Phenyl acrylamide compound as well as preparation method and application thereof

Abstract

The invention provides a phenylacrylamide compound and a preparation method and application thereof, the phenylacrylamide compound has a structure as shown in a formula I. The phenylacrylamide compound provided by the invention can be well combined with a plurality of target proteins, especially has good inhibitory activity on ACAT1 and 3-KAT targets, and can be used for preparing a medicine for treating cancer. Moreover, the compound has better bioavailability, and can be used for preparing drugs for preventing or treating tumors, cancer pain, radiotherapy and chemotherapy injury, autoimmune diseases, infection and drug-induced myocarditis, cytokine storm, ischemic heart disease, inflammatory diseases and neurodegenerative diseases or anti-aging drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and in particular relates to a phenylacrylamide compound and a preparation method and application thereof. Background Art

[0002] Medical data show that inflammation is closely related to the occurrence and development of tumors. For example, infection with Helicobacter pylori tends to increase the risk of gastric cancer; chronic hepatitis may also be a direct cause of liver cancer. Other factors, such as autoimmune intestinal disease, are closely related to colon cancer, and fine particulate matter in the air is also a black hand in inducing lung cancer. Tumors are defined as a non-infectious disease like heart disease, chronic respiratory disease or diabetes. Most of them are long-term chronic diseases that progress relatively slowly. 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] Phenyl acrylamide compounds are widely present in active natural products, especially natural products with hydroxyl substitution on the benzene ring, which have multiple biological activities, such as antibacterial, antioxidant, anti-inflammatory, anti-tumor, etc., and have very broad research prospects. Structural modification of such natural products will help to discover lead compounds with stronger tumor inhibitory activity, activity in treating autoimmune diseases and various infectious and non-infectious inflammatory diseases, and lower toxicity than the original natural products, and further become anti-tumor, neurodegenerative disease and anti-inflammatory immune drugs.

[0004] ACAT1 belongs to the superfamily of specific thiolase, and its Chinese name is acetyl CoA sulfidase, also known as acetyl CoA acetyltransferase (ACAT). Many reports show that ACAT1 expression in tumor cells is usually abnormal and plays a vital role in the occurrence and development of tumors. The upregulation mechanism of ACAT1 activity in different human cancer cells is worth studying. High expression of ACAT1 reduces its overall survival rate; ACAT1 overexpression in MDA-MB-231 human breast cancer cells leads to ketone body recycling, which drives tumor progression and metastasis. Therefore, the study of phenylacrylamide derivatives targeting ACAT1 may discover new anti-tumor drugs.

[0005] 3-KAT, also known as 3-ketoacyl-CoA thiolase, has the main biological function of catalyzing the complete cleavage of medium-chain and short-chain fatty acids into acetyl CoA in mitochondria; 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 chain is shortened, and the product enters the mitochondria for further oxidation. Inhibiting 3-KAT can also inhibit the growth of tumors, especially melanoma and glioma.

[0006] In addition, natural products based on this type of structure have multi-target characteristics, and other possible targets are summarized as follows:

[0007] Caspase-3 is a caspase protein that interacts with caspase-8 and caspase-9. It is encoded by the casp3 gene. Increased levels of caspase-3, p17 fragments in the blood are a sign of recent myocardial infarction. Studies have shown that caspase-3 may play an important role in embryonic and hematopoietic stem cell differentiation and apoptosis. Studies have shown that the expression of caspase-3 is abnormally increased in patients with head and neck cancer and breast cancer. Therefore, reducing its expression is one of the ways to reduce the occurrence and development of cancer to a certain extent.

[0008] Epidermal growth factor receptor (EGFR): The mutation and activation of EGFR is an important factor leading to abnormal biological activities of tumor cells. The T790M mutation in EGFR is a base pair change from cytosine (C) to thymine (T), that is, the threonine at position 790 in the EGFR tyrosine kinase function is replaced by methionine. This mutation can reactivate EGFR, thereby leading to resistance to tyrosine kinase inhibitors (TKI). 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 T790M mutations.

[0009] Histone deacetylase (HDAC): A class of proteases that play an important role in the structural modification of chromosomes and the regulation of gene expression. In general, the acetylation of histones is conducive to the dissociation of DNA from the histone octamer and the relaxation of the nucleosome structure, so that various transcription factors and co-transcription factors can bind specifically to the DNA binding site and activate gene transcription. In the cell nucleus, histone acetylation and histone deacetylation processes are in a dynamic equilibrium and are jointly regulated by histone acetyltransferase (HAT) and histone deacetylase (HDAC).

[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 to encode different isoforms; gene sequence changes are fundamentally different from epigenetic modification abnormalities. Because once the DNA sequence mutates, 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 related chromatin modification enzymes. Therefore, it is very important to clarify the mechanism of action of epigenetic modification enzymes in tumor cells, and then provide corresponding treatment means to prevent epigenetic modification mutations. At present, there are no EZH2 inhibitors on the market, and a total of five drugs are in clinical phase I / II research.

[0011] Src kinase: 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 cell apoptosis. The research on Src inhibitors has become a hot topic in the research of anti-tumor drugs. Currently, a series 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 many immune checkpoints are treasures waiting to be discovered. Since the efficacy of CTLA-4 and PD-1 / PD-L1 monoclonal antibodies has been confirmed, there has been fierce competition in this monoclonal antibody track. Therefore, more attention should be paid to other immune checkpoint targets, whose functions determine the efficacy of immunotherapy.

[0013] The survival and development of T cells are affected by TCR signals, and the TCR signaling pathway depends on Src family kinases (SFKs). Lck is an important member of SFKs and is expressed throughout most of the life cycle of T cells. In addition, Lck plays an important role in activating the TCR signaling pathway to activate T cells. CSK is a key regulator of SFKs, and its phosphorylation of Lck (Tyr505) inactivates Lck, which inhibits T cell activation through TCR. Therefore, CSK and p-Lck (Tyr505) may be effective targets for future immunomodulatory therapies.

