Phenylacrylamide compound, and preparation method therefor and use thereof

Abstract

Provided in the present application are a phenylacrylamide compound, and a preparation method therefor and the use thereof. The phenylacrylamide compound has a structure represented by formula I. The phenylacrylamide compound provided in the present application can exhibit good binding to a plurality of target proteins, particularly exhibiting a good inhibitory activity against ACAT1 and 3-KAT targets, and has good bioavailability. The compound can be used in the preparation of a drug for preventing or treating tumors, cancer pain, radiotherapy- and chemotherapy-induced injuries, autoimmune diseases, infections, drug-induced myocarditis, cytokine storms, ischemic heart disease, inflammatory diseases and neurodegenerative diseases, or for anti-aging.

Description

A phenylacrylamide compound, its preparation method and application Technical Field

[0001] This application belongs to the field of pharmaceutical technology, specifically relating to a phenylacrylamide compound, its preparation method, and its application. Background Technology

[0002] Medical data indicates a close relationship between inflammation and the development of tumors. For example, Helicobacter pylori infection tends to increase the risk of stomach cancer; chronic hepatitis may also be a direct cause of liver cancer. Other factors, such as autoimmune enteropathy, are closely related to colon cancer, and fine particulate matter in the air is also a contributing factor to lung cancer. Cancer is defined as a non-infectious disease, similar to heart disease, chronic respiratory diseases, or diabetes. These are mostly long-term chronic diseases with slow progression. The correlation between inflammation and tumors was first proposed by Galenus 1800 years ago, and many studies have confirmed that persistent inflammation can cause lesions to progress from infection or autoimmune inflammation to tumors.

[0003] Phenylacetamide compounds are widely found in bioactive natural products, especially those with hydroxyl substitutions on the benzene ring. These natural products possess a variety of biological activities, such as antibacterial, antioxidant, anti-inflammatory, and antitumor effects, and have a very broad research prospect. Structural modification of these natural products can help discover lead compounds with stronger tumor-suppressive activity, therapeutic activity for autoimmune diseases and various infectious and non-infectious inflammatory diseases, and lower toxicity than the original natural products. Furthermore, these lead compounds can be developed into antitumor drugs, drugs for treating neurodegenerative diseases, and anti-inflammatory immunomodulatory drugs.

[0004] ACAT1 belongs to the specific thiolase superfamily and is known as acetyl-CoA thiotransferase (ACAT). Many reports indicate that ACAT1 expression is often aberrant in tumor cells and plays a crucial role in tumorigenesis and development. The upregulation mechanisms of ACAT1 activity in different human cancer cells warrant further investigation. High ACAT1 expression reduces overall survival; in MDA-MB-231 human breast cancer cells, ACAT1 overexpression leads to ketone body reusability, driving tumor progression and metastasis. Therefore, research on phenylacrylamide derivatives targeting ACAT1 may lead to the discovery of novel antitumor drugs.

[0005] 3-KAT, also known as 3-ketoyl-CoA thiolytic enzyme, has its main biological function in mitochondria where it catalyzes the complete cleavage of long-chain and short-chain fatty acids into acetyl-CoA. In peroxisomes, its substrates are very long-chain fatty acids (≥ C22), long-chain fatty acids, and branched-chain fatty acids. After a limited β-oxidation cycle, the carbon chains are shortened, and the products then enter the mitochondria for further oxidation. Inhibiting 3-KAT can also inhibit the growth of tumors, especially melanoma and glioma.

[0006] Furthermore, natural products based on this structure exhibit multi-target characteristics, and other potential targets are summarized below:

[0007] Caspase-3 is a caspase protein that interacts with caspase-8 and caspase-9. It is encoded by the casp3 gene. Elevated levels of caspase-3, specifically the p17 fragment, in the blood are a hallmark of recent myocardial infarction. Current research suggests that caspase-3 may play an important role in embryonic and hematopoietic stem cell differentiation and apoptosis. Studies have shown abnormally high caspase-3 expression in patients with head and neck cancer and breast cancer; therefore, reducing its expression may be one way to mitigate cancer development and progression.

[0008] Epidermal growth factor receptor (EGFR): Mutant activation of EGFR is an important factor leading to abnormal biological activities of tumor cells. Among them, the T790M mutation in EGFR is a change in a base pair from cytosine nucleoside (C) to thymine (T), that is, the threonine at position 790 of the EGFR tyrosine kinase is replaced by methionine. This mutation can reactivate EGFR, thereby leading to resistance to tyrosine kinase inhibitors (TKIs). Some derivatives containing phenylacrylic acid and amide have been reported to have EGFR inhibitory activity. AIDD has also verified that such compounds also have inhibitory activity against the T790M mutation.

[0009] Histone deacetylases (HDACs) are a class of proteases that play a crucial role in chromosome structural modification and gene expression regulation. Normally, histone acetylation facilitates the dissociation of DNA from histone octamers, relaxes nucleosome structure, and allows various transcription factors and co-transcription factors to specifically bind to DNA binding sites, activating gene transcription. Within the cell nucleus, histone acetylation and deacetylation are in dynamic equilibrium, jointly regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs).

[0010] Histone-lysine N-methyltransferase EZH2: EZH2 is an enzyme encoded by the human EZH2 gene. Two transcript variants transcribed from this gene have been identified, encoding different subtypes. Gene sequence alterations are fundamentally different from epigenetic modification abnormalities. Once a DNA sequence mutation occurs, the gene is difficult to repair, or the mutated gene product is difficult to eliminate. However, epigenetic modification abnormalities can potentially be reversed by inhibitors of their associated chromatin-modifying enzymes. Therefore, clarifying the mechanism of action of epigenetic modifying enzymes in tumor cells is crucial, thereby providing corresponding therapeutic approaches to prevent epigenetic modification mutations. Currently, there are no marketed EZH2 inhibitors, and five drugs are in Phase I / II clinical trials.

[0011] Src kinase is a non-receptor protein kinase that is widely present in cancer cells and plays an important role in various processes of cell growth and proliferation, such as gene transcription, cell differentiation, migration, angiogenesis, and prevention of apoptosis. Research on Src inhibitors has become a hot topic in anti-tumor drug research. Currently, a number of Src inhibitors are in the clinical research stage.

[0012] Cancer immunotherapy is an increasingly effective cancer treatment strategy. T cells play a key role in immunotherapy, and numerous immune checkpoints are a treasure trove waiting to be explored. Since the efficacy of CTLA-4 and PD-1 / PD-L1 monoclonal antibodies has been confirmed, competition in this monoclonal antibody track has become fierce. Therefore, more attention should be paid to other immune checkpoint targets, whose functions determine the efficacy of immunotherapy.

