, the term "heterocyclic group" refers to a 4-10 membered heterocyclic group containing 1, 2, or 3 heteroatoms selected from N, O, and S. It may be used alone or in combination with other terms to refer to a saturated or partially saturated cyclic group consisting of 4-10 (4, 5, 6, 7, 8, 9, or 10) ring atoms, wherein 1, 2, or 3 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur atoms may optionally be oxidized (i.e., C(=O), NO, SO, and SO2). It includes monocyclic, bicyclic, and tricyclic systems, wherein bicyclic and tricyclic systems include spirocyclic, fused, and bridged rings. Furthermore, heteroatoms may occupy the connection positions between the heterocyclic group and the rest of the molecule; heterocyclic groups include saturated and partially unsaturated heterocyclic groups. Non-limiting examples of heterocyclic groups include azirrobutyl, oxacyclobutyl, thioheterobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuranyl-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperidinyl and 2-piperidinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, hexahydropyridazinyl, homopiperidinyl, and homopiperidinyl.

[0071] In this invention, the terms "aromatic ring" or "aryl" have the same meaning, and are preferably "C6-C10 aryl". The term "C6-C10 aryl" refers to an aromatic cyclic group with 6-10 carbon atoms that does not contain heteroatoms on the ring, such as phenyl, naphthyl, etc.

[0072] In this invention, the terms "aromatic heterocycle" or "heteroaryl" have the same meaning, referring to a heteroaromatic group containing one or more heteroatoms. For example, "5-10-membered heteroaryl" refers to an aromatic heterocycle containing 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen, and 2-7 carbon atoms. Non-limiting examples include: furanyl, thiophene, pyridinyl, pyrazolyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. The heteroaryl group may be optionally substituted or unsubstituted.

[0073] In this invention, the term "substitution" refers to the substitution of one or more hydrogen atoms on a specific group by a specific substituent. The specific substituent is the substituent described accordingly above, or the substituent appearing in the various embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substituted site of that group, and the substituents may be the same or different at each position. Those skilled in the art will understand that the combinations of substituents contemplated in this invention are those that are stable or chemically feasible. Such substituents include, but are not limited to: halogens, hydroxyl groups, carboxyl groups (-COOH), cyano groups (-CN), C1-C6 alkyl groups, C2-C6 alkenyl groups, C3-C8 cycloalkyl groups, 3- to 12-membered heterocyclic groups, aryl groups, heteroaryl groups, C1-C8 aldehyde groups, C2-C10 acyl groups, C2-C10 ester groups, amino groups, C1-C6 alkoxy groups, C1-C10 sulfonyl groups, etc.

[0074] In this invention, the terms 1-6 refer to 1, 2, 3, 4, 5, or 6. Other similar terms each have a similar meaning independently. The term "multiple" refers to 2-6 or more, such as 2, 3, 4, 5, or 6.

[0075] It should be understood that when a certain group exists simultaneously at multiple different positions in a compound, its definition at each position is independent of each other; they can be the same or different.

[0076] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, ( R )-and( S - Enantiomers, diastereomers, ( D )-Isomer, ( LEnantiomers, racemic mixtures thereof, and other mixtures thereof, such as mixtures of enantiomers enriched in diastereomers, are all within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0077] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by the compounds of the present invention with an acid or base that is suitable for use as a medicine. Pharmaceutically acceptable salts include both inorganic and organic salts. A preferred class of salts are those formed by the compounds of the present invention with an acid. Suitable acids for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

[0078] Another preferred class of salts are salts formed by the compounds of the present invention with a base, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (such as lower alkanol ammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0079] Pharmaceutical Compositions and Administration The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably, 5-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0080] "Pharmaceutically acceptable carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween®), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0081] The pharmaceutical composition is an injection, capsule, tablet, pill, powder, or granule.

[0082] There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and local administration.

[0083] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0084] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0085] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0086] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0087] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0088] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0089] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0090] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0091] The treatment method of the present invention can be used alone or in combination with other treatment methods or drugs.

[0092] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 5–1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician.

[0093] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0094] The raw materials and equipment used in this invention are all known products, obtained by purchasing commercially available products.

[0095] The "room temperature" or "room temperature" referred to in this invention is 25±10℃.

[0096] Example 1: Synthesis of Compound D1 The synthetic route for compound D1 is as follows: Dissolve a1 (1 g, 5.05 mmol, 1 eq) in acetonitrile, add methylpyridine (934 mg, 10.1 mmol, 2 eq), react overnight at room temperature, precipitate a solid, filter, and dry to give intermediate salt c1 as a white solid (1.1 g, 4.04 mmol, 80%). Me₂SO₄ and an equimolar amount of DMF were stirred at 80 °C for 3 h to obtain a mixed suspension. The cooled Me₂SO₄-DMF suspension (16 ml) was added to a DMF solution (20 ml) of c1 and stirred at room temperature for 15 min. Then, triethylamine (20 ml) was added under ice bath conditions, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction solution was quenched in ice water (100 ml). After standing, a large amount of yellow solid precipitated out. The solid was washed with water, dried under vacuum, and purified by column chromatography with petroleum ether / ethyl acetate (10:1) to obtain compound d1 as a yellow solid (598 mg, 2.71 mmol, 68%).

[0097] A mixture of compound d1 (270 mg, 1.22 mmol, 1 eq), palladium acetate (13 mg, 0.06 mol, 5 mol%), iodobenzene acetate (590 mg, 1.83 mmol, 1.5 eq), and acetic acid (146 mg, 2.44 mmol, 2 eq) in acetonitrile was refluxed for 6 h. After the reaction was completed as monitored by TLC, the solvent was removed under reduced pressure, and the target product D1 (196 mg, 0.71 mmol, 58%) was obtained by column chromatography with petroleum ether / ethyl acetate (10:1). The structural characterization data of compound D1 are as follows: 3-Benzoylindolizin-1-yl acetate (D1). Yellow solid with a yield of 58%. 1 H NMR (400 MHz, Chloroform-d) δ 9.95 (d, J= 7.1 Hz, 1H), 7.78 (d, J = 6.7 Hz, 2H), 7.56 – 7.43 (m, 4H), 7.29 (s, 1H),7.22 – 7.14 (m, 1H), 6.94 (t, J = 6.9 Hz, 1H), 2.34 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 184.58, 168.82, 140.44, 130.97, 130.19, 128.91, 128.27,128.20, 128.13, 124.10, 118.45, 116.87, 115.50, 114.47, 20.88. HRMS(ESI) m / z:[M+H] + calcd for C 17 H 13 NO3, 280.0968; found, 280.0961. Purity: 99.88%. Example 2: Synthesis of Compound D2 Compound D2 was prepared using a synthesis method similar to that in Example 1.

[0098] The structural characterization data of compound D2 are as follows: 3-(4-Fluorobenzoyl)indolizin-1-ylacetate (D2). Yellow solid with a yield of 18%. 1 H NMR (400 MHz, DMSO- d 6) δ9.81 (dt, J = 7.2, 1.2 Hz, 1H), 7.85 – 7.77 (m, 2H), 7.69 (dt, J= 9.0, 1.2Hz, 1H), 7.41 – 7.31 (m, 3H), 7.15 (td, J = 6.9, 1.4 Hz, 1H), 2.33 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 182.31, 169.39, 165.44, 162.97, 136.79 (d, 4 J C-F =3 Hz), 131.69(d, 3 J C-F = 9 Hz), 129.14 (d, 1 J C-F = 276 Hz), 128.28, 125.22,117.88, 117.09, 116.45, 115.87 (d, 2 J C-F = 22 Hz), 115.62, 21.02. 19 F NMR (376MHz, Chloroform-d) δ -108.76. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 12 FNO3,298.0874; found, 298.0861. Purity: 99.85%. Example 3: Synthesis of Compound D3 Compound D3 was prepared using a synthesis method similar to that in Example 1.

[0099] The structural characterization data of compound D3 are as follows: 3-(4-Chlorobenzoyl)indolizin-1-ylacetate (D3). Yellow solid with a yield of 60%. 1 H NMR (400 MHz, DMSO- d 6) δ9.82 (dt, J = 7.1, 1.1 Hz, 1H), 7.76 (d,J = 8.5 Hz, 2H), 7.70 (d, J = 8.8Hz, 1H), 7.61 (d, J = 8.5 Hz, 2H), 7.38 (ddd, J = 8.8, 6.8, 1.1 Hz, 1H), 7.25(s, 1H), 7.18 (td, J = 7.0, 1.4 Hz, 1H), 2.34 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 182.27, 169.39, 138.98, 136.37, 130.93, 130.71, 128.98, 128.38, 127.83,125.45, 117.83, 117.16, 116.49, 115.76, 21.03. HRMS(ESI) m / z: [M+H] + calcdfor C 17 H 12 ClNO3, 314.0579; found, 314.0566. Purity: 98.80%. Example 4: Synthesis of Compound D4 Compound D4 was prepared using a synthesis method similar to that in Example 1.

[0100] The structural characterization data of compound D4 are as follows: 3-(4-Methylbenzoyl)indolizin-1-ylacetate (D4). Yellow solid with a yield of 51%. 1 H NMR (400 MHz, DMSO- d 6) δ9.81 (d, J = 7.1 Hz, 1H), 7.70 – 7.61 (m, 3H), 7.37 – 7.29 (m, 3H), 7.24 (s,1H), 7.13 (td, J = 6.9, 1.4 Hz, 1H), 2.40 (s, 3H), 2.33 (s, 3H). 13 C NMR (101MHz, DMSO-d 6) δ 183.57, 169.37, 141.67, 137.56, 130.17, 129.40, 129.18,128.13, 127.66, 124.90, 118.09, 116.92, 116.38, 115.42, 21.52, 21.04. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO3, 294.1125; found, 294.1130. Purity:99.73%. Example 5: Synthesis of Compound D5 Compound D5 was prepared using a synthesis method similar to that in Example 1.

[0101] The structural characterization data of compound D5 are as follows: 3-(4-Ethylbenzoyl)indolizin-1-ylacetate (D5). Yellow solid with a yield of 61%. 1 H NMR (400 MHz, DMSO- d 6) δ9.82 (d, J = 7.1 Hz, 1H), 7.67 (d, J = 8.2 Hz, 3H), 7.40 – 7.29 (m, 3H), 7.25(s, 1H), 7.13 (td, J = 6.9, 1.4 Hz, 1H), 2.69 (q, J = 7.6 Hz, 2H), 2.33 (s,3H), 1.23 (t, J = 7.6 Hz, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 183.57, 169.37,147.79, 137.83, 130.18, 129.27, 128.23, 128.14, 127.67, 124.91, 118.10,116.97, 116.37, 115.42, 28.58, 21.04, 15.83. HRMS(ESI) m / z: [M+H]+ calcd forC 19 H 17 NO3, 308.1281; found, 308.1268. Purity: 99.46%. Example 6: Synthesis of Compound D6 Compound D6 was prepared using a synthesis method similar to that in Example 1.