[0014] CD73 is a 5-major nucleotide hydrolase that can hydrolyze extracellular adenosine monophosphate (AMP) into adenosine, a powerful immunosuppressive molecule that can inhibit the activation of CD8-positive T cells, thereby helping cancer cells escape the "hunt" of T cells. NK cells infiltrating tumors upregulate CD73 expression, and the frequency of these CD73+NK cells is related to the size of tumors in breast cancer patients. The results support that tumors can hijack NK cells for immune evasion, and CD73 expression defines an inducible NK cell population with immunomodulatory properties in the tumor microenvironment.

[0015] KIR, 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 class molecules on the cell surface, thereby exerting immune regulatory function.

[0016] LAG-3, lymphocyte activation 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-II class molecules. It has the function of maintaining the stability of the internal environment 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 ligand are another important co-stimulatory molecule besides the CD28 / B7 co-stimulatory signaling pathway.

[0018] In terms of antioxidant and anti-inflammatory immunity, it has been associated with:

[0019] Since the phenylacrylamide structure is widely present in the secondary metabolites of polyphenols in plants, it has strong antioxidant and anti-inflammatory properties. Its activity in scavenging 1,1-diphenyl-2-trinitrophenylhydrazine free radicals is 2 to 3 times that of vitamin C and E, and its activity in scavenging superoxide anion free radicals is 10 to 30 times that of vitamin C and E. In terms of anti-inflammatory effects, it has anti-inflammatory activity in the carrageenan-induced rat plantar swelling inflammation model. Its mechanism can inhibit 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 playing an anti-inflammatory role. It inhibits the activation of the NF-κB inflammatory pathway caused by bacterial or viral infection and the massive production and release of inflammatory factors such as interleukin-1, TNF-a, and interleukin-6 caused by p65 acetylation, thereby playing an anti-inflammatory role in the treatment of infectious myocarditis and cytokine storm.

[0020] Pathological studies of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease have shown that chronic oxidative stress and inflammatory responses can lead to neuronal damage. Based on the strong antioxidant and anti-inflammatory effects of phenylacrylic acid, it was found to have a good protective effect on the nervous system. Some clinical and preclinical studies have shown that phenylacrylamide natural products have a good therapeutic effect on Alzheimer's disease and Parkinson's disease. It can improve memory loss and hippocampal cell death after transient global ischemia. It prevents saturated free fatty acid-induced lipotoxicity by activating SIRT1-regulated mitochondrial function. Its mechanism is to reduce oxidative stress and mitochondrial dysfunction by reducing the production of ROS and increasing mitochondrial mass and mitochondrial membrane potential; significantly reduce 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 wide range of active effects as mentioned above, their exact mechanism of action and structure-activity relationship require further study. It is necessary to synthesize their structural derivatives and study their drug action mode and the diseases that can be used to treat them, especially inflammation-related (infectious and non-infectious), tumors, autoimmune diseases, neurodegenerative diseases and aging. Summary of the invention

[0022] In view of the deficiencies of the prior art, the object of the present invention is to provide a phenylacrylamide compound and a preparation method and application thereof. The phenylacrylamide compound provided by the present invention can have good binding with multiple target proteins and has good bioavailability, and can be used to prepare drugs for preventing or treating 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 drugs.

[0023] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0024] In one aspect, the present invention provides a phenylacrylamide compound having a structure shown in the following formula I:

[0025]

[0026] Among them, R 1 , R 4 , R 5 , R 6each independently selected from hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkoxycarbonyl, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkylcarbonyloxy, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkyl, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) , C4, C5, C6, etc.) alkoxy, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkoxymethyleneoxy, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkanoyl, trihalogenated C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkyl or trihalogenated C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkoxy; or, R 5 , R 6 are independently selected from -O(CH 2 ) n O-, and connected to the phenyl group substituted by it to form a ring, and n is selected from 1, 2 or 3;

[0027] The R 2 , R 3 Each is independently selected from hydrogen, hydroxy-C0-C6 (for example, C0, C1, C2, C3, C4, C5, C6, etc.) alkyl, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkylamino, cyano, nitro, C1-C6 (for example, C1, C2, C3, C4, C5, C6, etc.) alkoxycarbonyl, methanesulfonyl, dimethylaminosulfonyl, amino, C1- Any one of 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;

[0028] X is selected from CH 2 , CO or CH 2 CONH.

[0029] The anti-tumor targets of the phenylacrylamide derivatives designed in the present invention are mainly ACAT1 (acetyl CoA acetyltransferase 1) and 3-KAT (3-ketoacyl-CoA thiolase, 3-ketoacyl-CoA thiolase), and the therapeutic effect is achieved through dual inhibition.

[0030] In the present invention, the trihalogenated C1-C6 alkyl or trihalogenated C1-C6 alkoxy group refers to a group containing three halogens, including F 3 C1-C6 alkyl, Br 3 C1-C6 alkyl, Cl 3 C1-C6 alkyl, I 3 C1-C6 alkyl, F 2 Br C1-C6 alkyl, F 2 Cl C1-C6 alkyl, F 2 I C1-C6 alkyl, FBr 2 C1-C6 alkyl, FCl 2 C1-C6 alkyl, FI 2 C1-C6 alkyl, ClBr 2 C1-C6 alkyl, ICl 2 C1-C6 alkyl, ClI 2 C1-C6 alkyl; F 3 C1-C6 alkoxy, Br 3 C1-C6 alkoxy, Cl 3 C1-C6 alkoxy, I 3 C1-C6 alkoxy, F 2 Br C1-C6 alkoxy, F 2 Cl C1-C6 alkoxy, F 2 I C1-C6 alkoxy, FBr 2 C1-C6 alkoxy, FCl 2 C1-C6 alkoxy, FI 2 C1-C6 alkyl, ClBr 2 C1-C6 alkoxy, ICl 2 C1-C6 alkoxy, ClI 2 C1-C6 alkoxy.