[0013] T cell survival and development are influenced by TCR signaling, which in turn depends on Src family kinases (SFKs). Lck is an important member of the SFK family and is expressed throughout most of the T cell lifecycle. Furthermore, Lck plays a crucial role in activating the TCR signaling pathway to activate T cells. CSK is a key regulator of SFKs; its phosphorylation of Lck (Tyr505) inactivates Lck, which in turn inhibits T cell activation via the TCR. Therefore, CSK and p-Lck (Tyr505) may be effective targets for future immunomodulatory therapies.

[0014] CD73 is a 5-nucleotide hydrolase that hydrolyzes extracellular adenosine monophosphate (AMP) into adenosine. Adenosine is a potent immunosuppressive molecule that inhibits the activation of CD8-positive T cells, thereby helping cancer cells evade T cell attacks. Tumor-infiltrating NK cells upregulate CD73 expression, and the frequency of these CD73+ NK cells is correlated with tumor size in breast cancer patients. These findings support the idea that tumors can hijack NK cells for immune evasion, and that CD73 expression defines an induced NK cell population with immunomodulatory properties within the tumor microenvironment.

[0015] KIR, or killer cell immunoglobulin-like receptor, is a receptor expressed on the surface of NK cells and some T cells. It can specifically recognize MHC-I molecules on the cell surface, thereby exerting an immunomodulatory function.

[0016] LAG-3, lymphocyte activating factor 3 protein, is an immune negative regulatory molecule receptor distributed in activated T cells, NK cells, and dendritic cells that can bind to MHC class II molecules. It has the functions of maintaining homeostasis and participating in immune regulation, and is closely related to the occurrence and development of tumors.

[0017] 4-1BB, also known as CD137, is a member of the TNF family expressed on the surface of activated T cells. It is an inducible T cell surface receptor. 4-1BB and its ligands are another important co-stimulatory molecule in addition to the CD28 / B7 co-stimulatory signaling pathway.

[0018] It is associated with the following in terms of antioxidant and anti-inflammatory immune effects:

[0019] Because phenylacrylamide structures are widely found in polyphenolic secondary metabolites of plants, it possesses strong antioxidant and anti-inflammatory properties. Its activity in scavenging 1,1-diphenyl-2-trinitrophenylhydrazine free radicals is 2-3 times that of vitamins C and E, and its activity in scavenging superoxide anion free radicals is 10-30 times that of vitamins C and E. Regarding its anti-inflammatory effects, it exhibits anti-inflammatory activity in a carrageenan-induced rat paw edema inflammation model. Its mechanism involves inhibiting the production of interleukin-8 (IL-8) by scavenging intracellular ROS, inhibiting p38 cascade phosphorylation, and upregulating the nuclear factor NF-κB signaling pathway, thereby exerting an anti-inflammatory effect. It also inhibits the activation of the NF-κB inflammatory pathway and p65 acetylation caused by bacterial or viral infections, leading to the massive production and release of inflammatory factors such as interleukin-1, TNF-α, and interleukin-6, thus contributing to the treatment of infectious myocarditis and cytokine storms.

[0020] Pathological studies of neurodegenerative diseases Alzheimer's and Parkinson's diseases have shown that chronic oxidative stress and inflammation lead to neuronal damage. Based on the strong antioxidant and anti-inflammatory effects of phenylacrylic acids, they have been found to have good neuroprotective effects. Some clinical and preclinical studies have shown that phenylacrylamide natural products exhibit good therapeutic effects against Alzheimer's and Parkinson's diseases. They can improve memory loss and hippocampal cell death following transient global cerebral ischemia. They prevent saturated free fatty acid-induced lipotoxicity by activating SIRT1-regulated mitochondrial function. The mechanism involves reducing ROS production and increasing mitochondrial mass and mitochondrial membrane potential, thus alleviating oxidative stress and mitochondrial dysfunction; and significantly reducing the expression of the pro-apoptotic protein Bax, thereby reducing mitochondrial-mediated caspase-dependent apoptosis.

[0021] Although phenylacrylamide derivatives have been shown to have a broad range of activities as described above, their exact mechanisms of action and structure-activity relationships require further investigation. It is necessary to synthesize their structural derivatives and study their drug action and the diseases they can treat, especially inflammation-related (infectious and non-infectious), tumors, autoimmune diseases, neurodegenerative diseases, and aging. Summary of the Invention

[0022] This application provides a phenylacrylamide compound, its preparation method, and its applications. The phenylacrylic acid compound provided in this application can bind well to multiple target proteins and has good bioavailability. It can be used to prepare drugs for the prevention or treatment of tumors, cancer pain, radiotherapy and chemotherapy damage, autoimmune diseases, infectious and drug-induced myocarditis, cytokine storms, ischemic heart disease, inflammatory diseases, neurodegenerative diseases, or anti-aging.

[0023] In a first aspect, this application provides a phenylacrylamide compound having the structure shown in Formula I:

[0024] ;

[0025] Wherein, R1, R4, R5, and R6 are each independently selected from hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxycarbonyl, C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxycarbonyl, C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkyl ...4, C5, C6, etc.) alkyl, C1-C6 (e.g., C1, C2, C4, C5, C6, etc.) alkyl, C1-C6 (e.g., C1, C2, C4, C5, C6, R5 and R6 are any one of the following: C4, C5, C6, etc. alkoxy; C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxymethyleneoxy; C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkyl; or C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) trihalogenated alkyl; or trihalogenated C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxy; or R5 and R6 are independently selected from -O(CH2). n O-, and linked to its substituted phenyl group to form a ring, where n is selected from 1, 2 or 3;

[0026] R2 and R3 are each independently selected from hydrogen, hydroxyl-C0-C6 (e.g., C0, C1, C2, C3, C4, C5, C6, etc.) alkyl, C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkylamino, cyano, nitro, C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxycarbonyl, methanesulfonyl, dimethylaminesulfonyl, amino, Any one of the following: C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkyl, C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxy, trihalogenated C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkyl or trihalogenated C1-C6 (e.g., C1, C2, C3, C4, C5, C6, etc.) alkoxy;

[0027] X is selected from CH2, CO, or CH2CONH.

[0028] The phenylacrylamide derivatives designed in this application primarily target ACAT1 (acetyl-CoA acetyltransferase 1) and 3-KAT (3-ketoacyl-CoA thiolase) in their antitumor activity, achieving therapeutic effects through dual inhibition.