[0102] The structural characterization data of compound D6 are as follows: 3-(4-Methoxybenzoyl)indolizin-1-yl acetate (D6). Yellow solid with a yield of 37%. 1 H NMR (400 MHz, DMSO- d 6)δ 9.77 (dd, J = 7.2, 1.1 Hz, 1H), 7.75 (d, J = 8.8 Hz, 2H), 7.67 – 7.63 (m,1H), 7.30 (ddd, J = 8.9, 6.7, 1.1 Hz, 1H), 7.25 (s, 1H), 7.13 – 7.04 (m, 3H), 3.84 (s, 3H), 2.33 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 182.82, 169.40,162.23, 132.58, 131.24, 129.91, 128.00, 127.59, 124.60, 118.11, 116.67,116.35, 115.20, 114.19, 55.90, 21.04. HRMS(ESI) m / z: [M+H] + calcd forC 18 H 15 NO4, 310.1074; found, 310.1059. Purity: 99.46%. Example 7: Synthesis of Compound D7 Compound D7 was prepared using a synthesis method similar to that in Example 1.

[0103] The structural characterization data of compound D7 are as follows: Methyl 4-(1-acetoxyindolizine-3-carbonyl)benzoate (D7). Yellow solid with a yield of54%. 1 H NMR (400 MHz, DMSO-d6) δ 9.85 (d, J = 7.1 Hz, 1H), 8.09 (d, J = 8.4Hz, 2H), 7.85 (d, J = 8.3 Hz, 2H), 7.71 (d, J = 8.8 Hz, 1H), 7.40 (ddd, J =8.8, 6.8, 1.1 Hz, 1H), 7.24 – 7.16 (m, 2H), 3.90 (s, 3H), 2.33 (s, 3H). 13 CNMR (101 MHz, DMSO- d 6) δ 182.56, 169.35, 166.23, 144.33, 131.93, 130.95,129.69, 129.28, 128.54, 127.93, 125.74, 117.88, 117.27, 116.53, 115.95,52.91, 21.01. HRMS(ESI) m / z: [M+H] + calcd for C 19 H 15 NO5, 338.1023; found, 338.0923. Purity: 97.18%. Example 8: Synthesis of Compound D8 Compound D8 was prepared using a synthesis method similar to that in Example 1.

[0104] The structural characterization data of compound D8 are as follows: 3-(4-Cyanobenzoyl)indolizin-1-ylacetate (D8). Orange solid with a yield of 29%. 1H NMR (400 MHz, DMSO-d6) δ9.84 (d, J = 7.1 Hz, 1H), 8.01 (d, J = 8.1 Hz, 2H), 7.87 (d, J = 8.1 Hz, 2H), 7.72 (dt, J = 8.9, 1.3 Hz, 1H), 7.46 – 7.37 (m, 1H), 7.25 – 7.17 (m, 2H), 2.33 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 181.80, 169.37, 144.24, 132.95,131.18, 129.67, 128.65, 127.97, 126.00, 118.89, 117.70, 117.37, 116.56,116.11, 113.69, 21.02. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 12 N2O3, 305.0921;found, 305.0926. Purity: 99.61%. Example 9: Synthesis of Compound D9 Compound D9 was prepared using a synthesis method similar to that in Example 1.

[0105] The structural characterization data of compound D9 are as follows: 3-(4-Nitrobenzoyl)indolizin-1-ylacetate (D9). Orange solid with a yield of 71%. 1 H NMR (400 MHz, DMSO- d 6) δ9.85 (d, J = 7.1 Hz, 1H), 8.38 – 8.33 (m, 2H), 7.98 – 7.93 (m, 2H), 7.73 (dt,J = 8.9, 1.3 Hz, 1H), 7.43 (ddd, J = 8.9, 6.8, 1.1 Hz, 1H), 7.25 – 7.20 (m,2H), 2.33 (s, 3H). 13 C NMR (101 MHz, DMSO- d6) δ 181.41, 169.32, 149.09,145.89, 131.30, 130.25, 128.75, 128.04, 126.13, 124.08, 117.75, 117.38,116.58, 116.19, 20.99. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 12 N2O5, 325.0819;found, 325.0787. Purity: 98.56%. Example 10: Synthesis of Compound D10 Compound D10 was prepared using a synthesis method similar to that in Example 1.

[0106] The structural characterization data of compound D10 are as follows: 3-(2-Fluorobenzoyl)indolizin-1-ylacetate (D10). Yellow solid with a yield of 21%. 1 H NMR (400 MHz, DMSO- d 6) δ9.85 (d, J = 7.1 Hz, 1H), 7.72 (dt, J = 8.8, 1.3 Hz, 1H), 7.63 – 7.54 (m,2H), 7.42 (ddd, J = 8.9, 6.8, 1.1 Hz, 1H), 7.35 (q, J = 8.8, 7.6 Hz, 2H), 7.22 (td, J = 7.0, 1.3 Hz, 1H), 7.03 (d, J = 1.6 Hz, 1H), 2.31 (s, 3H). 13 CNMR (101 MHz, DMSO- d 6) δ 179.38, 169.32, 160.26, 157.80, 132.69 (d, 3 J C-F = 8Hz), 131.13, 130.43 (d, 4J C-F = 3 Hz), 128.62 (d, 2 J C-F = 16 Hz), 127.28 (d, 1 J C-F = 254.5 Hz) , 127.77 , 125.01 (d, 4 J C-F = 3 Hz), 118.54, 117.12 (d, 4 J C-F =2 Hz), 116.60 (d, 2 J C-F = 21 Hz), 116.58, 116.19, 21.01. 19 F NMR (376 MHz, Chloroform- d ) δ -112.37. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 12 FNO3, 298.0874;found, 298.0878. Purity: 99.78%. Example 11: Synthesis of Compound D11 Compound D11 was prepared using a synthesis method similar to that in Example 1.

[0107] The structural characterization data of compound D11 are as follows: 3-(2-Chlorobenzoyl)indolizin-1-ylacetate (D11). Yellow solid with a yield of 64%. 1 H NMR (400 MHz, DMSO- d 6) δ9.85 (d, J = 7.1 Hz, 1H), 7.72 (d, J = 8.9 Hz, 1H), 7.59 (d, J= 7.8 Hz, 1H),7.57 – 7.50 (m, 2H), 7.50 – 7.38 (m, 2H), 7.23 (td, J = 6.9, 1.4 Hz, 1H), 6.85 (s, 1H), 2.29 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 181.51, 169.27,139.55, 131.53, 131.20, 130.40, 130.21, 129.60, 128.63, 127.83, 127.57,125.97, 118.17, 117.06, 116.62, 116.24, 21.01. HRMS(ESI) m / z: [M+H] + calcdfor C 17 H 12 ClNO3, 314.0579; found, 314.0562. Purity: 98.84%. Example 12: Synthesis of Compound D12 Compound D12 was prepared using a synthesis method similar to that in Example 1.

[0108] The structural characterization data of compound D12 are as follows: 3-(2-Methylbenzoyl)indolizin-1-ylacetate (D12). Yellow solid with a yield of 70%. 1 H NMR (400 MHz, Chloroform- d ) δ 10.00 (d, J = 7.2 Hz, 1H), 7.46 (dt, J = 8.9, 1.3 Hz, 1H), 7.40 – 7.31(m, 2H), 7.25 – 7.15 (m, 3H), 7.02 – 6.91 (m, 2H), 2.37 (s, 3H), 2.31 (s,3H). 13 C NMR (101 MHz, Chloroform- d) δ 186.34, 168.74, 140.14, 136.01, 130.76,130.41, 129.43, 128.29, 128.11, 128.03, 125.12, 124.29, 119.23, 116.97,115.57, 114.57, 20.81, 19.64. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO3,294.1125; found, 294.1101. Purity: 99.69%. Example 13: Synthesis of Compound D13 Compound D13 was prepared using a synthesis method similar to that in Example 1.

[0109] The structural characterization data of compound D13 are as follows: 3-(2-Methoxybenzoyl)indolizin-1-yl acetate (D13). Yellow solid with a yield of 80%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.83 (d, J = 7.1 Hz, 1H), 7.67 (d, J = 8.9 Hz, 1H), 7.52 – 7.44 (m,1H), 7.39 – 7.29 (m, 2H), 7.16 (dd, J = 7.8, 5.3 Hz, 2H), 7.04 (t, J = 7.4Hz, 1H), 6.86 (s, 1H), 3.73 (s, 3H), 2.29 (s, 3H). 13 C NMR (101 MHz, DMSO- d6)δ 183.17, 169.31, 156.54, 131.49, 130.27, 130.22, 128.98, 128.18, 127.48,125.07, 120.63, 119.07, 116.89, 116.44, 115.71, 112.34, 55.93, 21.01. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO4, 310.1074; found, 310.1074. Purity:99.64%. Example 14: Synthesis of Compound D14 Compound D14 was prepared using a synthesis method similar to that in Example 1.

[0110] The structural characterization data of compound D14 are as follows: 3-(2-Trifluoromethylbenzoyl)indolizin-1-yl acetate (D14). Yellow solid with a yield of 60%. 1 HNMR (400 MHz, Chloroform- d ) δ 9.96 (d, J = 7.1 Hz, 1H), 7.78 – 7.74 (m, 1H), 7.63 – 7.46 (m, 4H), 7.28 – 7.21 (m, 1H), 6.99 (t, J = 6.9 Hz, 1H), 6.91 (s, 1H), 2.31 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 182.14, 169.24, 138.90,138.88, 132.72, 129.51, 129.48 (q, 1 J C-F = 271 Hz), 127.03 (q, 3 J C-F = 4 Hz), 126.67 (q, 2J C-F = 30 Hz), 125.95, 125.72, 122.99, 118.15, 117.15, 116.64,116.24, 20.99. 19 F NMR (376 MHz, Chloroform- d ) δ -58.21. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 12 F3NO3, 348.0842; found, 348.0829. Purity: 98.54%. Example 15: Synthesis of Compound D15 Compound D15 was prepared using a synthesis method similar to that in Example 1.

[0111] The structural characterization data of compound D15 are as follows: 3-(2-Nitrobenzoyl)indolizin-1-ylacetate (D15). Yellow solid with a yield of 57%. 1 H NMR (400 MHz, DMSO- d 6) δ9.78 (d, J = 7.0 Hz, 1H), 8.20 (dd, J = 8.2, 1.2 Hz, 1H), 7.89 (td, J = 7.5, 1.3 Hz, 1H), 7.80 (td, J = 7.8, 1.5 Hz, 1H), 7.77 – 7.70 (m, 2H), 7.43 (ddd, J = 8.9, 6.8, 1.1 Hz, 1H), 7.23 (td, J = 6.9, 1.4 Hz, 1H), 6.98 (s, 1H), 2.30 (s, 3H). 13 C NMR (101 MHz, DMSO- d6) δ 180.18, 169.26, 147.41, 135.74, 134.60,131.50, 131.17, 130.17, 128.62, 127.78, 125.95, 125.17, 117.76, 116.69,116.45, 116.26, 20.98. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 12 N2O5, 325.0819;found, 325.0805. Purity: 99.52%. Example 16: Synthesis of Compound D16 Compound D16 was prepared using a synthesis method similar to that in Example 1.