[0031] Preferred F 3 C1-C4 alkyl, Br 3 C1-C4 alkyl, Cl 3 C1-C4 alkyl, I 3 C1-C4 alkyl, F 2 Br C1-C4 alkyl, F 2 Cl C1-C4 alkyl, F 2 I C1-C4 alkyl, FBr 2 C1-C4 alkyl, FCl 2 C1-C4 alkyl, FI 2 C1-C4 alkyl, ClBr 2 C1-C4 alkyl, ICl 2C1-C4 alkyl, ClI 2 C1-C4 alkyl; F 3 C1-C4 alkoxy, Br 3 C 1-4 Alkoxy, Cl 3 C1-C4 alkoxy, I 3 C1-C4 alkoxy, F 2 Br C1-C4 alkoxy, F 2 Cl C1-C4 alkoxy, F 2 I C1-C4 alkoxy, FBr 2 C1-C4 alkoxy, FCl 2 C1-C4 alkoxy, FI 2 C1-C4 alkyl, ClBr 2 C1-C4 alkoxy, ICl 2 C 1-4 Alkoxy, ClI 2 C1-C4 alkoxy.

[0032] The best choice is F 3 C. F 3 CCH 2 Br 3 C.Br 3 CCH 2 , Cl 3 C, Cl 3 CCH 2 ,I 3 C.I 3 CCH 2 、F 2 BrC、F 2 BCH 2 、F 2 ClC、F 2 CCH 2 、F 2 IC, F 2 ICCH 2 , FBr 2 C. FBr 2 CCH 2 、FCl 2 C、FCl 2 C CH 2 , FI 2 C.FI 2 CCH 2 , ClBr 2 C、ClBr 2 CCH 2 、ICl 2 C.ICl 2 CCH2 , ClI 2 C. ClI 2 CCH 2 ; F 3 CO, F 3 CCH 2 O, Br 3 CO、Br 3 CCH 2 O、Cl 3 CO、Cl 3 CCH 2 O.I 3 CO、I 3 CCH 2 O.F 2 BrCO、F 2 BCH 2 O.F 2 ClCO、F 2 CCH 2 O.F 2 ICO, F 2 ICCH 2 O、FBr 2 CO、FBr 2 CCH 2 O、FCl 2 CO、FCl 2 CCH 2 O.FI 2 CO、FI 2 CCH 2 O、ClBr 2 CO、ClBr 2 CCH 2 O、ICl 2 CO、ICl 2 CCH 2 O、ClI 2 CO or ClI 2 CCH 2 O.

[0033] In the present invention, the phenylacrylamide compound is selected from any one of the following formula I:

[0034]

[0035] Among them, R 1 , R 4 , R 5 , R 6each independently selected from hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1-C4 (for example, C1, C2, C3, C4, etc.) alkoxycarbonyl, C1-C4 (for example, C1, C2, C3, C4, etc.) alkylcarbonyloxy, C1-C4 (for example, C1, C2, C3, C4, etc.) alkyl, C1-C4 (for example, C1, C2, C3, C4, etc.) , C2, C3, C4, etc.) alkoxy, C1-C4 (for example, C1, C2, C3, C4, etc.) alkoxymethyleneoxy, C1-C4 (for example, C1, C2, C3, C4, etc.) alkanoyl, trihalogenated C1-C4 (for example, C1, C2, C3, C4, etc.) alkyl or trihalogenated C1-C4 (for example, C1, C2, C3, C4, etc.) alkoxy; or, R 5 , R 6 are independently selected from -O(CH 2 ) n O-, and connected to the phenyl group substituted by it to form a ring, and n is selected from 1, 2 or 3;

[0036] The R 2 , R 3 Each is independently selected from any one of hydrogen, hydroxyl C0-C4 (for example, C0, C1, C2, C3, C4, etc.) alkyl, C1-C4 (for example, C1, C2, C3, C4, etc.) alkylamino, cyano, nitro, C1-C4 (for example, C1, C2, C3, C4, etc.) alkoxycarbonyl, methylsulfonyl, dimethylaminosulfonyl, amino, C1-C4 (for example, C1, C2, C3, C4, etc.) alkyl, C1-C4 (for example, C1, C2, C3, C4, etc.) alkoxy, trihalogenated C1-C4 (for example, C1, C2, C3, C4, etc.) alkyl or trihalogenated C1-C4 (for example, C1, C2, C3, C4, etc.) alkoxy;

[0037] X is selected from CH 2 , CO or CH 2 CONH.

[0038] In the present invention, R 1 , R 4 , R 5 , R 6Each is independently selected from any one of hydrogen, halogen, hydroxyl, dimethylamino, cyano, nitro, methylamino, methanesulfonyl, dimethylaminosulfonyl, amino, carboxyl, C1 alkoxycarbonyl, C2 alkoxycarbonyl, C1 alkylcarbonyloxy, C2 alkylcarbonyloxy, C1 alkyl, C2 alkyl, C1 alkoxy, C2 alkoxy, C1 alkoxymethyleneoxy, C2 alkoxymethyleneoxy, C1 alkanoyl, C2 alkanoyl, trihaloC1 alkyl, trihaloC2 alkyl or trihaloC1 alkoxy, trihaloC2 alkoxy; or, R 5 , R 6 are independently selected from -O(CH 2 ) n O-, and connected to the phenyl group substituted by it to form a ring, and n is selected from 1, 2 or 3.

[0039] The R 2 , R 3 Each is independently selected from any one of hydrogen, hydroxyl, hydroxy-C1 alkyl, C1 alkylamino, C2 alkylamino, cyano, nitro, C1 alkoxycarbonyl, C2 alkoxycarbonyl, methanesulfonyl, dimethylaminosulfonyl, amino, C1, C2 alkyl, C1, C2 alkoxy, trihalogenated C1 alkyl, trihalogenated C2 alkyl or trihalogenated C1 alkoxy, trihalogenated C2 alkoxy.

[0040] In the present invention, the R 1 , R 4 , R 5 , R 6 Any one selected from hydrogen, fluorine, chlorine, bromine, hydroxyl, dimethylamino, cyano, nitro, methoxycarbonyl, methylamino, methylsulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, acetoxy or methoxymethyleneoxy; or R 5 and R 6 Selected from -O(CH 2 ) n 'O-, and connected to the phenyl group substituted by it to form a ring; n' is selected from 1, 2 or 3;

[0041] The R 2 , R 3 Each is independently selected from any one of hydrogen, hydroxy, hydroxymethyl, dimethylamino, cyano, nitro, methoxycarbonyl, methylamino, methylsulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl or trifluoromethoxy;

[0042] X is selected from CH 2 , CO or CH 2 CONH.