[0029] In this application, trihalogenated C1-C6 alkyl or trihalogenated C1-C6 alkoxy refers to a compound containing three halogens, including F. 3 C1-C6 alkyl groups, Br3 C1-C6 alkyl groups, Cl 3 C1-C6 alkyl, I 3 C1-C6 alkyl, F2Br C1-C6 alkyl, F2Cl C1-C6 alkyl, F2I C1-C6 alkyl, FBr 2 C1-C6 alkyl groups, FCl 2 C1-C6 alkyl groups, FI 2 C1-C6 alkyl groups, ClBr 2 C1-C6 alkyl groups, ICl 2 C1-C6 alkyl groups, ClI 2 C1-C6 alkyl; F 3 C1-C6 alkoxy groups, Br 3 C1-C6 alkoxy groups, Cl 3 C1-C6 alkoxy groups, I 3 C1-C6 alkoxy group, F2Br C1-C6 alkoxy group, F2Cl C1-C6 alkoxy group, F2I C1-C6 alkoxy group, FBr 2 C1-C6 alkoxy groups, FCl 2 C1-C6 alkoxy groups, FI 2 C1-C6 alkyl groups, ClBr 2 C1-C6 alkoxy groups, ICl 2 C1-C6 alkoxy groups, ClI 2 C1-C6 alkoxy groups.

[0030] Preferred F 3 C1-C4 alkyl groups, Br 3 C1-C4 alkyl groups, Cl 3 C1-C4 alkyl, I 3 C1-C4 alkyl groups, F2Br C1-C4 alkyl groups, F2Cl C1-C4 alkyl groups, F2I C1-C4 alkyl groups, FBr 2 C1-C4 alkyl groups, FCl 2 C1-C4 alkyl, FI 2 C1-C4 alkyl groups, ClBr 2 C1-C4 alkyl groups, ICl 2 C1-C4 alkyl groups, ClI 2 C1-C4 alkyl; F 3 C1-C4 alkoxy groups, Br 3 C 1-4 Alkoxy, Cl 3 C1-C4 alkoxy groups, I 3 C1-C4 alkoxy groups, F2Br C1-C4 alkoxy groups, F2Cl C1-C4 alkoxy groups, F2I C1-C4 alkoxy groups, FBr 2 C1-C4 alkoxy groups, FCl 2 C1-C4 alkoxy groups, FI 2 C1-C4 alkyl groups, ClBr 2 C1-C4 alkoxy groups, ICl2C 1-4 Alkoxy, ClI 2 C1-C4 alkoxy groups.

[0031] The optimal choices are F3C, F3CCH2, Br3C, Br3CCH2, Cl3C, Cl3CCH2, I3C, I3CCH2, F2BrC, F2BrCCH2, F2ClC, F2ClCCH2, F2IC, F2ICCH2, FBr2C, FBr2CCH2, FCl2C, FCl2C CH2, FI2C, FI2CCH2, ClBr2C, ClBr2CCH2, ICl2C, ICl2CCH2, ClI2C, ClI2CCH2; F3CO, F3CCH2O, Br3CO, Br3CCH2O, Cl3CO, Cl3CCH2O, I3CO, I3CCH2O, F2BrCO, F2BrCCH2O, F2ClCO, F2ClCCH2O, F2ICO, F2ICCH2O, FBr2CO, FBr2CCH2O, FCl2CO, FCl2CCH2O, FI2CO, FI2CCH2O, ClBr2CO, ClBr2CCH2O, ICl2CO, ICl2CCH2O, ClI2CO or ClI2CCH2O.

[0032] In this application, the phenylacrylamide compound is selected from any one of the following formula I:

[0033] ;

[0034] Wherein, R1, R4, R5, and R6 are each independently selected from hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxycarbonyl, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxycarbonyl, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkyl ... R5 and R6 are any one of the following: C2, C3, C4 (e.g., C1, C2, C3, C4, etc.) alkoxy, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxymethyleneoxy, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkyl, or C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxy; or R5 and R6 are independently selected from -O(CH2). n O-, and linked to its substituted phenyl group to form a ring, where n is selected from 1, 2 or 3;

[0035] R2 and R3 are each independently selected from any one of hydrogen, hydroxy C0-C4 (e.g., C0, C1, C2, C3, C4, etc.) alkyl, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkylamino, cyano, nitro, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxycarbonyl, methanesulfonyl, dimethylaminesulfonyl, amino, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkyl, C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxy, trihalogenated C1-C4 (e.g., C1, C2, C3, C4, etc.) alkyl, or trihalogenated C1-C4 (e.g., C1, C2, C3, C4, etc.) alkoxy.

[0036] X is selected from CH2, CO, or CH2CONH.

[0037] In this application, R1, R4, R5, and R6 are each independently selected from hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1 alkoxycarbonyl, C2 alkoxycarbonyl, C1 alkoxycarbonyloxy, C2 alkoxycarbonyloxy, C1 alkyl, C2 alkyl, C1 alkoxy, C2 alkoxy, C1 alkoxymethyleneoxy, C2 alkoxymethyleneoxy, C1 alkyl, C2 alkyl, trihaloC1 alkyl, trihaloC2 alkyl, or trihaloC1 alkoxy, trihaloC2 alkoxy; or, R5 and R6 are independently selected from -O(CH2). n O-, which is linked to a substituted phenyl group to form a ring, where n is selected from 1, 2, or 3.

[0038] R2 and R3 are each independently selected from any one of hydrogen, hydroxyl, hydroxy-C1 alkyl, C1 alkylamino, C2 alkylamino, cyano, nitro, C1 alkoxycarbonyl, C2 alkoxycarbonyl, methanesulfonyl, dimethylaminesulfonyl, amino, C1, C2 alkyl, C1, C2 alkoxy, trihaloC1 alkyl, trihaloC2 alkyl, or trihaloC1 alkoxy, trihaloC2 alkoxy.

[0039] In this application, R1, R4, R5, and R6 are selected from any one of hydrogen, fluorine, chlorine, bromine, hydroxyl, dimethylamino, cyano, nitro, methoxycarbonyl, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, acetoxy, or methoxymethyleneoxy; or R5 and R6 are selected from -O(CH2). n 'O-, and linked to its substituted phenyl group to form a ring; n' is selected from 1, 2 or 3;

[0040] R2 and R3 are each independently selected from any one of hydrogen, hydroxyl, hydroxymethyl, dimethylamino, cyano, nitro, methoxycarbonyl, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, or trifluoromethoxy.

[0041] X is selected from CH2, CO, or CH2CONH.

[0042] In this application, the phenylacrylamide compound is selected from any one of the following M1-M10:

[0043] .

[0044] Secondly, this application provides a tautomer of the phenylacrylamide compound described in the first aspect, or a pharmaceutically acceptable salt thereof.

[0045] Preferably, the pharmaceutically acceptable salt comprises any one or a combination of at least two of the following: hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, p-toluenesulfonate, acetate, trifluoroacetate, salicylate, amino acid salt, citrate, maleate, tartrate, fumarate, citrate, lactate, sodium salt, potassium salt, calcium salt, magnesium salt, lithium salt, ammonium salt, or salt of an organic base that can provide a physiologically acceptable cation.