[0112] The structural characterization data of compound D16 are as follows: 3-(3-Fluorobenzoyl)indolizin-1-ylacetate (D16). Yellow solid with a yield of 20%. 1 H NMR (400 MHz, Chloroform- d ) δ 9.94 (d, J = 7.1 Hz, 1H), 7.56 (d, J = 7.3 Hz, 1H), 7.53 – 7.47 (m, 2H), 7.47 – 7.40 (m, 1H), 7.29 (s, 1H), 7.26 – 7.19 (m, 2H), 6.98 (t, J = 6.9 Hz, 1H), 2.36 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 182.50 (d, J = 1.8 Hz), 168.54, 162.27 (d, J = 247.5 Hz), 142.29 (d, J = 6.2 Hz), 130.41, 129.71 (d, J= 7.8 Hz), 128.15, 128.06, 124.41 (d, J = 3.1 Hz), 124.32, 117.86, 117.68(d, J = 21.3 Hz), 116.54, 115.59 (d, J = 22.6 Hz), 115.39, 114.54, 20.65. 19 FNMR (376 MHz, Chloroform- d ) δ -113.99. HRMS(ESI) m / z: [M+H] + calcd forC 17 H 12 FNO3, 298.0874 found, 298.0865. Purity: 98.98%. Example 17: Synthesis of Compound D17 Compound D17 was prepared using a synthesis method similar to that in Example 1.

[0113] The structural characterization data of compound D17 are as follows: 3-(3-Chlorobenzoyl)indolizin-1-ylacetate (D17). Yellow solid with a yield of 81%. 1 H NMR (400 MHz, Chloroform- d ) δ 9.92 (d, J = 6.9 Hz, 1H), 7.75 (s, 1H), 7.65 (dd, J = 7.6, 1.4 Hz, 1H), 7.54 – 7.46 (m, 2H), 7.44 – 7.37 (m, 1H), 7.26 (s, 1H), 7.25 – 7.19 (m, 1H), 7.01 – 6.94 (m, 1H), 2.35 (s, 3H). 13 C NMR (101 MHz, DMSO- d6) δ 181.80,169.37, 142.26, 133.66, 131.31, 130.89, 130.84, 128.54, 128.48, 127.87,127.68, 125.61, 117.73, 117.23, 116.49, 115.85, 21.04. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 12 ClNO3, 314.0579; found, 314.0563. Purity: 99.60%. Example 18: Synthesis of Compound D18 Compound D18 was prepared using a synthesis method similar to that in Example 1.

[0114] The structural characterization data of compound D18 are as follows: 3-(3-Nitrobenzoyl)indolizin-1-ylacetate (D18). Yellow solid with a yield of 65%. 1 H NMR (400 MHz, Chloroform- d ) δ 9.96 (d, J = 7.1 Hz, 1H), 8.63 (s, 1H), 8.39 (ddd, J = 8.2, 2.3, 1.1 Hz,1H), 8.11 (dd, J = 7.7, 1.4 Hz, 1H), 7.68 (t, J = 7.9 Hz, 1H), 7.54 (dt, J =8.9, 1.2 Hz, 1H), 7.34 – 7.26 (m, 2H), 7.08 – 7.00 (m, 1H), 2.37 (s, 3H). 13 CNMR (101 MHz, Chloroform- d) δ 181.08, 168.64, 148.03, 141.94, 134.55, 131.14,129.51, 128.78, 128.38, 125.38, 125.16, 123.74, 117.69, 116.53, 115.75,115.20, 20.87. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 12 N2O5, 325.0819; found, 325.0819. Purity: 96.98%. Example 19: Synthesis of Compound D19 The synthetic route for compound D19 is as follows: D15 (72 mg, 0.22 mmol, 1 eq) and SnCl2·2H2O (250 mg, 1.1 mmol, 5 eq) were added to a 50 mL double-necked flask and dissolved in ethyl acetate (5 mL). The mixture was heated at 70 °C for 7 h. TLC monitoring showed that the reaction was complete. Then, 5 mL of ethyl acetate and 10% sodium bicarbonate were added until no more precipitate formed. The mixture was filtered, and the filtrate was collected. The filtrate was extracted three times with H2O / ethyl acetate. The combined organic solutions were concentrated under vacuum. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 1:1 to obtain D19 (27 mg, 0.09 mmol, 42%).

[0115] The structural characterization data of compound D19 are as follows: 3-(2-Aminobenzoyl)indolizin-1-ylacetate (D19). Yellow solid with a yield of 42%. 1H NMR (400 MHz, DMSO-d6) δ9.70 (d, J = 7.2 Hz, 1H), 7.64 (dt, J = 8.9, 1.3 Hz, 1H), 7.43 (dd, J = 7.8,1.6 Hz, 1H), 7.29 (ddd, J = 8.8, 6.7, 1.1 Hz, 1H), 7.22 (ddd, J = 8.5, 7.2,1.6 Hz, 1H), 7.12 (s, 1H), 7.09 (td, J = 7.0, 1.4 Hz, 1H), 6.79 (dd, J = 8.3,1.1 Hz, 1H), 6.64 – 6.56 (m, 1H), 6.08 (s, 2H), 2.33 (s, 3H). 13 C NMR (101MHz, Chloroform- d ) δ 185.63, 168.78, 148.28, 132.07, 131.79, 129.95, 128.05,123.60, 122.44, 119.23, 116.85, 116.74, 116.43, 115.56, 114.05, 20.81. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 14 N2O3, 295.1077; found, 295.1064. Purity:98.19%. Example 20: Synthesis of Compound D20 The synthetic route for compound D20 is as follows: D18 (129 mg, 0.39 mmol, 1 eq) and SnCl2·2H2O (450 mg, 2 mmol, 5 eq) were added to a 50 mL double-necked flask and dissolved in ethyl acetate (5 mL). The mixture was heated at 70 °C for 7 h. After the reaction was complete by TLC, 5 mL of ethyl acetate and 10% sodium bicarbonate were added until no more precipitate formed. The mixture was filtered, and the filtrate was collected. The filtrate was extracted three times with H2O / ethyl acetate, and the combined organic solutions were concentrated under vacuum. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 1:1 to obtain D19 (80 mg, 0.27 mmol, 70%).

[0116] The structural characterization data of compound D20 are as follows: 3-(3-Aminobenzoyl)indolizin-1-ylacetate (D20). Yellow solid with a yield of 70%. 1 H NMR (400 MHz, DMSO- d 6) δ9.81 (d, J = 7.1 Hz, 1H), 7.66 (dd, J = 8.9, 1.4 Hz, 1H), 7.36 – 7.29 (m,1H), 7.24 (s, 1H), 7.19 – 7.09 (m, 2H), 6.94 (d, J = 2.0 Hz, 1H), 6.87 – 6.82(m, 1H), 6.76 (dd, J = 7.7, 2.3 Hz, 1H), 5.36 (s, 2H), 2.33 (d, J = 1.4 Hz, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 184.56, 169.41, 149.22, 141.04, 130.08,129.21, 127.93, 127.66, 124.76, 118.15, 116.97, 116.95, 116.60, 116.38,115.34, 114.02, 21.04. HRMS(ESI) m / z: [M+H] + calcd for C 17 H 14 N2O3, 295.1077; found, 295.1063. Purity: 96.60%. Example 21: Synthesis of Compound D21 Compound D21 was prepared using a synthesis method similar to that in Example 1.

[0117] The structural characterization data of compound D21 are as follows: 3-(Cyclopropanecarbonyl)indolizin-1-ylacetate (D21): Yellow solid with a yield of 65%.1 H NMR (400 MHz, DMSO- d 6) δ9.74 (d, J = 7.2 Hz, 1H), 7.88 (s, 1H), 7.61 (d, J = 8.9 Hz, 1H), 7.24 (ddd, J = 8.9, 6.7, 1.1 Hz, 1H), 7.06 – 7.00 (m, 1H), 2.71 (ddd, J = 12.5, 7.8, 4.6Hz, 1H), 2.37 (s, 3H), 1.02 – 0.96 (m, 2H), 0.91 (dt, J = 8.0, 3.1 Hz, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 188.38, 169.59, 129.43, 127.88, 127.38, 124.02,118.92, 116.30, 115.05, 114.91, 21.01, 17.66, 10.08. HRMS(ESI) m / z: [M+H] + calcd for C 14 H 13 NO3,244.0968; found, 244.0973. Purity:99.56%. Example 22: Synthesis of Compound D22 Compound D22 was prepared using a synthesis method similar to that in Example 1.

[0118] The structural characterization data of compound D22 are as follows: 3-(4-(Benzyloxy)benzoyl)indolizin-1-yl acetate (D22): Yellow solid with a yield of 45%. 1 H NMR (400MHz, DMSO- d 6) δ 9.78 (d, J = 7.1 Hz, 1H), 7.75 (d, J= 8.7 Hz, 2H), 7.66 (d, J = 9.0 Hz, 1H), 7.52 – 7.46 (m, 2H), 7.42 (t, J = 7.3 Hz, 2H), 7.38 – 7.28(m, 2H), 7.26 (s, 1H), 7.16 (d, J = 8.8 Hz, 2H), 7.11 (td, J = 6.9, 1.4 Hz,1H), 5.21 (s, 2H), 2.34 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 182.76, 169.42,161.33, 137.12, 132.78, 131.24, 129.95, 128.99, 128.48, 128.30, 128.02,127.60, 124.65, 118.09, 116.69, 116.37, 115.24, 114.99, 69.93, 21.06. HRMS(ESI) m / z: [M+H] + calcd for C 24 H 19 NO4,386.1386; found, 386.1365. Purity:99.89%. Example 23: Synthesis of Compound D23 Compound D23 was prepared using a synthesis method similar to that in Example 1.

[0119] The structural characterization data of compound D23 are as follows: 6-Bromo-3-(2-methylbenzoyl)indolizin-1-yl acetate (D23): Yellow solid with a yield of 43%. 1 H NMR (400MHz, DMSO- d6) δ 10.03 (s, 1H), 7.74 – 7.68 (m, 1H), 7.54 – 7.47 (m, 1H), 7.46 – 7.37 (m, 2H), 7.36 – 7.27 (m, 2H), 6.89 (s, 1H), 2.29 (s, 3H), 2.28 (s,3H). 13 C NMR (101 MHz, DMSO- d 6) δ 186.08, 169.17, 139.74, 135.79, 131.16,130.23, 128.99, 128.70, 128.23, 127.65, 127.06, 125.80, 119.24, 117.80,117.17, 109.94, 21.02, 19.61. Purity: 99.75%. Example 24: Synthesis of Compound D24 Compound D24 was prepared using a synthesis method similar to that in Example 1.

[0120] The structural characterization data of compound D24 are as follows: 8-Methyl-3-(2-methylbenzoyl)indolizin-1-yl acetate (D24): Yellow solid with a yield of 43%. 1 H NMR (400 MHz, Chloroform- d ) δ 9.92 (d, J = 7.0 Hz, 1H), 7.42 – 7.31 (m,2H), 7.26 – 7.19 (m, 2H), 6.93 (d, J = 6.8 Hz, 1H), 6.90 (s, 1H), 6.85 (t, J = 7.0 Hz, 1H), 2.56 (s, 3H), 2.37 (s, 3H), 2.29 (s, 3H). 13 C NMR (101 MHz, Chloroform- d) δ 186.24, 169.51, 140.32, 135.97, 130.72, 130.18, 129.35,128.99, 127.99, 127.18, 126.17, 125.10, 124.88, 119.14, 117.76, 114.36,20.91, 19.63, 18.18. Purity: 99.89%. Example 25: Synthesis of Compound D25 Compound D25 was prepared using a synthesis method similar to that in Example 1.