[0043] In the present invention, the phenyl acrylamide compound is selected from any one of the following M1-M10:

[0044]

[0045] In a second aspect, the present invention provides a tautomer of the phenylacrylamide compound according to the first aspect or a pharmaceutically acceptable salt thereof.

[0046] Preferably, the pharmaceutically acceptable salt includes any one or a combination of at least two of 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 a salt of an organic base that can provide a physiologically acceptable cation.

[0047] Preferably, the salt of the organic base capable of providing a physiologically acceptable cation includes any one of methylamine salt, dimethylamine salt, trimethylamine salt, piperidine salt, morpholine salt or tris(2-hydroxyethyl)amine salt, or a combination of at least two thereof.

[0048] In the present invention, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0049] If cis / trans isomers exist, the invention relates to both the cis form and the trans form as well as mixtures of these forms, and if a single isomer is desired it can be separated according to conventional methods or prepared by stereoselective synthesis.

[0050] All salts of the present invention can be prepared by conventional methods. In addition, during the preparation of the solvate of the compound represented by Formula I and its salt, polycrystals or co-crystals may appear under different crystallization conditions.

[0051] In a third aspect, the present invention provides a method for preparing the phenylacrylamide compound according to the first aspect, the preparation method comprising the following steps:

[0052] (1) reacting the compound represented by formula a with the compound represented by formula b (condensation reaction or substitution reaction) to obtain the compound represented by formula c. The reaction formula is as follows:

[0053]

[0054] (2) Deprotecting the compound represented by formula c to obtain the compound represented by formula d. The reaction formula is as follows:

[0055]

[0056] (3) reacting the compound represented by formula d with the compound represented by formula e to obtain the compound represented by formula I. The reaction formula is as follows:

[0057]

[0058] ; Wherein, the R' is selected from any one of tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, trimethylsilyloxy, triethylsilyloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy;

[0059] R 1 'Selected from any one of acetyl, tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, trimethylsilyloxy, triethylsilyloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy;

[0060] X' is selected from CHO, TsOCH 2 , COOH or BrCH 2 CONH;

[0061] X is selected from CH 2 , CO or CH 2 CONH;

[0062] Y is selected from chloro, methoxy or hydroxy.

[0063] In the present invention, in step (1), the reaction is carried out under alkaline conditions.

[0064] Preferably, in step (1), the reaction is carried out in the presence of a condensing agent, and the condensing agent comprises EDCI and / or DMAP.

[0065] Preferably, in step (1), the reaction temperature is 0-50°C (for example, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C or 50°C, etc.), and the reaction time is 2-15h (for example, 2h, 5h, 7h, 9h, 11h, 13h or 15h, etc.).

[0066] Preferably, in step (1), the molar ratio of the compound represented by formula a to the compound represented by 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.

[0067] Preferably, in step (2), the deprotection is carried out by a hydrolysis reaction or a hydrogenation reaction.

[0068] Preferably, in step (2), the deprotection is carried out by a hydrolysis reaction, and the hydrolysis reaction is carried out under acidic conditions or alkaline conditions.

[0069] Preferably, in step (2), the temperature of the deprotection reaction is 0-50°C (for example, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, etc.), and the reaction time is 1-24h (for example, 1h, 4h, 8h, 12h, 16h, 20h, 24h, etc.).

[0070] Preferably, in step (2), the deprotection reaction is carried out in a solvent, and the solvent is tetrahydrofuran and / or methanol.

[0071] Preferably, in step (3), the reaction is carried out under alkaline conditions.

[0072] Preferably, in step (3), the reaction is carried out in the presence of a condensing agent, and the condensing agent comprises EDCI and / or DMAP.

[0073] Preferably, in step (3), the molar ratio of the compound represented by formula d to the compound represented by 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.

[0074] Preferably, in step (3), the reaction temperature is 0-50°C (for example, it can be 0°C, 10°C, 20°C, 30°C, 40°C, 50°C, etc.), and the reaction time is 1-15h (for example, it can be 1h, 3h, 5h, 7h, 9h, 11h, 13h, 15h, etc.).

[0075] In a fourth aspect, the present invention 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.

[0076] 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% and the like.

[0077] The pharmaceutical composition provided by the present invention can be prepared according to methods known in the art, and can be prepared 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.

[0078] The phenylacrylamide compound described in the first aspect of the present invention, the tautomer of the phenylacrylamide compound described in the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the fourth aspect can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0079] The dosage form can be a liquid dosage form, a solid dosage form or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w types, w / o types and multiple emulsions), suspensions, injections (including water injections, powder injections and infusions), eye drops, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), sprays, etc.; semisolid dosage forms can be ointments, gels, pastes, etc.

[0080] The phenylacrylamide compounds described in the first aspect of the present invention, the tautomers of the phenylpropionamide compounds described in the second aspect or their pharmaceutically acceptable salts, or the pharmaceutical composition described in the fourth aspect can be prepared into ordinary preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle delivery systems.

[0081] In order to prepare the phenylacrylamide compound described in the first aspect of the present invention, the tautomer of the phenylacrylamide compound described in the second aspect or the pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the fourth aspect of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, and glidants. The diluent may be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent may be water, ethanol, isopropanol, etc.; the binder may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0082] The tablets can be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

[0083] In order to prepare capsules from the phenylacrylamide compound described in the first aspect, the tautomer of the phenylacrylamide compound described in the second aspect or its pharmaceutically acceptable salt, or the pharmaceutical composition described in the fourth aspect, it can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. It can also be first prepared into granules or pellets with a diluent, a binder, and a disintegrant, and then placed in a hard capsule or a soft capsule. The diluents, binders, wetting agents, disintegrants, and glidants used to prepare tablets can also be used to prepare capsules of the compounds of the present invention.