[0046] Preferably, the salt of the organic base that provides a physiologically acceptable cation includes any one or a combination of at least two of the following: methylamine salt, dimethylamine salt, trimethylamine salt, piperidine salt, morpholine salt, or tri(2-hydroxyethyl)amine salt.

[0047] In this application, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0048] If cis / trans isomers are present, this application relates to the cis and trans forms and mixtures thereof, and if a single isomer is required, it can be isolated by conventional methods or prepared by stereoselective synthesis.

[0049] All salts in this application can be prepared using conventional methods. Furthermore, during the preparation of the solvates of the compounds shown in Formula I and their salts, polycrystalline or eutectic crystals may occur under different crystallization conditions.

[0050] Thirdly, this application provides a method for preparing the phenylacrylamide compound according to the first aspect, the method comprising the following steps:

[0051] (1) The compound shown in formula a is reacted with the compound shown in formula b (condensation reaction or substitution reaction) to obtain the compound shown in formula c, and the reaction formula is shown below:

[0052] ;

[0053] (2) The compound shown in formula c is deprotected to obtain the compound shown in formula d, and the reaction formula is shown below:

[0054] ;

[0055] (3) Reacting the compound shown in formula d with the compound shown in formula e yields the compound shown in formula I, as shown in the following reaction formula:

[0056] ;

[0057] Wherein, R' is selected from any one of tert-butyldimethylsiloxy, tert-butyldiphenylsiloxy, trimethylsiloxy, triethylsiloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy;

[0058] R1' is selected from any one of acetyl, tert-butyldimethylsiloxy, tert-butyldiphenylsiloxy, trimethylsiloxy, triethylsiloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy.

[0059] X' is selected from CHO, TsOCH2, COOH, or BrCH2CONH;

[0060] X is selected from CH2, CO, or CH2CONH;

[0061] Y is selected from chlorine, methoxy, or hydroxyl.

[0062] In this application, in step (1), the reaction is carried out under alkaline conditions.

[0063] Preferably, in step (1), the reaction is carried out in the presence of a condensing agent, which includes EDCI and / or DMAP.

[0064] Preferably, in step (1), the reaction temperature is 0-50℃ (e.g., 0℃, 5℃, 10℃, 15℃, 20℃, 25℃, 30℃, 35℃, 40℃, 45℃ or 50℃, etc.), and the reaction time is 2-15 h (e.g., 2h, 5h, 7h, 9h, 11h, 13h or 15h, etc.).

[0065] Preferably, in step (1), the molar ratio of the compound shown in formula a to the compound shown in formula b is 1:(0.8-1.5); for example, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, etc.

[0066] Preferably, in step (2), the deprotection reaction is carried out by hydrolysis or hydrogenation.

[0067] Preferably, in step (2), the deprotection is carried out by a hydrolysis reaction, which is carried out under acidic or alkaline conditions.

[0068] Preferably, in step (2), the temperature of the deprotection reaction is 0-50℃ (e.g., 0℃, 5℃, 10℃, 15℃, 20℃, 25℃, 30℃, 35℃, 40℃, 45℃, 50℃, etc.), and the reaction time is 1-24 h (e.g., 1h, 4h, 8h, 12h, 16h, 20h, 24h, etc.).

[0069] Preferably, in step (2), the deprotection reaction is carried out in a solvent, wherein the solvent is selected from tetrahydrofuran and / or methanol.

[0070] Preferably, in step (3), the reaction is carried out under alkaline conditions.

[0071] Preferably, in step (3), the reaction is carried out in the presence of a condensing agent, which includes EDCI and / or DMAP.

[0072] Preferably, in step (3), the molar ratio of the compound shown in formula d to the compound shown in formula e is 1:(0.8-2); for example, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2, etc.

[0073] Preferably, in step (3), the reaction temperature is 0-50℃ (e.g., 0℃, 10℃, 20℃, 30℃, 40℃, 50℃, etc.), and the reaction time is 1-15 h (e.g., 1 h, 3 h, 5 h, 7 h, 9 h, 11 h, 13 h, 15 h, etc.).

[0074] Fourthly, this application provides a pharmaceutical composition comprising an active ingredient and a pharmacodynamically acceptable carrier, wherein the active ingredient comprises a tautomer of the phenylacrylic acid compound described in the first aspect or the phenylacrylamide compound described in the second aspect, or a pharmaceutically acceptable salt thereof.

[0075] Preferably, the mass percentage of the active ingredient in the pharmaceutical composition is 0.1-95%, for example, it can be 0.1%, 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, etc.

[0076] The pharmaceutical compositions provided in this application can be prepared according to methods known in the art and can be formulated into any dosage form suitable for human or animal use by combining the active ingredient with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants.

[0077] The phenylacrylamide compounds described in the first aspect of this application, the tautomers of the phenylacrylamide compounds described in the second aspect, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described in the fourth aspect, can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0078] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including water injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0079] The phenylacrylamide compounds described in the first aspect of this application, the tautomers of the phenylpropionamide compounds described in the second aspect, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described in the fourth aspect, can be formulated as ordinary formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0080] In order to formulate the phenylacrylamide compounds described in the first aspect of this application, the tautomers of the phenylacrylamide compounds described in the second aspect, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described in the fourth aspect, into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0081] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0082] To formulate the phenylacrylamide compounds described in the first aspect, the tautomers of the phenylacrylamide compounds described in the second aspect, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described in the fourth aspect into capsules, they can be mixed with diluents and gliding agents, and the mixture can be placed directly into hard or soft capsules. Alternatively, they can be first formed into granules or microspheres with diluents, binders, and disintegrants, and then placed into hard or soft capsules. Various diluents, binders, wetting agents, disintegrants, and gliding agents used for preparing tablets can also be used to prepare capsules of the compounds of this application.

[0083] To prepare injectable formulations of the phenylacrylamide compounds described in the first aspect, the tautomers of the phenylacrylamide compounds described in the second aspect, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described in the fourth aspect of this application, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure regulators can be added. Solubilizers or co-solvents may include poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters may include phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; and osmotic pressure regulators may include sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing lyophilized powder injections, mannitol, glucose, etc., may also be added as supporting agents.

[0084] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0085] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of this application may be administered by any known method of administration.

[0086] Fifthly, this application provides the use of the phenylacrylamide compound according to the first aspect, the tautomer of the phenylacrylamide compound according to the second aspect, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to the fourth aspect in the preparation of a drug for the prevention or treatment of tumors, cancer pain, radiotherapy and chemotherapy damage, autoimmune diseases, infections and drug-induced myocarditis, cytokine storms, ischemic heart disease, inflammatory diseases, neurodegenerative diseases, or anti-aging.

[0087] Preferably, the tumor is selected from any one or a combination of at least two of the following: glioma, melanoma, gastric cancer, lung cancer, breast cancer, kidney cancer, liver cancer, oral epithelial cancer, head and neck tumors, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, rectal adenocarcinoma, leukemia, or lymphoma.