[0121] The structural characterization data of compound D25 are as follows: 7-Methyl-3-(2-methylbenzoyl)indolizin-1-yl acetate (D25): Yellow solid with a yield of 36%. 1 H NMR (400 MHz, Chloroform- d ) δ 9.90 (d, J = 7.2 Hz, 1H), 7.37 (d, J = 7.5Hz, 1H), 7.32 (td, J = 7.5, 1.5 Hz, 1H), 7.26 – 7.17 (m, 3H), 6.91 (s, 1H), 6.80 (dd, J = 7.2, 1.8 Hz, 1H), 2.42 (s, 3H), 2.37 (s, 3H), 2.30 (s, 3H). 13 CNMR (101 MHz, Chloroform- d ) δ 185.72, 168.80, 140.29, 135.97, 135.72, 131.00,130.69, 129.27, 128.00, 127.87, 127.29, 125.07, 118.72, 117.29, 117.13,114.07, 21.44, 20.77, 19.62. Purity: 99.13%. Example 26: Synthesis of Compound D26 Compound D26 was prepared using a synthesis method similar to that in Example 1.

[0122] The structural characterization data of compound D26 are as follows: 3-(2-Methylbenzoyl)indolizin-1-ylpropionate (D26): Yellow solid with a yield of 29%. 1 H NMR (400 MHz, Chloroform- d ) δ 10.01 (d, J = 7.1 Hz, 1H), 7.46 (d, J = 8.9 Hz, 1H), 7.39 (dd, J = 7.5, 1.4 Hz, 1H), 7.34 (td, J = 7.5, 1.4 Hz, 1H), 7.27 (s, 1H), 7.22(td, J = 8.9, 8.5, 7.3 Hz, 2H), 6.98 (d, J = 8.4 Hz, 2H), 2.62 (q, J = 7.5Hz, 2H), 2.38 (s, 3H), 1.27 (t, J = 7.5 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 186.33, 172.22, 140.21, 135.97, 130.73, 130.40, 129.38, 128.28, 128.22,127.99, 125.12, 124.16, 119.24, 116.93, 115.57, 114.51, 27.46, 19.60, 9.10.Purity: 98.62%. Example 27: Synthesis of Compound D27 Compound D27 was prepared using a synthesis method similar to that in Example 1.

[0123] The structural characterization data of compound D27 are as follows: 3-(2-Methylbenzoyl)indolizin-1-yl isobutyrate (D27): Yellow solid with a yield of 39%. 1 H NMR (400 MHz, Chloroform- d ) δ 10.02 (d, J = 7.2 Hz, 1H), 7.44 (dt, J = 8.9, 1.3 Hz, 1H),7.41 – 7.37 (m, 1H), 7.34 (td, J = 7.5, 1.5 Hz, 1H), 7.26 – 7.17 (m, 3H), 6.99 (s, 1H), 6.96 (dd, J = 6.9, 1.4 Hz, 1H), 2.84 (hept, J = 7.0 Hz, 1H), 1.33 (s, 3H), 1.32 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 186.33, 174.86,140.22, 135.96, 130.73, 130.45, 129.37, 128.30, 127.98, 125.14, 124.19,119.24, 116.93, 115.52, 114.52, 34.08, 19.60, 19.03. Purity: 98.98%. Example 28: Synthesis of Compound D28 Compound D28 was prepared using a synthesis method similar to that in Example 1.

[0124] The structural characterization data of compound D28 are as follows: 3-(2-methylbenzoyl)indolizin-1-yl benzoate (D28): Yellow solid with a yield of 51%. 1 H NMR (400 MHz, DMSO- d 6) δ 9.93 (d, J= 7.2 Hz, 1H), 8.21 – 8.15 (m, 2H), 7.83 (d, J = 8.9 Hz,1H), 7.78 – 7.73 (m, 1H), 7.61 (t, J = 7.7 Hz, 2H), 7.45 – 7.38 (m, 3H), 7.37– 7.28 (m, 2H), 7.22 (td, J = 7.0, 1.4 Hz, 1H), 7.09 (s, 1H), 2.30 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 185.62, 164.30, 140.35, 135.62, 134.63, 131.06,130.53, 130.39, 129.95, 129.46, 128.92, 128.30, 128.13, 127.77, 125.78,125.53, 118.88, 116.87, 116.63, 115.95, 19.62. Purity: 99.81%. Example 29: Synthesis of Compound D29 Compound D29 was prepared using a synthesis method similar to that in Example 1.

[0125] The structural characterization data of compound D29 are as follows: 3-(2-trifluoromethylbenzoyl)indolizin-1-yl propionate (D29): Yellow solid with a yield of 8%. 1 HNMR (400 MHz, Chloroform- d ) δ 9.97 (d, J = 7.1 Hz, 1H), 7.76 (d, J = 6.8 Hz, 1H), 7.60 (dt, J = 9.2, 6.5 Hz, 2H), 7.55 – 7.46 (m, 2H), 7.28 – 7.21 (m,1H), 6.99 (td, J= 7.0, 1.4 Hz, 1H), 6.93 (s, 1H), 2.61 (q, J = 7.5 Hz, 2H), 1.26 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 182.64, 172.02, 139.04(d, 3 J C-F = 3 Hz), 131.30, 130.88, 129.40, 129.02, 128.56, 128.39, 128.05(d, 2 J C-F = 32Hz), 126.63(d, 3 J C-F = 5 Hz), 124.74, 123.81 (d, 1 J C-F = 272 Hz), 118.46, 116.63,115.68, 114.93, 27.45, 9.04. 19 F NMR (376 MHz, Chloroform- d δ -58.21. Purity: 99.88%. The following are comparative compounds and their synthetic methods. Comparative Example 1: Synthesis of compounds A1-A3 α-Halocarbonyl compound 1 (196 mg, 1 mmol, 1.0 equiv) was heated with pyridine (84 μl, 1 mmol, 1.0 equiv) in DMF at 60 °C for 2 h. Then, ethyl acrylate (163 μl, 1.5 mmol, 1.5 equiv), potassium dichromate (220 mg, 0.75 mmol, 0.75 equiv), and DMF were added to the reaction mixture. The mixture was then heated at 80 °C for 8 h, and the reaction was monitored by TLC. The mixture was cooled to room temperature, filtered through diatomaceous earth, washed with ethyl acetate, and the solvent was removed by concentration under reduced pressure. The residue was extracted with dichloromethane and brine to obtain the organic layer. The organic layer was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 10:1) to give a yellow solid A1 (161 mg, 55% yield). The synthesis of compound A2 followed the same procedure as A1.

[0126] Characterization data for A1: 1 H NMR (400 MHz, Chloroform- d ) δ 9.98 (d, J = 7.1 Hz, 1H), 8.40 (d, J = 9.0 Hz, 1H), 7.85 – 7.78 (m, 3H), 7.58 (t, J = 7.3 Hz, 1H), 7.51 (t, J = 7.3 Hz, 2H), 7.48 – 7.42 (m, 1H), 7.13 – 7.04 (m, 1H), 4.38 (q, J = 7.1 Hz, 2H), 1.40 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ185.63, 164.13, 139.91, 131.52, 129.24, 129.10, 129.00, 128.44, 127.77,122.52, 119.52, 115.35, 106.29, 60.17, 29.73, 14.58. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO3, 294.1128; found, 294.1116. Purity: 99.71%. Characterization data for A2: 1 H NMR (400 MHz, Chloroform- d ) δ 10.07 (d, J = 7.0 Hz, 1H), 8.37 (d, J = 8.9 Hz, 1H), 7.58 (s, 1H), 7.51 – 7.42 (m, 2H), 7.38 (dd, J = 7.5, 1.7 Hz, 1H), 7.14 – 7.07 (m, 1H), 7.07 – 6.99 (m, 2H), 4.35 (q, J =7.1 Hz, 2H), 3.81 (s, 3H), 1.37 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 185.03, 164.15, 156.86, 139.89, 131.30, 129.61, 129.46,129.23, 127.86, 123.52, 120.25, 119.49, 115.41, 111.49, 106.34, 60.12, 55.73,14.58. HRMS(ESI) m / z: [M+H] + calcd for C 19 H 17 NO4, 324.1230; found, 324.1219.Purity: 99.87%. A1 (60 mg, 0.2 mmol, 1.0 equiv) was dissolved in THF / MeOH 2:1 (v / v). 2NNaOH solution (5.0 equiv) was added to the reactant solution at 0°C, and the mixture was heated to 60°C and stirred for 16 h, monitored by TLC. After the reaction was complete, the mixture was concentrated under reduced pressure to remove most of the THF / MeOH, and the remaining solvent was diluted with water. The solution was acidified to pH 3-4 with 1M HCl to produce a solid precipitate, which was filtered and dried to give a white solid A3 (15 mg, 30% yield).

[0127] Characterization data for A3: 1 H NMR (400 MHz, DMSO- d 6) δ 12.62 (s, 1H), 9.86 (d, J=7.0 Hz, 1H), 8.34 (d, J = 8.9 Hz, 1H), 7.78 (d, J = 7.4 Hz, 2H), 7.69 – 7.48(m, 5H), 7.31 (t, J = 7.0 Hz, 1H). Comparative Example 2: Synthesis of Compounds A4 and B1-B3 Chromone (438 mg, 3 mmol, 1.0 equiv), 3 (913 μl, 6 mmol, 2.0 equiv), 1,4-dichlorobenzene (17 mL), elemental iodine (53 mg, 0.42 mmol, 7 mol%), and di-tert-butyl peroxide (DTBP, 1.1 mL, 6 mmol, 2.0 equiv) were stirred at 100 °C for 14 h. After the reaction was complete, the resulting solution was extracted with dichloromethane and brine. The organic layer was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 5:1) to give a yellow solid I-1 (690 mg, 74% yield).

[0128] I-1 (604 mg, 1.5 mmol, 1.0 equiv) was dissolved in THF / MeOH 2:1 (v / v). 4N NaOH solution (5.0 equiv) was added to the reactant solution at 0°C, and the mixture was heated to 60°C and stirred for 4 h, monitored by TLC. After the reaction was complete, the mixture was concentrated under reduced pressure to remove most of the THF / MeOH, and the remaining solvent was diluted with water. The solution was acidified to pH 3-4 with 1M HCl to produce a solid precipitate, which was filtered and dried to give a yellow solid A4 (320 mg, 74% yield).

[0129] Under a nitrogen atmosphere, DCM (dry) and DIPEA (105 μl, 0.6 mmol, 3.0 equiv) were sequentially added to a mixture containing A4 (56 mg, 0.2 mmol, 1.0 equiv) and HATU (114 mg, 0.3 mmol, 1.5 equiv). The mixture was stirred at room temperature for 1 h. Ethanolamine (13 μl, 0.22 mmol, 1.1 equiv) was then added to the reaction mixture, and the reaction was continued at room temperature for 4 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel column chromatography (DCM:MeOH = 10:1) to give a yellow solid B1 (36 mg, yield 58%). Compounds B2-B3 were prepared by the same synthetic method as B1.