[0084] In order to prepare the phenylacrylamide compound described in the first aspect of the present invention, the tautomer of the phenylacrylamide compound described in the second aspect or its pharmaceutically acceptable salt, or the pharmaceutical composition described in the fourth aspect into an injection, water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent and an appropriate amount of a solubilizer, a cosolvent, a pH adjuster, and an osmotic pressure regulator commonly used in the art can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH adjuster can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. If a lyophilized powder injection is prepared, mannitol, glucose, etc. can also be added as a support agent.

[0085] Furthermore, if necessary, colorants, preservatives, perfumes, flavoring agents or other additives may be added to the pharmaceutical preparations.

[0086] In order to achieve the purpose of medication and enhance the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method.

[0087] In a fifth aspect, the present invention provides a 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 preventing or treating 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.

[0088] Preferably, the tumor is selected from any one or a combination of at least two of 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.

[0089] Preferably, the autoimmune disease comprises any one of rheumatoid arthritis, systemic lupus erythematosus, ulcerative colitis, psoriasis, Sjögren's syndrome, dermatomyositis or spinal lateral sclerosis, or a combination of at least two thereof.

[0090] Preferably, the inflammatory disease includes any one or a combination of at least two of myocarditis, cytokine storm, non-infectious polyarteritis, phlebitis, and reflux esophagitis caused by infectious bacteria or viruses.

[0091] Preferably, the neurodegenerative disease includes Alzheimer's disease and / or Parkinson's disease.

[0092] The dosage of the phenylacrylamide compound of the first aspect of the present invention, the tautomer of the phenylacrylamide compound of the second aspect or its pharmaceutically acceptable salt or the pharmaceutical composition of the fourth aspect may vary widely according to the nature and severity of the disease to be prevented or treated, the individual conditions of the patient or animal, the route of administration and the dosage form. Generally speaking, the suitable daily dosage range of the phenylacrylamide compound of the first aspect of the present invention, the tautomer of the phenylacrylamide compound of the second aspect or its pharmaceutically acceptable salt or the pharmaceutical composition of 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, 0.6 mg / Kg body weight, 0.7 mg / Kg body weight, 0.8 mg / Kg body weight, 0.9 mg / Kg body weight, 10 mg / Kg body weight, 20 mg / Kg body weight, 30 mg / Kg body weight, 40 mg / Kg body weight, 50 mg / Kg body weight, 60 mg / Kg body weight, 70 mg / Kg body weight, 80 mg / Kg body weight, 90 mg / Kg body weight, 100 mg / Kg body weight, 150 mg / Kg body weight, 100 mg / Kg body weight, 150 mg / Kg body weight, 200 mg / Kg body weight, 300 mg / Kg body weight, 400 mg / Kg body weight, 50 mg / Kg body weight, 60 mg / Kg body weight, 70 mg / Kg body weight, 80 mg / Kg body weight, 90 mg / K The dosage may be in the range of 1-60 mg / kg body weight, 0.1-100 mg / kg body weight, 1-70 mg / kg body weight, and 2-60 mg / kg body weight. The dosage may be in the range of 1-60 mg / kg body weight, 0.1-100 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 dosage may be in the range of 1-60 mg / kg body weight, 0.1-100 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.

[0093] The phenylacrylamide compound of the first aspect of the present invention, the tautomer of the phenylacrylamide compound of the second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the fourth aspect can be taken alone or used in combination with other therapeutic drugs or symptomatic drugs. When other therapeutic drugs have synergistic effects, their dosages should be adjusted according to actual conditions.

[0094] Compared with the prior art, the present invention has the following beneficial effects:

[0095] The phenylacrylamide compounds provided by the present invention have good binding to multiple target proteins, especially have good inhibitory activity on ACAT1 (acetyl CoA acetyltransferase 1) and 3-KAT (3-ketoacyl-CoA thiolase, 3-ketoacyl coenzyme A thiolase) targets, and have good bioavailability, and can be used to prepare drugs for preventing or treating 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 drugs, wherein the tumor diseases are gliomas, melanomas, 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. BRIEF DESCRIPTION OF THE DRAWINGS

[0096] Figure 1 This is a test result of the effect of M1 compound on the body weight of LPS-induced cytokine storm model mice;

[0097] Figure 2A This is a test result diagram of the effect of M1 compound on the cardiac index of LPS-induced cytokine storm model mice;

[0098] Figure 2B This is a test result diagram of the effect of M1 compound on the spleen index of LPS-induced cytokine storm model mice;

[0099] Figure 2C This is a test result diagram of the effect of M1 compound on the lung index of LPS-induced cytokine storm model mice;

[0100] Figure 3 This is a test result diagram of the effect of M1 compound on the cytokine content in the LPS-induced mouse cytokine storm model. DETAILED DESCRIPTION

[0101] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0102] The reaction materials in the following examples are commercially available conventional products or known compounds reported in the literature.

[0103] Example 1

[0104] This embodiment provides a compound M1, and the preparation method of the compound M1 comprises the following steps:

[0105] 1.5g a-1 (1.05eq), 1.8g b-1 (1eq), 1.6g EDCI (1.5eq), 9mL dichloromethane were added to a 50mL single-mouth bottle, and 1.05g DIPEA (1.5eq) was slowly added dropwise. After the addition was completed, the reaction was allowed to react overnight at room temperature. TLC (DCM / MeOH volume ratio = 20 / 1) showed that the reaction was complete, and 10mL saturated sodium bicarbonate was added to quench the reaction, and then dichloromethane was added to extract. The organic phase was dried over anhydrous sodium sulfate, then mixed with silica gel and dried by column chromatography (DCM / MeOH = 20 / 1) to obtain 2g of the intermediate, and then 0.8g NaHCO was added to 50ml methanol. 3 The mixture was reacted at 50°C for 1.5 h, and then dried by spin drying. 1.2 g of the product was obtained by preparative liquid separation with a yield of 50%.

[0106]

[0107] Chemical formula: C 23 H 28 N 2 O 6， Calculated value: 428.19, [M+H] + Test value 429.70.