[0088] Preferably, the autoimmune disease includes any one or a combination of at least two of the following: rheumatoid arthritis, systemic lupus erythematosus, ulcerative colitis, psoriasis, Sjögren's syndrome, dermatomyositis, or amyotrophic lateral sclerosis.

[0089] Preferably, the inflammatory disease includes any one or a combination of at least two of the following: myocarditis caused by infectious bacteria or viruses, cytokine storm, non-infectious polyarteritis, phlebitis, and reflux esophagitis.

[0090] Preferably, the neurodegenerative disease includes Alzheimer's disease and / or Parkinson's disease.

[0091] The dosage of the phenylacrylamide compounds described in the first aspect of this application, the tautomers of the phenylacrylamide compounds described in the second aspect or their pharmaceutically acceptable salts, or the pharmaceutical compositions described in the fourth aspect may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual circumstances of the patient or animal, the route of administration, and the dosage form. Generally, the suitable daily dose range for the phenylacrylamide compounds described in the first aspect of this application, the tautomers of the phenylacrylamide compounds described in the second aspect or their pharmaceutically acceptable salts, or the pharmaceutical compositions described in the fourth aspect, is 0.001-150 mg / kg body weight (e.g., 0.001 mg / kg body weight, 0.005 mg / kg body weight, 0.008 mg / kg body weight, 0.01 mg / kg body weight, 0.05 mg / kg body weight, 0.1 mg / kg body weight, 0.5 mg / kg body weight, 0.8 mg / kg body weight, 1 mg / kg body weight, 3 mg / kg body weight, 5 mg / kg body weight, 10 mg / kg body weight, 30 mg / kg body weight, 50 mg / kg body weight, 80 mg / kg body weight, 100 mg / kg body weight, 120 mg / kg body weight, 140 mg / kg body weight or 150 mg / kg body weight), preferably 0.1-100 mg / kg body weight, more preferably 1-70 mg / kg body weight, and most preferably 2-60 mg / kg body weight. The above dosage can be administered as a single unit or divided into several units, depending on the physician's clinical experience and the dosing regimen, including the use of other treatment methods.

[0092] The phenylacrylamide compounds described in the first aspect of this application, the tautomers of the phenylacrylamide compounds described in the second aspect, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described in the fourth aspect, may be taken alone or in combination with other therapeutic or symptomatic medications. When other therapeutic agents have a synergistic effect, their dosage should be adjusted according to the actual situation.

[0093] Compared with the prior art, this application has the following advantages:

[0094] The phenylacrylamide compounds provided in this application exhibit good binding to multiple target proteins, particularly showing good inhibitory activity against ACAT1 (acetyl-CoA acetyltransferase 1) and 3-KAT (3-ketoacyl-CoA thiolase). They also possess good bioavailability and can be used to prepare drugs for the prevention or treatment of tumors, cancer pain, radiotherapy and chemotherapy damage, autoimmune diseases, infectious and drug-induced myocarditis, cytokine storms, ischemic heart disease, inflammatory diseases, neurodegenerative diseases, or anti-aging. The tumors mentioned include glioma, melanoma, gastric cancer, lung cancer, breast cancer, kidney cancer, liver cancer, oral epithelial cancer, head and neck tumors, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, rectal adenocarcinoma, leukemia, or lymphoma, etc. Attached Figure Description

[0095] Figure 1 shows the results of the test on the effect of compound M1 on the body weight of mice in an LPS-induced cytokine storm model.

[0096] Figure 2A shows the test results of the effect of compound M1 on the cardiac index of LPS-induced cytokine storm model mice.

[0097] Figure 2B shows the test results of the effect of compound M1 on the spleen index of LPS-induced cytokine storm model mice.

[0098] Figure 2C shows the test results of the effect of compound M1 on the lung index of LPS-induced cytokine storm model mice.

[0099] Figures 3A-3D show the test results of the effect of compound M1 on the cytokine content of an LPS-induced mouse cytokine storm model. Detailed Implementation

[0100] The technical solution of this application will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely to help understand this application and should not be regarded as specific limitations on this application.

[0101] In the following examples, the reaction raw materials are commercially available conventional products or known compounds reported in existing literature.

[0102] Example 1

[0103] This embodiment provides compound M1, and the preparation method of compound M1 includes the following steps:

[0104] 1.5 g a-1 (1.05 eq), 1.8 g b-1 (1 eq), 1.6 g EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide) (1.5 eq), and 9 mL dichloromethane (DCM) were added to a 50 mL single-necked flask. Then, 1.05 g DIPEA (N,N-diisopropylethylamine) (1.5 eq) was slowly added dropwise. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. After the reaction was monitored by TLC (thin-layer chromatography) (DCM / MeOH volume ratio = 20 / 1), the reaction was quenched by adding 10 mL of saturated sodium bicarbonate, followed by extraction with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and then evaporated to dryness with silica gel. The crude product was purified by column chromatography (DCM / MeOH = 20 / 1) to obtain 2 g of intermediate. The intermediate was placed in 50 mL of methanol, and 0.8 g of NaHCO3 was added. The reaction was carried out at 50 °C for 1.5 h. After removing the solvent by rotary evaporation, the product was separated by preparative liquid phase separation, yielding 1.2 g of product, with a yield of 50%.

[0105] ;

[0106] Chemical formula: C 23 H 28 N₂O₆, calculated value: 428.19, [M+H] + Test value: 429.70.

[0107] Example 2

[0108] This embodiment provides compound M2, prepared from 150 mg (1.00 eq) of compound a-1 and 193 mg (1.05 eq) of compound b-2, with the addition of 163 mg (1.50 eq) of condensing agent EDCI and 110 mg (1.50 eq) of organic base DIPEA. The preparation method of compound M2 is the same as that of compound M1, ultimately yielding 100 mg of a yellow solid, with a yield of 42.7%. The reaction formula for compound M2 is shown below:

[0109] ;

[0110] Chemical formula: C 22 H 24 N₂O₆; Calculated value: 412.16, [M+H] + Test value: 413.55.

[0111] Example 3

[0112] This embodiment provides compound M3, which is prepared by reacting compound a-1 (150 mg, 1.00 eq) and compound b-3 (201 mg, 1.05 eq) with condensing agent EDCI 163 mg (1.50 eq) and organic base DIPEA 110 mg (1.50 eq). The preparation method of compound M3 is the same as that of compound M1, yielding 156 mg of yellow solid M3 in a yield of 64.2%. The reaction formula for compound M3 is shown below:

[0113] ;

[0114] Chemical formula: C 23 H 28 N₂O₆; Calculated value: 428.19, [M+H] + Test value: 429.65.