[0130] Characterization data for I-1:1 H NMR (400 MHz, Chloroform- d ) δ 11.32 (s, 1H), 9.68(d, J = 7.1 Hz, 1H), 8.40 (d, J = 8.9 Hz, 1H), 7.94 (s, 1H), 7.86 (dd, J =7.9, 1.7 Hz, 1H), 7.53 – 7.41 (m, 2H), 7.12 – 7.03 (m, 2H), 6.99 (t, J = 7.6Hz, 1H), 4.40 (q, J = 7.1 Hz, 2H), 1.42 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz,Chloroform- d ) δ 187.11, 163.98, 161.71, 140.14, 134.85, 131.59, 129.24,128.96, 127.87, 121.76, 120.82, 119.71, 119.06, 118.19, 115.33, 106.82,60.27, 14.58. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO4, 310.1074; found,310.1059. Purity: 99.90%. Characterization data of A-4: 1 H NMR (400 MHz, DMSO-d6) δ 9.87 (d, J = 6.9 Hz, 1H),8.33 (d, J = 9.1 Hz, 1H), 7.60 (t, J = 7.9 Hz, 1H), 7.43 (s, 1H), 7.37 (d, J= 7.3 Hz, 2H), 7.29 (t, J = 7.1 Hz, 1H), 6.99 (d, J = 8.3 Hz, 1H), 6.93 (t, J= 7.6 Hz, 1H). 13 C NMR (101 MHz, DMSO- d6) δ 185.20, 165.25, 155.61, 139.39,132.09, 129.67, 128.98, 128.90, 128.47, 127.56, 122.95, 119.58, 116.97,116.31, 107.42. HRMS(ESI) m / z: [M+H] + calcd for C 16 H 11 NO4, 282.0761; found, 282.0766. Purity: 99.92%. Characterization data for I-1: 1 H NMR (400 MHz, Chloroform- d ) δ 11.32 (s, 1H), 9.68 (d, J = 7.1 Hz, 1H), 8.40 (d, J = 8.9 Hz, 1H), 7.94 (s, 1H), 7.86 (dd, J =7.9, 1.7 Hz, 1H), 7.53 – 7.41 (m, 2H), 7.12 – 7.03 (m, 2H), 6.99 (t, J = 7.6Hz, 1H), 4.40 (q, J = 7.1 Hz, 2H), 1.42 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 187.11, 163.98, 161.71, 140.14, 134.85, 131.59, 129.24,128.96, 127.87, 121.76, 120.82, 119.71, 119.06, 118.19, 115.33, 106.82,60.27, 14.58. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO4, 310.1074; found, 310.1059. Purity: 99.90%. Characterization data for B-1: 11H NMR (400 MHz, DMSO-d6) δ 10.00 (s, 1H), 9.85 (d, J =7.0 Hz, 1H), 8.53 (d, J = 8.9 Hz, 1H), 8.31 (t, J = 5.6 Hz, 1H), 7.86 (s,1H), 7.55 – 7.47 (m, 1H), 7.39 (d, J = 7.6 Hz, 2H), 7.23 (t, J = 6.9 Hz, 1H),6.99 (d, J = 8.1 Hz, 1H), 6.94 (t, J = 7.4 Hz, 1H), 4.71 (t, J = 5.5 Hz, 1H),3.46 (q, J = 6.0 Hz, 2H), 3.27 (q, J = 6.0 Hz, 2H). 13 13C NMR (101 MHz, DMSO-d6)δ 185.18, 163.86, 155.96, 139.25, 131.98, 129.90, 128.48, 127.52, 127.49,125.95, 122.43, 120.13, 119.24, 117.01, 115.98, 109.95, 60.45, 41.93. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 16 N2O4, 325.1183; found, 325.1185. Purity:99.63%. Characterization data of B-2: 1 1H NMR (400 MHz, DMSO- d 6) δ 10.14 (s, 1H), 9.83 (d, J J =7.0 Hz, 1H), 7.91 (d, J J = 8.9 Hz, 1H), 7.54 – 7.45 (m, 1H), 7.42 – 7.34 (m,2H), 7.25 (d, J J = 7.7 Hz, 2H), 7.02 (d, J J = 8.2 Hz, 1H), 6.92 (t, J J = 7.4 Hz,1H), 2.31 (s, 3H), 1.36 – 1.14 (m, 8H). 13C NMR (101 MHz, DMSO- d 6) δ 184.77,164.51, 155.87, 138.39, 132.24, 130.02, 128.39, 127.37, 127.16, 126.57,122.11, 119.31, 119.25, 117.06, 115.99. HRMS(ESI) m / z: [M+H] + calcd forC 21 H 21 N3O3, 364.1656; found, 364.1663. Purity: 95.91%. Characterization data for B-3: 1 H NMR (400 MHz, DMSO-d6) δ 9.97 (s, 1H), 9.86 (d, J =7.0 Hz, 1H), 8.85 (t, J = 6.1 Hz, 1H), 8.56 (d, J = 9.0 Hz, 1H), 7.92 (s,1H), 7.53 (t, J = 7.9 Hz, 1H), 7.42 – 7.34 (m, 2H), 7.26 (m, J = 24.6 Hz, 6H), 7.00 – 6.91 (m, 2H), 4.43 (d, J = 5.9 Hz, 2H). 13 C NMR (101 MHz, DMSO- d 6)δ 185.20, 163.68, 155.95, 140.54, 139.36, 132.01, 129.94, 128.72, 128.54,127.76, 127.65, 127.48, 127.12, 125.92, 122.54, 120.14, 119.25, 117.00,116.07, 109.64, 42.35. HRMS(ESI) m / z: [M+H] + calcd for C 23 H 18 N2O3, 371.1390; found, 371.1372. Purity: 99.17%. Comparative Example 3: Synthesis of Compound C1 A mixture of 4 (388 μl, 3 mmol, 1.0 equiv) and an equal volume of iodine (755 mg, 3 mmol, 1.0 equiv) was refluxed in pyridine at 140 °C for 5 h. The mixture was cooled to room temperature, and a precipitate was formed. The precipitate was washed with petroleum ether and dried to give a brown solid 5 (550 mg, 51% yield). 1,4-Dioxane was added to 5 (540 mg, 1.5 mmol, 1.1 equiv) and chromone (240 mg, 1.4 mmol, 1.0 equiv), and DBU (360 μl, 2.8 mmol, 2.0 equiv) was added to the mixture. The mixture was stirred at 80 °C for 8 h, and the reaction was monitored by TLC. After completion, the resulting solution was extracted with EA and brine. The organic layer was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 5:1) to give a yellow solid C1 (82 mg, 18% yield).

[0131] Characterization data for C-1: 1 H NMR (400 MHz, Chloroform-d) δ 11.93 (s, 1H), 9.99 (d, J = 7.0 Hz, 1H), 8.50 (d, J = 8.9 Hz, 1H), 7.80 – 7.72 (m, 3H), 7.68 (s,1H), 7.58 (t, J = 7.9 Hz, 1H), 7.47 (dd, J = 10.4, 7.7 Hz, 3H), 7.22 (t, J =7.0 Hz, 1H), 7.06 (d, J = 8.4 Hz, 1H), 6.93 – 6.87 (m, 1H). 13 C NMR (101 MHz, Chloroform- d ) δ 193.17, 184.28, 162.34, 140.96, 138.16, 137.96, 135.20,131.51, 130.39, 129.79, 129.24, 129.14, 128.86, 122.43, 120.74, 120.20,118.85, 118.44, 116.57, 113.24. HRMS(ESI) m / z: [M+H] + calcd for C 22 H 14 ClNO3,376.0735; found, 376.0734. Purity: 99.86%. Comparative Example 4: Synthesis of Compound E1 An emulsion of benzaldehyde (205 μl, 2 mmol, 1.0 equiv) was prepared using 10 mL of cold pure water. An equimolar amount of 9 (225 μl, 2 mmol, 1.0 equiv) was added under vigorous stirring, followed by the dropwise addition of 10% KOH aqueous solution until the solution changed color. The emulsion was stirred at 4°C for 4 h, filtered, washed with water, and dried to obtain crude product 10, which was directly used for sampling. Acetic anhydride (1.5 mL) and 1,2-dichloroethane (10 mL) were added to 10, and the mixture was stirred at 110°C for 22 h, then cooled to room temperature. After completion, the resulting solution was extracted with petroleum ether and brine. The organic layer was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 10:1) to obtain a green oily substance E1 (36 mg, yield 19%).

[0132] Characterization data for E-1: 1 H NMR (400 MHz, Chloroform-d) δ 8.08 (d, J = 7.2 Hz, 1H), 7.46 (dd, J = 8.2, 1.3 Hz, 2H), 7.36 (t, J = 7.7 Hz, 2H), 7.24 (t, J =7.4 Hz, 1H), 7.21 – 7.14 (m, 1H), 6.73 (s, 1H), 6.54 (ddd, J = 9.1, 6.4, 1.0Hz, 1H), 6.36 – 6.31 (m, 1H), 2.28 (s, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ169.51, 131.87, 129.01, 128.20, 127.35, 127.27, 123.10, 122.08, 121.68,116.59, 116.35, 110.98, 106.65, 21.00. HRMS(ESI) m / z: [M+H] + calcd forC 16 H 13 NO2, 252.1019; found, 252.1013. Purity: 98.69%. Comparative Example 5: Synthesis of compounds F1-F3 Pyridine (241 μl, 3 mmol, 1.0 equiv) and 11 (363 μl, 3.3 mmol, 1.1 equiv) were heated in ethyl acetate at 50 °C for 12 h, filtered, and washed with EA to obtain salt 12 (600 mg, 82% yield). 1,4-Dioxane was added to 12 (438 mg, 3 mmol, 1.0 equiv) and chromone (808 mg, 3.3 mmol, 1.1 equiv), and DBU (896 μl, 6 mmol, 2.0 equiv) was added to the mixture, which was stirred at 80 °C for 8 h. After the reaction was complete, the resulting solution was extracted with EA and brine. The organic layer was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 10:1) to give a yellow solid F1 (461 mg, 50% yield).

[0133] F1 (155 mg, 0.5 mmol, 1.0 equiv) was dissolved in methanol, and CeCl3 (270 mg, 1.1 mmol, 2.2 equiv) and sodium borohydride (30 mg, 0.8 mmol, 1.6 equiv) were rapidly added under ice bath conditions, and the mixture was stirred at room temperature for 22 h. The reaction was monitored by TLC. After the reaction was complete, a saturated aqueous solution of ammonium chloride was added, and the solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate and washed successively with a saturated aqueous solution of ammonium chloride and saturated brine. The organic layer was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 3:1) to give a purple solid F2 (11 mg, 7% yield). The synthesis of compound F3 followed the same procedure as A3.

[0134] Characterization data for F-1: 1 H NMR (400 MHz, Chloroform- d ) δ 12.03 (s, 1H), 9.60 (d, J = 7.0 Hz, 1H), 8.47 (d, J = 8.9 Hz, 1H), 7.90 (d, J = 9.1 Hz, 2H), 7.53– 7.46 (m, 1H), 7.46 – 7.38 (m, 1H), 7.14 – 7.04 (m, 2H), 6.98 (t, J = 7.6Hz, 1H), 4.41 (q, J = 7.1 Hz, 2H), 1.42 (t, J= 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform- d ) δ 193.10, 162.23, 161.16, 140.20, 134.86, 131.86, 128.09,127.12, 125.63, 120.95, 120.34, 118.76, 118.24, 115.61, 115.19, 112.13,60.60, 14.54. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 15 NO4, 310.1074; found, 310.1059. Purity: 98.99%. Characterization data for F-2: 1 H NMR (400 MHz, Chloroform-d) δ 9.40 (d, J = 7.2 Hz,1H), 7.48 (d, J = 8.9 Hz, 1H), 7.35 (s, 1H), 7.19 – 7.11 (m, 2H), 7.00 – 6.95(m, 1H), 6.89 (td, J = 7.5, 1.2 Hz, 1H), 6.82 – 6.76 (m, 2H), 4.83 (s, 1H), 4.34 (q, J = 7.1 Hz, 2H), 4.10 (s, 2H), 1.37 (t, J = 7.1 Hz, 3H). 13 C NMR (101MHz, Chloroform- d ) δ 160.28, 152.76, 129.36, 126.74, 126.37, 125.58, 120.42,120.13, 119.92, 116.08, 114.74, 111.76, 111.00, 58.70, 25.68, 13.57. HRMS(ESI) m / z: [M+H] + calcd for C 18 H 17 NO4, 312.1230; found, 312.1236. Purity:98.76%. Characterization data for the F-3: 1 H NMR (400 MHz, DMSO- d6) δ 12.99 (s, 1H), 10.41 (s, 1H), 9.56 (d, J = 7.1 Hz, 1H), 8.43 (d, J = 8.9 Hz, 1H), 7.56 (s, 2H), 7.51 –7.31 (m, 2H), 7.27 (s, 1H), 7.09 – 6.81 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ190.84, 162.41, 156.81, 138.95, 132.69, 130.05, 128.43, 128.17, 126.86,126.06, 120.00, 119.43, 117.21, 116.32, 115.86, 113.06. HRMS(ESI) m / z: [M+H] + calcd for C 16 H 11 NO4, 282.0761; found, 282.0759. Purity: 99.82%. The following biological test examples demonstrate the beneficial effects of the present invention.