[0108] Example 2

[0109] This example provides compound M2, using compound a-1 150mg (1.00eq) and compound b-2 193mg (1.05eq) as raw materials, condensation agent EDCI 163mg (1.50eq), organic base DIPEA 110mg (1.50eq). The preparation method of compound M2 is the same as compound M1, and compound M2 is obtained as a yellow solid 100mg, with a yield of 42.7%. The reaction formula of compound M2 is as follows:

[0110]

[0111] Chemical formula: C 22 H 24 N 2 O 6 Calculated value: 412.16, [M+H] + Test value 413.55.

[0112] Example 3

[0113] This example provides compound M3, using compound a-1 (150 mg, 1.00 eq) and compound b-3 (201 mg, 1.05 eq) as raw materials, condensation agent EDCI 163 mg (1.50 eq), organic base DIPEA 110 mg (1.50 eq). The preparation method of compound M3 is the same as compound M1, and compound M3 is obtained as a yellow solid 156 mg, with a yield of 64.2%. The reaction formula of compound M3 is as follows:

[0114]

[0115] Chemical formula: C 23 H 28 N 2 O 6 Calculated value: 428.19, [M+H] + Test value 429.65.

[0116] Example 4

[0117] This embodiment provides a compound M4, wherein the preparation method of the compound M4 comprises the following steps:

[0118] Add 5 mL of MeCN into a 25 mL three-necked flask, add 720 mg of piperazine (trimetazidine hydrochloride, 2.12 mmol) and 1.1 mL of DIPEA (2.12 mmol) in sequence under stirring, and lower the internal temperature by 0-5°C; put the diluted solution of acyl chloride in acetonitrile (0.54 g, 2.12 mmol) into a constant pressure dropping funnel, and slowly drip it into the reaction solution, controlling the internal temperature not higher than 20°C; after the addition is complete, react at room temperature for 2 h. After the reaction is completed, the temperature is lowered by 5-10°C and water (5mL) is slowly dripped in to quench the reaction. Ethyl acetate (30mL) is added to dilute the reaction solution, and then water is added to stir the reaction solution, extract and separate the liquids. The aqueous phase obtained for the first time is extracted once more with ethyl acetate (20mL), the organic phases are combined, and the organic phases are washed with 10% brine, separated, added with anhydrous sodium sulfate to dry, filtered, and concentrated; methanol (60mL) is added to evaporate the solvent, and then methanol (8mL) is added, and the mixture is beaten in an oil bath at 55°C for 1h, and the temperature is lowered to 15-20°C within 2h for crystallization for 3h, filtered, and the filter cake is washed with a small amount of methanol until the filtrate is light yellow; the filter cake is collected and dried at 50°C to constant weight to obtain 1.0g of light yellow powdery solid, which is M4, with a yield of 97%.

[0119]

[0120] Chemical formula: C 23 H 28 N 2 O 6 Calculated value: 485.23, [M+H] + Test value 485.23.

[0121] Example 5

[0122] This embodiment provides a compound M5, and the preparation method of the compound M5 comprises the following steps:

[0123] 0.9g M4 (1.86mmol) was dissolved in 10ml methanol, and 465mg potassium bicarbonate (4.65mmol, 2.5eq) was added, and the mixture was reacted at room temperature for 2h. After the reaction was completed, water was added to terminate the reaction, and the mixture was extracted with EA and concentrated to obtain 0.6g M5 with a yield of 73.2%.

[0124]

[0125] Chemical formula: C 23 H 28 N 2 O 6 Calculated value: 443.22, [M+H] + Test value 443.22.

[0126] Example 6

[0127] This example provides compound M6, with compound a-1 75mg (1.00eq, 0.28mmol) and compound b-4 110mg (1.05eq) as raw materials, condensation agent EDCI 82mg (1.50eq), organic base DIPEA 55mg (1.50eq). The preparation method of compound M6 is the same as compound M1, and compound M6 is obtained as a yellow solid 93mg, with a yield of 71.5%. The reaction formula of compound M6 is shown below:

[0128]

[0129] Chemical formula: C 24 H 30 N 2 O 7 Calculated value: 458.21, [M+H] + Test value 459.80.

[0130] Example 7

[0131] This example provides compound M7, with compound a-1 50mg (1.00eq, 0.19mmol) and compound b-572mg (1.05eq) as raw materials, condensation agent EDCI 72mg (1.50eq), organic base DIPEA 97mg (4.0eq). The preparation method of compound M7 is the same as compound M1, and compound M7 is obtained as a yellow solid 30mg, with a yield of 33.7%. The reaction formula of compound M7 is as follows:

[0132]

[0133] Chemical formula: C 24 H 29 N 3 O 7 , calculated value: 471.20, [M+H] + Test value 472.40.

[0134] Example 8

[0135] This example provides compound M8, using compound a-3 50mg (1.00eq) and compound b-5 98mg (1.05eq) as raw materials, condensation agent EDCI 98mg (2.0eq), organic base DIPEA 133mg (4.0eq). The preparation method of compound M8 is the same as compound M1, and compound M8 is obtained as a yellow solid 50mg, with a yield of 40%. The reaction formula of compound M8 is shown below:

[0136]

[0137] Chemical formula: C 25 H 31 N 3 O 7 ,calculated value:485.22,[M+H] + Test value 486.50.

[0138] Example 9

[0139] This example provides compound M9, using compound a-4 50mg (1.00eq) and compound b-5 98mg (1.05eq) as raw materials, condensation agent EDCI 98mg (2.0eq), organic base DIPEA 133mg (4.0eq). The preparation method of compound M9 is the same as compound M1, and compound M9 is obtained as a yellow solid 65mg, with a yield of 52%. The reaction formula of compound M9 is shown below:

[0140]

[0141] Chemical formula: C 25 H 31 N 3 O 7 ,calculated value:485.22,[M+H] + Test value 486.35.

[0142] Example 10

[0143] This example provides compound M10, with compound a-1 50mg (1.00eq) and compound b-6 72mg (1.05eq) as raw materials, condensation agent EDCI 72mg (2.0eq), organic base DIPEA 97mg (4.0eq). The preparation method of compound M10 is the same as compound M1, and compound M9 is obtained as a yellow solid 65mg, with a yield of 52%. The reaction formula of compound M10 is as follows:

[0144]

[0145] Chemical formula: C 24 H 29 N 3 O 7 Calculated value: 471.20, [M+H] + Test value 472.80.