[0115] Example 4

[0116] This embodiment provides compound M4, and the preparation method of compound M4 includes the following steps:

[0117] Add 5 mL of acetonitrile (MeCN) to a 25 mL three-necked flask. While stirring, add 720 mg of substituted piperazine (trimetazidine hydrochloride, 2.12 mmol) and 1.1 mL of DIPEA (2.12 mmol). Cool the reaction system to 0–5 °C. Dilute the acyl chloride (0.54 g, 2.12 mmol) with an appropriate amount of acetonitrile and slowly add it dropwise through a constant-pressure dropping funnel, controlling the reaction temperature not to exceed 20 °C. After the addition is complete, allow the reaction system to react at room temperature for 2 h. After the reaction is complete, cool the reaction system to 5–10 °C and slowly add 5 mL of water to quench the reaction. Add ethyl acetate (30 mL) to the system to dilute the reaction solution, then add water. Stir the reaction solution, perform extraction and separation. Extract the aqueous phase obtained in the first extraction with ethyl acetate (20 mL) for a second extraction. Combine the organic phases, wash the organic phase with 10% saline solution, separate the organic phase, dry it with anhydrous sodium sulfate, filter, and concentrate. Methanol (60 mL) was added to the concentrated product and evaporated to remove solvent. Then, methanol (8 mL) was added, and the mixture was stirred in an oil bath at 55 °C for 1 h. The temperature was then lowered to 15–20 °C over 2 h, and crystallization was allowed to occur for 3 h. The mixture was filtered, and the filter cake was washed with a small amount of methanol until the filtrate was pale yellow. The filter cake was collected and dried at 50 °C to constant weight to obtain 1.0 g of pale yellow powdery solid, which is M4, with a yield of 97%.

[0118] ;

[0119] Chemical formula: C 23 H 28N₂O₆; Calculated value: 485.23, [M+H] + Test value: 485.23.

[0120] Example 5

[0121] This embodiment provides compound M5, and the preparation method of compound M5 includes the following steps:

[0122] 0.9 g of M4 (1.86 mmol) was dissolved in 10 mL of methanol, and 465 mg of potassium bicarbonate (4.65 mmol, 2.5 eq) was added. The mixture was reacted at room temperature for 2 h. After the reaction was completed by TLC monitoring, water was added to terminate the reaction. The mixture was extracted with ethyl acetate (EA) and concentrated to give 0.6 g of M5, with a yield of 73.2%.

[0123] ;

[0124] Chemical formula: C 23 H 28 N₂O₆; Calculated value: 443.22, [M+H] + Test value: 443.22.

[0125] Example 6

[0126] This embodiment provides compound M6, which was prepared by reacting compound a-1 75 mg (1.00 eq, 0.28 mmol) and compound b-4 110 mg (1.05 eq) with condensing agent EDCI 82 mg (1.50 eq) and organic base DIPEA 55 mg (1.50 eq). The preparation method of compound M6 is the same as that of compound M1, ultimately yielding 93 mg of compound M6 as a yellow solid, with a yield of 71.5%. The reaction formula for compound M6 is shown below:

[0127] ;

[0128] Chemical formula: C 24 H 30 N₂O₇; Calculated value: 458.21, [M+H] + Test value: 459.80.

[0129] Example 7

[0130] This embodiment provides compound M7, which was prepared by reacting compound a-1 50 mg (1.00 eq, 0.19 mmol) and compound b-5 72 mg (1.05 eq) with condensing agent EDCI 72 mg (1.50 eq) and organic base DIPEA 97 mg (4.0 eq). The preparation method of compound M7 is the same as that of compound M1, ultimately yielding 30 mg of compound M7 as a yellow solid, with a yield of 33.7%. The reaction formula for compound M7 is shown below:

[0131] ;

[0132] Chemical formula: C 24 H 29 N3O7, calculated value: 471.20, [M+H] + Test value: 472.40.

[0133] Example 8

[0134] This embodiment provides compound M8, which is prepared by reacting compound a-3 50 mg (1.00 eq) and compound b-5 98 mg (1.05 eq) with condensing agent EDCI 98 mg (2.0 eq) and organic base DIPEA 133 mg (4.0 eq). The preparation method of compound M8 is the same as that of compound M1, ultimately yielding 50 mg of a yellow solid, compound M8, in a yield of 40%. The reaction formula for compound M8 is shown below:

[0135] ;

[0136] Chemical formula: C 25 H 31 N3O7, calculated value: 485.22, [M+H] + Test value: 486.50.

[0137] Example 9

[0138] This embodiment provides compound M9, which is prepared by reacting compound a-4 50 mg (1.00 eq) and compound b-5 98 mg (1.05 eq) with condensing agent EDCI 98 mg (2.0 eq) and organic base DIPEA 133 mg (4.0 eq). The preparation method of compound M9 is the same as that of compound M1, ultimately yielding 65 mg of compound M9 as a yellow solid, with a yield of 52%. The reaction formula for compound M9 is shown below:

[0139] ;

[0140] Chemical formula: C 25 H 31 N3O7, calculated value: 485.22, [M+H] + Test value: 486.35.

[0141] Example 10

[0142] This embodiment provides compound M10, which is prepared by reacting compound a-1 50 mg (1.00 eq) and compound b-6 72 mg (1.05 eq) with condensing agent EDCI 72 mg (2.0 eq) and organic base DIPEA 97 mg (4.0 eq). The preparation method of compound M10 is the same as that of compound M1, ultimately yielding compound M9 65 mg as a yellow solid, with a yield of 52%. The reaction formula for compound M10 is shown below:

[0143] ;

[0144] Chemical formula: C 24 H 29 N3O7; Calculated value: 471.20, [M+H] + Test value: 472.80.

[0145] Pharmacological experiments

[0146] Experimental Example 1

[0147] IIC values ​​of phenylacrylamide derivatives targeting ACAT1 and 3-KAT 50 Measurement

[0148] (I) Experimental Methods

[0149] Screening methods for ACAT1 small molecule inhibitors

[0150] ACAT1 catalyzes the reversible formation of acetyl-CoA from two molecules of acetyl-CoA. Its activity is measured by the reaction of ACAT1 with the substrate acetyl-CoA and CoA to produce the product acetyl-CoA. Since acetyl-CoA exhibits specific absorption in a particular spectrum, the increase or decrease in absorption in this specific spectrum reflects the effect on ACAT1 enzyme activity.

[0151] Recombinant ACAT1 protein was expressed and purified using *E. coli*, yielding a concentration of 1 mg / mL. For screening ACAT1 small molecule inhibitors, a buffer solution of 50 mM Tris-HCl (pH 8.1), 20 mM MgCl2, and 40 mM KCl was used. In a 200 μL enzyme catalysis system, 1 μL of recombinant ACAT1 protein, acetyl-CoA substrate to a final concentration of 25 μM, CPM probe to a final concentration of 100 μM, and different concentrations of small molecule inhibitors were added. Detection was performed using a Biotek Synergy H1 microplate reader with excitation light at 355 nm and emission light at 460 nm. Inhibitor IC50 analysis was performed using Prism 7.0 software. 50 The calculation.