[0135] Example 1: Study on the activity of compound D1 of the present invention and comparative compounds A1-A4, B1-B3, C1, D1, E1 and F1-F3 against ferroptosis. 1. Experimental Methods The ferroptosis screening model primarily employs the MTT assay for cell viability. First, PC-12 (highly differentiated) rat pheochromocytoma cells are cultured in dishes. Cells in logarithmic growth phase are seeded at 5000 cells per well (100 μL per well) into 96-well plates and incubated at 37°C with 5% CO2 to allow cell adhesion. After 24 h, 100 μL of a serially diluted mixture of the compound and the ferroptosis inducer RSL3 (final concentration 1 μM) prepared in complete culture medium is added. Four replicates are used for each compound to ensure accuracy. A positive control group (treated with 1 μM RSL3), a negative control group (treated with 500 nM fer-1 and 1 μM RSL3), and a blank control group are also included. After drug addition, the plates are incubated for 24 h. On the day of the MTT assay, prepare the MTT test solution (5 mg / ml MTT solution dissolved in PBS, stored at 4°C in the dark) in advance. Add 20 μL of MTT solution to each well and incubate for 4 h. Then, add 150 μL of DMSO to each well in the dark, mix well, and measure the absorbance at 490 nm using a microplate reader to calculate the inhibitory rate of ferroptosis. After obtaining the absorbance values ​​(denoted as A), calculate the average value of the replicates and calculate the cell viability using the following formula: Cell viability % = (A 实验组 / A 空白对照组 )*100%.

[0136] A survival rate histogram was fitted using GraphPad Prism software.

[0137] 2. Experimental Results The results are as follows Figure 1 As shown.

[0138] 3. Results analysis, by Figure 1 We used Fer-1 and DFP as positive controls to test the protective effect of each compound against ferroptosis when combined with RSL3 at two concentrations: 6.25 μM and 12.5 μM. The results showed that at 12.5 μM, only compounds D1 and F2 exhibited superior activity compared to 100 μM DFP (Figure A). In the RSL3 (1 μM)-induced PC12 cell ferroptosis model, the half-maximal effective concentrations (EC50) of D1 and F2 were 9.07 μM and 13.73 μM, respectively (Figures B and C), indicating moderate inhibitory activity against ferroptosis.

[0139] Example 2 of biological testing: Cytotoxicity study of compound D1 of the present invention and comparative compound F2. 1. Experimental Methods PC12 cells were planted at 1 × 10⁶ cells per well. 4 Cells were seeded at a density of [number] cells per well in culture plates. After 24 h of routine culture, the target compound was added at a final concentration of 3–100 μM for intervention treatment, and incubated for 48 h. Cell viability was assessed using the MTT assay to evaluate the in vitro biosafety of the compound within this concentration range for PC12 cells. After obtaining absorbance values ​​(denoted as A), the average value of replicates was calculated, and cell viability was calculated using the following formula: Cell viability % = (A 实验组 / A 空白对照组 )*100%.

[0140] A survival rate histogram was fitted using GraphPad Prism software.

[0141] 2. Experimental Results The results are as follows Figure 2 As shown 3. Results analysis, by Figure 2 The cytotoxicity of D1 and F2 after 48 hours of exposure was assessed using the MTT assay. The results showed that, within the test concentration range of 3-100 μM, compound D1 prepared in this invention had no significant cytotoxicity to PC12 cells, and its cytotoxicity was significantly lower than that of compound F2.

[0142] Biological Testing Example 3: Study on the activity of the compound of the present invention against ferroptosis 1. Experimental Methods The ferroptosis screening model primarily employs the MTT assay for cell viability. First, PC-12 (highly differentiated) rat pheochromocytoma cells are cultured in dishes. Cells in logarithmic growth phase are seeded at 5000 cells per well (100 μL per well) into 96-well plates and incubated at 37°C with 5% CO2 to allow cell adhesion. After 24 h, 100 μL of a serially diluted mixture of the compound and the ferroptosis inducer RSL3 (final concentration 1 μM) prepared in complete culture medium is added. Four replicates are used for each compound to ensure accuracy. A positive control group (treated with 1 μM RSL3), a negative control group (treated with 500 nM fer-1 and 1 μM RSL3), and a blank control group are also included. After drug addition, the plates are incubated for 24 h. On the day of the MTT assay, prepare the MTT test solution (5 mg / ml MTT solution dissolved in PBS, stored at 4°C in the dark) in advance. Add 20 μL of MTT solution to each well and incubate for 4 h. Then, add 150 μL of DMSO to each well in the dark, mix well, and measure the absorbance at 490 nm using a microplate reader to calculate the inhibitory rate of ferroptosis. After obtaining the absorbance values ​​(denoted as A), calculate the average value of the replicates and calculate the cell viability using the following formula: Cell viability % = (A 实验组 / A 空白对照组 )*100%.

[0143] A survival rate histogram was fitted using GraphPad Prism software.

[0144] 2. Experimental Results The results of tests performed on the series of compounds of the present invention are as follows: Figure 3 As shown.

[0145] 3. Results analysis, by Figure 1 It is evident that the compounds prepared in this invention all exhibit good inhibitory activity against ferroptosis. Among them, compound D21 shows slightly lower inhibitory activity; compounds D1, D2, D5-8, D13, D16, D19, and D20 all show lower activity than 100 μM DFP at low concentrations (6.25 μM); compounds D3, D4, D9-12, D14, D15, D17, D18, and D22 demonstrate superior antiferroptosis activity compared to 100 μM DFP at both administered concentrations, showing potential as good antiferroptosis inhibitors. Compounds D23-D29, structurally modified based on compounds D12 and D14, all show good antiferroptosis activity, except for compound D28, which shows weak activity. Compounds D24 and D25 show better activity than D12 at 50 nM.

[0146] Example 4 of biological testing: Further study on the activity against ferroptosis. 1. Experimental Methods Similar to Experiment 1, the ferroptosis model was used, but cell viability was tested using a series of concentration gradients with lower concentration ranges. The survival rate variation curve was fitted using GraphPad Prism 5 software, and the EC was calculated. 50 .

[0147] 2. Experimental Results The results of tests performed on compounds with activity better than 100 μM DFP are shown in Table 1. Table 1. Results of in vitro bioactivity tests of compounds D1-D22 3. Results Analysis: As shown in Table 1, compounds D10, D15, D12, D17, D18, D3, D14, and D22 are the best-performing ECs with nanomolar activity among the substitutions at each site on the benzene ring. 50 The compounds, in which D12, D14, D15, and D22 are all less than 100 nM.

[0148] Example 5 of biological testing: Study on the safety of compounds to cells 1. Experimental Methods Cell safety was primarily evaluated using the MTT assay. First, PC-12 (highly differentiated) rat pheochromocytoma cells and 293T human embryonic kidney cells were cultured in dishes. Cells in logarithmic growth phase were seeded at 10,000 cells per well (100 μL per well) into 96-well plates and incubated at 37°C with 5% CO2 to allow cell adhesion. After 24 h, 100 μL of the target concentration of the compound prepared in complete culture medium was added. Four replicates were performed for each compound to ensure accuracy. A solvent control group (equal volume of DMSO), an experimental group, and a blank control group were also included. After drug addition, the plates were incubated for 24 h. On the day of the MTT assay, prepare the MTT test solution in advance (5 mg / mL MTT solution dissolved in PBS, stored at 4°C in the dark). Add 20 μL of MTT solution to each well and incubate for 4 h. Then, add 150 μL of DMSO to each well in the dark, mix well, and measure the absorbance at 490 nm using a microplate reader to calculate cell viability. After obtaining the absorbance values ​​(denoted as A), calculate the average of the replicates and use the following formula to calculate cell viability: Cell viability % = [(A) 实验组 - A溶剂对照组 ) / (A 空白对照组 - A 溶剂对照组 )*100%.

[0149] A survival rate histogram was fitted using GraphPad Prism 5 software.

[0150] 2. Experimental Results The preferred compounds were subjected to toxicity tests, and the results are as follows: Figure 4 As shown.

[0151] 3. Results analysis, by Figure 4 It can be seen that all compounds have excellent cell safety.

[0152] Biological Testing Example 6: Study on the Effects of the Compounds of the Invention on Cell Morphology 1. Experimental Methods First, PC-12 (highly differentiated) rat adrenal pheochromocytoma cells were cultured in a dish. Cells in the logarithmic growth phase were seeded into the culture dish at a density of approximately 15 × 10⁶ cells / mL. 5 Cells were placed in a dish with complete culture medium and incubated at 37°C with 5% CO2 for 24 h to allow cell adhesion and 70%-80% confluence. After 24 h, a mixture of compound D1 (final concentration 12.5 μM) and ferroptosis inducer RSL3 (final concentration 1 μM) was added. A negative control group (treated with 1 μM RSL3) and a blank control group were also set up. After adding the drugs, the cells were placed in an incubator and incubated for 8 h. The culture medium was discarded, and the cells were gently washed twice with pre-cooled PBS. The cells were then digested with 0.25% trypsin for 1 min, and the digestion was immediately stopped by adding complete culture medium. The cells were centrifuged at low speed (700 r, 4 min), the supernatant was discarded, and electron microscopy fixative (the fixative should be brought to room temperature beforehand) was added. The cell clumps were dispersed and resuspended. The cells were fixed at room temperature in the dark for 30 min, and then transferred to 4°C for storage and delivery.

[0153] 2. Experimental Results Tests performed on cells treated with compound D1 yielded the following results: Figure 5 As shown.

[0154] 3. Results analysis, by Figure 3 It can be seen that after treatment with the ferroptosis inducer RSL3, the mitochondria of cells atrophied and the cristae were missing. When RSL3 and D1 were co-treated, the mitochondrial morphology of the cells was significantly improved. The Flameng score also showed that the mitochondrial damage of the cells was enhanced after treatment with the ferroptosis inducer RSL3. When RSL3 and D1 were co-treated, the mitochondrial damage of the cells was significantly improved.

[0155] Biological Testing Example 7: Study on the chelating iron and free radical scavenging effects of the compounds of the present invention This experiment uses compounds D1, D12, D14, and D15 as examples to study the chelating iron and free radical scavenging effects of the compounds of this invention.