[0146] Pharmacological experiments

[0147] Experimental Example 1

[0148] IC of phenylacrylamide derivatives against ACAT1 and 3-KAT targets 50 Determination

[0149] 1. Experimental methods

[0150] Screening methods for small molecule inhibitors of ACAT1

[0151] ACAT1 can catalyze the reversible formation of acetoacetyl-CoA from two molecules of acetyl-CoA. Its activity is measured by ACAT1 catalyzing the reaction of substrate acetoacetyl-CoA with CoA to generate product acetyl-CoA. Since acetoacetyl-CoA has specific absorption in a specific spectrum, the effect on the ACAT1 enzyme activity is reflected by detecting the increase or decrease in absorption of the specific spectrum.

[0152] The ACAT1 recombinant protein was expressed and purified by E. coli, and the concentration of the obtained ACAT1 recombinant protein was 1 mg / mL. When screening for ACAT1 small molecule inhibitors, the buffer used was 50 mM Tris-HCl (pH 8.1), 20 mM MgCl 2 , 40mM KCl. In the 200μl enzyme catalysis system, 1μL of ACAT1 recombinant protein, the final concentration of substrate acetyl-CoA was 25μM, the CPM probe was added to a final concentration of 100μM, and small molecule inhibitors of different concentrations were added. The detection was performed by using an ELISA reader (Biotek Synergy H1) with an excitation light of 355nm and an emission light of 460nm, and the inhibitor IC was performed using the software Prism 7.0. 50 Calculation.

[0153] Screening methods for small molecule inhibitors of 3-KAT

[0154] Ketoacyl-CoA thiolase uses fatty acids of various chain lengths as substrates, catalyzing the final reaction of fatty acid β-oxidation, releasing acetyl-CoA and producing acyl-CoA that is short of two carbon groups. In eukaryotes, ketoacyl-CoA thiolase is divided into mitochondrial type and peroxisomal type. Under the catalysis of long-chain thiolase I, long-chain ketoacyl-CoA (e.g., ketohexadecanoyl-CoA; 3-ketohexadecanoyl-CoA) undergoes cleavage of magnesium-enolate, resulting in a reduction in the long-chain ketoacyl-CoA substrate. Under a spectrophotometer (303nm), the absorbance value decreases, which is used to quantitatively analyze the activity of long-chain ketoacyl-CoA thiolase.

[0155] 10 μg of mitochondrial protein from B16F10 cells was extracted, 50 μM Palmityl CoA was added, and 2 mM NAD was added after reaction at 37°C for 15 min. The reaction was continued at 37°C for 15 min to obtain 3-KAT protein. When screening for 3-KAT small molecule inhibitors, the buffer used was 50 mM Tris-HCl (pH 8.1), 20 mM MgCl 2 , 40mM KCl. In the 200μL enzyme catalytic system, 90μL of 3-KAT recombinant protein, the final concentration of substrate coenzyme A was 25μM, and small molecule inhibitors of different concentrations were added. The spectrophotometer (Flex A-200) was used for detection with an absorbance of 303nm, and the software Prism7.0 was used for inhibitor IC 50 Calculation.

[0156] (II) Experimental results

[0157] The results of the screening for the effects on ACAT1 and 3-KAT activities in vitro are shown in Table 1 .

[0158] Table 1

[0159]

[0160]

[0161] From the data in Table 1 above, it can be seen that 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.

[0162] Experimental Example 2

[0163] Pharmacodynamic study of M1 on LPS-induced cytokine storm model in mice

[0164] 1. Experimental methods

[0165] Pharmacodynamics study methods in the LPS-induced cytokine storm model in mice

[0166] A total of 70 male Balb / c mice were randomly divided into solvent control group, model group (LPS, 10 mg / kg, ip), positive drug control group Tocilizumab (targeting IL-6) monoclonal antibody group (Tocilizumab, 5 mg / kg, iv), prednisone acetate group (Prednisone, 5 mg / kg, po, qd), M1 intravenous injection group (3 mg / kg, iv, qd), M1 low-dose group (30 mg / kg, po, qd), M1 high-dose group (60 mg / kg, po, qd), 10 mice in each group. Except for the Tocilizumab group, the drug-treated group started to be treated on day 0 for 7 consecutive days. The Tocilizumab group was treated 1 hour before modeling. One hour after the last administration, 100 μL of LPS solution was injected into each mouse by intraperitoneal injection. Except for the animals in the solvent control group, the animals in the other model groups and drug-treated groups were modeled, with a total of 60 models. The animals were weighed every day, and 24 hours after modeling, the animals were processed and the organ index and cytokine levels (48 in total) were tested.

[0167] (II) Experimental results

[0168] The effect of M1 compound on the body weight of LPS-induced cytokine storm model mice is shown in Figure 1 .

[0169] Depend on Figure 1 It can be seen that the body weights of mice in the two oral administration groups of M1 were better than those in the model group, indicating that M1 has a significant protective effect on LPS-induced cytokine storm model mice.

[0170] The effects of M1 compound on organ indexes of LPS-induced cytokine storm model mice are shown in Figure 2A-2C .

[0171] Depend on Figure 2A-2C It can be seen that the 30 mg / kg M1 administration group can significantly reduce the mouse lung index, and the heart index is significantly lower than that of the model group, indicating that 30 mg / kg M1 can significantly reduce the mouse heart and lung inflammatory organ index caused by LPS-induced cytokine storm, and has a significant anti-inflammatory cytokine storm effect on heart and lung damage.

[0172] The effects of M1 compound on cytokine content in LPS-induced mouse cytokine storm model are shown in Figure 3 .

[0173] Depend on Figure 3 It can be seen that 48 cytokines were detected in the LPS-induced mouse cytokine storm model, of which 30 mg / kg The levels of 27 cytokines in the M1-treated group, 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, Rankl, ST2 (IL-33R), IL-7, IL-25, BTC, BAFF, Leptin, IL-7R, IL-2RA, and IL-3, were significantly lower than those in the model group at 2 h or 24 h. The M1-treated group was superior to the positive control drug prednisone acetate in reducing the levels of some cytokines, indicating that 30 mg / kg M1 plays a significant role in inhibiting inflammatory factors in the LPS-induced cytokine storm model, thereby reducing heart and lung damage and playing a protective role, and has the potential to be used in the treatment of cytokine storm and infectious myocarditis.