[0152] Screening methods for 3-KAT small molecule inhibitors

[0153] Ketoacyl-CoA thiolytic enzymes catalyze the final reaction of fatty acid β-oxidation, using various long-chain fatty acids as substrates, releasing acetyl-CoA and producing acyl-CoA with a two-carbon deficiency. In eukaryotes, ketoacyl-CoA thiolytic enzymes are further divided into mitochondrial and peroxidosomal types. Long-chain ketoacyl-CoA (e.g., ketohexadecyl-CoA; 3-ketohexadecanoyl-CoA) undergoes cleavage of its magnesium-enolate under the catalysis of long-chain thiolytic enzyme I, resulting in a reduction in the long-chain ketoacyl-CoA substrate. This reduction is observed as a decrease in absorbance at a spectrophotometer (303 nm), allowing for quantitative analysis of the activity of long-chain ketoacyl-CoA thiolytic enzymes.

[0154] Mitochondrial protein was extracted from 10 μg of B16F10 cells, and 50 μM Palmityl CoA was added. The mixture was reacted at 37℃ for 15 min, followed by the addition of 2 mM NAD, and then reacted again at 37℃ for another 15 min to obtain 3-KAT protein. For screening 3-KAT small molecule inhibitors, the buffer solution used was 50 mM Tris-HCl (pH 8.1), 20 mM MgCl2, and 40 mM KCl. In a 200 μL enzyme catalysis system, 90 μL of recombinant 3-KAT protein, a final concentration of coenzyme A of 25 μM, and different concentrations of small molecule inhibitors were added. Detection was performed using a spectrophotometer (Flex A-200) at an absorbance of 303 nm, and IC50 analysis of the inhibitors was conducted using Prism 7.0 software. 50 The calculation.

[0155] (II) Experimental Results

[0156] The screening results of the in vitro effects on ACAT1 and 3-KAT activities are shown in Table 1.

[0157]

[0158] As shown in Table 1 above, compounds M1, M2, and M3 have inhibitory activity against both ACAT1 and 3-KAT targets, M6, M8, and M10 have inhibitory activity against ACAT1, and M4, M7, and M9 have strong inhibitory activity against 3-KAT.

[0159] Experiment Example 2

[0160] Pharmacodynamic study of M1 in a mouse model of LPS-induced cytokine storm

[0161] (I) Experimental Methods

[0162] Pharmacodynamic study methods in a mouse LPS-induced cytokine storm model

[0163] Seventy male Balb / c mice were used in the experiment and randomly divided into four groups: a solvent control group, a model group (LPS, 10 mg / kg, intraperitoneal injection (ip)), a positive control group (tocilizumab, 5 mg / kg, intravenous injection (iv)), a prednisone acetate group (prednisone, 5 mg / kg, po, once daily (qd)), an M1 intravenous injection group (3 mg / kg, iv, qd), a low-dose M1 group (30 mg / kg, oral po, qd), and a high-dose M1 group (60 mg / kg, po, qd), with 10 mice in each group. Except for the tocilizumab group, all other treatment groups were administered medication starting on day 0 for 7 consecutive days. The tocilizumab group was administered medication 1 hour before modeling. One hour after the last administration, 100 μL of LPS solution was injected into each mouse via intraperitoneal injection. Modeling was performed on all mice except the solvent control group, resulting in a total of 60 mice with the model. Animal weight was measured daily, and animals were processed 24 hours after modeling for organ index and cytokine levels (a total of 48 types).

[0164] (II) Experimental Results

[0165] The effect of compound M1 on body weight in a mouse model of LPS-induced cytokine storm is shown in Figure 1.

[0166] As shown in Figure 1, the body weight of mice in both M1 gavage groups was better than that in the model group, indicating that M1 has a significant protective effect on LPS-induced cytokine storm model mice.

[0167] The effects of compound M1 on organ indices in a mouse model of LPS-induced cytokine storm are shown in Figures 2A-2C.

[0168] As shown in Figures 2A-2C, the 30 mg / kg M1 administration group significantly reduced the lung index in mice, and the heart index was significantly lower than that in the model group. This indicates that 30 mg / kg M1 can significantly reduce the heart and pneumonia organ indices in mice caused by LPS-induced cytokine storm, and has a significant anti-inflammatory effect on heart and lung damage caused by cytokine storm.

[0169] The effect of compound M1 on cytokine levels in an LPS-induced mouse cytokine storm model is shown in Figures 3A-3D.

[0170] As shown in Figures 3A-3D, in the LPS-induced mouse cytokine storm model, 48 cytokines were detected. Among them, 27 cytokines, including IFN-gamma, IL-1beta, IL-12p70, IL-13, IL-4, IL-33, IL-10, IL-9, IL-5, GM-CSF, IL-27, IL-23, IFN-alpha, IL-15 / IL-15R, IL-31, IL-28, IL-1alpha, Rank1, ST2 (IL-33R), IL-7, IL-25, BTC, BAFF, Leptin, IL-7R, IL-2RA, and IL-3, were significantly lower in the 30 mg / kg M1 treatment group at 2h or 24h than in the model group. This group showed superior reduction in some cytokine levels compared to the positive control drug prednisolone acetate, indicating that the 30 mg / kg M1 treatment group... M1 exerted a significant inhibitory effect on inflammatory factors in an LPS-induced cytokine storm model, thereby alleviating cardiac and lung damage and playing a protective role, showing potential for the treatment of cytokine storms and infectious myocarditis.

[0171] The applicant declares that this application illustrates the process method through the above embodiments, but this application is not limited to the above process steps, that is, it does not mean that this application must rely on the above process steps to be implemented. Those skilled in the art should understand that any improvements to this application, equivalent substitutions of the raw materials used in this application, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this application.