[0156] 1. Experimental Methods The chelating properties of the compounds were studied using ultraviolet absorption spectroscopy. Fe was prepared in methanol to a final concentration of 100 μM. 2+ Solution (FeSO4·7H2O), Fe 3+ Solution (FeCl3·6H2O), DFP solution, and solution of the compound to be tested. A solvent blank control must be performed before the experiment to verify that the solvent has no absorption or negligible absorption at 510 nm. The UV absorption of 100 μM compound or DFP in the wavelength range of 200 nm to 600 nm is measured using a UV spectrophotometer, with or without 100 μM Fe(II) or Fe(III) as the solvent.

[0157] The reducing properties were studied using the 2,2-diphenyl-1-pyridylhydrazine (DPPH) method. Stable free radical 1,1-diphenyl-2-pyridylhydrazine (DPPH) was dissolved in methanol to a concentration of 100 μM, and 100 μL was added to each well of a 96-well plate. Test compounds (D1, D12, D14, D15) were dissolved in DMSO to prepare serially diluted solutions, and 100 μL was added to each well, resulting in final concentrations of 5, 2.5, 1.25, and 0.625 mM. After thorough mixing, the solutions were incubated at room temperature for 30 minutes, and the absorbance at 517 nm was measured using a multi-microplate reader. The experiment was repeated three times, and the mean and standard deviation of the results were calculated.

[0158] 2. Experimental Results Use GraphPad Prism software to generate a co-op graph.

[0159] 3. Results analysis, results ( Figure 6 , 7 The results show that the compounds (D1, D12, D14, D15) prepared in this invention do not inhibit ferroptosis through free radical capture and iron chelation mechanisms.

[0160] Biological Test Example 8: Effect of the Compounds of the Present Invention on Intracellular Lipid Peroxidation Levels This experiment uses compounds D12, D14, and D15 as examples to study the potential intracellular mechanisms of the compounds of this invention.

[0161] 1. Experimental Methods Classical fluorescence imaging was used to investigate the reduction of lipid peroxidation levels by compounds. Before the experiment, the C11 BODIPY 581 / 591 probe was diluted 1:1000 with serum-free medium, prepared fresh and used immediately, and kept in the dark throughout the process. Adherent cells in the logarithmic growth phase were digested with trypsin and resuspended in serum-containing medium. 100 μL of cell suspension was added to each well of a 96-well plate to a cell concentration of 5 × 10⁶ cells / well. 3 Cells / wells were incubated at 37°C, 5% CO2 for 24 hours to allow cell adhesion and confluence to reach 70%-80%. A mixed solution of compounds Fer-1, D12, D14, D15 (final concentration 1 μM) and ferroptosis inducer RSL3 (final concentration 1 μM) was added. A negative control group (treated with 1 μM RSL3) and a blank control group were also set up. After adding the drugs, the cells were incubated for 8 hours. The old culture medium in the 96-well plate was discarded, and the cells were gently washed twice with PBS. 100 μL of pre-prepared C11 BODIPY 581 / 591 working solution was added to each well (ensuring the liquid completely covers the cells), and the cells were incubated at 37°C, 5% CO2 in the dark for 30 minutes. After incubation, discard the C11 BODIPY 581 / 591 working solution, wash cells three times with PBS, and finally add 100 μl of PBS to each well. Place the wells on an ImageXpressMicro 4 high-content imaging analysis system. For unoxidized probes (red fluorescence): excitation wavelength 581 nm, emission wavelength 591 nm (TRITC / Rhodamine channel); for oxidized probes (green fluorescence): excitation wavelength 488 nm, emission wavelength 510 nm (FITC channel). Observe the intracellular fluorescence distribution.

[0162] 2. Experimental Results Results analysis, results ( Figure 8 The results show that the compounds (D12, D14, D15) prepared in this invention can effectively restore the level of intracellular lipid peroxidation and all have good anti-ferroptosis potential.

[0163] Biological Testing Example 9: Effects of the Compounds of the Present Invention on Intracellular ROS This experiment uses compounds D12, D14, and D15 as examples to study the effects of the compounds of this invention on intracellular ROS.

[0164] 1. Experimental Methods Classical fluorescence imaging was used to investigate the reduction of intracellular ROS and lipid peroxidation levels by compounds. DCFH-DA powder was dissolved in sterile DMSO to prepare a 10 mM stock solution. Before the experiment, the stock solution was diluted to a final concentration of 10 μM with serum-free medium. The solution was prepared fresh and used immediately, and the entire process was conducted in the dark. Adherent cells in the logarithmic growth phase were digested with 0.25% trypsin and resuspended in serum-containing medium. 100 μL of cell suspension was added to each well of a 96-well plate to bring the cell concentration to 5 × 10⁶ cells / well. 3 Cells / wells were incubated at 37°C, 5% CO2 for 24 hours to allow cell adhesion and confluence to reach 70%-80%. A mixed solution of compounds Fer-1, D12, D14, D15 (final concentration 1 μM) and ferroptosis inducer RSL3 (final concentration 1 μM) was added. A negative control group (treated with 1 μM RSL3) and a blank control group were also set up. After drug addition, the cells were incubated for 8 hours. The old culture medium in the 96-well plate was discarded, and the cells were gently washed twice with PBS. 100 μL of pre-prepared DCFH-DA working solution was added to each well (ensuring complete cell coverage), and the cells were incubated at 37°C, 5% CO2 in the dark for 30 minutes. After incubation, the DCFH-DA working solution was discarded, and the cells were washed three times with PBS. Finally, 100 μl of basal culture medium was added to each well, and the cells were placed in a high-content imaging analysis system (ImageXpress Micro 4). The excitation wavelength was 488 nm and the emission wavelength was 525 nm (FITC channel) to observe the fluorescence distribution of DCF in the cells.

[0165] 2. Experimental Results The fluorescence intensity was quantified using ImageJ software, and the coherence plot was simulated using GraphPad Prism software.

[0166] Results analysis, results ( Figure 9 The results show that the compounds (D12, D14, D15) prepared in this invention can effectively reduce intracellular ROS levels, and D12 and D14 are superior to the classic ferroptosis inhibitor (Fer-1).

[0167] Biological Testing Example 10: Effects of the Compounds of the Present Invention on Intracellular Nrf and GPX4 This experiment uses compound D12 as an example to study the effect of the compound of the present invention on intracellular ROS.

[0168] 1. Experimental Methods Western blotting analysis was performed. PC12 cells were seeded in 60 mm culture dishes and cultured for 24 hours until 80% confluence. After treatment with RSL3 (1 μM) and the test compound (1 μM) for 6 hours, the cells were washed twice with PBS buffer and lysed on ice with RIPA lysis buffer containing 1× protease inhibitor mixture and 1 mM benzyl sulfonyl fluoride (PMSF) for 15–20 minutes. The cell supernatant was collected by centrifugation, and the total protein concentration was determined using a BCA protein quantification kit. Equal volumes of cell lysis buffer (30 µg per lane) were separated by 10% SDS-PAGE and then transferred to a PVDF membrane for 1 hour. The membrane was blocked in TBST buffer containing 5% skim milk powder for 3 hours, and then incubated overnight at 4°C with the following primary antibodies: anti-GPX4 antibody (1:1000), anti-Nrf2 antibody (1:1000), and anti-GAPDH antibody (1:10000). After washing three times with TBST, the membrane was incubated with secondary antibody at room temperature for 1 hour. Protein bands were developed using ECL chemiluminescent substrate (Beijing 4A Biotechnology Co., Ltd.), and images were recorded using the eBlot Touch Imager contact chemiluminescence imaging system (Touch Imager S).

[0169] 2. Experimental Results The integrated optical density of the strips was quantitatively analyzed using ImageJ software.

[0170] Results analysis, results ( Figure 10 To elucidate whether 1,3-disubstituted indoleazine compounds exert their effects by regulating core antioxidant pathways, we analyzed the protein expression levels of transcription factor Nrf2 and its key downstream effector protein GPX4. Nrf2 is a core regulator of the cellular antioxidant defense system, playing a crucial role in resisting oxidative stress and maintaining redox homeostasis. Its downstream target, GPX4, as a phospholipid hydroperoxidase, directly inhibits lipid peroxidation and is a key regulatory protein in ferroptosis. Western blot analysis showed (… Figure 10 RSL3 treatment significantly downregulated the protein expression of Nrf2 and GPX4 in PC12 cells; however, co-treatment with D12 effectively reversed this inhibitory effect, significantly restoring their protein levels. These results suggest that D12 may alleviate lipid peroxidation by activating the Nrf2-GPX4 signaling axis.

[0171] Biological Testing Example 10: Study on the solubility, lipid-water partition coefficient, and liver microsomal stability of the compounds of the present invention. This experiment uses compounds D12 and D14 as examples to study the solubility, lipid solubility, and liver microsomal metabolism of the compounds of this invention.

[0172] 1. Experimental Methods Standard curve creation Weigh 1 mg of the analyte into a 1.5 mL EP tube, prepare a 1 mg / mL stock solution using acetonitrile as solvent, and dilute twice with methanol to final concentrations of 0.005, 0.01, 0.02, 0.04, 0.05, 0.08, 0.1, 0.16, 0.31, 0.62, 1.25, and 5 μg / mL. Filter the sample through a 0.45 μm syringe filter before LC / MS analysis. Plot a standard curve based on the concentration and relevant peak areas.

[0173] Solubility determination Weigh 3 mg of the compound, add 2 ml of pure water, and place it in a shaker at 37 ℃ for 24 h. After filtering the sample through a 0.45 μm syringe filter, dilute it 1000 times and perform LC / MS analysis. Calculate the solubility based on the relevant peak areas and substitute them into the standard curve.

[0174] Lipid-water partition coefficient determination The partition coefficient (Log P) of the n-octanol / water system was determined using the shake-flask method. Before the experiment, pure water and analytical grade n-octanol were pre-saturated and shaken in a constant-temperature shaker for 24 h. Afterward, the mixture was allowed to stand for a sufficient time to completely separate the two phases, yielding water-saturated n-octanol and n-octanol-saturated water, respectively. 2 mg of the test compound was transferred to 2 ml of n-octanol (H₂O saturated) / H₂O (n-octanol saturated) (1:1), stirred for 4 h to reach equilibrium, and diluted 1000-fold. The equilibrium sample was then analyzed by LC / MS, and the concentration was calculated using the standard curve described when determining the degree of hydrolysis. Log P = Log(C O / C W C O The concentration of the test compound in the n-octanol phase is indicated by C. W This indicates the concentration of the test compound in the aqueous phase. Each sample was tested three times.

[0175] Liver microsomal stability The 20 mg / mL mixed liver microsomes were stored at -80°C before use. To initiate the reaction, 25 μL of NADPH stock solution (2 mM, in 0.1 M PBS with 3.5 mM MgCl, pH 7.4) was added to 25 μL of buffer containing the analyte compound (final concentration 2 μg / mL) and liver microsomes (final concentration 1 mg / mL). After incubation for 0, 5, 15, 30, 45, and 60 min, 100 μL of ice-cold acetonitrile was added to the corresponding well to stop the reaction. The mixture was then shaken for 10 min and centrifuged at 10,000 rpm for 15 min. 100 μL of supernatant was collected from each well for LC / MS analysis. The half-life and clearance rate were calculated based on the peak areas and substituted into the standard curve.