[0174] The applicant declares that the present invention illustrates the process method of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned process steps, that is, it does not mean that the present invention must rely on the above-mentioned process steps to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials selected by the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A phenylacrylamide compound, characterized in that: The phenylacrylamide compound has a structure shown in the following formula I: Wherein, 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)alkylcarbonyloxy, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxymethyleneoxy, C1-C6 alkanoyl, trihalogenated C1-C6 alkyl or trihalogenated C1-C6 alkoxy; or, R5 and R6 are independently selected from -O(CH2) n O-, and connected to the phenyl group substituted by it to form a ring, and 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, dimethylaminosulfonyl, amino, C1-C6 alkyl, C1-C6 alkoxy, trihalogenated C1-C6 alkyl or trihalogenated C1-C6 alkoxy; X is selected from CH2, CO or CH2CONH.

2. The phenylacrylamide compound according to claim 1, characterized in that 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 alkylcarbonyloxy, C2 alkylcarbonyloxy, C1 alkyl, C2 alkyl, C1 alkoxy, C2 alkoxy, C1 alkoxymethyleneoxy, C2 alkoxymethyleneoxy, C1 alkanoyl, C2 alkanoyl, trihaloC1 alkyl, trihaloC2 alkyl or trihaloC1 alkoxy, trihaloC2 alkoxy; or, R5 and R6 are independently selected from -O(CH2) n O-, and connected to the phenyl group substituted by it to form a ring, and n is selected from 1, 2 or 3; The 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, dimethylaminosulfonyl, amino, C1, C2 alkyl, C1, C2 alkoxy, trihalogenated C1 alkyl, trihalogenated C2 alkyl or trihalogenated C1 alkoxy, trihalogenated C2 alkoxy.

3. The phenylacrylamide compound according to claim 1 or 2, characterized in that: The 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 connected to the phenyl group substituted by it 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, methylsulfonyl, dimethylaminosulfonyl, amino, methyl, ethyl, methoxy, ethoxy, trifluoromethyl or trifluoromethoxy; X is selected from CH2, CO or CH2CONH.

4. The phenylacrylamide compound according to any one of claims 1 to 3, characterized in that: The phenyl acrylamide compound is selected from any one of the following M1-M10:

5. A tautomer of a phenylacrylamide compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof; Preferably, the pharmaceutically acceptable salt includes any one or a combination of at least two of 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 a salt of an organic base that can provide a physiologically acceptable cation; Preferably, the salt of the organic base capable of providing a physiologically acceptable cation comprises any one of methylamine salt, dimethylamine salt, trimethylamine salt, piperidine salt, morpholine salt or tris(2-hydroxyethyl)amine salt, or a combination of at least two thereof.

6. A method for preparing a phenylacrylamide compound according to any one of claims 1 to 4, characterized in that: The preparation method comprises the following steps: (1) reacting the compound represented by formula a with the compound represented by formula b to obtain the compound represented by formula c. The reaction formula is as follows: (2) Deprotecting the compound represented by formula c to obtain the compound represented by formula d. The reaction formula is as follows: (3) reacting the compound represented by formula d with the compound represented by formula e to obtain the compound represented by formula I, as shown in the following reaction formula: Wherein, the R' is selected from any one of tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, trimethylsilyloxy, triethylsilyloxy, benzyloxy, p-methoxybenzyloxy, methoxymethyleneoxy, benzyloxycarbonyloxy or tert-butyloxycarbonyloxy; R1' is selected from any one of acetyl, tert-butyldimethylsilyloxy, tert-butyldiphenylsilyloxy, trimethylsilyloxy, triethylsilyloxy, 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 chloro, methoxy or hydroxy.

7. The preparation method according to claim 6, characterized in that: 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, and the condensing agent comprises EDCI and / or DMAP; Preferably, in step (1), the reaction temperature is 0-50°C and the reaction time is 2-15h; Preferably, in step (1), the molar ratio of the compound represented by formula a to the compound represented by formula b is 1:(0.8-1.5); Preferably, in step (2), the deprotection is carried out by a hydrolysis reaction or a hydrogenation reaction; Preferably, in step (2), the deprotection is carried out by a hydrolysis reaction, and the hydrolysis reaction is carried out under acidic conditions or alkaline conditions; Preferably, in step (2), the temperature of the deprotection reaction is 0-50° C., and the reaction time is 1-24 h; Preferably, in step (2), the deprotection reaction is carried out in a solvent, and the solvent is 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, and the condensing agent comprises EDCI and / or DMAP; Preferably, in step (3), the molar ratio of the compound represented by formula d to the compound represented by formula e is 1:(0.8-2); Preferably, in step (3), the reaction temperature is 0-50°C, and the reaction time is 1-15h.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises an active ingredient and a pharmacodynamically acceptable carrier, wherein the active ingredient comprises a phenyl acrylic acid compound according to any one of claims 1 to 4 or a tautomer of a phenyl acrylamide compound according to claim 5 or a pharmaceutically acceptable salt thereof; Preferably, the mass percentage of the active ingredient in the pharmaceutical composition is 0.1-95%.

9. Use of the phenylacrylamide compound according to any one of claims 1 to 4, the 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 preventing or treating 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.

10. The use according to claim 9, characterized in that: The tumor is selected from any one or a combination of at least two of 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 comprises any one of rheumatoid arthritis, systemic lupus erythematosus, ulcerative colitis, psoriasis, Sjögren's syndrome, dermatomyositis or spinal lateral sclerosis, or a combination of at least two thereof; Preferably, the inflammatory disease includes any one or a combination of at least two of myocarditis, cytokine storm, non-infectious polyarteritis, phlebitis, and reflux esophagitis caused by infectious bacteria or viruses; Preferably, the neurodegenerative disease includes Alzheimer's disease and / or Parkinson's disease.

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