Claims

A phenylacrylamide compound having the structure shown in Formula I: ； in, R1, R4, R5, and R6 are each independently selected from any one of hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1-C6 alkoxycarbonyl, C1-C6 alkoxycarbonyloxy, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxymethyleneoxy, C1-C6 alkylyl, trihaloC1-C6 alkyl, or trihaloC1-C6 alkoxy; or, R5 and R6 are independently selected from -O(CH2). n O-, and linked to its substituted phenyl group to form a ring, where n is selected from 1, 2 or 3; R2 and R3 are each independently selected from any one of hydrogen, hydroxy-C0-C6 alkyl, C1-C6 alkylamino, cyano, nitro, C1-C6 alkoxycarbonyl, methanesulfonyl, dimethylaminesulfonyl, amino, C1-C6 alkyl, C1-C6 alkoxy, trihaloC1-C6 alkyl, or trihaloC1-C6 alkoxy. X is selected from CH2, CO, or CH2CONH. According to claim 1, the phenylacrylamide compound, wherein, R1, R4, R5, and R6 are each independently selected from hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1 alkoxycarbonyl, C2 alkoxycarbonyl, C1 alkoxycarbonyloxy, C2 alkoxycarbonyloxy, C1 alkyl, C2 alkyl, C1 alkoxy, C2 alkoxy, C1 alkoxymethyleneoxy, C2 alkoxymethyleneoxy, C1 alkyl, C2 alkyl, trihaloC1 alkyl, trihaloC2 alkyl, or trihaloC1 alkoxy, trihaloC2 alkoxy; or, R5 and R6 are independently selected from -O(CH2). n O-, and linked to its substituted phenyl group to form a ring, where n is selected from 1, 2 or 3; R2 and R3 are each independently selected from any one of hydrogen, hydroxyl, hydroxy-C1 alkyl, C1 alkylamino, C2 alkylamino, cyano, nitro, C1 alkoxycarbonyl, C2 alkoxycarbonyl, methanesulfonyl, dimethylaminesulfonyl, amino, C1, C2 alkyl, C1, C2 alkoxy, trihaloC1 alkyl, trihaloC2 alkyl, or trihaloC1 alkoxy, trihaloC2 alkoxy. According to claim 1 or 2, the phenylacrylamide compound, wherein, R1, R4, R5, and R6 are selected from any one of hydrogen, fluorine, chlorine, bromine, hydroxyl, dimethylamino, cyano, nitro, methoxycarbonyl, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, acetoxy, or methoxymethyleneoxy; or R5 and R6 are selected from -O(CH2). n 'O-, and linked to its substituted phenyl group to form a ring; n' is selected from 1, 2 or 3; R2 and R3 are each independently selected from any one of hydrogen, hydroxyl, hydroxymethyl, dimethylamino, cyano, nitro, methoxycarbonyl, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, or trifluoromethoxy. X is selected from CH2, CO, or CH2CONH. The phenylacrylamide compound according to any one of claims 1-3, wherein, The phenylacrylamide compound is selected from any one of the following M1-M10: 。 A tautomer of a phenylacrylamide compound according to any one of claims 1-4, or a pharmaceutically acceptable salt thereof; Preferably, the pharmaceutically acceptable salt includes any one or a combination of at least two of the following: hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, p-toluenesulfonate, acetate, trifluoroacetate, salicylate, amino acid salt, citrate, maleate, tartrate, fumarate, citrate, lactate, sodium salt, potassium salt, calcium salt, magnesium salt, lithium salt, ammonium salt, or salt of an organic base that can provide a physiologically acceptable cation; Preferably, the salt of the organic base that provides a physiologically acceptable cation includes any one or a combination of at least two of the following: methylamine salt, dimethylamine salt, trimethylamine salt, piperidine salt, morpholine salt, or tri(2-hydroxyethyl)amine salt. A method for preparing a phenylacrylamide compound according to any one of claims 1-4, comprising the following steps: (1) The compound shown in formula a is reacted with the compound shown in formula b to obtain the compound shown in formula c. The reaction formula is shown below: ； (2) The compound shown in formula c is deprotected to obtain the compound shown in formula d, and the reaction formula is shown below: ； (3) Reacting the compound shown in formula d with the compound shown in formula e yields the compound shown in formula I, as shown in the following reaction formula: ； Wherein, R' is selected from any one of tert-butyldimethylsiloxy, tert-butyldiphenylsiloxy, trimethylsiloxy, triethylsiloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy; R1' is selected from any one of acetyl, tert-butyldimethylsiloxy, tert-butyldiphenylsiloxy, trimethylsiloxy, triethylsiloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy. X' is selected from CHO, TsOCH2, COOH, or BrCH2CONH; X is selected from CH2, CO, or CH2CONH; Y is selected from chlorine, methoxy, or hydroxyl. According to the preparation method of claim 6, wherein, In step (1), the reaction is carried out under alkaline conditions; Preferably, in step (1), the reaction is carried out in the presence of a condensing agent, which includes EDCI and / or DMAP; Preferably, in step (1), the reaction temperature is 0-50℃ and the reaction time is 2-15 h; Preferably, in step (1), the molar ratio of the compound shown in formula a to the compound shown in formula b is 1:(0.8-1.5); Preferably, in step (2), the deprotection is carried out by hydrolysis or hydrogenation. Preferably, in step (2), the deprotection is carried out by a hydrolysis reaction, which is carried out under acidic or alkaline conditions; Preferably, in step (2), the temperature of the deprotection reaction is 0-50℃ and the reaction time is 1-24 h; Preferably, in step (2), the deprotection reaction is carried out in a solvent, wherein the solvent is selected from tetrahydrofuran and / or methanol; Preferably, in step (3), the reaction is carried out under alkaline conditions; Preferably, in step (3), the reaction is carried out in the presence of a condensing agent, which includes EDCI and / or DMAP; Preferably, in step (3), the molar ratio of the compound shown in formula d to the compound shown in formula e is 1:(0.8-2); Preferably, in step (3), the reaction temperature is 0-50℃ and the reaction time is 1-15 h. A pharmaceutical composition comprising an active ingredient and a pharmacodynamically acceptable carrier, said active ingredient comprising a phenylacrylic acid compound of any one of claims 1-4 or a tautomer of a phenylacrylamide compound of claim 5 or a pharmaceutically acceptable salt thereof; Preferably, the active ingredient in the pharmaceutical composition has a mass percentage content of 0.1-95%. The use of a phenylacrylamide compound according to any one of claims 1-4, a tautomer of the phenylacrylamide compound according to claim 5, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8 in the preparation of a drug for the prevention or treatment of tumors, cancer pain, radiotherapy and chemotherapy damage, autoimmune diseases, infections and drug-induced myocarditis, cytokine storms, ischemic heart disease, inflammatory diseases, neurodegenerative diseases, or anti-aging. According to the application of claim 9, wherein, The tumor is selected from any one or a combination of at least two of the following: glioma, melanoma, gastric cancer, lung cancer, breast cancer, kidney cancer, liver cancer, oral epithelial cancer, head and neck tumors, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, rectal adenocarcinoma, leukemia, or lymphoma. Preferably, the autoimmune disease includes any one or a combination of at least two of the following: rheumatoid arthritis, systemic lupus erythematosus, ulcerative colitis, psoriasis, Sjögren's syndrome, dermatomyositis, or amyotrophic lateral sclerosis. Preferably, the inflammatory disease includes any one or a combination of at least two of the following: infectious myocarditis caused by bacteria or viruses, cytokine storm, non-infectious polyarteritis, phlebitis, and reflux esophagitis; Preferably, the neurodegenerative disease includes Alzheimer's disease and / or Parkinson's disease.

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