[0176] 2. Experimental Results Table 2. Solubility, lipid-water partition coefficient, and liver microsomal stability of compounds D12 and D14 3. Results Analysis: The results (Table 2) show that compounds D12 and D14 prepared in this invention have better lipid-water partition coefficients than Fer-1, and D12 has better solubility (5 μg / ml). The metabolic stability of compounds D12 and D14 is slightly higher than that of Fer-1, with D12 being superior.

[0177] Biological Testing Example 11: Study of the Compounds of the Invention in an Animal Model of Liver Injury This animal experiment was approved by the Ethics Committee of Southwest Medical University (No. SWMU20250074).

[0178] Acetaminophen (APAP) overdose is a significant cause of drug-induced liver injury. Numerous studies have found that ferroptosis is involved in drug-induced acute liver injury. This experiment uses D12 as an example to investigate the therapeutic effect of the compounds of this invention on liver injury.

[0179] 1. Experimental Methods The in vivo effects of compound D12 were evaluated using an APAP-induced acute liver injury model.

[0180] The group processing for each experiment is as follows: Experimental (APAP+D12 10mg / kg) group: Male mice were given 10mg / kg of compound D12, followed by 500mg / kg of APAP.

[0181] Experimental (APAP+D12 20mg / kg) group: Male mice were given 20mg / kg of compound D12, followed by 500mg / kg of APAP.

[0182] Experimental (APAP+Fer1 10mg / kg) group: Male mice were given 10mg / kg of Fer1, followed by 500mg / kg of APAP.

[0183] Blank group: No processing is given.

[0184] Control (APAP+Vehicle) group: Male mice were given APAP at a dose of 500 mg / kg.

[0185] The solvent used in this experiment was: 5% DMSO + 30% PEG400 + 2% TWEEN + 67% physiological saline.

[0186] C57 BL / 6 mice were fasted for 12 hours prior to the experiment. The drug was administered 1 hour before modeling. Physiological saline was preheated to 70°C to prepare a 500 mg / kg APAP solution, which was maintained at this preheated temperature during the experiment to prevent precipitation. Modeling was initiated 1 hour later via intraperitoneal injection. Six hours after APAP treatment, liver tissue and blood were collected for hematoxylin and eosin (H&E) staining or biochemical analysis.

[0187] 2. Experimental Results The results are as follows Figure 11 As shown.

[0188] 3. Results Analysis Visual imaging of mouse livers showed severe liver lesions in the control group mice, including coagulative necrosis and congestion in the peripheral regions of the liver lobules. Treatment with Fer-1 and compound D12 significantly improved these lesions, with compound D12 and Fer-1 showing comparable improvement. Figure 11 A). H&E staining also showed that the control group exhibited liver structural damage, irregular hepatocyte arrangement, inflammatory cell infiltration, and increased necrosis. Administration of Fer-1 and compound D12 effectively reduced APAP-induced damage, preserved the natural liver structure, and reduced the area of ​​necrosis. Figure 11 B). Serum aspartate aminotransferase (ALT), alanine aminotransferase (AST) Figure 11 C, D) and elevated liver MDA levels ( Figure 11 E) Decreased liver GSH levels ( Figure 11F) indicates that the acute liver injury model was successfully established. Conversely, administration of compound D12 at a concentration of 10 mg / kg reduced liver tissue damage, manifested by inhibiting hepatocyte death, decreasing serum ALT, AST, and hepatic MDA levels, and increasing hepatic GSH; when compound D12 was administered at a concentration of 20 mg / kg, the therapeutic effect was further enhanced, showing a certain dose-dependent effect. These results indicate that the compounds of this invention can be applied to the treatment of APAP-induced drug-induced liver injury.

[0189] Biological Testing Example 12: Study of the Compounds of the Invention in a Brain Ischemia-Reperfusion Model This animal experiment was approved by the Ethics Committee of Southwest Medical University (No. 20241023-052).

[0190] Cerebral ischemia-reperfusion injury (CIRI) is a common secondary injury following the revascularization of ischemic stroke (such as cerebral infarction). Its core mechanisms involve oxidative stress, inflammatory response, and apoptosis. Ferroplasmosis has been proven to be one of the key pathways for neuronal and glial cell death in CIRI.

[0191] This experiment uses compound D12 as an example to study the preventive and therapeutic effects of the compound of the present invention on cerebral ischemia-reperfusion.

[0192] 1. Experimental Methods The in vivo effects of compound D12 were evaluated using a focal cerebral ischemia-reperfusion model (suture occlusion method).

[0193] The experimental groups and treatments are as follows: Sham surgery group: only blood vessels were exposed, without inserting sutures.

[0194] Control group: 2 hours of ischemia followed by 2 hours of reperfusion, no drug administration. Experimental group: D12 (10mg / kg) was injected via the tail vein 1 hour before surgery. The solvent used in this experiment was: 5% DMSO + 30% PEG400 + 5% TWEEN80 + 63% physiological saline.

[0195] Fasting was required for 12 hours prior to administration, and administration was administered 1 hour before surgery. SD rats were deeply anesthetized and fixed on a temperature-controlled surgical board. The hair in the anterior neck region was thoroughly shaved using an electric shaver. A longitudinal incision of approximately 2.5 cm was made along the midline of the anterior neck to expose the trachea and the right carotid artery sheath. The common carotid artery (CCA) was carefully dissected, and the distal end of the external carotid artery (ECA) was double-ligated and cut in the middle. A suture plug was inserted into the ECA to completely block CCA blood flow, and the onset time of ischemia was recorded. The incision was sutured, and observation was performed. After 2 hours of ischemia, the suture plug was removed, and reperfusion was initiated. The animals were returned to their cages and observed in an indoor environment at 25±2℃.

[0196] 2. Experimental Results: Neurological deficits were scored according to the Longa scoring system (Table 3). Neurological deficits were assessed under blinded conditions after the rats were fully awakened from anesthesia and 24 hours after reperfusion to ensure the objectivity of the scoring. Higher scores indicate greater severity of injury.

[0197] Table 3 Neurological deficit scores The volume of cerebral infarction was determined using the TTC staining method. After 24 h of reperfusion, rats were deeply anesthetized, their hearts were exposed via thoracotomy, and intact brain tissue was harvested on ice after perfusion with physiological saline. The blood was gently rinsed with pre-cooled physiological saline, and the tissue was flash-frozen in liquid nitrogen for 20 min, then stored at -80°C for TCC staining. Normal brain tissue, due to normal mitochondrial enzyme activity, was stained red; infarcted brain tissue, due to impaired mitochondrial function, could not reduce TTC and appeared white.

[0198] Normal tissue (red) and infarcted area (white) were analyzed using ImageJ to calculate the percentage of cerebral infarction volume.

[0199] 2. Experimental Results The results are as follows Figure 12 As shown.

[0200] 3. Results Analysis ( Figure 12 In the sham-operated group, both cerebral hemispheres of the rats were uniformly deep red, and the brain tissue morphology was intact; in the model group, the ischemic side (right) cerebral hemisphere of the rats showed obvious ischemic necrosis, which was pale white, while the contralateral side was uniformly deep red; in the experimental group (compound D12, 10 mg / kg), the area of ​​the pale white necrotic area on the ischemic side was significantly reduced, and the contralateral hemisphere was deep red with no abnormalities. Figure 12 B). Statistical analysis showed that the infarct volume in the control group was significantly higher than that in the sham surgery group, while the infarct volume in the experimental group was significantly lower than that in the control group. Figure 12 C). The neurological deficit score in the control group was also higher than that in the sham surgery group, while the score in the experimental group was significantly lower than that in the control group. Figure 12 D). These results indicate that the compounds of the present invention can be used for the prevention and treatment of cerebral ischemia-reperfusion.

[0201] In summary, this invention provides a novel indoleazine skeleton compound, its preparation method, and its use in ferroptosis. The compound prepared by this invention exhibits excellent inhibitory effects on ferroptosis, and its inhibition of ferroptosis does not occur through traditional free radical capture or iron chelation mechanisms, belonging to a class of unique non-chelating, non-free radical capture ferroptosis inhibitors. It can be used to prepare non-chelating, non-free radical capture ferroptosis inhibitors. Experimental results show that this type of compound exhibits significant protective effects in ferroptosis-related pathological models, especially showing potential for prevention and treatment of cerebral ischemia-reperfusion injury, and can effectively alleviate acetaminophen-induced acute liver injury. Therefore, the compound of this invention can be used for tissue ischemia-reperfusion injury and acute organ injury, and has broad market prospects.

[0203] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. The above description is merely a preferred embodiment of the invention and is not intended to limit the invention. Those skilled in the art can make various modifications and improvements within the scope of the invention's concept, and such modifications and improvements should also be considered to fall within the protection scope of the invention.

Claims

1. Use of a compound of Formula I or a pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug thereof in the preparation of a medicament for inhibiting ferroptosis: Equation I in: X is -C(O)-; R 1 Selected from: C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, 3, or 4 substituents selected from the group consisting of: halogen, amino, hydroxyl, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy; R 2 Selected from: hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, carboxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; R 3 R 4 R 5 R 6 Each is independently selected from: hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, -C(O)OR′, wherein R′ is independently hydrogen or C1-C3 alkyl, and R 3 R 4 R 5 R 6 At least two of them are hydrogen; A is selected from: C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, or 3 substituents selected from Ra; each Ra is independently selected from: hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, -C(O)OR b -O-CH2-phenyl; wherein R b It is a C1-C4 alkyl group.

2. The use according to claim 1, characterized in that, A is selected from: phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

3. The use according to claim 1, characterized in that, The compound has the structure shown in Formula II: II n is 0, 1, 2, or 3; R 1 R 2 R 3 R 4 R 5 R 6 X and Ra are as defined in claim 1.

4. The use according to claim 1, characterized in that, A is selected from: , , The definition of Ra is as described above.

5. The use according to claim 1, characterized in that, The compound is selected from the following structures: 。 6. The compound represented by Formula I or a pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug thereof: Equation I in: X is -C(O)-; R 1 Selected from: C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 alkylamino, C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, 3, or 4 substituents selected from the group consisting of: halogen, amino, hydroxyl, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy; R 2 It is hydrogen; R 3 R 4 R 5 R 6 Each is independently selected from: hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, -C(O)OR′, wherein R′ is independently hydrogen or C1-C3 alkyl, and R 3 R 4 R 5 R 6 At least two of them are hydrogen; A is selected from: C3-C8 cycloalkyl, 4-10 heterocyclic, C6-C10 aryl, 5-10 heteroaryl; the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally substituted by 1, 2, or 3 substituents selected from Ra; each Ra is independently selected from: hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, -C(O)OR b -O-CH2-phenyl; wherein R b It is a C1-C4 alkyl group; An additional condition is that the compound represented by Formula I is not selected from the following compounds: 。 7. The compound according to claim 6, characterized in that, The compound has the structure shown in Formula II: II n is 0, 1, 2, or 3; R 1 R 2 R 3 R 4 R 5 R 6 X and Ra are defined as above.

8. The compound according to claim 6, characterized in that, The compound is selected from the following structures: 。 9. A pharmaceutical composition comprising a therapeutically effective amount of the compound of any one of claims 6-8, or a pharmaceutically acceptable salt, solvate, stereoisomer, isotope label, or prodrug thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

10. The use as described in claim 1, characterized in that, The ferroptosis referred to is ferroptosis associated with ischemic brain injury or drug-induced liver injury.

11. The use as described in claim 1, characterized in that, The drug is used to treat or prevent acetaminophen (APAP)-induced liver injury.

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